WE WELCOME ALL OUR SPEAKERS AND
PARTICIPANTS AND WE WOULD ALSO
LIKE TO WELCOME THE 175
PARTICIPANTS WHO SIGNED UP TO
ATTEND ONLINE.
I'M SURE THEY'RE NOT HERE QUITE
YET BUT WE DO EXPECT A LARGE
TURN OUT ONLINE AS WELL.
WE WILL REMIND ALL ONLINE
PARTICIPANTS TO REMAIN MUTED
DURING THE DURATION OF THE
MEETING.
SO ANIMAL MODELS PLAY CRUCIAL
ROLES IN BASIC STUDIES OF
RESPIRATORY I HAVE USS AIMED AT
UNDERSTANDING PATHOGENESIS,
TRANSMISSION AND IMMUNITY.
--AND AN MOLL MODELS PROVIDED A
FOUNDATION FOR MANY NEW VACCINE
APPROACHES.
HOWEVER TO IMPROVE OUR
APPROACHES TO VACCINE AND
THERAPY DEVELOPMENT WE FEEL
THERE A STRONG NEED TO
UNDERSTAND DETAILS FOR
RESPIRATORY PATHOLOGY AND IMMUNE
RESPONSES.
SO THIS WORKSHOP WILL BE FOCUSED
ON ANIMAL MODELS USED TO STUDY
BASIC ASPECTS OF INFLUENZA AND
SARS-COV-2 INFECTION IN HUMANS
AND THIS TEMS FROM THE CAPACITY
FOR SUBSTANTIAL ANTIGENIC DRIFT
AND THE PANDEMIC POTENTIAL OF
BOTH OF THESE VIRUSES, AS WELL
AS NATURE OF THE RESEARCH BEING
PERFORMED IN NIAID PORTFOLIOS.
ALTHOUGH IT MAY BE ATTEMPTING
TODAY AND TOMORROW TO DISCUSS
THE STATUS OF THE H5 N1
INFLUENZA OUTBREAK IN COW XTION
OTHER DAIRY ANIMALS WE WANT THIS
WORKSHOP TO TRY TO REMAIN
FOCUSED ON BETTER UNDERSTANDING
THE HUMAN HEALTH OUTCOMES AND
NOT STRAY TOO FAR INTO ANIMAL
HEALTH OUTCOMES.
SO WE ASKED SPEAKERS TO ADDRESS
THE EXPERIMENT AMILLIO BENEFITS
OF LIMITATIONS OF THE ANIMAL
MODELS THEY USE TO STUDY VIRUS
INFECTIONS.
WE ENCOURAGE RESPECT OF
DISCOURSE ON APPROPRIATE ANIMAL
MODELS FOR STUDYING VIRAL
PATHOGENESIS AND TRANSMISSION.
AND PERHAPS IDENT AREAS WHERE
NOVEL APPROACHES ARE NEEDED TO
IMPROVE OUR UNDERSTANDING OF
INFLUENZA AND SARS-COV-2
INFECTIONS AND VIRUS HOST
INTERACTIONS.
THROUGHOUT THE WOWRS OF THE
WORKSHOP, WE WOULD LIKE ALL
PARTICIPANTS TO NOT ONLY THINK
ABOUT THE IMPACT OF WHAT IS
DISCUSSED ON YOUR OWN RESEARCH
BUT ALSO TO THE FIELD AS A
WHOLE.
AND SO SOME OF THE QUESTIONS WE
WANT YOU TO THINK ABOUT ARE WHAT
ARE SOME OF THE REAGENT NEEDS
THAT ARE NEEDED TO ADVANGS YOUR
RESEARCH.
WHAT ARE SOME OF THE MOST
APPROPRIATE VIRUS STRAINS TO BE
USING FOR THE STUDIES IN YOUR
LAB AND IN THE FIELD.
AND WHAT ANIMAL MODELS ARE MOST
SUITED TO ANSWER SPECIFIC
EXPERIMENTAL QUESTIONS.
AND ARE THERE IMPROVEMENTS TO
THESE ANIMAL MODELS THAT ARE
NEEDED SO THAT THEY BETTER
REFLECT HUMAN PHYSIOLOGY.
SO TO GUIDE YOU IN THINKING
ABOUT THESE QUESTIONS AND TO
ALLOW YOU TO OFFER CONCRETE
FEEDBACK TO US WE'VE INCLUDED 3
QUESTIONS FOR YOU TO CONSIDER
DURING THE COURSE OF THIS
WORKSHOP.
THESE QUESTIONS ARE POSTED HERE.
WHAT REAGENTS ARE NEEDED TO
ADVANCE THE RESEARCH IN YOUR
FIELD?
IS THERE A NEED FOR THE
CONSISTENT, VALIDATED USE OF
CONSISTENT VALIDATED CHALLENGED
STRAINS OF VIRUS TO BE ABLE TO
COMPARE STUDIES IN ANIMAL MODELS
AND IF SO WHAT VIRUS STRAINS
WORK BROADLY IN DIFFERENT ANIMAL
MODELS?
AND A BIT CHALLENGING QUESTION
IS WHAT CRITERIA SHOULD BE
CONSIDERED WHEN COMPARING ANIMAL
MODELS TO HUMAN DATA TO ENSURE
THE MODELS ARE CLINICALLY
RELEVANT?
SO THE WAY WE WILL COLLECT
FEEDBACK IS USING THE
CONFERENCE'S I. O. APP.
SO IF YOU WANT TO ENTER THIS WEB
LINK, IT WILL TAKE YOU THERE OR
YOU CAN USE THE QR CODE.
AND OUR GOAL HERE IS WE WILL
START TO LOOK AT INITIAL
RESPONSES AT THE END OF THE
TODAY BUT WE WILL ALSO
INCORPORATE THESE RESULTS INTO
SESSIONS FOR TOMORROW WHEN WE
DISCUSS SMALL ANIMAL MODEL
RESOURCES AND DEVELOPMENT AND WE
WILL ALSO TRY TO POST THIS LINK
INTO THE ZOOM CHAT AS WELL FOR
THOSE WHO ARE ONLINE.
BEFORE WE GET STARTED I HAVE
JUST A FEW HOUSEKEEPING ITEMS.
IF YOU WOULD LIKE TO ORDER LUNCH
FOR TODAY PLEASE USE THE QR CODE
OR LINK PROVIDED ON THE
AYEPPEDDA TO PLACE AN ORDER WITH
CHOPT.
THERE ARE FLYERS ON EACH TABLE
WITH ORDERING INSTRUCTIONS.
YOU HAVE TO PLACE THESE ORDERS
BY 9:30 AMIN ORDER FOR THEM TO
MAKE IT TODAY SO MAKE YOUR LUNCH
PLANS AS SOON AS POSSIBLE.
FOR EACH SESSION, WE HAVE AN
ASSIGNED MODERATOR WHO WILL HAVE
SOME MAJOR TESTS, FIRST IS
KEEPING THE TALKS ON TIME
BECAUSE WE DO HAVE A RELATIVELY
TIGHT PROGRAM AND THEP MAKING
SURE THAT THE DISCUSSION STAYS
ON TOPIC.
WE DO HAVE NIAID THROW A
FOOTBALL MEMBERS WHO WILL HELP
MONITOR ZOOM QUESTIONS BUT WE
WILL TRY TO PRIORITIZE THE
IN-PERSON DISCUSSIONS.
SO NOW I WOULD LIKE TO INTRODUCE
DR. MICHAEL ISON, WITH THE NIAID
FOR A WELCOME.
>> WELL, THANK YOU ALL FOR
JOINING US TODAY.
FOR THOSE THAT DON'T KNOW ME,
I'VE BEEN HERE FOR ABOUT A YEAR
AND HALF NOW.
I COME FROM A BACKGROUND OF
ANTIVIRALS AND ANIMALS, AND I
MOST REEBTLY WORKED ON THE GSK
RSV VACCINE, AS YOU ULTIMATELY
KNOW WORKING WITH SMALL ANIMALS,
NONE OF THE CLINICAL IMPACT OF
ALL THE MEDICAL MEASURES WE KNOW
WOULD BE POSSIBLE WITHOUT SMALL
ANIMAL MODELS.
I'M REALLY EXCITED TO LISTEN TO
THE DISCUSSION TODAY AND I WANT
TO THANK ALL OF YOU FOR COMING
TO THE MEETING EITHER IN PERSON
OR VIRTUALLY, CONTRIBUTING TO
THE DISCUSSION.
OUR TEAM REALLY IS COMMITTED TO
TAKING THESE HOME AS LESSONS TO
SEE HOW WE CAN HELP FACILITATE
THE WORK OF THE RESEARCH
COMMUNITY.
SO PLEASE SHARE YOUR IDEAS, BE
THOUGHTFUL, IF THINGS COME UP OR
ISSUES, AN IDEA COMES TO YOU
AFTER YOU LEAVE THE MEETING,
PLEASE DON'T HESITATE TO REACH
OUT TO ANY OF US.
I WANT TO THANK OUR TEAM,
PARTICULARLY MICHELLE AND STEVEN
WHO HAVE DONE INCREDIBLE WORK
AND WHILE IT MAY NOT FEEL LIKE
IT FOR YOU THIS IS THE SECOND
MEET THANKSGIVING WEEK THAT
MICHELLE HAS BEEN HEAVILY
INVOLVED WITH IT AND THE THIRD
BIG MEETING IN THE LAST MONTH
ORIGINAL SO SO SHE'S BEEN
WORKING EXTRA HARD SO
CONGRATULATIONS MICHELLE AND
AGAIN, THANKS TO THE ENTIRE
VIRAL SECTION OF THE RESPIRATORY
DISEASE BRANCH WHO HAD BEEN
DOING INCREDIBLE AMOUNT OF WORK
BOTH WITH THE H5 RESPONSE, AND
WORK TO HELP SUPPORT YOUR
RESEARCH ENDEAVORS.
THANKS FOR COMING AND WELCOME TO
NAID.
>> HI, EVERYONE, I AM
DISCIPLINARY 18 POST AND I AM
THE SECTION CHIEF FOR THE NAID,
AND IT IS MY PLEASURE TO
INTRODUCE DR. STACEY CHERRY,
ST. JUDE CHILDREN'S RESEARCH
HOSPITAL, STUDIESOT ECOLOGY OF
INFLUENZA AND ANIMALS AND BIRDS,
THE SENIOR ASSOCIATE DEAN FOR
ACADEMIC AFFAIRS FOR THE
TBRADUATE SCHOOL OF BIOMEDICAL
SCIENCES AND COPI OF OUR CENTERS
OF EXCELLENCE FOR INFLUENZA
RESEARCH AND RESPONSE AS WELL AS
1 OF OUR CIVICS PROGRAMS AND
STACEY HAS A LOT OF EXPERTISE
AND A LOT OF DIFFERENT ANIMAL
MODELS AND WITH MAKING REAGENTS
A LOT OF EXPERTISE IN THAT AREA,
SO WE'RE VERY HAPPY TO HAVE HER
AS OUR KEY NOTE SPEAKER TODAY.
SO WELCOME, STACEY.
>> GOOD MORNING.
THANK YOU FOR THAT INTRODUCTION.
THIS IS NICE TALKING TO A
SMALLER GROUP, I FEEL LIKE I'M A
LAB MEETING OR DEPARTMENT, SO
IT'S A LITTLE LESS DOWN THING
WHEN I AM PREACHING TO THE CHOIR
INSTEAD OF THE EXPERTS.
SO MICHELLE SAID WHEN SHE
SEPTORS OUT THE SCHEDULE I'M
LIKE OH MY GOD.
OKAY, SO, WHAT I'M HOPING WE CAN
DO IS HAVE A CONVERSATION, MAYBE
THE ONLINE PEOPLE, MOSTLY US AND
I'M LOOKING AT YOU BECAUSE I
THINK THERE ARE AREAS WHERE WE
NEED TO DISCUSS HOW TO MOVE
FORWARD AND TO MOVE FORWARD IN
THE RIGHT WAY.
SO LET ME GO AHEAD AND GET
STARTED.
I DON'T THINK THERE'S A POINTER.
THERE'S THAT.
SO AS WE KNOW THERE WHEN WE'RE
EXPOSED TO A RESPIRATORY
INFECTION, THERE'S A SERIES OF
EVENTS THAT NEED TO HAPPEN
STARTING WITH THE MUCOSAL RATE
DEALING WITH THE IMMUNE RESPONSE
AND THE MICROBIAL DEPARTMENT
PLAYS A BIG ROLE IN WHAT'S
HAPPENING AND WHAT HAPPENS THEN
IS YOU ILLICIT THAT SYSTEM
TEMMIC INNATE IN THE ADAPTIVE
IMMUNE RESPONSES AND IN MOST
PEOPLE, IT WORKS BEAUTIFULLY.
BUT WE KNOW IT DOES NOT WORK THE
SAME WAY IN ALL PEOPLE AND THIS
STARTS TO BECOME SOMETHING WE
HAVE TO THINK ABOUT WHEN WE ARE
TRYING TO PLAN WHAT ANIMAL
MODELS ARE WE GOING TO USE, WHO
ARE WE TRYING TO MODEL BECAUSE
IT'S VERY COMPLEX AND IT DEPENDS
ON WHAT THE QUESTION IS YOU'RE
ASKING AND WHO YOU'RE ASKING IT
IN.
WE ALSO HAVE TO THINK ABOUT WHAT
VIRUSES WE'RE WORKING WITH AND
MOST OF US THINK ABOUT
MONOINFECTIONS, BUT IN REALITY
IS THAT TRULY WHAT'S HAPPENING?
OR ARE THERE A LOT MORE
CO-INFECTIONS, AND HERE WE'RE
JUST FOCUSING ON THE BACTERIA
BECAUSE THE BACTERIA ARE WELL,
THE BACTERIA BUT THEY PLAY AN
IMPORTANT ROLE, THE UNDERLYING
COMMENSAL OR MIKE ROY BIOME AS
WELL AS THOSE MORE OPPORTUNISTIC
BACTERIA.
AND WE ALSO HAVE TO THINK ABOUT
WHEN WE'RE LOOKING AT ANIMAL
MODELS HOW EACH OF RESPIRATORY
VIRUSES MAY BEHAVIOR DIFFERENTLY
AND DIFFERENT MODELS MIGHT BE
IMPORTANT.
SO WHAT WE WANT TO TALK ABOUT OR
WHAT I WANT TO TALK ABOUT TODAY
ARE A COUPLE OF THINGS.
THIS IS--I TYPICALLY DON'T DO AN
OUTLINE BUT THIS WAS HELPFUL FOR
ME TO THINK ABOUT THE BIG TOPICS
I THOUGHT WE MAYBE NEEDED TO
DISCUSS.
ONE IS YOU HAVE TO TART WITH
JUST COMPARING AND CONTRASTING
THE DIFFERENT MODELS AND THIS IS
WHERE YOU GUYS ARE THE EXPERTS
SO I'M HOPING YOU WILL YUSM IN
AND TELL ME WHERE I GOT THINGS
WRONG OR WHERE WE NEED TO THINK
MORE ABOUT IT.
AND THEN, WHAT QUESTION FOR YOU
GETTING AT?
WHAT ARE YOU ACTUALLY TRYING TO
MODEL IN THE ANIMAL AND WHO ARE
YOU TRYING TO MODEL IT?
IF IT'S AN INFANT USING NAIVE
MICE, PERFECT, IF IT'S AN ADULT,
MAYBE YOU NEED TO USE IMMUNE
HISTORY OR DIRTY MICE LIKE WE'LL
HEAR ABOUT FROM RYAN.
CAN WE IMPROVE THESE MODELS AND
IF SO, HOW DO WE DO HA AND WHAT
ARE THE WAYS FORWARD FOR THE
FIELD.
SO THESE ARE THE TOPICS I WILL
TRY AND HINT ON AND AGAIN I'M
HOPING IT'S MORE OF A DISCUSSION
SO FEEL FREE TO JUMP IN.
SO THERE'S A LOT OF GREAT LARGER
ANIMAL MODELS, YOU KNOW,
NONHUMAN PRIMATES FOR BASICALLY
ALL THE VIRUSES, FOR FLU,
CHICKENS, PIGS, [INDISCERNIBLE]
VIRUS, SHEEP, ALL FANTASTIC
MODELS, OF COURSE THOSE OF US
DOING FLU ARE NOW THINKING ABOUT
COWS IN PARTICULAR, IN
PARTICULAR COW UTTERS AND
MAMMARY EPITHELIUM BUT WE WILL
NOT FOCUS ON ANY OF THAT TODAY
AS MICHELLE SAID, WE ARE REALLY
GOING TO FOCUS ON THE SMALL
ANIMAL MODELS OR THE 1 WE'RE
MORE USE THE TO WORKING WITH THE
IN THE LABORATORY PRIMARILY
RODENTS AND FERRETS.
I DIDN'T INCLUDE ANYTHING WITH
RABBITS AND I DON'T KNOW OF
ANYBODY OTHER THAN THE PEOPLE
DOING PNEUMOCOCCUS AND EAR
INFECTIONS THAT ACTUALLY USE
CHINCHILLA.
DOES ANYBODY ELSE KNOW OF
ANYBODY WHO WORKS IN VIRUSES IN
CHINCHILLA.
THAT'S GOOD.
THEY'RE HARD TO FINE EMPLOY
LTD.'S JUST START.
WE WILL HIT THIS ON THE HEAD.
IF WE WERE TALKING ABOUT LARGER
REMODELS, WE WOULD HAVE
FANTASTIC REAGENTS, THERE ARE
FANTASTIC REAGENTS FOR PIGS,
COWS, NONHUMAN PRIMATES LESS SO
FOR SMALL ANIMAL MODELS, MICE
FANTASTIC BUT WE NEED TO GET
THAT OUT THERE, THEY ARE
IMPROVING AND IT'S SOMETHING WE
MEDE TO TALK ABOUT IN THE WAYS
FORWARD IS WHAT KIND OF TOOL
BOXES DO WE HAVE FOR THIS
SPECIES, HOW DO WE PRIORITIZE
AND HOW DO WE GET OUT THAT THESE
ARE AVAILABLE.
SO FIRST COMPARE XG CONTRASTING,
LET'S THINK ABOUT THE DIFFERENT
SMALL ANIMAL MODELS, AGAIN, THIS
IS WHAT I'VE SEEN IN THE
LITERATURE FOR THE NONINFLUENZA
VIRUSES SO EXPERTS PLEASE
CORRECT ME WHERE IT'S WRONG.
MICE, WE ALL USE MICE.
WE SAVE MILLIONS OF MICE EACH
YEAR FROM RESPIRATORY INFECTION,
AND THEY ARE FANTASTIC.
THEY CAN HAVE ANYTHING FROM
SUBCLINICAL INFECTION TO DEATH.
CAN YOU MAKE GREAT SCORING
CHARTS, YOU CAN MONITOR THINGS,
THERE'S A PROBLEM, THEY'RE NOT
REALLY SUSCEPTIBLE TO MOST OF
THE RESPIRATORY VIRUSES WE LIKE
TO USE.
THIS BECOMES AN ISSUE RIGHT?
FOR THINGS LIKE STUDY SECTION, I
WILL BE THE FIRST TO ADMIT FOR
BLASTING PEOPLE FOR SAYING THEY
USE PR8, BUT PR8 IS THE BEST
VIRUS TO LOOK AT WHAT YOU'RE
MODELING.
SHOULD IT BE OKAY?
RSV, LOTS OF WORK IN MICE BUT
YOU WILL SEE A STUDY SECTION,
YOU SHOULDN'T USE MICE FOR RSV,
SO THESE ARE JUST THINGS AS A
FIELD WE NEED TO THINK ABOUT,
YOU KNOW THE AIRWAY ANATOMY HOW
MUCH IS IT LIKE HUMAN?
IT'S OKAY.
SITES OF REPLICATION, TALKING
ABOUT A PNEUMONIA MODEL WHILE
YOU WILL GET SOME RESPIRATORY
TRACT, IT IS LOWER, TRANSMISSION
AT LEAST FOR FLU, SARS, SOME OF
THOSE WILL TRANSMIT IN A MOUSE.
I HAVE SEEN REPORTS IN INFANT
MICE THAT FLU WILL TRANSMIT.
REPEATED SAMPLING, NOT SO MUCH.
THERE ARE THINGS THAT ARE EASY
TO GET FROM THE SAME MOUSE OVER
AND OVER, BUT YOU'RE NOT GOING
TO GET A LUNG.
LOTS OF REAGENTS, RIGHT?
COSTS ARE REASONABLE BUT THEY
ARE INCREASING, I DON'T KNOW IF
ANYBODY ELSE HAD PROBLEMS
GETTING MICE RECENTLY FROM JACKS
LABS.
THAT'S A REAL THING WHETHER
THEY'RE BACK ORDERED OR THEY'RE
JUST GETTING SORT OF PRICED UP.
WHEN YOU GO TO PUBLISH, WHEN YOU
GO TO STUDY SECTION, THE
EXPECTATIONS, YOU WILL BE ABLE
TO DO STATISTICALLY POWERED
STUDIES, YOU WILL ADDRESS
BIOLOGICAL SEX, YOU'RE GOING TO
DO FANCY KNOCK OUTS, YOU KNOW
THE EXPECTATIONS ARE VERY HIGH
WHEN YOU'RE USING MOUSE MODELS
BECAUSE THE REAGENTS ARE THERE.
IT STARTS GETTING A LITTLE BIT
DIFFERENT WHEN YOU LOOK AT OTHER
MODELS.
FOR EXAMPLE, HAMSTERS, NEVER
THOUGHT ABOUT A HAMSTER BEFORE
ALTHOUGH THAT WAS SOMETHING THAT
[INDISCERNIBLE]'S GROUP HAS BEEN
PUSHING FOR YEARS FOR FLU, BUT
AGAIN ASYMPTOMATIC TO DEATH AND
DISEASE, SUSCEPTIBILITY IS
BETTER.
SARS-COV-2 WAS THE FIRST TIME
WE'VE EVER HAD TO WORK WITH A
HAMSTER, BUT RSV WILL AFFECT
THEM AND THEIR AIRWAY ANATOMY IS
CLOSER TO A HUMAN.
SITES OF REPLICATION, YOU CAN
GET UPPER AND LOWER RESPIRATORY
TRACT AND THAT IS SOMETHING WE
NEED TO CONSIDER MOVING FORWARD
IS THAT MORE AND MORE PEOPLE ARE
LOOKING AT MUCOSAL IMMUNITY AND
WILL THEY BE THE ANIMAL MODELS
TO APPROPRIATELY DO THOSE
STUDIES AND COMPARE THEM TO
HUMANS.
TRANSMISSION, YES.
AGAIN REPEATED SAMPLING, IT
DEPENDS ON THE SAMPLE THAT
YOU'RE COLLECTING.
THE REAGENTS ARE UNLIMITED BUT
THEY ARE IMPROVING.
COSTS ARE TILL REASONABLE, AND I
DON'T KNOW FOR THE PEOPLE THAT
DO HAMSTER MODELS, YOU KNOW, CAN
YOU DO A LOT OF MODELS, ARE YOU
EXPECTED TO DO MALE AND FEMALE,
THERE'S REALLY NO FANCY MODELS I
COULD FIND ALTHOUGH I DO THINK
WE HAVE A TALK ON GENERATION OF
MAKING TRANSGENIC HAMSTERS WHICH
I THINK WILL BE REALLY EXCITING.
RAT, HMM, OKAY, SO, AGAIN, FOR
SOME RESPIRATORY PATHOGENS
THEY'RE FANTASTIC.
RSV, THOSE [INDISCERNIBLE] ARE
THE GOLD STANDARD IN THE FIELD.
THEY'RE ALSO TERRIFYING AND AS
WE'RE GETTING MODEL CITIZENNER
INTO RSV, THAT IS 1 I WOULD
RATHER DO A PIG IN HIGH
CONTAINMENT THAN A COTTON RAT IN
BL2.
SARS, FLU, THERE ARE A NUMBER OF
RESPIRATORY THAT WILL REPLICATE
IN THE RAT, GOT A BETTER AIRWAY
ANATOMY AND AGAIN YOU CAN GET
UPPER AND LOWER RESPIRATORY
TRACT AND FOR RSV, THIS IS THE
MODEL WHERE PEOPLE DO A LOT WITH
ASTHMA AND LOOK AT A LOT OF
THERAPEUTICS.
YOU CAN GET TRANSMISSION, YEAH,
WE'RE LIMITED WITH THE REAGENTS,
RIGHT?
THE COSTS ARE REASONABLE, BUT
THE EXPECTATIONS ARE NOT AS FAR
AS I KNOW QUITE WHAT THEY ARE
WITH THE MOUSE WHERE YOU HAVE TO
HAVE 20 MICE PER GROUP.
THINGS LIKE THAT.
I PUT TERRIFYING BECAUSE I FIND
THE COTTON RATS SCARY.
GINNY PIGS, THIS HAS BEEN AN
EXCITING MODEL TO WATCH DEVELOP.
DENEEZ LOHAN REALLY PIONEERED
THIS SEVERAL YEARS AGO NOW FOR
LOOKING AT TRANSMISSION WITH
INFLUENZA VIRUS AND DISEASE
VARY, FOR FLEW WE THINK ABOUT
THEM BEING SUBCLINICAL BUT WITH
OTHER RESPIRATORY VIRUSES YOU DO
GET CLINICAL DEC.
AIRWAY ANATOMY, AGAIN CLOSER TO
HUMAN ANDS YOU DO GET BOTH UPPER
AND LOWER RESPIRATORY TRACT.
REAGENTS ARE JUST AS LIMITED,
BUT WE'RE DOING MORE AND MORE
STUDIES ESPECIALLY FOR
TRANSMISSION BECAUSE THEY ARE
SOMEWHAT EASIER TO WORK WITH
THAN A FERRET AND AGAIN, WE WILL
TALK LATER ABOUT FANCY NEW GINNY
PIG MODELS THAT ARE BEING
DEVELOPED.
FERRETS, THESE ARE GREAT.
A LOT DEPENDS ON HOW YOU
MANIPULATED THAT MODEL, SO WITH
A HUMAN SEASONAL INFLUENZA
STRAIN FOR THIS PART, WE GET A
LOT OF DISEASE--IT DEPENDS ON
THE STRAIN, IF YOU PUT H5 IN
THERE, YOU MAY NOT GET LUNG
DISEASE, IT'S GOING IT GO
NEUROTROAPIC AND IT'S GOING TO
BE FATAL.
SARS WOULD INFECT, TRANSMIT
BEAUTIFULLY, SO THESE ARE GOOD
MODELS, YOU DO HAVE THAT NICE
AIRWAY ANATOMY MORE SIMILAR TO A
HUMAN AND AGAIN YOU WILL GET
UPPER AND LOWER RESPIRATORY
TRACT REPLICATION.
REPEATED SAMPLING, THEY GET
EASIER AS THEY GET LARGER, CAN
YOU GET SALIVA, NAISAL SWABS,
NAISAL WASHES, WHATEVER YOU WANT
AND LIKE THE OTHERS REAGENTS ARE
LIMITED BUT THEY ARE IMPROVING.
WE WILL TALK ABOUT THAT AT THE
END.
WITH ALL OF THESE WITH MORE
PEOPLE USING THESE MODELS, THERE
IS MORE OF A CONCERTED EFFORT TO
MAKE THE TOOLS THAT WE NEED TO
BASICALLY MAKE THESE MODELS
LARGE MICE IS THE HOPE.
THEIR EXPENSIVE, THESE ARE NOT
INEXPENSIVE ANIMALS, HOUSING IS
MORE EXPENSIVE AND THEY CAN BE
DIFFICULT TO WORK WITH.
WHEN YOU GO TO PUBLISH, IT'S A
LITTLE BIT EASIER TO--I WON'T
SAY GET AWAY WITH BUT IT'S
EASIER TO JUSTIFY NOT DOING 20
FERRETS PER GROUP AND 5 TIME
POINTS.
BIOLOGICAL SEX, YOU HAVE TO
ADDRESS IT BUT WORKING WITH
FEMALE FERRETS CAN BE DIFFICULT,
RIGHT?
ESPECIALLY WHEN THEY HIT SEXUAL
MATURITY, IT CAN BE TRICKY.
FANCY MODELS, THERE ARE ACTUALLY
TRANSGENIC FERRETS.
THERE'S A GROUP IN IOWA THAT
SPENDS A LOT OF TIME
PARTICULARLY FOR CYSTIC FIBROSIS
AND OTHER LUNG DISEASE IS MAKING
VERY MUCH CELL BIOLOGY BASED
REAGENTS LOOKING AT LUNG REPAIR,
LUNG INJURY AND HE HAS THE
CAPABILITIES OF MAKING
TRANSGENIC FERRETS FOR PEOPLE
AND WILL HOUSE THEM THERE.
AND THEN WE HAVE THE PEN
ULTIMATE ANIMAL MODEL WHICH IS
HUMANS WHICH IS HUMANS RIGHT.
SO THERE, DEC, ASYMPTOMATIC TO
DEATH, JUST LIKE AN ANIMAL
MODEL, YOU CAN DO GOOD SCORING
CHARTS IN PEOPLE, THEY CAN EVEN
HELP YOU.
SUSCEPTIBILITY, GOOD, DEPENDING
ON THE PERSON, DEPENDING ON THE
STRAIN, YOU HAVE AIRWAY ANATOMY
THAT LOOKS LIKE A HUMAN AND YOU
HAVE SITES OF VIRAL REPLICATION,
UPPER, LOWER AND WE'RE ALL
GIGGLING BUT THERE ARE PEOPLE
THAT ARE PUSHING TO GET AWAY
FROM ANIMAL MODELS AND IF YOU
WANT TO STUDY INFECTION IN A
HUMAN, YOU STUDY IT IN A HUMAN
SO I THINK IT NEEDS TO BE HERE
BECAUSE WE NEED TO THINK ABOUT
HOW CAN WE DO THIS AND CAN IT
CAN BE DONE AND CAN IT BE DONE
IN SUCH A WAY YOU CAN BRING THAT
INFORMATION BACK TO IMPROVE OUR
ANIMAL MODELS AS WELL.
WILL YOU GET TRANSMISSION IN
ABSOLUTELY GIVEN THE RIGHT
PEOPLE.
REPEATED SAMPLING, YEAH, IF THAT
PERSON WILL LET YOU DO IT, YOU
CAN GET LOTS OF DIFFERENT
THINGS.
AGAIN, YOU'RE NOT GOING TO GET A
LUNG SAM SAMPLE FROM THEM BUT
MAYBE THEY'LL LET YOU DO 1
BOWEL.
REAGENTS, LOTS OF REAGENTS FOR
HUMANS, RIGHT?
NOW THEY'RE GOING TO BE PRICEY
AND DIFFICULT TO DO, AND THEN
YOU'RE REALLY GETTING INTO THE
COMPLEXITY THAT I THINK ISN'T
PRESENT IN MOST FOR ANIMAL
MODELS THAT ALLOWS US TO BETTER
UNDERSTAND THINGS
MECHANISTICALLY WITHOUT THE
COMPLICATIONS OF HOST GENETICS
AND UNDERLYING INFECTIONS, BUT
THAT'S ALSO I THINK WHAT THE
ARGUMENT IS ABOUT WHY THE ANIMAL
MODELS WILL NEVER COMPLETELY
RECAPITULATE WHAT WE SEE IN
HUMAN, SO IS IT REALITY TO THINK
THAT WE WOULD MOVE TOWARDS JUST
DOING EVERYTHING IN AWE HUMAN.
I DON'T THINK SO, BUT THERE ARE
PLENTY OF PEOPLE THAT DO THINK
THIS IS POSSIBLE ESPECIALLY WITH
THE NEW HUMAN CHALLENGE VIRUSES
IN HUMAN CHALLENGE MODELS.
SO WE HAVE A LOT OF DIFFERENT
ANIMAL MODELS, WE POTENTIALLY
HAVE PEOPLE, BUT THE FIRST THING
WE HAVE TO THINK ABOUT IS WHAT
ARE THE QUESTIONS THAT YOU'RE
ASKING, WHAT ARE YOU TRYING TO
MODEL AND THAT WILL HELP YOU
THINK ABOUT WHAT'S THE BEST WAY
TO DO THIS.
SO IF YOU'RE DOING DISEASE
SEVERITY, REALLY LOOKING AT
PNEUMONIA, LUNG, YOU KNOW, YOU
KNOW THAT YOU CAN DO THESE
STUDIES IN MICE, BUT THERE ARE
OTHER MODELS THAT WILL ALLOW YOU
TO DO THIS, DEPENDING ON THE
VIRAL STRAIN AND DEPENDING ON
WHAT YOU'VE DONE TO THAT HOST,
IF YOU MADE THAT HOST IMMUNO
COMPROMISED IN SOME WAY,
MALNOURISHED, PREGNANT.
IF YOU'RE USING MALE VERSUS
FEMALE, THERE ARE ALL THINGS CAN
YOU DO TO MODULATE THAT SO THAT
YOU DO GET MORE OF A SEVERE
DISEASE.
IF YOU'RE TRYING TO MODEL WHAT
HAPPENS MUCOSAL AT THE UPPER
RESPIRATORY TRACT, THIS GETS
TRICKIER, SPECIALLY IN A MOUSE,
WHERE IT MAY BE HARDER TO DO,
SOMETHING WITH AN INFLUENZA
VIRUS WHERE IT WAS SET UP TO BE
THE PNEUMONIA MODEL, IT CAN BE
DONE BUT YOU'RE HAVING TO THINK
ABOUT WHAT YOU'RE TRYING TO
MODEL AND HOW YOU'RE GOING TO DO
IT, WHAT STRAIN OF VIRUSES ARE
YOU GOING TO USE, AND THEN YOU
HAVE TO THINK ABOUT THOSE
CO-INFECTIONS, I DON'T KNOW IF
ANYBODY ELSE HAS HAD SOMETHING
RUN THROUGH THEIR ANIMAL
FACILITY THAT ALL OF A SUDDEN,
YOUR WANING STRAIN OF VIRUS IS
KILLING ALL OF YOUR ANIMALS
BECAUSE SOMETHING HAS GONE
THROUGH THE FACILITY, NORO
VIRUS, ASTRO VIRUS, PIN WORMS,
WHATEVER SO WE KNOW THESE CAN
INFLUENCE THE MODELS THAT WE'RE
USING.
IF YOU'RE DOING VACCINE STUDIES,
I THINK THIS IS WHERE IT ALSO
BECOMES REALLY IMPORTANT.
AGAIN, WE DO A LOT OF VACCINE
WORK IN MICE AND WHAT WE'RE
LOOKING AT IS A BABY RIGHT?
IF YOU'RE USING THIS NAIVE MOUSE
THAT HAS NO IMMUNE HISTORY,
YOU'RE BASICALLY MODELING WHAT
WOULD HAPPEN IN AN INFANT.
IS THAT WHAT YOU'RE TRYING TO
MODEL?
BECAUSE IF YOU GO INTO A HUMAN
TRIAL, THAT'S THOT WHERE IT'S
GOING TO GO.
IT'S GOING TO GO IN AN ADULT.
SO DO YOU NEED TO USE AN ANIMAL,
FERRET HAMSTER, WHATEVER YOU
WANT THAT HAS MORE OF AN ADULT
IMMUNOLOGICAL RESPONSE OR HAS
THAT IMMUNE HISTORY AND I THINK
MOST OF US ARE GOING TOWARDS
THAT IMMUNE HISTORY, IT'S
IMPORTANT FOR FLU, IT'S NOW
IMPORTANT FOR SARS THAT YOU HAVE
A COMPLEX SARS-COV-2 IMMUNE
HISTORY AND IT'S GETTING THAT
WAY FOR RSV AS WELL.
AND I CAN SAY FOR THE PAST 12,
15 YEARS BEFORE WE WOULD ORDER
FERRETS, WE WOULD PUT A LOT OF
TIME AND EXPENSE INTO SCREENING
EVERY FERRET.
WE ORDER SERIES POINTSA FROM
THESE FERRETS, HERE'S THE 1S WE
WILL CHOOSE, WE WILL CHOOSE 20%
MORE THAN WHAT WE NEED, WE WILL
SCREEN THEM AND FIEBD FIND THET
ARE NAIVE, I DON'T KNOW IF
ANYBODY ELSE DOES THAT BUT THAT
IS WHAT WE WOULD DO, WE DO
HHIs, AND I THINK THAT WAS
COMPLETELY THE WRONG WAY TO GO.
BECAUSE NOW WE'RE GETTING THESE
FERRETS AND HAVING TO GIVE THEM
AN IMMUNE HISTORY WHERE WE COULD
HAVE HAD THEM RIGHT FROM THE
BEGINNING.
SO AGAIN, WE HAVE TO THINK ABOUT
THAT IMMUNE RESPONSE, WHAT ARE
YOU TRYING TO LOOK AT AND IS IT
THE RIGHT MODEL AND THEN WHAT I
MEAN IS THEY'RE CLEAN, RIGHT?
THEY'RE GETTING A MONOINFECTION,
THEY HAVE UNDERLYING THINGS BUT
NOT--WE KNOW THEY DON'T HAVE
THINGS THAT WE DON'T WANT THEM
TO HAVE, THEY DON'T HAVE FLU,
THEY DON'T HAVE RSV, THEY DON'T
HAVE SARS-COV-2.
DOES THAT MAKE A DIFFERENCE?
YEAH.
SO, THIS IS SOME OF MY
INDIGENOUS PEOPLE ENROLLED IN
OUR STUDIES IN COLUMBIA AND WHAT
WE DID IS WE WENT IN AND JUST
OFFERED THE GOVERNMENT FLU
VACCINE BECAUSE THEY DIDN'T TAKE
IT BECAUSE THEY DIDN'T HAVE
ACCESS TO IT.
SO NOT A CLINICAL TRIAL, JUST
BRING IT TO THE COMMUNITIES IF
THEY WANT TO TAKE IT, THEY CAN,
SO HERE YOU HAVE THESE PEOPLE,
REALLY COMPLICATED IMMUNE
HISTORIES, RIGHT, ALL FROM
NATURAL INFECKIONS, THEY HAVE
WORMS, THEY HAVE PARASITES, THEY
HAVE A LOT OF ZOANNOTTIC
DISEASES.
WHAT WE DIDN'T EXPECT IS THIS IS
JUST A SAMPLE IS A NUMBER OF
THESE PEOPLE, I WOULD SAY ABOUT
HALF OF THEM IN THIS FIRST PHASE
OF STUDY WHEN THEY CAME IN, AT
DAY 0 TO GET THEIR FLU SHOT,
THEY HAD SARS AND YOU CAN SEE
WHAT THE CT VALUES LOOK LIKE.
I MEAN 18.
THAT'S NOT NOTHING.
NO CLINICAL DISEASE WHATSOEVER,
THAT'S VERY TYPICAL, IT'S
COMPLETELY THE OPPOSITE OF WHAT
WE HAD HYPOTHESIZED WHEN WE
START DOG THESE STUDY SYSTEM
THEY'RE GOING TO GET REALLY
SEVERE DISEASE.
THESE GUYS DON'T GET SICK FROM
RESPIRATORY VIRUSES AND THEN 28
DAYS LATER WE GET INTO, WE ARE
STARTING TO SEE FLU BUT NOT
SOMETHING WE THOUGHT ABOUT FOR A
VACCINE STUDY IN A MOUSE OR
FERRET IS THAT OOH, WHAT
HAPPENED TO THIS PERSON ACTUALLY
HAS SARS-COV-2 AT THE SAME TIME
SO THIS IS THE REAL WORLD AND
THIS IS 1 GROUP YOU CAN SAY IT'S
INDIGENOUS, I DON'T THINK SO,
IT'S JUST DO WE EVER LOOK,
RIGHT?
TRANSMISSION.
THIS IS FUN AND SUSCEPTIBILITY
TO TRANSMISSION, THIS IS WHERE
HAVING REALLY CLEAN ANIMALS
COULD CHANGE WHAT YOU'RE SEEING
RIGHT?
YOU'RE USED TO WORKING WITH
CLEAN ANIMALS BUT IF THEY HAVE
UPDATER LYING BACTERIA INFECS,
WE KNOW CERTAIN THINGS LIKE
STREPT O COCCUS PNEUMONIA, IF
THEY GET FLU, IT CAN HELP IT
TRANSMIT.
IT CAN HELP IT BIND TO CELLS, WE
KNOW IF IT HAS RSV, THAT CAN
COMPLETELY CHANGE SO THAT
INFECTION HISTORY ESPECIALLY AT
THE MUCOSAL SURFACE, HOW WILL
THAT CHANGE THAT SUSCEPTIBILITY
TOO AND TRANSMISSION FROM THE
ANIMAL AND THERE'S DIFFERENT
TYPES OF CONTACT AND WE WILL
HEAR MORE ABOUT THAT BECAUSE I
THINK THAT'S SOME OF THE REALLY
EXCITING STUDIES COMING OUT.
SEEMA'S GROUP IS DOING A LOT BUT
ALSO THE MECHANICS OF
TRANSMISSION, BEING IN A CAGE
TOGETHER, DOES THAT REALLY
MIMIC, ALTHOUGH TIMES YOU FEEL
LIKE THAT AT HOME WITH YOUR KIDS
BUT IT DOES REALLY MIMIC HOW WE
WOULD GET EXPOSED IN NATURE?
THE OTHER THING WE HAVE TO THINK
ABOUT, WHEN YOU'RE DOING
AN ANIMAL MODEL IS WHAT ARE YOUR
DOWN STREAM APPLICATIONS?
WHAT ARE YOU GOING TO DO WITH
THOSE TISSUES, IF YOU ARE GOING
TO DO TYPOLOGY, VIRAL
EMONSTRATIONS AND CYTOKINE ANDS
CHEMOKINES, ANY MODEL WILL DO,
IF YOU WANT TO GET FANCY AND YOU
WANT TO DO TRANSCRIPTOMICS OR
SINGLE CELL SEQUENCING IT GETS
TRICKIER IF THE ANIMAL IS LARGE,
SO THIS IS 1 EXAMPLE, THIS IS
ACTUALLY TAKING A MOUSE LUNG, 3
DAYS POST INFLUENZA INFECTION
AND DOING THIS TISSUE CLEARING
LIGHT PHASE MICROSCOPY AND ALL
THE FLU IS INFLUENZA VIRUS
NUCLEIC PROTEIN SO THIS IS SUPER
ARE EASY TO DO WITH THE MOUSE.
THE MOUSE LUNG, THE BRAIN, THE
HEART, WE'VE DONE THEM ALL NOW,
WE'VE DONE UPPER RESPIRATORY
TRACT, A LITTLE BIT TRICKIER IN
THE A FERRET BECAUSE THE TISSUE
IS SO MUCH BIGGER BECAUSE YOU
DON'T REALLY GET TO MODEL IT THE
SAME WAY AND THAT'S SOMETHING
WE'VE HAD TO THINK ABOUT DOING
THIS SPATIAL IS WE DON'T WANT TO
DO IT IN MICE BUT CAN WE DO IT
IN ANOTHER MODEL BECAUSE THE
TISSUE IS SO BIG.
WHO'S YOUR TARGET?
WHAT IS YOUR TARGET AUDIENCE,
RIGHT?
IS IT ADULTS, IS IT HEALTHY
ADULTS, IS IT THE ELDERLY?
DOES IT MAKE A DIFFERENCE?
ABSOLUTELY I MEAN WE KNOW WHEN
YOU THINK ABOUT VIRULENCE, WHEN
YOU THINK ABOUT TRANSMISSION,
WHEN YOU THINK ABOUT
THERAPEUTICS, IT'S REALLY DEPEND
EPT ON THE STRAIN, THE HOST AND
AGAIN WHAT THEY'VE SEEN IN TERMS
OF BEING ABLE TO PREVENT AND
CONTROL AND WHEN IT COMES TO THE
ANIMAL MODEL, WHEN WE KNOW IS IT
HAPPENS IN AN ANIMAL MODEL.
ABSOLUTELY.
CYBER KLEIN GROUPS HAS SHOWN THE
IMPORTANCE OF BIOLOGICAL SEX, WE
ALL DO A STUDY NOW AND WE HEAR
SABRA, IN OUR HEAD WHAT HAPPENS
TO THE FEMALES AND YOU DO IT
BECAUSE IT CAN BE VERY, VERY
DIFFERENT.
SARS IN INFANTS, WE WILL HEAR A
TALK ABOUT THAT AND STANLEY
PERLMAN'S WORK WITH SARS AND
AGED, SHOWING IN THE AGED MICE,
THATTA WHERE YOU CAN GET
DISEASE, YOU KNOW WE HAVE MATT
FRIEMAN WHO WORKED A LOT WITH
DIABETES AND OTHER UNDERLYING
CONDITIONS AND MIKE WILLA
WORKING WITH OBESITY IN SARS,
ALL OF THESE MAKE A DIFFERENCE
IN PEOPLE AND NOT PRIZINGLY IT
DOES MAKE A DIFFERENCE IN YOUR
ANIMAL MODEL, EVEN USING THE
SAME STRAIN OF VIRUS.
IT CAN BE A GAME CHANGER IN WHAT
YOU'RE ACTUALLY MODELING AND
WE'VE SPENT A LOT OF TIME FOR
FLU WORKING OUT NEW MODELS,
PARTICULARLY AROUND OBESITY
WHERE IN MICE, WE HAVE LOTS OF
DIFFERENT MODELS, DIET INDUCED
OBESITY, I WILL SHOW YOU A BIT
ABOUT A NEW CHILDHOOD OBESITY
MODEL THAT WE'RE WORKING ON,
PREGNANCY, TIME PREGNANCY,
MALNOURISHED, SO GOING THE OTHER
DIRECTION BECAUSE WE KNOW IT'S
NOT THAT OBESITY PER SE BUT MORE
THE METABOLIC SYNDROME.
WE'RE CALLING IT OZEMPIC, BUT
ACTUALLY OZEMPIC VERUS METFORMIN
VERSUS DIET AND EXERCISE.
EFFICACY RESPONSES, DONE THE
SAME THING IN FERRETS TO BE ABLE
TO UNDERSTAND HOW THESE USHED
LYING CONDITIONS OR
CO-MORBIDITIES IMPACT
TRANSMISSION OF THE VIRUS AS
WELL AS SUSCEPTIBILITY TO
GETTING INFECTED.
AND THEN, YOU KNOW TRYING TO
TRANSLATE WHAT WE'RE FINDING,
THERE'S GREAT NEW WAYS TO DO
THAT, EVEN EXVIVO.
USING PRIMARY RESPIRATORY CELLS,
USING THINGS LIKE THE
ORGANELLESOT CHIP MODEL SYSTEMS
WHICH ARE GETTING RETINAL
LOCATION REALLY GOOD AND
ULTIMATELY BEING ABLE TO DO
HUMAN CORHOTTERS AND THEN
PULLING THAT DATA TOGETHER.
SO WE THINK ABOUT GOING 1 WAY,
TAKING OUR ANIMALS, TRANSLATING
IT TO HUMANS BUT I THINK WE
SHOULD ALSO BE GOING THE OTHER
WAY AND THINK ABOUT WHAT WE SEE
IN HUMANS AND CAN WE
RECAPITULATE THAT IN THE ANIMAL
MODELS.
SO JUST TO SHOW YOU SOME.
DOES IT MATTER?
SO THESE ARE OUR DIET INDUCED
OBESE FERRETS, OR FURY
FOOTBALLS, THEY WEIGH ABOUT 1.5
KILOGRAMS SORE THEY'RE HONKING
HUGE BEASTYS, SO WE GET THEM IN,
WE PUT THEM ON A HIGH FAT DIET
FOR UP TO 12 WEEKS.
YOU HAVE TO MAKE IT BACKON
FLAVORED OR THEY WON'T EAT IT,
YOU HAVE TO GIVE THEM WET CAT
FOOD, SO THERE ARE THESE LITTLE
TRICKS YOU HAVE TO PLAY AND IF
WE GIVE THEM SEASONAL INFLUENZA
VIRUS INCLUDING FLEW B IT'S A
MUCH MORE SEVERE DISEASE.
SO IF YOU ARE USE A REPORTER
VIRUS, WE'VE USED
[INDISCERNIBLE], IF I CAN GET
THIS TO WORK, MAYBE NOT, WE'VE
USED [INDISCERNIBLE] AND
[INDISCERNIBLE], WHAT YOU CAN
SEE HERE IS THAT THE OBESE YOU
HAVE MORE VIRUS IN THE LUNG.
SO IT DOESN'T STAY AN UPPER
RESPIRATORY TRACT INFECTION NOW.
YOU'VE LOST THE BARRIER AND IT'S
GETTING DEEPER INTO THE LUNGS OF
THESE ANIMALS.
AND WHEN YOU LOOK AT YOUR TCID50
OR TISSUE INFECT CULTURE DOSE 50
AND YOU LOOK AT THE DIFFERENT
LOBES OF THE LUNG, CAN YOU SEE
CLEARLY WE WE HAVE A LOT MORE
VIRUS IN NEEZ OBESE ANIMALS.
THE OTHER THING THAT WILL HAPPEN
IS WE WILL TRULY DEPENDOT STRAIN
AND HOW BIG THESE ANIMALS ARE
GET INTO ACUTE RESPIRATORY
DISTRESS SYNDROME, AGAIN THIS,
IS ABOUT THESE ANIMALS HAVING
METABOLIC SYNDROME.
ANYBODY WHO ORDERS ANIMALS FROM
TRIPLE F, PRE AND POST PANDEMIC
YOU'RE GETTING THESE FERITS AND
THEY'RE A LOT BIGGER THAN THEY
WERE PREVIOUSLY BUT THEY'RE
STILL METABOLICALLY HEALTHY.
SO THAT'S THE KEY OBESITY STARTS
SEEING VIRUS IN THE HEART.
IT'S TRULY REPLICATING IN THE
HEART.
THIS IS WHAT THE LUNGS LOOK LIKE
WHERE YOU CAN SEE IN THE TOP AT
3 DAYS POST INFECTION, THE
PATHOLOGY THAT'S HAPPENING THAT
EXPLAINS THE INCREASED CLINICAL
DISEASE, INCREASE INFLAMMATORY
RESPONSE AND THAT'S WHAT THE
LUNGS END UP LOOKING LIKE.
AND WE DEVELOPED, I THINK WE'RE
OVER A HUNDRED NOW, VALIDATED
PRIMER PROBES TO LOOK AT LUNG
REPAIR INJURY, INFLAMMATORY
RESPONSES, SO IF ANYBODY WANTS
THOSE, THOSE ARE AVAILABLE TO
YOU EMPLOY DOES IT MATTER BEYOND
DISEASE SEVERITY?
ABSOLUTELY, WHEN YOU LOOK AT
TRANSMISSION OR SUSCEPTIBILITY,
TOO, THOSE METABOLICALLY
UNHEALTHY FERRETS ARE MORE
LIKELY TO GET INFECTED EVEN WITH
ZOANNOTTIC STRAINS OF VIRUS,
THEY SPREAD MORE.
THEY SHRED MORE MORE VIRUS, SHS
LONGER, AND IT'S IT DOESN'T
MATTER THE STRAIN OR FLU.
YOU ALSO GET CHANGES TO THE
VIRUS ITSELF, WHICH MAKES SENSE.
HAVE YOU A LOT MORE VIRUS
REPLICATING LONGER IN NISSAN
MALS AND WHAT IT LEADS TO IS A
CHANGE INLET VIRAL QUASI
SPECIES.
THERE'S FEWER DEFECTIVE
PARTICLES, THERE'S ACTUALLY MORE
CHANGES WHEN YOU LOOK AT SINGLE
NUCLEOTIDE VARIANTS INCLUDING IN
AREAS OF THE GENOME THAT WE KNOW
ARE IMPORTANT FOR ANTIGENIC
DRIFT.
WE ALSO SEE MORE RESORT ANTS
HAPPEN IN THESE ANIMALS AND THIS
IS NOT FERRET SPECIFIC, IT
HAPPENS ALSO IN MICE.
WE KNOW THE SAME WAS TRUE WITH
SARS.
MICHAEL HAD DONE SARS AND
OBESITY IN MICE, SHOWED IT'S
WORSE.
WE REPEATED IT AND SAW EXACTLY
THE SAME THING INCLUDE NOTHING
OUR OBESE FERRETS, THEN WE
DECIDED TO DO RSV, FIGURED THIS
HAS TO BE THE SAME THING, THIS
IS GOING TO BE A GENERAL
PRINCIPAL OF OBESITY AND AT
LEAST PREEMILYINARLY THAT IS NOT
WHAT WE'RE SEEING, WHAT WE'RE
ACTUALLY SEE SUGGEST RSV IN THE
FERRET, THERE'S ACTUALLY--AND
THIS IS PRELIMINARY--BUT APPEARS
TO BE LESS VIRUS GETTING TO THE
LUNGS OF THESE ANIMALS.
AND MORE VIRUS STAYING IN THE
UPPER RESPIRATORY TRACT AND
ACTUALLY SHEDDING IT A BIT
LATER.
SO WE'RE NOT DOING COTTON RATS
THERE, 'S NO WAY IN--BUT WE WILL
DO OBESE MICE, AND WE DID THOSE
THINKING OKAY, THIS IS WHERE
RUBBER HITS THE ROAD, THESE GUYS
WILL DROP DEAD, NOT SO MUCH.
ACTUALLY IN OBESE MICE, RSV IS
BETTER.
SO IF YOU LOOK AT THE PERCENT
WEIGHT CHANGE, DAYS POST
INFECTION, HERE'S YOUR LEAN
ANIMALS WHERE WE'RE USING 10 TO
THE SEVENTH RSV WHERE WE CAN GET
DISEASE AND THIS HAS BEEN
REPRODUCIBLE IS THE DIET INDUCED
OBESE ARE NOT AS SICK AND WE SEE
THIS, THERE'S LESS VIRUS IN THE
LUNGS, IT'S LOW BUT WE'RE
REPEATING THIS LOOKING AT
DIFFERENT TIME POINTS AND THE
SPREAD, BECAUSE THAT'S THE KEY
IS GETTING WITH RSV IS GETTING
DOWN TO THE ALVEOLAR SPACES AND
LISTING THAT IMMUNO PATHOLOGY
AND WE'RE LOOKING AT SPREAD BUT
WE KNOW FROM FLOW CYTOMETRY
WE'RE SEEING LESS IMMUNO
PATHOLOGY IN THESE MICE BECAUSE
IT'S DELAYED AND FOR FLU THAT'S
BAD, FOR SARS THAT'S BAD, FOR
RSV IT'S ACTUALLY GOOD.
SO THIS MAKE ITS REALLY
INTERESTING THAT IT MAY NOT JUST
BE THE BROAD STROKE.
SO IT'S SOMETHING WE'RE HAVING
TO LOOK AT AND THINK ABOUT.
AND THIS IS JUST SHOWING YOU IN
THE MICE, YOU CAN SEE JUST A LOT
LESS PATHOLOGY, AND AGAIN IT'S
BECAUSE WE'RE GETTING LESS OF
THOSE IMMUNE CELL INFILTRATES,
PARTICULARLY NUTRIFILLS, MA
ROUGH ATOM PHAGES AND T-CELLS.
SO, THIS HAS BEEN A LITTLE BIT
SURPRISING AND WE DON'T KNOW
ANYTHING, AT LEAST I HAVEN'T
SEEN ANYTHING ABOUT OBESITY AND
RSV, BUT WE'RE DIGGING THROUGH
ACTUALLY CLINICAL INFORMATION
AND METADATA.
SO ALSO WHEN YOU THINK ABOUT
INFECTION, DO YOU HAVE ANOTHER
VIRUS IN THERE?
AND THE ANSWER IS ABSOLUTELY.
WE KNOW IN OUR FERRETS IF THEY
HAVE RSV, THEY CAN STILL GET FLU
AND BOY IS IT BAD.
IT'S BAD, BAD.
EVEN IF THE RSV ARE 3 OR 4
MONTHS BEFORE, IT REALLY MAKES A
BIG DIFFERENCE.
CONTRAST IF THE FERRETS HAVE FLU
OR HAD FLU, THEY DON'T GET RSV.
IT'S ALMOST BLOCKED AND WE KNOW
THAT THERE'S INTERACTION BETWEEN
RSV AND SARS, AND WE NOW HAVE
DATA ON SOME OF THESE OTHER
RESPIRATORY VIRUSES, BUT ALSO
BACTERIA, WE'VE DONE WORK WITH
SEEMA, ON BACTERIA AND WE HAVE A
GROUP AT ST. JUDE THAT WILL HELP
US LOOK AT FUNGUS, SO WHATEVER
YOU SEE IN HUMAN IN THOSE
HOUSEHOLD TRANSMISSION MODELS,
CAN YOU THEORETICALLY MODEL THIS
IN THE FERRETS.
PREGNANCY IS ANOTHER 1 AND WE
THINK THIS IS REALLY
INTERESTING, IS, YOU KNOW
PREGNANT WOMEN, HIGH, HIGH, RISK
FOR FLU, HIGH RISK FOR
SARS-COV-2.
ENCOURAGE THEM TO GET VACCINATED
AND WHEN WE VACCINATE WE HAVE
LOTS OF DIFFERENT FLU VACCINES,
WE HAVE LIVE VIRUS, THE
ENACTIVATED FLU MIST,
RECOMBUPENORPHINE ANT HA BASED
AND WHEN YOU TAKE A PREGNANT
MOUSE AND YOU VACCINATE IT, LET
THAT MOM GIVE BIRTH AND THEN YOU
CHALERG HER, YOU THINK THEY'RE
ALL GOING TO BE PROTECTED
BECAUSE IF IT'S NOT A PREGNANT
MOUSE, JUST A FEMALE, CAN YOU
PROTECT THEM WITH ALL THESE
VACCINES, NOT THE CASE IF THAT
MOM IS PREGNANT WHEN SHE GETS
VACCINATED.
OUR ENACTIVATED VACCINE DOES NOT
WORK WELL AT ALL.
WHERE THE OTHER 1S, AND YOU HAVE
TO USE A REALLY LOW DOSE OF
FLEW, WHEN YOU CHALLENGE THEM,
IT DOESN'T WORK WELL, SO THERE'S
SOMETHING VERY DIFFERENT ABOUT
THAT PREGNANCY STATE THAT
CHANGES HOW YOUR VACCINE
PLATFORMS BEHAVE AND THAT HOLES
TRUE THEN WHEN YOU LOOK AT HOW
DO THOSE PUPS BORN TO THAT
VACCINATED MOM, HOW DO THEY
SURVIVE THAT CHALLENGE.
THAT PLATFORM MAKES A DIFFERENCE
IN HOW THAT MOM RESPONDS AND
THIS WAS SOMETHING WE HADN'T
REALLY EXPECTED TO SEE, WE KNEW
THERE WOULD BE MORE SUSCEPTIBLE
TO INFECTION BUTEE DIDN'T EXPECT
TO SEE THAT THE VACCINE PLATFORM
WAS MAKE SUCH A BIG DIFFERENCE
IN A VACCINATED--IN A PREGNANT
VERSUS A NONPREGNANT FEMALE
MOUSE.
SO I THINK THE NEXT QUESTION IS,
CAN WE IMPROVE, OR WHAT CAN WE
DO TO IMPROVE?
AND THERE'S A LOT OF THINGS WE
CAN DO.
THERE'S MANY, MANY NEW FUN
MODELS COMING OUT.
RYAN'S GOING TO TALK ABOUT HIS
DIRTY MICE, WHICH I THINK HAVE
BEEN A GAME CHANGER IN A LOT OF
WAYS FOR UNDERSTANDING MAYBE
SOME OF THE ISSUES WE NEED TO
CONSIDER IF YOU'RE USING SPF
MOUSE AND WHAT YOU'RE REALLY
MODELING IN TERMS OF THE
IMMUNITY.
AND OF COURSE I LIKE IT BECAUSE
SHE'S SHOWN THAT A LOT OF THOSE
SORT OF INDEMMIC MOUSE ENTERIC
PATHOIENS END UP BEING REALLY
IMPORTANT WITH THE RESPIRATORY
VIRUS INFECTIONS.
WE CAN MANIPULATE OUR
TRANSMISSION MODELS, AND SEEMA
WILL TALK ABOUT, I ASSUME YOUR
DAY CARE TRANSMISSION MODEL
WHICH IS REALLY COOL AND FORCES
YOU TO THINK ABOUT HOW WE'RE
DOING THIS BECAUSE HISTORICALLY
WITH FLU THE WAY WE'VE DONE
TRANSMISSION, DIRECT CONTACT OR
RESPIRATORY, IS YOU INFECT THAT
INDEX, 24 HOURS LATER, YOU PUT
THE OTHER, THE NAIVE FERRET IN
CONTACT AND THEY JUST SIT THERE
FOR OVER LONG THAT YOU ARE GOING
TO BE THERE, WHICH I GUESS IT
COULD BE A HOUSEHOLD MODEL, BUT
YOU ARE IN CLOSE INTIMATE
CONTACT WITH THAT OTHER FERRET
FOR 14 DAYS.
AND THEN ZONDA WANG WILL TELL US
MORE ABOUT THE FANCY GINY PIGS
AND AN INTERESTING MOUSE MODEL
SHE'S MAKING.
WE'VE ALSO WORKED ON A NUMBER OF
IMPROVED TRANSMISSION MODELS,
FIRST WE'RE CALLING THE SLEEP
OVER WHICH IS BASIC BASIC LYE E
THAT INFECTED INDEX AND WE DO IT
FOR FLU AND RSV.
ALTHOUGH RSV IS A VERY DIFFERENT
TRANSMISSION TIMING, MAKING IT A
BIT MORE CHALLENGING, MAKE IT 1
DAY YOU HAVE TO WAIT 3 OR 4 DAYS
BEFORE YOU PUT THAT CONTACT IN,
BUT THE SLEEP OVER IS YOU HAVE
YOUR INFECTED INDEX YOU PUT YOUR
CONTACT IN, YOU LET THEM SPEND
THE NIGHT TOGETHER AND YOU PULL
THEM OUT.
SO SLEEP OVER.
WE ALSO HAVE 1 WE CALL IT
KROGER, BECAUSE THAT'S OUR
FAVORITE GROCERY STORE, CAN YOU
CALL IT PUBLIC, WHEREVER YOU
ARE, PIGGLY WIGGLY, THAT'S A
SHORTER TRANSMISSION PERIOD,
THAT'S REALLY FORCING THE ISSUE
WHETHER IT'S DIRECT OR
RESPIRATORY CONTACT, YOU'RE
TALKING ABOUT A LESS THAN 4 HOUR
EXPOSURE TIME SO YOUR REALLY
FORCING THAT VIRUS OR THAT HOST
TO TRANSMIT ASK TELL YOU IT
WORKS BEAUTIFULLY WITH THE OBESE
FERRETS WITH ALL THE STRAINS
WE'VE TESTED.
AND THEN WE HAVE 1 THAT WE
AFFECTATELY IN THE LAB CALL
THIS, WHICH I WON'T SAY.
IT'S BASICALLY IF YOU ARE OUT IN
PUBLIC AND YOU'VE BEEN
VACCINATED OR YOU'RE ON TAMIVIR,
OR--YOU KNOW, WHATEVER YOU WANT,
AND YOU WERE EXPOSED TO SOMEBODY
WHETHER AT A SLEEP OVER OR AT
YOUR LOCAL GROCE REAR STORE THAT
HAS NOT BEEN VACCINATED OR
TREATED OR WEARING A MASK, ARE
YOU GOING TO BE PROTECTED WITH
THAT THERAPEUTIC IF THAT PERSON
IS SHEDDING SOMETHING?
AND THAT'S REALLY INTERESTING
BECAUSE YOU WOULD ASSUME, YES,
IF THINGS ARE WORKING RIGHT BUT
SURPRISINGLY IT'S NOT QUITE THE
CASE AT LEAST IN THE ANIMAL
MODELS.
THE OTHER THINGS WE'VE BEEN
WORKING ON ARE REALLY AROUND
OBESITY AND THAT HAS BEEN MAKING
A METABOLICALLY HEALTHY OBESE
MOUSE AND A METABOLICALLY
UNHEALTHY LEAN MOUSE, THE
UNHEALTHY LEAN A LITTLE BIT
EASIER, YOU CAN BASICALLY DO
FOOD WITHDRAW FOR ABOUT 24-48
HOURS IF YOUR [INDISCERNIBLE]
WILL LET YOU AND YOU GET INTO A
MALNOURISHED STATE PRETTY
QUICKLY.
THIS IS INTERESTING, THIS IS
DBA, BALB-C, WE'VE DONE 6 J AND
6 N, MALE ANDS FEMALES YOU CAN
SEE ON A HIGH FAT DIET THEY ALL
GAIP WEIGHT, IN FACT THE DBAs
ARE REALLY FAST, THEY TRIP
NELSON CEASAR SIZE WITHIN ABOUT
A MONTH.
BUT UNLIKE THE 6 J AND UNLIKE
THE C57 BLACK 6 J, I DON'T CARE
HOW BIG WE MAKE THESE GUYS, AND
THEY'RE LIKE 80 GRAPS THEY'RE
LIKE WATER BUFFALO, THEY'RE
HEALTHY, THEY CONTROL THEIR
GLUCOSE, AND IF YOU WERE TO GO
IN AND INFECT THEM YOU SAW NO
DIFFERENCE IN DISEASE SEVERITY,
IF YOU VACCINATE THEM, THE
VACCINES STILL WORK BEAUTIFULLY,
WHICH IS NOT TRUE IN THE
METABOLICALLY UNHEALTHY ANIMAL.
WE--PEOPLE IN THE LAB, THEY WANT
TO DO JAMBA JUICE DIETS, THEY
WANT TO DO ATKINS DIETS, THEY
WANT TO DO LIPO SECTION, WHICH,
NO, WE'RE NOT DOING THAT BUT WE
HAVE GOTTEN INTO HOW DOES DIET
OR OZEMPIC, OR AND METFORMIN,
YOU CAN HAVE THE BIGGEST
HEAVIEST MOUSE ON THERE AND YOU
PUT THEM ON METFORMIN, AND IT
WORKS BEAUTIFULLY.
SO JUST WORKING OUT THE NEW
MODELS AND THIS IS THE 1 WE'VE
BEEN THINKING ABOUT A LOT IS
THIS CHILDHOOD OBESITY BECAUSE
THIS IS THE GROUP THAT YOU'RE
SEEING THE MOST DRAMATIC
INCREASE IN OBESITY AND THERE'S
DEFINITELY HEALTH DISPARITY
ASPECTS TO THIS, SOCIOECONOMIC
ISSUES, BUT THIS IS THE GROUP
THAT REALLY IS, BUT HOW DO YOU
MODEL THAT IN A MOUSE, RIGHT?
OR IN A FERRET BECAUSE IN ALL OF
THESE YOU'RE STARTING WITH 3-6
WEEK OLD, PUTTING THEM ON DIETS
FOR 12 + WEEKS SO YOU'RE GETTING
INTO ADULT OR YOUNG ADULT.
SO WHAT WE'VE DECIDED TO DO IS
LET'S HAVE THEM BE BORN TO
PARENTS OF DIFFERENT AND AGAIN
IT'S METABOLIC STATE, SO
TYPICALLY YOU HAVE A
METABOLICALLY HEALTHIER LEAN MOM
AND DAD, WHAT HAPPENS IN BOTH OF
THEM ARE METABOLICALLY
UNHEALTHY, OR 1 OR THE OTHER
PARENT HAS AN ISSUE.
AND THESE PUPS AT BIRTH, EVEN AT
WEANING, THERE ARE DRAMATIC
DIFFERENCES IN THESE ANIMALS.
SO MOST--AND WE'RE DOING IT IN
C57 BLACK 6, MOST WEANLINGS ARE
14, 15, CANE GRAMS, SOME OF
THESE ESPECIALLY IF THE MOM IS
METABOLICALLY UNHEALTHY, AT THAT
3 WEEKS OF AGE, THEY CAN BE
30-GRAMS AND METABOLICALLY
UNHEALTHY.
AND SO WE THINK WE WILL ARE A
MODEL WHERE WE WILL BE ABLE TO
WORK WITH THEM RIGHT AWAY AND
THERE IS FOR--THERE IS HUGE SEX
BIAS WHEN YOU TART LOOKING AT
THIS.
I MEAN IT'S REALLY DRAMATIC HOW
THE FEMALE OFFSPRING BEHAVE
COMPARED TO THE MALES, AND
REALLY TRACKS BACK TO WHAT THAT
MOM'S METABOLICITATE WAS.
FUN NEW MODELS, HUMANS, RIGHT?
WE HAVE A LOT OF COLLEAGUES
DOING HUMAN COHORTS, WE HAVE
HIP-C CENTERS, THAT'S ALL THEY
DO IS HUMAN IMMUNOLOGY.
AND THE QUESTION BECOMES CAN WE
PARTNER UP WITH THEM AND LEARN
MORE ABOUT WHAT THEY'RE SEEING
ESPECIALLY UPPER MUCOSAL, YOU
KNOW MUCOSAL, AND CAN WE TAKE
THAT INFORMATION BACK AND TRY
AND MODEL IT IN THE ANIMALS TO
REALLY UNDERSTAND WHICH ANIMAL
OR ANIMALS WILL BE THE BEST
MODEL TO ACTUALLY TRY AND
RECAPITULATE WITH WHAT THEY'RE
SEEING, EVEN IN THE COMPLEXITY
OF A HUMAN AND I THINK THAT'S
SOMETHING THAT MAY BE
COMMUNICATIONS BETWEEN THESE
DIFFERENT GROUPS, PEOPLE FOCUS
SOLELY ON HUMAN IMMUNITY AND
THOSE OF US DOING MUCH MORE OF
THE ANIMAL MODELS, GETTING US
TOGETHER GETTING IN THE SAME
ROOM TO SAY, HOW DO WE PULL THIS
TOGETHER BECAUSE AGAIN, RATHER
THAN JUST GOING FORWARD, IS THIS
TRANSLATABLE OR HOW CLOSE ARE WE
TO CLINIC, CAN WE BRING THAT
BACK?
AND THEN I WOULD ARGUE IF YOU
REALLY WANT TO LOOK AT THE
COMPLEXITY OF PATHOGENESIS,
IMMUNE RESPONSE, YOU HAVE TO DO
IT IN A WHOLE ANIMAL.
YOU KNOW WE HAVE THESE GREAT
MORE AND MORE TIPS, 2 Ds, AND
3D CULTURES.
LOTS OF COOL ENGINEERING
ORGANOIDS, ANAROIDS, ORGANS ON A
CHIP, WHICH WE JUST GOT IN THE
SYSTEMS AND THEY ARE INCREDIBLE,
AND YOU CAN MAKE THEM FOR ANY
ORGAN, YOU CAN MAKE THEM FOR
HUMANS, YOU CAN MAKE THEM FOR
ANIMALS, YOU JUST NEED THOSE
PRIMARY CELLS, CAN YOU FLOW IN
IMMUNE CELLS BUT IT'S NEVER
GOING TO BE THE SAME AS A WHOLE
ANIMAL.
IT WILL HELP US GET MUCH, MUCH
MORE MECHANISTIC ESPECIALLY
ABOUT THOSE NONIMMUNE CELLS AND
THOU THEY RESPOND TO INFECTION
EMPLOY SO I THINK THINGS LIKE
THE HUMAN LUNG ON A CHIP IS
GOING TO BE GREAT TO SEE HOW
MIGHT LIVE ATTENUATE VACCINE
THAT SHOULDN'T GO TO THE LUNG,
BUT WHAT IF IT DID, WHAT'S IT
GOING TO DO, HOW ARE SOME OF
THESE NEW THERAPEUTICS GOING TO
WORK IN THE HUMAN LUNG BECAUSE
THESE LUNG ON A CHIP, THEY
BREATHE, YOU HAVE WHE
KAN--KANAICAL STRESS, HAVE YOU
BLOOD GOING THROUGH THEM SO THEY
WILL BE GOOD FOR THINGS LIKE
THAT YOU ABOUT WILL THEY
RECAPITULATE WITH AN ANIMAL
MODEL?
MAYBE SOMEDAY BUT NOT THERE YET,
BUT ALSO HAPPY FOR FRIENDLY
DISCOURSE.
WAYS FORWARD.
THIS IS A BIGGIE, REAGENTS,
RIGHT?
WE'RE VERY MUCH LIMITED IN A LOT
OF WAYS BY THE REAGENTS IF WE
WANT TO LOOK AT IMMUNE RESPONSE,
IF YOU WANT TO DO PATHOGENESIS,
VIRAL TITERS, HISTOPATHOLOGY,
THEY'RE ALL GREAT BUT IF YOU
REALLY WANT TO START PULLING
APART MECHANISM, WE NEED A LOT
MORE REAGENTS AND THAT CAN
INCLUDE OBVIOUSLY ANTIBODIES BUT
PRIMER PROBES, IF WE CAN GET THE
RIGHT SETS OF REAGENTS, WORKING
WITH SOMEBODY LIKE MILLI PLEX TO
MAKE MORE AND MORE OF THESE
LUNINEX PANELS TO HAVE HIGH
THROUGH PUT.
HAVING THESE CELLS FROM
DIFFERENT SPECIES WILL HELP A
LOT SO IF ANYBODY WANTS ANY COW,
RESPIRATORY OR MAMMARY
EPITHELIAL CELLS, LET US KNOW,
WE HAVE PLENTY OF THEM AND EVEN
FROM PEOPLE.
SO THERE'S A LOT OF PEOPLE
WORKING ON REAGENTS, SO THIS NOT
A NEW INITIATIVE, RIGHT?
WE HAVE DMID, WE HAVE DATE
PEOPLE, WE HAVE PEOPLE,A ROUND
THE WORLD WORKING ON REAGENTS,
BUT WE DON'T HAVE A CONSOLIDATED
WAY OF KNOWING WHO'S WORKING ON
WHAT, WHAT REAGENTS ARE ACTUALLY
AVAILABLE.
THERE ARE SOME THROUGH BEI,
THAT'S GREAT BUT HAVE THEY BEEN
VALIDATED, I CAN TELL YOU IN
MOST CASES THOSE FERRET REAGENTS
ARE NOT GREAT.
YOU KNOW TRYING TO GET REAGENTS
FROM OTHER PEOPLE, ESPECIALLY
OTHER PARTS OF THE WORLD, YOU'RE
REALLY STUCK BY MTAs, LICENSE,
YOU CAN'T GET A LOT OF THESE
REAGENTS THAT HAVE BEEN CREATED
BECAUSE THEY'RE LOCKED UNDER
TECH TRANSCRIPT.
SO I THINK WOO WE NEED TO THINK
ABOUT WHAT NEEDED AND I THINK
THAT'S SOMETHING I HOPE WE CAN
GET FROM THIS IS MORE DISCUSSION
FROM YOU GUYS ABOUT WHAT DO WE
REALLY NEED AND HOW DO WE FOCUS
ON THAT?
WHERE SOME THEY BE STORED?
THAT'S A HUGE 1.
I THINK THEY SHOULD ALL BE
STORED AT BEI, I THINK BEI
THINKS OTHERWISE, RIGHT IN THIS
IS A LOT.
YOU HAVE TO THINK ABOUT WHO'S
GOING TO HAVE FREEZERS THAT THEY
CAN STORE ALL OF THESE REAGENTS
AND BASICALLY BE AMAZON AND
WE'VE DONE THIS FOR OUR PRIMARY
CELLS AND IT REALLY IS LIKE
BEING AMAZON, WE WILL GET
REQUESTS DAILY AND SO WE'VE
TALKED TO ERIC AND MICHELLE
ABOUT CAN WE PLEASE PUT THESE IN
BEI SO WE DON'T HAVE TO DROP
EVERYTHING AND SHIP CELLS EVERY
OTHER DAY.
AND THAT'S A LOT, JUST THE
STORAGE.
HOW SHOULD THE INFORMATION BE
COMMUNICATED?
THE ONLY WAY PEOPLE KNEW ABOUT
OUR PRIMARY CELLS WAS THROUGH
PUBLICATIONS, RIGHT IN AND THEY
WERE LOOKING FOR IT, THERE'S GOT
TO BE A BETTER WAY, IT CAN BE
THE JUST BE BASED ON
PUBLICATIONS BECAUSE A LOT OF
THESE YOU WILL NOT PUBLISH, WHAT
ABOUTA ALL THE COMMERCIAL
ANTIBODIES, ALL OF US HAVE
TESTED THAT DON'T WORK EMPLOY
RIGHT?
YOU SAY, OH THEY'RE SUPPOSED TO
WORK AGAINST EVERY SPECIES, WELL
THEY DON'T WORK AGAINST MY
FERRET WHEN YOU DO PBMCs, THAT
INFORMATION IS CRITICAL AND
NEEDS TO BE SOMEWHERE SO WE'RE
NOT ALL CONSTANTLY REINVENTING
THE WHILE.
BECAUSE THEN YOU TALK TO
SOMEBODY AND SAY, OH YEAH, TRIED
THAT 3 YEARS AGO AND IT DOESN'T
WORK, SO YOU TYPICALLY DON'T
PUBLISH THINGS LIKE THAT EITHER.
ARE REAGENTS BEING VALIDATED?
AND WHAT DO WE MEAN BY
VALIDATION?
WHAT DOES THE COMMUNITY WANT TO
SEE, WHAT ARE YOU GOING TO BE
COMFORTABLE WITH IF WE SAY THIS
IS A VALIDATED REAGENT.
IS IT SIMPLY THAT FLOW CYTOMETRY
WORKS WITH PBMCs OR DO YOU
WANT MASS SPEC OR AMINO ACID
SEQUENCING, DO YOU WANT TO KNOW
WHAT THIS THING IS BINDING TOO.
AND WHAT ARE THE MODELS NEEDED.
WE WILL HEAR A LOT THAT DURING
THIS WORKSHOP AND IT'S REALLY
EXCITING BUT IF WE'RE THINKING
OUTSIDE OF THE BOX, WHAT ARE THE
NEW MODELS THAT WE NEED TO BE,
WORKING ON.
AND THEN COMING BACK TO THOSE
VIRAL STRAINS THAT WE ALL GET A
LITTLE BIT POOPY AT STUDY
SECTION SAYING I DON'T WANT TO
SEE PR8, I DON'T WANT TO SEE
X31, I DON'T WANT TO SEE RSV IN
A MOUSE, WHAT'S ACCEPTABLE AND
HOW DO THOSE NEED TO BE VALIDATE
EXCLUDE IF IT'S ACCEPT TO USE
PR8, THEN MAYBE WE DON'T NEED TO
BE POOPY AT STUDY SECTION, WE
SAY, AH, PR8 IS THE BEST 1 SO I
THINK THAT ALSO NEEDS TO BE
SOMETHING WE THINK ABOUT MORE
FOR THE WHOLE COMMUNITY BECAUSE
IF IT'S NOT ACCEPT, THEN THERE
NEEDS TO BE A CLEARER MESSAGE,
YOU KNOW, THERE IS A TIME AND
PLACE FOR THIS, BUT WE'VE MOVED
ON FROM THERE AND DO WE NEED
VALIDATED MAMMALIAN ADAPTED
VIRUSES OF SOME KIND.
SO THE BIG THING IS WE REALLY
NEED TO WORK TOGETHER AS A
COMMUNITY TO FIGURE OUT THE WAYS
FOR IT ESPECIALLY FOR THESE
REAGENTS AND HOW DO WE, HOW DO
WE FIGURE OUT WHAT'S NEEDED AND
GET IT SOME PLACE WHERE PEOPLE
CAN GET IT AND THAT THEY KNOW
IT'S AVAILABLE.
AND THEN OF COURSE, APOLOGIES
FOR ANYBODY'S STUFF THAT I
MISSED OR I MISSPOKE, THAT'S ON
ME.
AND WITH THAT, THANK THE LAB AND
THE FERRETS AND THE MICE AND ALL
OF YOU GUYS FOR LISTENING TO
STUFF YOU PROBABLY ALREADY
ANYHOW AND HAPPY TO ANSWER ANY
QUESTIONS AND HOPEFULLY HAVE
SOME DISCUSSION.
>> THAT WAS REALLY GREAT.
THANKS IT'S A WONDERFUL OVERVIEW
AND FUN TO SEE THE NEW STUFF.
I'M CURIOUS ABOUT SINGLE CELL
RNA SEQUENCING IN THE LESS
TRADITIONAL ANIMAL MODELS, THAT
SHOULD DEMOCRATIZE THE STUDY
ESPECIALLY FOR IMMUNOLOGY, ARE
THE GENOMES GOOD ENOUGH FOR
FERRETS, GINNY PIGS, HAMSTERS TO
GET HIGH QUALITY ANALYSIS.
>> YEAH, THAT'S A GREAT
QUESTION, RYAN, I THINK AND
STANLEY, OTHER PEOPLE, HAMSTER
HAS BEEN SEQUENCED BOTH SYRIAN,
GOLDEN.
FOR THE FERRET, YEAH, THERE'S
GENOMES OUT THERE BUT WE STILL
HAVE AN ANNOTATION PROBLEM.
IT'S GETTING BETTER, IT'S
GETTING A LOT BETTER BECAUSE YOU
HAVE PEOPLE THAT ARE FOCUSED ON
ANNOTATING IT APPROPRIATELY, SO
YEAH, WE FEEL MORE COMFORTABLE
NOW GOING IN AND WE'RE DOING
SINGLE CELL SEQUENCING ON UPPER
VERSUS LOWER RESPIRATORY TRACT,
THE PROBLEM COMES WHEN YOU WANT
TO VALIDATE ANYTHING SO WE GET A
LOT OF HITS AND A LOT OF DATA
BUT THEN YOU HAVE TO DEVELOP THE
TOOLS TO BE ACTUALLY BE ABLE TO
VALIDATE WHAT YOU'RE SIGHING BY
THE SINGLE CELL SEQUENCING
MAKING PRIMER PROBES EASY BUT
WHEN YOU GOET INTO PROTEINS OR
ANTIBODIES IT GETS MUCH, MUCH
TRICKIER AND THAT'S WHERE YOU
KIND OF GET STUCK IF YOU GO TO
PUBLISH THAT, OKAY, WELL WE
CAN'T VALIDATE THIS.
SO THE I THINK THE GENOMES ARE
BETTER--DOES ANYBODY KNOW ABOUT
THE HAMSTER OR ANY OF THE OTHER
1S GO,A HEAD?
YEAH, YOU'RE PRETTY MUCH SPOT ON
WITH IT, WE'RE WORKING ON OUR
SECOND ANNOTATION FOR THE
HAMSTER BUT THEY'RE GETTING
BETTER ABOUT YOU THERE ARE CHRIS
CREPANCYS BETWEEN THE 2
ANNOTATIONS THAT ARE EXISTING
AND HOPEFULLY SOME MORE DATA
WILL RESULT IN ANNOTATIONS AND
IT'S GETTING BETTER AND I THINK
IN SINGLE CELL HAMSTERS SO FAR,
YOU CAN IDENTIFY CELL TYPES AND
GET NICE USEFUL INFORMATION OUT
OF IT.
NI'VE NEVER DONE THIS BEFORE, SO
I WILL MAKE MORE OF A COMMENT
THAN A QUESTION, I REALLY HATE
ALL THE PR8 THAT HAPPENS BECAUSE
THERE ARE NO NATURAL ORTHOTHOUGH
NECKS O VIRUSES, IN US SO IN MY
OPINION, CAL9 OR UDON OR
WHATEVER IS YOUR FAVORITE FLU IS
NO DIFFERENT THAN PR8, AND PR8
HAS ADVANTAGES OVER THESE OTHER
STRAINS SO I REALLY--IT ALWAYS
MAKES ME CRINGE DEEPLY WHEN
THOSE CONCERNS COME UP AT STUDY
SECTION OR REVIEWER CONCERNS THE
PAPER.
>> SO NOTE TO EVERYBODY ONLINE
MAKE SURE RYAN'S ON STUDY
SECTION IF YOU'RE GOING TO
SUBMIT YOUR PR8.
>> CAN I RESPOND TO THAT.
>> I CAN COMMENT.
>> YEAH, NO, I AM A PR8 HATER AS
WE HAVE HAD MANY DISCUSSIONS ON
AND I--I WILL SAY WHY I THINK
PR8, WE NEED TO MOVE ON FROM.
IT IS NOT BECAUSE IT IS A MOUSE
ADAPTED VIRUS, IT IS BECAUSE IT
DOES NOT RECAPITULATE THE THINGS
THAT WE'RE TRYING TO MODEL IN
HUMANS, RIGHT, SO IF YOU WANT TO
USE A MOUSE ADAPTED STRAIN OF
INFLUENZA, FABULOUS, USE 1 THAT
IS RELEVANT TO WHAT IS
CIRCULATING IN HUMANS AND THAT
HAS A SIMILAR SORT OF PHENOTYPIC
MANIFESTATION.
PEOPLE ARE NOT DROPPING DEAD
FROM 50 PFU OF AN INFLUENZA
VIRUS, AND AT SOME POINT FORWARD
THEY MAY BE DEAD NTHEY MAY BE
DEAD.
OTHER PEOPLE I'VE INFECTED DO
NOT DIE SO I WILL SAY IT TAKES A
LOT MORE THAN THAT BUT--NO, BUT
THAT'S MY POINT.
MY POINT IS THAT--WE SHOULDN'T
USE PR8 BECAUSE OF THE MOUSE
INFECTED VIRUS BUT THAT SHE
SHOULD BE MORE THOUGHTFUL ON
WHAT IS THE QUESTION AND WHAT IS
THE BEST SYSTEM TO ANSWER THAT
QUESTION.
IF YOU'RE USING MICE, LET'S USE
A MORE RELEVANT STRAIN, WELL
THE'S THINK ABOUT SOMETHING
BETTER.
>> ISICAL 09 A PR8.
>> SUPER QUICKLY, BUT PR8 CAN
MODEL A LOT OF THINGS THAT DO
HAPPEN IN PEOPLE, WE CAN DISCUSS
THIS OFFLINE BUT I DO THINK
THERE'S A LOT OF VALUE IN THAT
MODEL SYSTEM.
NMORE DISCOURSE.
>> I SECOND THAT, I THINK PR8 IS
GREAT, BUT ALSO IT DEPENDS ON
THE QUESTION, AND I THINK THE
QUESTION IS MORE IMPORTANT TO
FOCUS ON SO IF YOU'RE QUESTION
IS ABOUT HUMAN STRAINS, CLEARLY
YOU CAN'T USE A MOUSE TO MODEL
THOSE.
BUT IF YOU'RE QUESTION IS ABOUT
HOST FACTORS AND UNDERSTANDING
THE CONTRIBUTION TO PATHOGENESIS
OR TRANSMISSION, AT THIS MOMENT
WE DON'T HAVE GOOD REAGENTS FOR
FERRETS OR GINNY PIGS SO WE RELY
ON MICE TO UNDERSTAND THOSE
MECHANISMS, AND UNFORTUNATELY WE
DO HAVE TO USE STRAINS THAT WORK
IN THAT MODEL.
SO I THINK FOCUSING ON THE
QUESTION IS A MORE IMPORTANT
ASPECT.
>> GOOD QUESTION.
I'M STILL SENDING MY GRANTS TO
YOU THAT INCLUDE PR8.
>> SO I HAVE A QUESTION, YOU
WORK AT A HOSPITAL WITH IMMUNO
COMPROMISED CHILDREN, DO YOU
EVER OR LET ME CHANGE THAT--A
GOOD WAY TO DO THINGS THAT WOULD
BE TO CORRELATE FINDINGS THAT WE
FIND IN OUR ANIMAL MODELS THAT
WE TRY TO MAKE THOSE IMMUNO
COMPROMISED WITH WHAT YOU'RE
SEEING IN PATIENT WHO IS HAVE
DECS WHETHER IT BE SARS 2 OR
WHATEVER ELSE, INFLUENZA,
WHATEVER, DO YOU HAVE ANY--DO
HAVE A WAY TO SET THAT UP AT
ST. JUDE SO CAN YOU TEST SOME OF
THE THINGS YOU FIND AND THEN
INFORM WHAT YOU SHOULD BE
LOOKING FOR IN YOUR ANIMAL
MODELS.
NYEAH, THAT'S A GREAT QUESTION.
SO I'M AT ST. JUDE'S CHILDREN'S
RESEARCH HOSPITAL WHICH IS
CATATROPHIC CHILDHOOD ILLNESS
ALWAYS BEEN KNOWN FOR CANCER BUT
NOW THERE'S A LOT OF NEUROLOGIC
AND SICKLE CELL ANEMIA, THINGS
LIKE THAT, HIV, SO THE ANSWER IS
YES, WE DO.
AND IT'S SORT OF--IT SOUNDS
WEIRD BUT IT'S SORT OF A
DISCONNECT FROM MOST OF THE WERE
WE DO BECAUSE WHAT WE IN
ST. JUDE KIDS IS SO UNUSUAL
RIGHT BECAUSE THEY'RE SO IMMUNO
COMPROMISED THAT YOU WILL GET
PERSONS WITH INFECTIONS, CAN YOU
DO A LOT OF REALLY COOL, WELL,
NOT FOR THE KIDS, VIRAL SHEDDING
STUDIES BECAUSE THEY'RE SHEDDING
VIRUS FOR A LONG TIME SO TO BE
ABLE TO TAKE THOSE AND MOODLE
THINGS SO THAT WE'RE 1-2 STEPS
REMOVED FROM THE CLINIC, WE CAN
AND SO WE HAVE MOUSE MODELS THAT
ARE IMMUNO COMPROMISED, THEY'RE
TREATED WITH MEX O TREKSATE,
WE'RE FOLLOWING THE CHEMO
THERAPEUTIC THAT THE KIDS ARE ON
SO THEY'RE IMMUNO COMPROMISED IN
THE SAME WAY, THEY WILL SEE
THESE THINGS AND WE CAN DO THE
SAME THING WITH FERRETS IS YOU
CAN DO [INDISCERNIBLE] DID THIS
ORIGINALLY USING MEXA TREKSATE
TO MAKE IMMUNO COMPROMISED
FERRETS THAT WILL RECAPITULATE
THAT WE SEE IN THE KIDS.
SO YEAH, WE DO THAT FREQUENTLY
ESPECIALLY WHEN LOOKING AT MOUSE
MODELS.
SO THEY'RE AVAILABLE AND THEY'RE
PRETTY EASY ACTUALLY WHAT WE DID
FOR THE MICE ADAPTED REALLY WELL
IN THE FERRETS, I WOULD IMAGINE
YOU CAN DO THE SAME THING OR
SOMETHING SIMILAR FOR HASMSTERS
IF WE WANT TO LOOK AT THEM IN
SARS AS WELL.
>> TEEING OFF THAT THOUGH, I
THINK THERE'S A LOT OF FOCUS ON
MEX O TREKSATE OR CYCLOFOSTER
NURSED MID FOR THE THAT WHICH IS
GREAT FOR THE CANCER PATIENTS
BUT DOES FOCUS ON THE REFLECT
THE SOLID ORGAN DEFECTSAs
WELL, SO AGAIN SOME OF THE
LESSONS MAY BE HELPFULFUL BUT
THERE MAY BE DIFFERENCES
PARTICULARLY WHEN IT COMINGS TO
VACCINES.
>> YEAH XSINGLE CELL WE
ACTUALLY--OR SICKLE CELL WE
ACTUALLY HAVE SICKLE CELL MICE,
SO WE CAN MODEL THAT PRETTY EASY
LE BUT EVEN DOING THINGS LIKE
THOO ZETAMIN, YOU KNOW THE
CANCER PATIENTS ARE ON SO MANYY
ANTIBIOTICS RIGHT TO PREVENT
REJECTION, CAN YOU EVEN DO THOSE
MODELS WHERE YOU TREAT THEM WITH
ANTIBIOTIC ANDS THAT WILL CHANGE
THE PATHOGENESIS OF YOUR VIRAL
INFECTION FOR SURE, JUST BEING
ON THESE CONSTANT ANTIBIOTICS.
>> SO DO YOU HAVE ACCESS-DO THE
PHYSICIANS TAKING CARE OF THOSE
KID ANDS DO THE PARENTS CONSENT
FOR ACTUALLY BEING ABLE TO GET
SPECIMENS THAT AGAIN CAN CROSS
VALIDATE WITH YOUR FINDING IN
ANIMAL MODELS?
>> YEAH, WE DO AND THEY DO THAT
A LOT FOR ACTUALLY LIKE XEIN,
O-TRANSPLANTS SO THEY CAN GET
THAT TUMOR FROM THE PATIENT AND
TRANSPLATE IT INTO A SKID OR NOD
MOUSE AND BE ABLE TO DO THAT SO
IT'S EASIER FOR US TO INCREASE
IN BODY ABLE TO TAKE THE
INFORMATION WE'RE SEEING, YOU
KNOW IF THEY'RE LOOKING
IMMUNOLOGICALLY OR THEY HAVE THE
RIEROUSS FROM THEM, WE CAN BRING
THOSE INTO THE ANIMAL MODELS.
NI JUST WANTED TO KIND OF RAISE
THIS POINT ABOUT THE UNDERLYING
GENETICS OF KIND OF THE FINE O
TYPES WE'RE LOOKING AT IN A LOT
OF THE ANIMAL MODEL STUFF IS
PHENOTYPE FOCUSED, RIGHT IN LIKE
DID MY MOUSE GET SICK, DID MY
FERRET GET SICK, BUT ALL OF THE
ANIMAL MODELS ARE TO A LARGE
EXTENT GENETICALLY SIMILAR, EVEN
OUTBRED ANIMALLINGS LIKE FERRETS
ARE COMING FROM A TRIPLE F
FOUNDER STRAIN OR WHATEVER.
>> THEY HAVE 8 FOUNDER STRAINS,
THAT'S IT, SO YEAH.
>> BUT I WONDER, HOW OFTEN ARE
THE DEC MODELS THAT WE'RE USING
RECAPITULATING A LARGE MAIORITY
OF WHAT WE WOULD SEE IN PEOPLE
VERSUS BEING KIND OF NARROW AND
WHAT YOUR THOUGHTS ARE, MOVING
FORWARD, HOW TO MAKE SURE THE
STUFF THAT WE'RE FINDING IN
THESE ANIMALS APPLIES TO HUMANS
AS MUCH AS POSSIBLE.
NYEAH, I THINK THAT'S A GREAT
QUESTION.
CLEARLY THE COLLABORATIVE COST
THEY HAVE AT UNIVERSITY OF NORTH
CAROLINA WOULD GO A LONG WAYS
TOWARDS LOOKING AT WHAT ROLE THE
GENETICS MIGHT PLAY.
SO THEY HAVE--AND I THINK THOSE
ARE NOW AVAILABLE TO PEOPLE.
>> YEAH, SO CAN YOU GET THEM
FROM THE SYSTEMS GENETICS CORE
FACILITY AT UNC BUT ALSO SOME OF
THE LINES HAVE BEEN SENT TO
JACKS AND YOU CAN GET THEM
THERE, THEY'RE EXPENSIVE BUT--
>> YEAH AND THERE'S A GROUP IN
THE UK THAT'S ALSO MADE
SOMETHING SIMILAR TO THE
COLLABORATIVE CROSS, SOMEBODY IN
GERMANY SO YOU CAN GET THOSE BUT
THAT WOULD BE LOOKING AT HOST
GENETICS, YOU VALID TO USE
SOMETHING LIKE THE COLLABORATIVE
CROSS AND FROM MARK'S WORK IT
DOES MAKE A DIG DIFFERENCE IN
HOW THOSE ANIMALS RESPOND WHEN
YOU'RE LOOKING FOR THE
PHENOTYPES.
HOW DO WE RECAPITULATE SOMETHING
LIKE THAT IN FERRETS.
THAT WILL BE A LITTLE BIT
HARDER, SHORT OF--THAT'S GOING
TO BE A LITTLE BIT HARDER.
BUT DEFINITELY AND I DON'T KNOW
HOW INBRED OUR HAMSTERS AND
COTTON RATS ARE PRETTY INBRED AS
WELL.
>> I THINK THEY'RE TECHNICAL
OUTBRED BUT COMING FROM THE SAME
COLONY, SAME STORY.
>> SO JUST TO COMPLETE THE LOOP
THERE, IS THERE ANY APPETITE FOR
EVALUATING COLLABORATIVE CROSS
MICE LIKE THAT AND SWINGING BACK
AND SAYING AS A FIELD WE DON'T
LIKE BULB C ANYMORE, OR WE DON'T
LIKE FERRETS FROM TRIPLE F, WE
LIKE FERRETS FROM WHEREVER ELSE
AND SAYING HERE'S WHERE WE THINK
A BETTER MODEL WITHIN THE MODEL
ACTUALLY IS, AND MAYBE IT'S
PHENOTYPE SPECIFIC FOR PREGNANCY
OR WHATEVER ELSE.
NI THINK THAT'S A GREAT
SUGGESTION IS TO ABLE TO GET THE
WHOLE SPAN OF COLLABORATIVE
CROPS AND ASK WHAT HAPPENS IN
ALL OF THESE?
WHAT DOES IT LOOK LIKE?
I THINK THAT WOULD BE REALLY
VALUABLE, I'M THINKING
ESPECIALLY SOMETHING LIKE, YOU
KNOW H5 INFLUENZA VIRUS WHERE
MAYBE THERE'S COLLABORATIVE
CROSS MICE THAT WON'T DROP DEAD
IN 4 DAYS WHEN THEY'RE INFECT
WIDE H5.
AND SO WE WOULD NEED TO KNOW
THAT BECAUSE IT WOULD GIVE US
INDICATION THAT THE HOST
GENETICS MAY BE REAL--WHICH WE
THINK IS TRUE ANYWAY WITH H5.
I THINK THERE WOULD BE A LOT OF
VALUE TO DOING SORT OF A GROUP
EFFORT.
MAYBE THERE CAN BE A NEW CENTER
FOR FOR SOMETHING LIKE THAT,
BECAUSE THEN WE CAN EVEN THINK
ABOUT HOW DO YOU THEN TRANSLATE
THAT TO NONMOUSE MODELS.
>> WE DO HAVE A FEW QUESTIONS
FROM ONLINE THAT I DON'T WANT TO
LET GO, THIS WAS FROM THE
BEGINNING OF YOUR TALK FROM RYAN
AT CORNELL, PERSUSCEPTIBILITY,
DO WE HAVE A STRONG OR SUMMARY
ANALYST OF EXPWRIEWKS IN
RESPIRATORY TRACTS FOR EACH
SMALL ANIMAL MODEL?
>> THAT'S A GREAT QUESTION.
I KNOW WE DO, SO YOSHI'S GROUP,
PETE'S SMILING, MICE, CATS,
DOGS, FERRETS, HAMSTERS, DO YOU
HAVE GINNY PIGS, PIGS, I KNOW
YOU HAVE PIGS, CHICKENS, WHAT
ELSE?
RATS?
SO THE ANSWER IS YES AT A LEVEL,
SO THIS IS SIELIC ACID
DISTRIBUTION FOR THE OTHER
VIRUSES AND RECEPTORS I DON'T
KNOW IF THERE IS SOMETHING
COMPREHENSIVE LIKE THAT.
>> YEAH, NEXT QUESTION FROM
AMELIA [INDISCERNIBLE] THIS IS
WHEN YOU WERE TALKING ABOUT THE
METABOLIC EXPERIMENTS, THE
DIFFERENT MOUSE MODEL STRAINS
DIFFER IN OTHER WAYS EG A
DIFFERENT SET OF NONCLASSICAL
MHC RECEPTORS, THIS MAY CONFOUND
STUDIES COMPARING THE IMPACT OF
METABOLIC STRAINS BETWEEN
DIFFERENT METABOLIC MOUSE
STRAINS.
>> AGREED NAND AS A QUESTION
ANYONE WHO ASKED A QUESTION
ONLINE IS WELCOME TO UNMUTE OR
ASK INSTEAD OF ME, BUT 1 MORE,
IT'S MORE OF A COMMENT FROM MARK
WILLIAMS AT NIAID, BECAUSE PR8
HAS BEEN USED IF ARE MANY YEARS
ARE THERE REFERENCE SEQUENCES OR
WELL CHARACTERIZED STOCKS SUCH
THAT HARMONIZED USE OF VALIDATED
PR8 WOULD MINIMIZE VARIABILITY
IN HOST RESPONSES AND/OR IMMUNO
PATHOLOGY CAUSED BY CHANGES IN
CRITICAL SEQUENCE ELEMENTS?
>> YEAH, THATIA SOMETHING I
THINK THE VALIDATED STRAINS IN
TOCK STYLES WILL BE SOMETHING WE
TALK ABOUT THROUGH THE CIVIC AND
THE SEER AND TRYING TO DO THAT.
SO YOU'RE SOMEWHAT COMPARING
APPLES TO APPLES WHEN YOU DO
THESE DIFFERENT ANIMAL
EXPERIMENTS SO I THINK SOMETHING
LIKE THAT WOULD BE AVAILABLE,
BUT THEN ONCE YOU GET IN YOUR
LAB, AND YOU PROPAGATE IT, 1
TIME IN A CELL, YOUR POPULATION
WILL CHANGE AND I THINK YOU JUST
NEED TO BE WILLING TO DEEP
SEQUENCE AND REALLY CLASSIFY HOW
YOUR VIRUS HAS CHANGED, NOT JUST
OKAY, THE CONSENSUS IS THE SAME,
OKAY WE HAVE THESE NEW SINGLE
NUCLEOTIDE VALID AND RELIABLE
YABTS POPPING UP THAT ARE
DIFFERENT THAN WHAT RYAN HAS IN
HIS FREEZER OR SEEMA HAS IN
HERS.
BUT THERE WOULD BE THE
POSSIBILITY OF HAVING THIS SORT
OF STOCK PILE BUT KNOWING THE
CAVEAT THAT WILL CHANGE IT AS
SOON AS YOU GET IT INTO YOUR
HANDS, YOU AMPLIFY IT 1 TIME TO
GROW UP STOCK BUT THE SECOND AND
THIRD AND FOURTH TIME.
OTHERS THOUGHTS ON THAT?
BECAUSE THERE'S BEEN LOTS OF
DISCUSSIONS ABOUT GETTING
VALIDATED VIRAL STOCKS.
>> CAN I JUST ADD TO THAT, YOU
KNOW AS WE TALK ABOUT STRAINS WE
WE TALK A LOT ABOUT IT FROM A
RISK ASSESSMENT PANDEMIC,
PREPAREDNESS, ANGLE, BUT I THINK
THAT'S SORT OF TRANSLATES INTO
JUST ALL OF IT.
AS WE THINK ABOUT STRAINS,
EITHER PR8 OR MOVING FORWARD
WITH 1S THAT ARE YOU KNOW MORE
CIRCULATED IN THE POPULATION,
YOU KNOW THE REVERSE GENETICS,
PLAZ MIDS MIDS AND NOW WITH THE
LICENSING EXPIRED, THERE IS AN
OPPORTUNITY HERE TO REALLY LEVEL
THE PLAYING FIELD SUCH THAT
ACCESSIBILITY WITH STRAINS WITH
THE SAME GENETIC SIGNATURE CAN
HAPPEN BY MAKING THOSE PLAZ MIDS
MIDS AVAILABLE THROUGH BEI OR
OTHER RESOURCES.
>> RIGHT, BUT YOU HAVE TO BE
CAREFUL WITH THOSE, TOO, RIGHT
BECAUSE A LOT OF TIMES, WE MAKE
A VIRUS FROM PLAZ MIDS MIDS,
DOES NOT BEHAVE THE SAME WAY
THAT THAT WILD-TYPE VIRUS.
AT LEAST IN OUR--I THINK THAT'S
A REALLY GOOD POINT IS THAT
AGAIN YOU WOULD HAVE TO KNOW
THIS IS PREPARED FROM THESE PLAZ
MIDS MIDS AND HERE YOU'RE STILL
GOING TO HAVE TO SEQUENCE TO
KNOW WHAT YOU'RE DEALING WITH.
>> I KNOW THE DISCUSSION ABOUT
VIRUS STRAINS HAS BYE-BYE A BIT
FLU SPECIFIC BUT I'M TARTING TO
SEE THE SAME THING OCCUR FOR
MOUSE MODELS IN FOR SARS-COV-2
ASK I'M CERTAIN WE WILL PROBABLY
TALK ABOUT THAT TODAY BUT, AT
SOME POINT THOSE OF US DOING
ANIMAL WORK, YOU JUST HAVE TO
HONE IN AND CHOOSE THE STRAIN
BECAUSE YOU CANNOT CONTINUE TO
DO AND ASK THE QUESTIONS YOU'RE
TRYING TO ASK WITH 3 AND 4
DIFFERENT STRAINS.
SO YOU'RE STARTING TO SO THAT
WE'RE AT THAT POINT SO THAT SOME
PEOPLE ARE STILL WORKING WITH
ANCESTRAL, OTHERS ARE WORKING
WITH MOUSE ADAPTED, OTHERS ARE
ON TO OMICRON.
>> I THINK THE SAME IS TRUE FOR
RSV.
>> SAME IS TRUE FOR FLU.
>> YEAH, YEAH, THANK YOU FOR
RAISING THAT.
NO THIS IS ACROSS THE BOARD.
THIS IS TRUE FOR THE RSV FIELD
AS WELL, WHERE IT MAKES A BIG
DIFFERENCE, NOBODY DOES ANYTHING
WITH RSVB, THIS IS NOT A FLU
SPECIFIC.
>> OKAY, WELL WE ARE AT TIME SO
I WOULD LIKE TO THANK STACEY FOR
A FANTASTIC PRESENTATION FOR ALL
OF THE GREAT DISCUSSION.
NEXT WE HAVE A BREAK, I THINK,
AND SO WE WILL BE BACK IN HERE
AT 10:00 O'CLOCK AND WE WILL
CONTINUE WITH THE AGENDA.
THANK YOU EVERYBODY.
COLLECTION OF TALKS GOING INTO
THE DATA, LIMITATIONS OF IT AND
THES OF DIFFERENT MODELS, AND
LOOKING ACROSS SARS-COV-2 ACUTE
INFECTION, LONG-TERM INFECTION
AS WELL AS INFLUENZA,--SORRY,
THEY'RE ALL CORONA VIRUS INFECS,
I ARKPOLOGYIZE.
SO OUR FIRST SPEAKER WILL BE
STANLEY PERLMAN FROM THE
UNIVERSITY OF IOWA WHO WILL TALK
TO STRENGTHS AND LIMITATIONS ON
MOUSE MODELS FOR ACUTE
SARS-COV-2 AND P A S-C.
>> THANK YOU AND THANK YOU TO
THE ORGANIZERS FOR INVITING ME.
SO I WILL TALK ABOUT TODAY IS
REALLY I FEEL LIKE A SUBCHAPTER
IN THE TALK THAT STACEY GAVE,
STACEY GAVE A NICE OVERVIEW OF
THE WHOLE FIELD OF HOW 1 THINKS
ABOUT THIS AND I'M GOING TO TALK
MOSTLY ABOUT SARS-COV-2.
OKAY SO THIS WILL REQUIRE FINE
MUTER SKILLS.
SO I WILL JUST START AND GIVE
BROAD VIEWS OF CORONA VIRUS
INFECTIONS IN COVID-19
INFECTION.
SO STARTING WITH SARS-COV-2 AND
MOSTLY DEALING WITH THAT AND SO
WE KNOW THAT MANY ANIMALS CAN BE
INFECTED BY SARS-COV-2, BUT FEW
OF THEM DEVELOP CLINICAL SIGNS
OR SYMPTOMS AND AS WE TALK ABOUT
THE END OF THE DISCUSSION AFTER
STACEY'S TALK, HOW IMPORTANT IS
THIS?
AND SOMEBODY'S USED TO TAKE CARE
OF BABY ANDS CHILDREN WHO ARE
REALLY SICK, MY TENDENCY WAS TO
STUDY ANIMALS WHO WERE REALLY
SICK, SO I WAS MORE INTERESTED
IN THE REALLY PATHOGENIC SIDE OF
WHAT THESE VIRUSES CAN DO AND WE
HAVE ALWAYS BEEN INTERESTED IN
LABORATORY MICE, THAT'S WHO WE
WORK WITH AND LABORATORY MICE
CAN BE INFECTED WITH THE
ANCESTRAL STRAIN OF SARS-COV-2,
THIS WAS THE PROBLEM FOR JUST A
FEW MONTHS BECAUSE AS EVERYONE
WAS AWARE, THE ALPHA STRAIN
WHICH AROSE THAT CAN INFECT MICE
BUT AT THE TIME WE WERE
DESPERATE TO FIND OUT WHY ARE
PEOPLE DYING?
WHY ARE PEOPLE GETTING SICK WITH
THESE VIRUSES, AND WHAT BECAME
CLEAR QUICKLY IS 1 THAT COULD
INFECT MICE AND AMINO ACIDS AND
THE MOUSE OR SPIKE PROTEIN.
SO THEY ALSO KNEW THAT MICE
WOULD BE USEFUL BECAUSE THERE'S
A VARIETY OF STRAINS WITH
VARYING SUSCEPTIBILITY TO
INFECTION AND WE HEARD ABOUT
THIS IN VARIOUS MEETINGS AND
SESSIONS WE HAVE ONLINE TALKING
ABOUT THESE VIRUSES AND THE
INFECTION, SO WE KNOW FOR
EXAMPLE, THAT BALB-C MICE ARE
READILY INFECTIBLE AT ALL AGES
WITH SARS-COV-2 AND SARS CAV,
AND OTHERS AT BALB-C, THERE'S
AGE DEPENDENCE IS THAT LEDITOUS
THINGS BEING INTERESTED IN AGE
DEPENDENCE IN SARS-COV-2 BECAUSE
WHAT WE KNOW FROM OTHER SARS
CORONA VIRUSES, WHETHER IT'S
SARS-COV-2, IS THAT OLDER
INDIVIDUALS ARE MORE SUSCEPTIBLE
TO INFECTION THAN OUR YOUNGER
1S.
SO SOME OF THE APPROACHES TO
MOUSE MODELS WHICH WE AND OTHERS
USE ARE AS FOLLOWS, BY
SENSITIZING MICE AGAIN, I'M
TALKING HERE ABOUT THE ANCESTRAL
STRAIN.
THIS WOULD HAVE BEEN DONE
DIFFERENTLY IF 1 OF THE STRAINS
THAT COULD DIRECTLY INFECT MICE
CAME UP ORIGINALLY AND WITH
SARS-COV, THAT WAS THE CASE, THE
VIRUS THAT WAS ORIGINALLY
ISOLATED COULD INFECT MICE CAUSE
ALMOST NO DISEASE BUT COULD
INFECT MICE DIRECTLY SO
SARS-COV-2 WAS JUST COMING OFF
THAT, SO HOW DO YOU DO THAT?
HOW DO YOU MAKE THEM ABLE TO
INFECT MICE?
THE FIRST APPROACH WAS 1 THAT WE
AND OTHERS HAD USED ACTUALLY
EARLY IN THE SARS PROJECTION
NEURONS OR PIONS DEMIC, SO WE
SENSITIZED MICE WITH ADENO-5
VECTORS EXPRESSING HUMAN ASE 2,
THE ADMIN RECORDS ANDAGE OF THIS
IS IN THEORY CAN YOU MAKE THE
CONSTRUCTS WITHIN A MONTH AND 1
HAS A MODEL NOT FOR SYMPTOMATIC
INFECTION OR NOT FOR VERY IMP
TOMATIC INFECTION, OR AT LEAST
FOR INFECTION, AND WE HAD AS
PART OF THE SARS EPIDEMIC WE
DEVELOP THESE K18 MICE AND THEY
BECAME USEFUL AND I WILL MENTION
THAT IN A BIT AND THEN WE MADE
MORE KNOCK-IN MICE AND 1 TO
CHANGE THE MOUSE IS TO
MANIPULATE THE VIRUS SO THAT IT
COMBINED AND AS I SAID THE
ANCESTRAL STRAIN OTHER THAN
DELTA THAT CAN INFECT MICE
DIRECTLY.
SO, GOING THROUGH THE STRAINS
SINGLE MUTATIONS OF THESE
DIFFERENT APPROACHES, SO FOR THE
A D5 SYSTEM WHICH IS DEVELOPED
BY MY COLLEAGUE PAUL MCCRAE.
IT'S QUICKLY MADE.
IT'S USEFUL, YOU INFECT MICE OR
TRANSDUCE MICE WITH THE ADEN O
VIRUS CONSTRUCT AND THEN YOU
INFECT THEM WITH SARS-COV-2.
YOU USE HIGH DOSES AND ANY
STRAIN OF SARS-COV-2 WILL WORK
IN THESE MICE BECAUSE HAVE YOU
THE HUMAN RECEPTOR AND THESE
CONTRACTS ARE AVAILABLE FROM BEI
IF 1 DOESN'T WANT TO MAKE THEM
THEMSELVES, AND THE ADVANTAGES
ARE NOT ONLY CAN YOU SENSITIZE
ANY STRAIN OF MICE BUT YOU CAN
USE THEM FOR DOING THINGS LIKE
IDENTIFYING T-CELL RESONS
SPONSES, ANTIBODY RESPONSES,
TEST VACCINES AND THERAPEUTICS
AND THIS WAS IMPORTANT IN THE
BEIMINNING OF THE PANDEMIC WHEN
WE REALLY DIDN'T HAVE VERY MUCH
TO USE IN TERMS OF MICE.
HAMSTERS FROM THE BEGINNING WERE
KNOWN TO BE INFECTIBLE AND WE
WILL HEAR ABOUT THAT SO THAT WAS
USEFUL.
THE LIMITATIONS OF THE SYSTEM IS
THAT YOU GET MINIMAL CLINICAL
DISEASE AND SLIGHTLY MORE WEIGHT
LOSS, FOR THE IMMUNO DEFICIENT
MICE ARE TRANSDUCED AND VIRUS
CLEARED VERY QUICKLY WITHIN A
WEEK AND 1 OF THE REASONS THAT I
THINK THAT MICE DO WELL IS
BECAUSE THE AT1 CELLS WHICH ARE
IMPORTANT FOR AIR EXCHANGE
ACTUALLY SURVIVE IN THESE MICE.
LET ME SEE IF I CAN--YES, SO,
THIS JUST BRIEFLY HOW DOES THE
ARROW WORK IN DO YOU KNOW?
YEAH, OKAY, SO, WE HAD MADE
THESE VIRUSES SARS-COV-2 THAT
EXPRESS CREE AND VENUS AND THE
VIRUS IS BASICALLY VIRULENT AS
WILD-TYPE VIRUS IN MICE AND WHAT
THIS DOES IS THE VENUS ALLOWS
YOU TO IDENTIFY A CELL INFECTED
AND EARLY TIMES OF INFECTION AND
CREE IF 1 INFECTS MICE THAT HAS
A FOX [INDISCERNIBLE] IT CAN
IDENTIFY THE CELLS THAT WERE
INFECTED AND IN THIS EXPERIMENT,
WE TOOK THE MICE, SHOWING
PREVIOUSLY INFECTED CELLS IN
ADEN O VIRUS, MOST OF THE CELLS
THAT ARE SURVIVING ARE AT1 CELLS
THATTA SHOWN HERE AND IS A FEW
AT2 CELLS WHILE IN THE MORE
VIRULENT STRAINS WHICH I WILL
TALK ABOUT IN A COUPLE MINUTES
ISSUE THE AT-2 CELLS ARE THE
ONLY 1S THAT ARE SURF VIEFING O
BY HAVING AT2 CELLS SURVIVE,
THESE CAN BE PRECURSORS, BUT IF
THE AT1 CELLS ARE DYING, THE
MICE DIE BECAUSE THEY NEED THAT
FOR OXYGEN EXCHANGE.
SO THIS SYSTEM SHOWS POSSIBLY 1
OF THE REASONS THAT ADENOVIRUS
MICE DO BUT WE KNOW THAT AD-5
MAY MODESTLY INDUCE AN IMMUNE
RESPONSE AND THAT'S WHY WE HAVE
WAYS TO TEST MICE AND WE KNOW
THE NUMBER OF SUSCEPTIBLE CELLS
IS LIMITED BECAUSE OF THE
TRANSDUCTION.
BUT STILL A BIT OF A MYSTERY FOR
THESE MICE TO SHOW RESISTANT TO
THE VIRUS.
THE SECOND APPROACH IS TO USE
THESE K18 MICE AND THESE ARE THE
1S THAT JACKSON LAB DID A GREAT
JOB IN RESUSCITATING OR RECOVERY
EARLY IN THE PANDEMIC, SO WE HAD
HAD THESE MICE, WE USED THEM IN
SARS COV AND WE DIDN'T LIKE THEM
VERY MUCH BECAUSE NICE ALL GOT
NEUROLOGICAL DISEASE AND THEY
DIED COMFORTABLY, BUT FOR
SARS-COV-2 IT SEEMS LIKE
NEUROLOGIC DEC NOT TOTALLY CLEAR
BECAUSE WE HADN'T HAD SIDE BY
SIDE COMPARISON, WITH SARS-COV-2
BUT SO IN THESE RESUSCITATED WE
WERE WORRIED THESE WERE NOT
GOING TO BE VERY USEFUL BUT THEY
TURNED OUT TO BE INCREDIBLY
USEFUL BECAUSE NOT ALL MICE USE
NEUROLOGICAL DISEASE BUT ALSO
BECAUSE IT WAS SO SUSCEPTIBLE TO
INFECTION.
AND SO JUST A COUPLE COMMENTS ON
THEM, THEY WERE AVAILABLE EARLY
IN THE PANDEMIC AND I THINK 1 OF
THE THINGS THAT BOTH JACKSON
LABS AND UNIVERSITY AND THE
UNIVERSITY OF IOWA DID IS THEY
MADE THESE MICE AVAILABLE AT
REALLY RELATIVELY LOW PRICES
OPPOSED TO DESIGNER MICE WHICH
WERE VERY EXPENSIVE.
THESE MICE COST INITIALLY TWICE
AS MUCH AS A B6 MOUSE SO IT WAS
AFFORDABLE AND SO, JACKSON LABS
ACTUALLY INCREASED THEIR
PRODUCTIVITY OF THESE HIGHLY AND
THEY BECAME WIDELY USED.
THESE HAVE CERTAIN ADVANTAGES
AND CERTAIN DISADVANTAGES THESE
MICE, SO THE STRENGTH IS THAT
THEY DEVELOP A CLINICAL DEC AND
DOSE DEPENDENT FASHION, MALE AND
FEMALE MICE WERE SUSCEPTIBLE TO
INFECTION, AND TELL BE
THE--THERE'S A FUNNY BUSINESS
WITH THESE MICE BECAUSE
NEUROLOGICAL DISEASE DEVELOPS
MOSTLY IN YOUNGER MICE BUT AT
THE SAME TIME BECAUSE THESE MICE
WERE ON A B6 BACKGROUND, THEY
BECOME MORE SENSITIVE, AS MICE
AGE.
SO EARLY ON, THEY DIVE FROM MICE
MAY DIE FROM RESPIRATORY OR
NEUROLOGICAL DISEASE, BUT LATER
ON THEY DIE FROM RESPIRATORY
DISEASE.
AND WE SEE A LOT OF THINGS THAT
ARE SIMILAR TO THE HUMAN
INFECTION, WE SEE INFECTION OF
SUSTEN TACTULAR CELLS IN THE
NAISAL CAVITY, WHICH I WILL TALK
ABOUT IF A GET TO THEM, AND THE
INFECTION OF ALVEOLAR CELLS AND
THROMBIOLOGICAL AND IT'S STUDIES
THAT PEOPLE USE FOR THERAPEUTICS
AND VACCINES, AND THE WEAKNESSES
ARE THAT THESE SOMETIMES FELT
ENCEPHALITIS, SARS KICKOFF
RARELY IF EVER AFFECTS THE HUMAN
BRAIN, THIS WAS VERY
CONTROVERSIAL IN THE BEGINNING
OF THE PANDEMIC AND I THINK
STILL IS SOMEWHAT CONTROVERSIAL
BUT NOT AS MUCH, BRAIN INFECTION
IN THESE MICE OCCURS IN THE
EPITHELIUM, PERHAPS THE BLOOD
AND PERSIEVEERAL NERVES.
AND IT'S BECAUSE BY BEING
EXPRESSED BEHIND THE CYTOKERATIN
PROMOTER, IT'S NOT THE SAME
EXPRESSION PATTERN AS YOU SEE IN
HUMANS OR MICE OF PHASE 2.
AND SO THE NEXT APPROACH IS 1
THAT WAS USED BY OUR LAB AND
OTHERS.
SO THE IDEA WAS HERE TO CHANGE
THE MOUSE, THE VIRUS RATHER SO
WE COULD BETTER USE MACE2, AND
BASED ON THIS ANALYSIS, THE
LAB,--DEVELOPED A VIRUS THAT CAN
INFECT MICE BY CHANGING 2
RESIDUES AND JOE'S LAB, ISOLATED
A RIEROUS THAT COULD INFECT MICE
DIRECTLY WITHOUT ANY FURTHER
MANIPULATION AND THIS WAS
CURIOSITY BECAUSE THE WAY HE DID
IT, THE WAY THE LAB DID IT IS
THEY WERE ABLE TO TAKE THE
NORMAL STOPS OF ANCESTRAL VIRUS
AND WE KNOW THAT AS WAS
MENTIONED ALL THESE VIRUSES OF
THESE SPECIES, SO HE WAS ABLE IN
THAT GROUP TO ISOLATE A VIRUS
THAT COULD DIRECTLY INFECT THE
MOUSE, MICE AND SO, WE DEVELOPED
A--YOU WILL USE IN 501 MUTATION
WHICH TURNED OUT TO BE CRITICAL
FOR THE HUMAN INFECTIONS AFTER
THAT, WE USE THIS TO MAKE A
REVERSE GENETIC--USE REVERSE
GENETICS TO MAKE A MOUSE THAT
COULD THEN INFECT MICE AND 1 OF
THE THINGS ABOUT ALL THOSE VIRUS
SYSTEM THAT THEY'RE ALL
ATTENUATE IN MICE, INITIALLY
EXCEPT THE BATTA VARIANT MAY
HAVE MORE VIRULENCE AND AFTER
PASSAGE THROUGH A MOUSE LUNGS,
FOR A PERIOD OF TIME, WE WILL
HEAR LATER ABOUT M810, WE
PASSAGE OURS THIRD TIMES AND WE
GOTIE VERY VIRULENT VIRUS CELL.
SO I ONLY SHOW THIS SLIDE FOR 1
POINT, SO THIS SLIDE MOSTLY
INFECTS THE LUNGS BUT 1 THING
THAT WAS CURIOUS IS THAT THEY
TOO HAD TITERS IN VARIOUS ORGANS
WHETHER IT'S THE BRAIN, HEART,
LIVER UAND THE REASON I SHOW
THIS IS BECAUSE 1 THING THAT'S
BEEN INTERESTING, I THINK
EVERYBODY IN THE FIELD IS HOW
EXACTLY DO PEOPLE GET THE
LONG-TERM SEQUEL I OF
SARS-COV-2, IS THERE ANYTHING
THAT OCCURRED EARLY IN THE
INFECTION THAT COULD BE DOING
THIS AND SO THIS--I DON'T KNOW
WHERE VIRUS IN 2 DAYS, IT GOES
AWAY, THERE'S NO PUREOLOGICAL
EVIDENCE, AND CONTRIBUTE ARING
COULD GET LONG-TERM EFFECTS AND
WE LOOK INNED BLOOD HERE AND
THERE'S VERY, VERY LITTLE VIRUS
IN THE BLOOD, SO IT'S NOT--I
CAN'T BELIEVE IT'S JUST BLOOD
CONTAMINATION SO THIS IS STILL
CURIOSITY IS THE VIRUS DOING
SOMETHING WITHIN THE ENDOTHELIAL
CELLS, I DON'T KNOW THE ANSWER
TO THAT, SO THE ADVANTAGES OF
MOUSE ADAPTED SET THE SEVERAL
VARIATIONS DONE, THIS IS DONE IN
SEVERAL LABS, PEOPLE EVEN
PASSAGED OTHER STRAINS LIKE THE
BETA STRAIN THAT'S BEEN
PASSAGED, AND 1 GETS A VIRULENT
VIRUS THAT'S A LITTLE DIFFERENT,
THIS IS ALL CAUSE OF A
RESPIRATORY INFECTION IN A DOSE
DEPENDENT FACTION, THESE VIRUSES
HAVE THE AT1 AND 2 CELLS AS
OPPOSE IN PATIENTS, THERE'S NO
MENTION OF BRAIN OR PULMONARY
INFECTION, AND OFFICIALLY THIS
THE QUESTION THAT 1 HAS TO PUT
IN THE BIG PICTURE OF HOW
IMPORTANT THIS IS, THIS VIRUS,
THE 1 WE DEVELOPED HAS 7
MUTATIONS, 5 ARE IN THE SURFACE
FOR DPLIEK O PROTEIN, ALL 5 ARE
DOWN IN THE OMICRON VARIANTS, 2
ARE IN NONSTRUCTURAL PROTEINS
WE'VE DONE A SET OF THIS FOR 4
AND 8 IS ALL YOU NEED FOR MOUSE
ADAPTATION, WE DON'T KNOW WHY.
BUT VERY LITTLE IS NEEDED AND
NOT FOR MOUSE ADAPTATION, IT'S
TO MACK A VIRULENT VIRUS BECAUSE
THE PROTEINS BY--THE N501 Y WILL
GIVE YOU A VIRUS THAT WILL
INFECT MICE.
OKAY, SO IN THE LAST FEW MINUTES
I HAVE, I WILL TALK ABOUT OTHER
ADVANTAGES AND DISADVANTAGES OF
THESE MODELS.
SO WE'VE BECOME INTERESTED IN P
A SC, THE POST ACUTE SEQUEL I OF
COVID-19, AT THIS POINT WE READ
INTO THE NEWSPAPERS AND HEAR
ABOUT THE INCREASED NUMBER OF
PATIENTS THAT WE HAVE NOW
SARSARS-KICKOFF 2, PEOPLE ARE
NOT BEING HOSPITALIZED, THERE'S
STILL MORE HOSPITALIZATION AND
MORE DEATH THAN WE WOULD LIKE
PARTLY BECAUSE PEOPLE ARE NOT
UPTAKING THE VACCINE THE WAY WE
WOULD LIKE AND A LOT OF PEOPLE
WHO ARE DYING ARE ACTUALLY
THEY'RE AGED OR IMMUNO
COMPROMISED SO THOSE ARE
POPULATIONS THAT I THINK AS
STACEY WAS TALKING ABOUT, I
THINK THEY'RE REALLY FOR AT
LEAST A LOT OF THESE DISEASES,
REALLY THE POPULATIONS HAVE THAT
WE WANT TO THINK ABOUT BECAUSE
THOSE ARE THE PEOPLE WHO ARE
INFECTED AND GET SEVERE DISEASE.
THE THING ABOUT P A SC, OR
AFFECTS MANY ORGANS, WHO HAVE P
A SC ARE OFTEN FATIGUES AND A
LOT OF THESE GO AWAY WITH TIME.
I'M NOT GOING TO SHOW A SLIDE
BUT ROUGHLY MY CALCULATION IS
ABOUT 1% OF PEOPLE AFTER 2020
HAVE P A SC AFTER 1 YEAR, OTHER
ESTIMATES ARE HIGHER.
IF 1 LOOKS AT LARGE STUDY ANDS
THERE WAS JUST A BIG STUDY
PUBLISHED THAT OVER 3 YEARS
PEOPLE THAT HAD PUNISHING ASC,
IT'S REALLY DIMINISHED IN ALL
CATEGORIES WHETHER IT'S CARDIAC
OR NEUROLOGICAL SO THIS IS
SOMETHING THAT MAY
NOT--CERTAINLY NOT BE PERMANENT
FOR MOST PEOPLE AND I THINK
PEOPLE ADAPT, TO, BUT SOME WHO
ARE STILL ASHINGED SO WE WANT TO
LEARN SOMETHING ABOUT WHY THIS
IS HAPPENING, IT SEEMS TO BE
DECREASED BY VACCINATION AND IT
SEEMS TO BE LESSER AN EXTENT
AFTER INFECTION WITH THE OMICRON
VARIANTS.
SO THIS IS--I'M NOT GOING TO
MENTION THIS, EXCEPT THAT WE'VE
BECOME VERY INTERESTED IN THE
LOSS OF THE SENSE OF SMELL WHICH
IS A BIG DEAL, STILL.
I RECEIVED A CALL A MONTH AGO
WHO HAD BEEN INFEC INDEED
NOVEMBER 2022, SOMEBODY IN CHILE
AND SHE SAYS THAT RIGHT NOW HER
SMELL RANGE IS EVERYTHING SHE
TASTES RANGES FROM NOTHING TO
TERRIBLE.
SO THERE'S NOTHING THAT SHE
ENJOYS ANYMORE.
SO FOR SOME PEOPLE THIS IS A
HUGE DEAL AND THEN THE OTHER
THING THAT HAPPENS IS, YOU GET P
A RASMIA, IS XA'S PROBABLY WHAT
THIS WOMAN HAD, SHE COULD SMELL
THINGS BUT NONE OF THE SMELLS
SHOULD NOT HAVE BEEN WHAT THEY
SHOULD HAVE BEEN.
STRAWBERRIES SMELLED LIKE SEWAGE
OR WHATEVER, SO WE'VE BEEN
INTERESTED IN THAT, TOO, AND YOU
CAN INVESTIGATE THIS IN MICE AND
HAMSTERS, SPRMAL ANIMALS OFTEN
GET THE SAME SYMPTOMS, SO WE USE
THIS VIRUS OR M830 VIRUS BECAUSE
IT INFEBS THE CELLS AND 1 OF THE
THINGS ABOUT WHETHER IT BE
HUMANS OR MICE OR HAMSTERS, IS
THE ONLY THING THAT ARE INFECTED
ARE THESE CELLS, WHICH ARE THE
SUPPORTING CELLS OF THE NAISAL
EPITHELIUM, SO THE NEURONS ARE
CARRIED AT THE BACK OF THE BRAIN
BUT THESE CELLS ARE NOT INFECTED
BUT THEY'RE AFFECTED AND NEURONS
CAN'T BE INFECTED SO HOW THIS IS
EXACTLY WORKING IS I THINK A
REALLY IMPORTANT RESEARCH
QUESTION .
I DON'T HAVE A SOLID ANSWER
ABOUT WHAT'S GOING ON HERE BUT 1
CAN SEE HERE THIS IS THE WAY
OLFACTORY NEURONS WORK, EACH 1
HAS A SINGLE RECEPTOR, EACH 1
HAS A CILIA THAT HAS THE
RECEPTOR ON IT, THE CILIA ARE
NOT KILLED HERE, BUT YOU CAN SEE
IF THIS IS A NORMAL CILIA,
THEY'VE FALLEN OVER SO THEY'RE
NOT HAPPY CILIA.
AND WE THINK THAT TRIBUTES TO
ANOSMIA, BUT WE ALSO KNOW IT
RECOVERS VERY QUICKLY SO THESE
MICE HAD NORMAL CILIA BUT THEN
HAD ANASMIA, AND OTHERS THAT I
WILL FALL IN FOR THE LAST FEW
MINUTES.
SO THIS ANOTHER RESULT WHICH IS
KIND OF PUZZLING TO US BECAUSE
WHAT WE MEASURE SIDE OLFACTORY
RECEPTOR EXPRESSION.
WHAT WE FOUND IS--THIS WAS
REPORTED BY OTHERS AS WELL, IS
THAT WITH--SOON AFTER INFECTION,
THIS IS 2 DAY AFTER INFECTION,
THIS IS DECREASED IN OLFACTORY
RECEPTOR, EXPRESSION AT THE IN,
RNA LEVEL.
--MRNA LEVEL.
SO THIS IS UNEXPECTED SO WHY
WOULD MRNA BE AFFECTED OR
DECREASED BY SOMETHING GOING ON
IN THE NEIGHBORING SUPPORTING
CELLS AND REMEMBER, I DIDN'T
SHOW A FIGURE OF THIS BUT NOT
ALL THE CELLS ARE INFECTED, ONLY
A MINORITY ARE SO THERE'S
PROBABLY SOME INFLAMMATORY
RELATED MOLECULE THAT'S DOING
SOME OF THIS EMPLOY THE OTHER
THING THAT'S CURIOUS IS I THINK
THIS IS PROBABLY THE MOST
CURIOUS.
IS THAT AFTER OLFACTORY
RECOVERY, YOU CAN SEE THIS
HIGHER LEVELS IN HERE.
WHY DOES THIS MATTER IN 1 CAN
IMAGINE IF 1'S IMETTING A SIGNAL
FROM A NEURON AND THAT NEURON IS
NOW RESPONDING DIFFERENTLY OR
THE NEURONS BEHAVING DIFFERENTLY
IN TERMS OF HOW THEY'RE DOING
WITH ODORANTS BECAUSE EVERY
SMELL ACTUALLY A COMBINATION OF
THE DIFFERENT ODORANTS BINDING
TO DIFFERENT RECEPTORS.
SO 1 CAN SEE THAT IF SOME ARE
COMING BACK AT HIGHER LEVELS OR
LOWER LEVELS WHAT 1 WOULD HAVE
NORMALLY IMIVEN A CERTAIN SMELL
MIGHT GIVE A DIFFERENT SMELL
NOW.
O WE THINK THAT THAT CONTRIBUTES
ALSO TO ANASMIA.
SO WHAT HAPPENS AFTER THE
OLFACTORY NEURONS ARE DAMAGED?
SO OLFACTORY NEURONS CONNECK TO
THE OLFACTORY BULB AND THERE'S A
MOLECULE IN THE OLFACTORY BULB
VERY IMPORTANTLY WITH TYROVENE
HYDROX CLAYS FOR THE MIEWRONS
AND WHAT WE SEE IN THESE ANIMALS
IS THAT THE AMOUNT OF TYROSEEN
HYDROX LACE DECREASED AND THAT'S
SHOWN HERE SO THIS IS THE NUMBER
OF TH-POSITIVE CELLS SO THIS IS
NOT AT THE MRNA LEVEL, THIS IS
AT THE PROTEIN LEVEL.
AND WE SEE THE SAME THING AT 120
DAYS AFTER INFECTION, SO THIS IS
USEFUL FOR OUR STUDIES OF TRYING
TO UNDERSTAND LONG-TERM EFFECTS
OF ANASMIA, SO WE SEE DECREASED
TYROSEEN RIDROKS LACE AT THE
RTHIS,A AND PROTEIN LEVELS, AND
WE SEE INCREASED AMOUNTS OF
INTERFERON BETA AND SEVERAL
OTHER INFLAMMATION BASED
MOLECULES BUT NOT ALL.
CCL5 DECREASED CXCL10 IS
DECREASED, WHY DON'T KNOW WHAT
THIS MEANS, SOME GO UP AND SOME
GO DOWN BUT THE FACT THAT
THERE'S INFLAMMATORY MOLECULE
CHANGES, WE IT CONTRIBUTES TO
THE WHAT WE SEE IN OLYMPIC FAC.
SO THE NEXT THING WE WANTED TO
DEAL WITH, IS WHAT ABOUT OTHER
EFFECTS AND OTHER PARTS OF THE
BRAIN, SO AS TYROSEEN, OUR
NEURONS OF EXPRESSING TYROSINE
RIDROKS LACE, SO WE LOOKED IN
THE PART OF THE BRAIN CALLED THE
SUBSTANTIA NIAGARA, FOR THOSE
WHO THINK ABOUT NEUROLOGICAL
ISSUES KNOW THIS IS THE AREA
THAT'S INVOFFED IN PARKINSON'S
DEC AND THIS IS THE AREA THAT
WAS STRONGLY INFECT INDEED 1918
INFLUENZA EPIDEMIC.
AND THIS DATA IS SHOWING THAT
THE 1918 FLU EPIDEMIC ACTUALLY
OR PANDEMIC RESULTED IN
INCREASED CASES OF PARKINSON'S
DISEASE.
SO WHAT WE DID HERE IS WE LOOKED
ON THE SUBITANTIA NIAGARA AND WE
FOUND THAT HERE AS WELL AS IN
THE OLFACTORY BULB, IS DECREASED
NUMBERS OF TYROSINE HYDROX LACE
EXPRESSING CELLS.
BUT MORE IMPORTANTLY THESE CELLS
WERE ALL DECREASED IN THE
SUBSTANTIA NIGRA, AND THEY
APPEAR TO BE THE MOST COMMONLY
WE SEE ACROSS THE BOARD AND IN
THESE PARTS OF THE BRAIN WAS
INCREASED.
AND TO SEE IF THIS IS RELEVANT
FOR YOU HAD HANS, WE OBTAINED
AUTOPSIES SAMPLES AND WE SEE
AGAIN IN THE SUBSTANTIA NIGRA,
THERE'S INCREASED NUMBERS OF
POSITIVE CELLS.
AND SO WE DID TESTING HERE.
THIS A TEST--IT'S SUBSTANTIA
NIGRA, BUT WE PUT THESE ON RODS
AND MADE IT FASTER AND DEPENDING
ON HOW COMPETENT THE MICE ARE IN
DEALING WITH THE MUSCLE FUNCTION
THEN 1 SEES THE MICE FALLING OFF
THE ROD, SO 1 HERE DECREASED THE
ABILITY TO STAY ON THESE ROD IN
THESE MICE 100 DAYS AFTER
INFECTION.
THIS IS NOT INFECTED ANIMALS AND
THESE ARE INFECTED ANIMALS.
SO THE WHYED IS THAT WE'RE
SEEING DYSFUNCTION IN THE
OLFACTORY BULB BUT THE
SUBSTANTIA NIGRA IS HAVING
CLINICAL EFFECTS.
AND THEN IS ANOTHER OPEN FIELD
TEST AND WITHOUT LOOKING AT THE
GRAPHS 1 COULD SEE IF THE
FIGURES THAT INFECKED MICE RUN
DIFFERENT THAN THE MICE THAT
WERE MOCK INFECKED AND WE COULD
QUANTIFY THIS AS WELL, SHOWING
THESE CHANGES IN THE OPEN FIELD
TEST.
SO THE CONCLUSIONS HERE IS THAT
THESE MICE, DEAL WITH MANY SIGNS
OF ACUTE COVID-19 IN P A SC.
BUT IT'S IMPORTANT TO EMPHASIZE,
MICE DO NOT DEVELOP ALL THE
FINDINGS PRESENT IN PATIENTS AND
AT LEAST NOT TO THE SAME EXTENT.
EVEN THOUGH I SHOWED 1 SLIDE OR
I MENTIONED THAT
THROMBO-INVOLVED PHENOMENA COULD
BE SEEN IN SOME CASES, IT'S NOT
SEEN ENOUGH SO THAT CAN CAN
STUDY THEM HERE AND CERTAINLY A
LOT THINGS THAT THAT PEOPLE
COMPLAIN ABOUT, BRAIN FOG,
NONNEUROLOGICAL P A SC, BRAIN
FOG MICE, WE DON'T KNOW IF THEY
HAVE THAT NOT OR NOT AND FOR
NONNEUROLOGICAL WE HAVEN'T
INVESTIGATED THE HEART OR OTHER
NEUROLOGICAL TESTS AND FUNCTIONS
BUT SO DON'T KNOW.
BUT IN GENERAL, I THINK MICE
GIVEN ALL THE CAVEATS OF MICE,
THEY'RE PRETTY USEFUL FOR
STUDYING SOME OF THESE SEQUEL I,
WE HAVE TO BE CAREFUL ABOUT WHAT
WE EXPECT FROM THEM AND WHAT WE
COULD HOPEFULLY GET.
THE OTHER THING I SHOULD SAY,
WE'VE DONE EXPERIMENTS TREATING
THESE MICE EARLY ON WITH
ANTIVIRALS AND IT HAS NO EFFECT
ON THE DEVELOPMENT OF P A SC,
AND THE THING THAT'S INTERESTING
ABOUT THIS IS THAT WHEN WE WERE
DOING IT, THE SAME TRIALS WERE
BEING PUBLISHED BY HUMANS WHO
ALSO DIDN'T HAVE P A SC HELP
THEM WITH--DIDN'T HAVE
ANTIVIRALS HELP THEM ONCE THEY
DEVELOP P A SC WHICH IS NOT THE
SAME THING BUT IT MAY BE THAT
ANTIVIRALS HAVE REALLY NO ROLE
IN THIS, WHATEVER'S OCCURRING
AND IS VERY DIFFICULT TO
PREVENT.
SO WITH THAT I WILL TAKE ANY
QUESTIONS.
>> THANK YOU.
QUESTIONS NOW.
STACEY?
>> BEAUTIFUL PRESENTATION, SO
I'M SURE AS YOU KNOW IPT GREATER
FOMNIA IS 1 OF THE FIRST
INDICATORS OF NEURODEGENERATIVE
DISEASES SO I WONDER IF YOU
LOOKED AT MICROGLIA ACTIVATION
THAT'S QUESTION NUMBER 1.
QUESTION # 2 IS FOR ALZHEIMERS
THE PEANUT BUTTER SMELL TEST IS
A REAL THING ASK IT SHOWS YOU
EARLY INDICATIONS OF HOW IS
HAVING INSOMNIA FROM COVID GOING
TO IMPACT PEOPLE USING THOSE
TYPES OF TESTS FOR
NEURODEGENERATIVE DISEASES.
>> YEAH, IT'S AN IMPORTANT
QUESTION BECAUSE I THINK THAT IF
PEOPLE--I THINK ACUTELY, OF
COURSE YOU COULDN'T USE A PEANUT
BUTTER TEST.
YOU WOULD THINK A YEAR OR SOME
MONTHS LATER IT MIGHT BE USEFUL
AND PEOPLE HAVE PERSISTENT
NASMIA, BUT WHETHER PEOPLE USE
IT, THIS IS THE BIG CONCERN THAT
PEOPLE WILL DWEF SOME SORT OF
NEURODEGENERATIVE DISEASE AND
YES MICROGLIA ARE ACTIVATED.
WE HAVE ACTUALLY ANALYSIS OF
THOSE.
I DIDN'T HAVE TIME TO SHOW IT
BUT WE SEE THAT THEY'RE
ACTIVATED.
STANLEY I HAVE A QUESTION SO IN
TERMS OF EXTRA PULMONARY ORGAN
INFECTION, WHEN YOU PASSAGE
SARS-COV-2 IN MICE TO GET THAT
SEVERE DISEASE OR THE MOUSE 30
STRAIN, IS THERE A CORRELATION
BETWEEN DISEASE SEVERITY VIRUS
RUPLICATION IN THE LUNGS WITH
HOW MUCH REPLICATION YOU ARE
SEEING IN THE OTHER ORGANS
OUTSIDE OF THE LUNGS?
>> I DON'T THINK WE'VE DONE THAT
CAREFULLY BECAUSE WE HAVE DOSE
CURVES ON THE LUNGS.
I DON'T THINK WE WANT ALL THE
ORGANS AT DIFFERENT DOSES, MY
GUESS IS THERE WOULD BE BESES
BUT I DON'T KNOW THAT BECAUSE
IT'S ALL PROPORGAL TO THE LUNG
INFECTION.
>> I WAS TRYING TO FIGURE OUT
HOW MUCH HEMORRHAGES WAS
OCCURRING IN THE LUNGS IN THOSE
SEVERELY INFECTED MICE AND HOW
MUCH OF THAT OVER FROM THE LUNGS
BUT JUST ENOUGH TO GET THOSE
PERIPHERAL ORGANS SEATED MAY NOT
BE ENOUGH TO BE DETECTED BY PC R
OR PLAQUE ASSAY.
>> I DON'T THINK THERE'S ANY
VIRUS--I DON'T KNOW WHAT'S GOING
ON IN 2 DAYS, BECAUSE WHAT WE'VE
DONE IS WITH THAT VIRUS THAT I
SHOWED WHERE WE COULD LABEL
SURVIVING CELLS BECAUSE WE USE
THE VIRUSES EXPRESSING CREE, WE
SEE NO SURVERIFYING CELL OR I
SHOULD SAY WE DON'T SEE NONE.
IF WE LOOK AT 5 SLIDES EACH YOU
FIND 1 CELL OR 1.5 CELLS.
SO VERY, VERY LEGISLATES LITTLE
EXTRA PULMONARY INFECTION.
>> I WANT TO SMOTE NAWE WILL
HAVE ADDITIONAL TIME DURING
DISCUSSIONINGS DURING THE PANEL
DISCUSSION.
>> THANK YOU SO MUCH.
OUR NEXT SPEAKER WILL BE LISA
GRALINSKI, FROM UNIVERSITY OF
NORTH CAROLINA CHAPEL HILL, SHE
WILL TALK ABOUT MOUSE MODELS OF
HIGHLY PATHOGENIC HUMAN CORONA
VIRUS INFECTION.
NALL RIGHT, THANK YOU EVERYONE,
THIS WILL BE A NICE FOLLOW UP TO
STANLEY'S TALK.
SOME OF THE SAME THEMES BUT A
TOALTALLY GOOD NIGHT VIRUS
BECAUSE ALL THE CORONA I
HAVOLOGYIST WANTED TO MAKE TOOLS
TO DIVE INTO WHAT WAS HAPPENING
IN ANIMAL MODELS AS QUICKLY AS
POSSIBLE IN 2020 SO THIS TALK IS
REPRESENTING WORK FROM MY LAB
AND OUR BROADER GROUP AT UNC, SO
WE'VE NAMED OURSELVES THE
CAROLINA WERE GROUP, OR CDRF P
AND WE'RE ALL IN THE SAME BL3,
CONITANTLY HELPING EACH OTHER
ANYWAYS AS 1 DOES WHEN THERE ARE
A LOT OF MICE HAPPENING.
NALL RIGHT, SO I'M GOING TO
SPEND MOST OF THIS TALK SPEAKING
ABOUT SARS-COV-2 MOUSE MODELS
SINCE THAT'S OUR CURRENT ACTIVE
STILL CIRCULATING THE GLOBE
VIRUS SPEAKING ABOUT BOTH THE
ACUTE DEC OF THE PHASE AND
GETTING INTO THE CHRONIC DEC
BECAUSE FOLLOWING UP ON THESE
LONG-TERM SEQUEL I ARE A AN AREA
OF INTEREST ARE IF A LOT OF US
AND AN AREA THAT'S GROWING A LOT
AND A TOUCH ON SYSTEMIC CENTER
FOR EXCELLENCE ON AGINGS OF THE
DEC.
BUT THEN ALSO TALK ABOUT
VARIANTS OF CONCERN AND HOW
WE'RE UPDATING THESE MODELS AND
AT THE END REALLY BRIEFLY GET
INTO MERS AND I WILL NOT TALK
ABOUT ORIGINAL SARS BECAUSE OF
TIME AND NOT THAT MEAN PEOPLE
ARE WORKING ON THE VIRUS WITH
THE SELECT AGENT CONCERNS AS
WELL, BUT IT IS 1 OF MY FAVORITE
VIRUSES SO IF YOU LIKE IT, WE
CAN TALK AFTERWARDS.
SO WE ALREADY TALKED ABOUT THE
NEED FOR SMALL ANIMAL MODELS.
AT UNC WE'RE A MOUSE-CENTRIC
ORGANIZATION AND THAT'S WHERE
OUR EXPERTISE AND OUR BL3
EQUIPMENT WAS FOCUSED AND SO WE
WANTED TO GENERATE MOUSE MODELS
AS QUICKLY AS POSSIBLE INSTEAD
OF SWITCHING OVER TO A NEW
ANIMAL SPECIES AND HAVING TO
RESTART.
SO LUCKILY WITH THE HISTORY OF
RESEARCH ON HUMAN AND THAT
CORONA VIRUSES, THERE WAS A
REALLY GOOD AMOUNT OF
FOUNDATIONAL KNOWLEDGE ABOUT
WHAT THOSE CONTACT RESIDUES WERE
FOR BINDING WITH THE SPIKE
PROTEIN OF THE VIRUS AND THE ACE
2 RECEPTOR BECAUSE WE LEARNED
VERY QUICKLY THAT ACE 2 IS ALSO
THE RECEPTOR FOR SARS 2.
SO KENNY WAS A GRADUATE STUDENT
OF [INDISCERNIBLE] AND HE WORKED
WITH SEVERAL OF US ON THE
PROJECT BUT HE QUICKLY MODELED
THE CONTACT RESIDUES AND TAKING
ADVANTAGE OF THIS PREVIOUS
RESEARCH WAS ABLE TO IDENTIFY
THAT AT RESIDUE 498 BUT THERE
WAS A GLUTAMINE HERE INSTEAD OF
A PRETTY CONSERVED TYROSEEN AND
OTHER SPIKES THAT WAS ABLE TO
MEDIATE THAT CONTACT WITH ACE 2.
SO HE SPECIFICALLY MUTATED
RESIDUE 498 AND ADJACENT RESIDUE
499 BECAUSE THAT WAS A TYPE OF
PROLEAN AND HE THOUGHT IT WAS
INTERVIEWS A WEIRD KINK IN THE
STRUCTURE OF THE THAT WAS NOT
ABLE TO RESTORE BUT INTRODUCE
BINDING TO HOUSE ACE 2, THAT WAS
MORE SIMILAR TO WHAT'S SEEN WITH
HUMAN ACE 2, THESE HYDROGEN
BONDS WERE RESTORED AND WE'RE
ABLE TO DEVELOP A MODEL THAT
DOES INFACT INFECT MICE, THAT
WAS REALLY A REPLICATION ONLY
MODEL, HOWEVER, SO SUBSEQUENT
WORK WAS DONE PASSAGING THIS IN
VIVO, THROUGH THE LUNGS OF
BALB-C MICE AND AFTER 10
PASSAGES WE HAD A HIGHLY
VIRULENT VIRUS NAIN A DOSE AND
AGE DEPENDENT MANNER COULD
EITHER BE PRETTY MILD DISEASE OR
EXTREMELY LETHAL DISEASE SO
HERE'S WRUOF THE A SCHEMATIC OF
THAT WHOLE GENOME OF THE MATTEN
VIRUS WITH THE 2 MUTATIONS THAT
WE INTIEWS SPECIFICALLY, AND
THEN THE 5 ADDITIONAL MUTATIONS
THAT CAME UP THROUGH PASSAGE.
SO USING THAT VIRUS WE DID ALL
THE BASIC I HAVE ROW LONGICAL
OBSERVATION.
DOSE RESPONSE, TIME COURSE,
SETTLED IN ON PRETTY STANDARD
CHARACTERISTICS THAT HAVE ALL
BEEN PUBLISHED AND I DON'T WANT
TO SPEND TOO MUCH TIME ON THAT.
BUT WHAT I THINK IS MOST
INTERESTING TO A LOT OF US IS
DIVING IN ON WHAT IS REALLY
HAPPENING IN THE LUNG, IN THE
ORGANS THAT ARE INFECTED, MICE
LOSE WEIGHT THAT'S A GREAT
QUANTITATIVE MARKER BUT IT'S NOT
SUPER REFLECTIVE OF HUMAN DEC,
IT'S NOT THE MOST RELEVANT OF
THE PHENOTYPE 2, BUT WHAT'S
HAPPENING IN THE ORGANISM,
WHAT'S HAPPENING DISEASE WISE?
WE COULD SEE WHAT WOULD HAPPEN
FROM OTHER CORONA VIRUSES THAT
WE HAVE PEAK TITER EARLY, SO
STAIN FOR NUCLEIC CAPSID HERE IN
THOSE AIRWAY EPITHELIAL CELLS,
AND ALSO THE TYPE 2 SITES IN THE
LUNG, THIS REPLICATION DECREASES
OVERTIME BUT OUR LUNG PATHOLOGY
INCREASES EMPLOY A PRETTY
STANDARD STORY FOR A LOT OF
RESPIRATORY VIRUSES.
BUT GETTING BEYOND THESE INITIAL
CHARACTERIZATIONS, WE'VE DONE A
LOT OF WORK MORE RECENTLY WITH
DIFFERENT DIGITAL PARTICIPATE IN
A TRIALIAL FILING PLATFORMS
GETTING INTO MORE FUN MODERN
TECHNOLOGY AND WITH THESE ARK
PROACHS, I THINK WE CAN REALLY
LEARN A LOT ABOUT WHAT'S
HAPPENING IN THE LUNGS.
SO IN THIS WORK BY NICK
[INDISCERNIBLE] IN OUR GROUP,
HE'S DONE DIGIT AT SPATIAL
PROFILING.
THIS IS 1 OF MANY WE'RE TESTING
OUT TRYING TO GET AS MUCH DATA
AS WE CAN FROM ALL THESE
DEFINITE PLATFORMS BEFORE WE
SETTLE IN ON SOMETHING, BUT I
THINK IT'S ALSO REALLY
COMPLIMENTARY.
SO THIS IS LOOKING AT ACUTE
INFECTION, POWER DAYS POST
INFECTION.
YOU CAN SEE JUST THE H& E
STAINED IMAGES DOWN HERE AND
THEN LOOKING AT EXPRESSION OF
SEVERAL GENES THAT WERE
INTERESTED IN, SO KRT8 IS A
MARKER OF REPAIR OF THOSE
ALVEOLAR CELLS.
WE'RE LOOKING AT SURFACTIN
EXPRESSION, CLASSICALLY, THIS
GOES DOWN AFTER INFECTION, WE
SAW THIS IN SARS 1, WE SEE IT IN
THE TYPE 2 PNEUMOSIGHTS ARE
KILLED OFF, THEY'RE NOT ABLE TO
PRODUCE SURFACTIN ANYMORE AND
THIS GOES WITH OVERALL
DEPRIVATIONMENT OF LUNG DEC AND
THEN WE ALSO SEE INCREASED
EXPRESSION OF GENES LIKE HIP 1
ALPHA WHICH IS INDICATIVE OF
HYPOXIA THAT'S HAPPENING IN
THESE LUNGS AND GIVING US A NICE
COMPREHENSIVE PICTURE OF LUNG
DEC, YOU CAN SEE SPATIALLY WHERE
THIS IS HAPPENING RELATIVE TO
LIKE YOUR LARGE AIR WAYS, YOUR
VASCULATURE, AND WE WERE GOING
TO BE SPENDING A LOT MORE TIME
ON THIS.
IF ONLY WE CAN DO IT FOR EVERY
MOUSE BUT DEFINITELY NOT
POSSIBLE.
WE'VE ALSO BEEN WORKING WITH
BOARDS ON EMPLOY CAMPUS, GETTING
SET UP WITH A RECYCLER, LOOKING
AT RNA PROTEIN EXPRESSION AND
IT'S NICELY COMPLIMENTARY AND IF
ONLY WE HAD THE TIME AND
RESOURCES TO DO IT ON A
FULL-TIME COURSE OR AN ARRAY OF
GENETIC PACK GROUNDS BUT JUST
FROM BASIC CHARACTER EYATION
WITH THIS ACUTE IN INFECTION,
YOU CAN SEE THAT IN COMPARISON
TO THE MOCK INFECTED LOWNGOT
LEFT HERE AND ON THE RIGHT WHERE
THE MATTEN INFECS HAPPEN, IN
GREEN YOU CAN SEE THAT THE
SURFACTIN THAT'S NICELY
EXPRESSED ALL THROUGHOUT THE
LUNG HERE HAS REALLY DISAPPEARED
SPERKS LEECIALLY IN AREAS WHERE
YOU SEE THE RED, WE HAVE
INCREASED BLUE HERE, THIS IS
MUCIN, ALL IN ALL A REALLY NICE
PICTURE OF WHAT'S HAPPENING WITH
THE LUNGS SO WE'RE NOT JUST
LOOKING AT BULK RNA OR PROTEIN,
BUT YOU CAN SEE REALLY FOCUS
REGIONS OF DISEASE, SO UP HERE
THERE ISN'T THAT MUCH INFECTION
AND THE LUNG LOOKS PRETTY
NORMAL, DOWN HERE WE HAVE A
REALLY NICE PATCH OF VOCAL
DISEASE EMPLOY OF COURSE WE'VE
BEEN DOING A LOT OF RNAseq AND
IN THIS CASE IT'S NICE BECAUSE
WE CAN DIVE INTO WHAT'S
HAPPENING IN PARTICULAR AND
DISTINCT LUNG CELLS AND FIND OUT
HOW THEY'RE RESPONDING TO
INFECTION IN SO HERE'S THAT
CLASSIC UMAP PICTURE OF SNCHT
CELL POPULATIONS THAT WE'RE
SEEING IN THE LUNG.
LOOKING AT THAT POPULATION AND
SEE THAT INFECKED ANIMALS THERE
ARE A LOT OF ISGs BEING
EXPRESSED AS 1 WOULD EXEC, THERE
ARE A LOT OF MATURATION AND
ACTIVATION TELLING US HAY, IT
WILL BE NICE TO DIVE IN MORE
ABOUT WHAT'S HAPPENING ON
NUTRIFILLS, LET'S DO DEPLETION
SPRMS, LET'S FOLLOW UP WITH
WHAT'S HAPPENING HERE AND IT
WILL CONTINUE BEING A PRIMARY
RESEARCH OF FOCUS WE KNOW THERE
ARE NONPULMONARY CENTER FOR
EXCELLENCE ON AGINGS OF COVID-19
AND IT CAN BE DIFFICULT TO ASK
THE MICE HOW ARE YOU FEELING?
DO YOU HAVE BRAIN FOG?
ARE YOU ACHING, THOSE REALLY
NICE BEHAVIORIAL ASSAYS SO WE'RE
JEALOUS ABOUT THAT SO WE KNOW IN
HUMAN 19 COVID PATIENTS THIS CAN
BE AN EFFECT IN THE RESPIRATORY
SYSTEM AND THROUGHOUT THE REST
OF THE BODY AND HALLMARKS OF
THIS ARE THAT YOU HAVE INCREASED
FIEB RENO GENERATED GENERATION,
THROMBIN, ENHANCED PLAZ MIDS
--PLASMID, SO
THIS WHOLE SYSTEM IS HIGHLY,
HIGHLY ACTIVE.
SO YOU ARE GETTING A LOT OF
PLOTS BEING DEPOSITED AND
THEY'RE NOT BREAKING UP AS THEY
NORMALLY WOULD.
IN OUR SARS-COV-2 INFECTED MICE
WE SEE SIM LAWYER LIGFATTURES AS
MUCH AS WE CAN SO FAR AND THIS
IS JUST LOOKING AT FIEB RIN
DEPOSITION IN THE LUNGS AFTER
INFECTION.
IF WE ZOOM IN YOU CAN SEE THE
STAINING AND IT'S NOT JUST IN
THE VASCULAR SPACES SO THIS IS
NOT HEALTHY, THIS IS NOT
SOMETHING THAT THAT WE WANT TO
SEE, IF WE GO IN AND LOOK
SPECIFICALLY AT PLOTTING MARKERS
THAT YOU SEE INCREASED PLASMA
GENERATION, HIGHER PEEK AND
FASTER TIME TO THAT PEEK AND
ALSO INCREASE THROMBIN
GENERATION AND A FASTER TIME TO
THAT PEEK.
SO WHILE IT CAN BE INTERESTING
TO LOOK AT THESE FACTORS IN
MICE, WE ARE STILL SEEING
SIGNATURES OF WHAT HAPPENS IN
HUMANS.
AND THEN DIVING IN MORE DEEPLY
SOME OF THAT BECAUSE WE HAVE
NICE GENETIC MODELS AVAILABLE TO
US IN MOUSE SYSTEMS WE CAN LOOK
AT PLASMA DEFICIENT MICE OR
FIBROGENERATED DEFICIENT MICE
AND SO THAT THEY MUTATED IN
THOSE WAYS.
PLUS SEE THAT THE MICE ARE
PROTECTED FROM DISEASE COMPARED
TO THE HETEROZYGOUS ROLES AND WE
ALSO HAVE CHANGES IN MACROPHAGE
AND PERSISTENT IN THE LUNGS OUT
AT LATER TIME POINTS IN THE
DEFICIENT MICE, SO A LOT OF
AREAS WE CAN FOLLOW UP HERE AND
WE DISSECT OUT SPECIFIC CELL
TYPES EVEN FURTHER BUT I THINK
THESE ARE STRENGTHS OF OUR MOUSE
MODELS WE'VE ALSO WORKED ON
THESE AND THAT IS DONE IN
COLLABORATION WITH GREGORY VIX'S
LAB AT THE UNIVERSITY, AND SO
SHE HAS DONE A LOT OF INFECTIONS
AND SENT OFF CNS TISSUE TO THEIR
LAB FOR ANALYSIS.
AND WHAT'S REALLY INTERESTING
THAT THERE'S A DECREASE IN THE
TIGHT JUNCTION MARKERS SO
IMPLYING THAT THERE IS, POSSIBLY
SOME BREAK DOWN OF THAT BLOOD
BRAIN BIRRIER SURFACESSIER IN
THE MICE THAT ARE INFEC WIDE
MA-10 AND I KNOW THEY HAVE A LOT
MORE WORK PLANNED IN THIS AREA.
SO THAT'S MOSTLY BEEN ACUTE
DISEASE BUT WE'RE ALSO VERY
INTERESTED IN WHAT'S HAPPENING
LONG-TERM IN THESE MICE THAT
HAVE BEEN INFECTED.
SO THIS IS DATA THAT WAS
PUBLISHED A LITTLE BIT AGO, SO I
WILL GO THROUGH IT REALLY
QUICKLY, BUT WE SHOWED ACUTE
INFECTION BY MOST TRADITIONAL
MEASURES THAT WE CAN OBSERVE IN
THE LABORATORY.
THE MICE HAVE RECOVERED BY 15
DAYS POST INFECTION.
WHAT GETS INTERESTING IS--CAN WE
MAKE THE SLIDES COME BACK?
THANK YOU.
IT SHOULD BE ON THE NEXT SLIDE,
BUT WHEN WE LOOK AT THESE MOST
INFECTION, THIS IS A MEASURE OF
FIBROSIS THAT THESE LUNGS ARE
NOT NORMAL, THEY HAVE NOT COME
BACK TO BASE LINE AND IF WE DO
SPECIFIC STAINING, THIS IS AT 15
AND 30 DAYS POST INFECTION, CAN
YOU SEE THERE IS A LOT OF SMOOTH
MUSCLE ACTING, A LOT OF SERIOUS
RED STAINING, MARKERS OF
FIBROSIS.
WE DID SPATIAL PROFILING ON
THESE LONGER TERM MICE AS WELL,
AND QUANTIFIED MARKERS OF
FIBROSIS TO CONFIRM OUR INITIAL
SUSPICIONS.
A LOT OF COLLAGENS, FIEK ROUGH
ATOM NECTIN, SBP1 IS A MARKER OF
MACROPHAGES, ALL OF THESE GO UP
AND STAY UP IN THE LUNGS OF
SARS-COV-2 INFECTED MICE.
NOW THIS CAME UP IN STACEY'S
TALK SO I'M GLAD THIS SLIDE CAME
IN.
SO THESE ARE MICE FROM THE
CC POPULATION AT UHC, IT'S A
GENETIC POPULATION THAT WAS
DERIVED FROM 8 FOUNDER LINES,
THEY WERE BRED TOGETHER IN THESE
BREEDING SCHEMES OVER AND OVER
AND OVER AND THEN INBRED FOR 30
OR SO GENERATIONS BEFORE THEY
WERE RELATIVELY FIXED.
SO WHEN YOU LOOK AT THE MICE,
THEY HAVE BITS OF EACH OF THE 8
FOUNDERS, THEY'RE COLOR COATED
SO A BLACK BIT INDICATES THAT
CAME FROM B6 AND A YELLOW BIT
INDICATES IT CAME FROM AJ, AND
THE CC REPRESENT BOO 90% OF THE
GENETIC DIVERSITY IN THE GENOME,
MOST OF THAT GENETIC SIGNIFY
DIVERSITY CAME FROM THE PARENTS
THERE BUT THERE ARE 5 MORE
STANDARD LAB LINES THAT WERE
INVOLVED IN THIS AS WELL.
SO SARAH, ALEX AND JOHN POWERS
AND NICK, AND SPENT A LOT OF
TIME INFECTING A NUMBER OF
CC-LINES, WE DID SOMEWHAT
PRESLICT FOR MICE THAT WEREN'T
ALL GOING TO DIE, WHEN WE DID
SHORTER TERM INFECTIONS THERE
WERE SCREENING FOR ANIMALS THAT
WERE NOT IMPOSSIBLE TO WORK WITH
AS WELL BECAUSE IF YOU HAD THE
PLEASURE, YOU KNOW SOME OF THEM
RETAINED A LOT OF THEIR WILD
CHARACTERISTICS AND THAT'S NOT
AN ANIMAL YOU CAN HAVE IN 30
DAYS.
SO WHAT YOU CAN SEE FROM 12 OR
SO LINE SYSTEM THAT THERE'S A
HIGH AMOUNT OF VARIABILITY AND
RESPONSE TO INFECTION, THESE
WERE ALL INFECT WHD THEY WERE 12
WEEKS OLD, BUT A VERY WIDE RANGE
OF OUTCOMES BECAUSE OF THAT
GENETIC DIVERSITY.
SO 3 FELT LINES I WANT TO
HIGHLIGHT HERE, CC4 AND CC10, IN
THIS THE ORANGE AND GRAY HAD
MODERATE WEIGHT LOSS BUT CC25 IN
THE RED HAD MORE WEIGHT LOSS AND
THE MICE WITH INFEC.
WHEN WE TOOK THESE OUT TO 30
DAYS POST INFECTION AND REALLY
WERE FOCUSED ON FIBROSIS
PHENOTYPES, THE SLIDES THAT WERE
ORIGE NAWILLLY STAINED HERE WENT
OFF TO JOCK AT MICHIGAN STATE
UNIVERSITY FOR EXTENSIVE
PATHOLOGY SCWOAR SCORING AND HE
WAS REALLY FOCUSED ON A NUMBER
OF PARAMETERS ASSOCIATED WITH
HIEBROSIS.
SO LOOKING AT THE RANGE OF
SCORES HERE, JUST GENERAL
FIBROSIS, YOU CAN SEE THAT ON
CC10, AT DESCENT FIBROSIS WHERE
THEY HAD NOTHING EVEN THOUGH
THEY HAD SIMILAR WEIGHT LOSS,
CC25 HAD PRETTY EXTREME WEIGHT
LOSS WAS ALSO HIGH BUT NONE OF
THESE LINES HAD MUCH IN THE WAY
OF MUCUS CELL OR DISPLASSIA EVEN
THOUGH THERE WERE SOME LINES
THAT DO.
WHEN WE ELECTRIC AT WHAT'S
ACTUALLY HAPPENING ON H& E
STAINED SLIDES HERE YOU CAN SEE
THAT THE CC4 MICE AND THE CC10
VERY DIFFERENT LOOKING EACH
THOUGH THEY HAVE THAT SIMILAR
WEIGHT LOSS, SO WE HAVE SOME
TERTIARY LYMPHOID STRUCTURE
THERE, IN THE CC10S AS WELL AS A
LOT OF FOAMY MACROPHAGES AND A
LOT OF INTERESTING STUFF WE
WOULD LIKE TO DIVE IN MORE
DEEPLY THERE AND THEN IN THE
CC25S THERE ISN'T A LOT OF AND
SO OTHER ASPECTS OF FIBROSIS.
OKAY SO ALL OF THIS IS BUILT ON
THE MA-10 PLATFORM WHICH IS
BASED ON SEQUENCE WHICH WE
HAVEN'T SEEN IN A LONG, LONG
TIME AND SOMETHING--SO SOME WORK
THAT JOHN POWERS IN THE LAB HAS
LED, HAS REALLY TAKEN ADVANTAGE
OF THE FRAGMENT SYSTEM THAT WE
USE TO GENERATE OUR INFECTIOUS
CLONE, TYPICALLY WE USE 6 OR 7
FRAGMENTS AND THIS IS FROM AND
TOGETHER WITH THE TRANSCRIBED,
SO, WE DESIGNED FRAGMENT F TO
ENCODE THE ENTIRETY OF THE SPIKE
JOIN, AND SO THE REST OF THE
GENOME HAS THOSE AND THE SB4
MUTATIONS ARE THERE, BUT WE
SWAPPED OUT THE SPIKE OR
WHATEVER THE VARIANT OF SPIKE IS
THAT WE'RE INTERESTED IN, IN
THIS CASE ALL OF THESE MUTATIONS
WENT IN AND THEN A LOT OF TIMES
WE WILL MAKE A VERSION OF THE
IRS THAT HAS IT IN PLACE SO THAT
WE CAN HAVE A REPORTER VIRUS
TOOL AVAILABLE FOR LIVE VIRUS
NEUTRALIZATION ASSAYS.
SO THAT IN WORK HERE, CAN YOU
SEE THE OMICRON VIRUS CAUSES AGE
DEPENDENT DEC SO IN YOUNG,
THERE'S NOT MUCH GOING ON.
NO WEIGHT LOSS, THE VIRUS IS
REPLICATING JUST FINE AND WE
LOOK AT RESPIRATORY FUNCTION,
SIMPLE BODY TOMOGRAPHY, 10 H
SPOKES, LIKE DAY 2 OR 3, AGAIN
NOT MUCH OF A SIGNAL IN YOUNG
BAC1 INFECTED MICE.
WHEN WE GO TO A 12 MONTH OLD
BALC MODEL THOUGH, WE HAVE 2
DIFFERENT DOSES FOR MID10 TO THE
FOURTH AND FIFTH HERE.
THERE'S A LOT MORE WEIGHT LOSS
THAN THERE WAS WITH THE YOUNG SO
THERE'S ABSOLUTELY NOTHING BUT
STILL NOT AS MUCH DISEASE AS YOU
WOULD SEE IN THE MATTEN INFECTED
MICE.
OF COURSE WE'VE ALSO DONE THIS
WITH BA5 AND SUBSEQUENT FLAVORS
OF OMICRON, IN THIS CASE, BA5
HAS A SIMILAR DEC PENE O TYPE TO
MATTEN, WHERE THERE'S MORTALITY
THAT DEVELOPS AT A PRETTY
SIMILAR TIME POINT, RESPIRATORY
FUNCTIONS ARE SIMILAR SO THESE
TOOLS ARE REALLY IMPORTANT FOR
LOOKING AT BREDTH OF VACCINE
PROTECTION, DIFFERENT DISEASE
WHERE HAVING A ROBUST MODEL IS
REALLY IMPORTANT.
RALPH LIKES TO SAY IT'S EASY TO
SO YOUR VACCINE WORKS IF YOUR
CHALLENGE VIRUS DIDN'T MAKE THE
ANIMAL SICK TO BEGIN WITH SO WE
WANT A ROBUST VIRUS SO WE CAN BE
COMTENT IN WHATEVER WE'RE
TESTING.
>> YAWN HAS ALSO TAKEN THE BA5
INFEBTED MICE OUT TO LATER TIME
POINTS.
MOSTLY WITH THE LOWER DOSE SO
THAT THEY ACTUALLY SURVIVE THAT
LONG, AND WE ALSO SEE SIGNS OF
FIBROSIS, THIS IS SERIOUS RED
STAINING HERE AT LATER TIME
POINTS IN THOSE BA5 INFECTED
LUNGS.
SO SIMILAR PATHOLOGY TO WHAT WE
SEE IN THE ME10 INFECTED MICE
WHEREAS WITH THE BAC1 ANIMALS
NOT SO MUCH WAS HAPPENING EMPLOY
SO FOR THE LAST LITTLE BIT OF
THE TALK, I WANT TO TALK ABOUT
OTHER MODELS AND OTHER CORONA
VIRUSES BECAUSE GOING WITH A
TYPICAL MOUSE ADAPTED VIRUS
APPROACH IS MOST COMMON BUT
THERE ARE SOME UNEEK TOOLS WE
CAN USE.
SO 1 OF THOSE THINS WE DID
EARLIER IN THE PANDEMIC WAS TO
WORK WITH J.VICTOR GARCIA'S LAB
AT UNC BUT MOST RECENTLY MOVED
TO UNIVERSITY OF ALABAMA.
HE WAS AN HIV RESEARCHER BUT HIS
LAB DEVELOPED THESE HUMAN
ORGANOID PLANT AND THEY CAN GROW
IT UNDER THE SKIN AND GROW THEM
AS LUNG CELLS OR SOMETIMES THEY
ADD HUMANIZATION OF THE MOUSE
IMMUNE SYSTEM WITH BONE MARROW,
LIVER, THYMUS, BUT YOU GET THIS
MOUSE THAT AS A BIG LUMP
UNDERNEATH ITS FUR AND WHEN A
CROSS SECTION IS TAKEN HERE AND
H& E STAINED IT'S HIGHLY
VASCULARIZED.
THERE ARE APPROXIMATIONS OF WHAT
WOULD BE AN AIR WAY IF THIS WAS
ACTUALLY IN CONTACT WITH AIR.
SO YOU CAN SEE WHAT OUR
PREALVEOLAR SPACE IS, AND LUNG
STRUCTURE THAT IS IMPEDIMENTS
SIMILAR TO WHAT WOULD BE IN A
HUMAN.
SO THESE ARE HIGHLY INFECTIBLE,
AND YOU CAN SEE THAT IN THE LUNG
ONLY MICE THAT DON'T HAVE THE
RECONSITUTED IMMUNE SYSTEM, THAT
THIS INFECTION DOES PERSIST OVER
TIME.
WHEN WE STAIN, WHAT CELLS IN THE
LUNG ARE INFECTED HERE AND YOU
CAN SEE THE SARS NUCLEIC CAPSID
IS IN RED AND IT'S CO LOCALIZING
FOR MARKERS FOR EPITHELIAL CELLS
SO THIS IS CONSISTENT WITH WHAT
WE WOULD EXPECT.
AND SO YOU DO BASIC
CHARACTERIZATION HERE, IT IS
NICE YOU CAN TEST ANTIVIRALS IN
HUMAN CELLS IN A QUASI-IN VIVO
MODEL BUT WHAT I THINK IS REALLY
INTERESTING IS THAT IN THESE
MODELS WE'VE ALSO BEEN ABLE TO
GET INTO LONGER TERM INFECTION,
LOOKING AT LONG-TERM TOPOLOGY
AND EVEN IN THIS LONG-TERM
MEASURING SYSTEM, YOU SEE
MARKERS OF FIBROSIS MIMICKING
WHATY - SEE IN PEOPLE AND IN
MICE, SO I THINK BEING OPEN
MINDED TO NONTRADITIONAL MODELS
CAN BE PRETTY PRODUCTIVE.
AND THEN FINALLY I WANT TO TALK
ABOUT LONG-TERM MERS MODELS.
SO PREVIOUSLY WE HAD ADAM
COCKEREL WHO DEVELOPED A
TRANSGENIC 38 MODEL FOR
MERCOMPATTIBILITY AND THE
COMPATIBILITY WHERE IT CANNOT
BIEBD TO THE DP4454 RECEPTOR, HE
ENDED UP WITH A VIRUS THAT WAS
HIGHLY PATHOGENIC AND LETHAL IN
A MOUSE MODEL.
MORE RECENTLY HEATHER
[INDISCERNIBLE] HAS TAKEN THIS
AND INSTEAD OF LOOKING AT ONLY
ACUTE INFECTION, SHE WANTED TO
SEE IF IN A LONG-TERM MERS MODEL
TO WE SEE SIMILAR SIGNS OF DEC,
SIMILAR ASSPECS OF CHRONIC
INFECTION AS WE DO WITH SARS 2.
AND IT SEEMS LIKE THIS IS GOING
TO BE THE CASE.
WE HAVEN'T GOTTEN THAT DEEPLY
INTO IT AS STANLEY'S LAB HAS BUT
HEATHER IS DOING COOL STUFF
HERE.
SHE AND NICK HAVE DONE SINGLE
CELL ANALYSIS ON THESE MERS
INFECTED MICE, SEE ALL THE
DIFFERENT MAPPED POPULATIONS
HERE.
LOOKING AT 30 DAYS POST
INFECTION THEY DID, ON DAY 3 AND
30 TO CAPTURE EARLY AND LATE
TIME POINT AND LOOKING AT THE
TOP DIFFERENTIALLY EXPRESSED
GENES HERE AND COMPARING THEM TO
WHAT WE SEE IN A SARS-COV-2
INFECTION, SOME OF THE CLASSIC
CATEGORIES THAT ARE INTERESTED
AND ARE THERE MARKERS OF
FIBROSIS, COMPLEMENT COMES UP A
LOT IN SARS-COV-2 INFECTED
LUNGS, AND SO HERE, WE'RE
LOOKING AT SBP1 WHICH IS THE
MARKER OF FIBROTIC MACROPHAGES
AND YOU CAN SEE THAT IN THE
ALVEOLAR MACROPHAGE POPULATION
THAT EVEN IN ANIMALS THAT HAD
MODERATE DISEASE, AND ALSO MORE
SEVERE DISEASE AS MEASURED BY
THE WEIGHT LOSS THAT WE HAVE A
LOT OF THE SBP1 EXPRESSION IN
THIS POPULATION SO THIS ANALYSIS
IS REALLY PRELIMINARY BUT I KNOW
HEATHER IS EXCITED TO DO MORE
WITH IT.
SO WITH THAT, HOPEFULLY I'VE
SHOWN YOU THAT EVEN THOUGH HUMAN
CORONA VIRUSES DON'T ALWAYS
INFEJT MICE INITIALLY, WE CAN
KIND OF FIX THAT PROBLEM AND
WHILE THAT TAKES SOME
ENGINEERING WE END UP WITH
MODELS THAT ARE STILL HIGHLY
REFLECTIVE OF HUMAN DEC AND THAT
WE CAN USE TO TEASE APART SOME
FINELY MECHANISTIC DETAILS OF
WHAT'S HAPPENING, AND DISSECT
LUNG PATHOLOGY AND FUNCTION IN
WAYS WE CAN'T DO IN HUMAN
POPULATIONS AND 1 THING I LOVE
ABOUT THESE VIRUSES, SARS, MERS
AND SARS 2, ALL CAUSE PRETTY
SIMILAR DEC PATHWAYS AND THERE'S
A REAL OPPORTUNITY THERE I THINK
TO FIND COMMON PATH WAGHT IS,
COMMON THERAPEUTICS AND
HOPEFULLY BE PREPARED FORMENT
NEXT SARS VIRUS IS.
I WANT TO THANK ALL PEOPLE WHO
HELPED WITH THIS WORK.
I HIGHLIGHTED PEOPLE FROM THE
CBRG BUT THERE ARE A LOT MORE OF
US WHO ALSO AND SUPPORT IN
DIFFERENT WAYS, THE PATHOLOGIST
STEPHANIE AND JACK HAVE BEEN
REALLY, REALLY CRITICAL TO THIS.
THE MARSICO LUNG INSTITUTE,
COLLABORATORS, AND GARCIA LAB
COLLABORATORS, AND WITH THAT I
WOULD LOVE TO TAKE ANY
QUESTIONS.
>> THANK YOU SO MUCH.
WE HAVE TIME FOR QUESTIONS.
NNICE TALK, REALLY ENJOYED IT,
HOW ABOUT THE HOST STRAINS, YOU
FOUND THAT SUBLINEAGES OR
SUBSTRAIN THEY HAVE DIFFERENT
DEC SEVERITY, HAVE YOU
INVESTIGATE WITH WHAT KIND OF
GENETIC LOCI THAT CORRELATE WITH
THE DISEASE?
>> SO WE HAVE DONE A NUMBER OF
KREENS LOOKING FOR LOCI THAT ARE
ASSOCIATED WITH ACUTE INFECTION,
WE HAVEN'T DONE A FULL LONG-TERM
DISEASE ANALYSIS YET JUST
BECAUSE THE NUMBER OF MICE WE
WOULD NEED TO TAKE OUT TO DAY 30
WOULD BE PRETTY EXTREME.
WHAT WE'RE MORE INTERESTED IN
DOING IS TAKING HIGH AND LOW
OUTLIERS WITH THAT DAY 30
FIBROSIS PHENOTYPE AND DOING AN
F2 PROMPT SO YOU CAN HAVE
ANALYSIS THERE.
WE DO KNOW THAT THE GENETICS OF
EVERY CC MOUSE SINCE THEY'RE
INBRED AT THIS POINT SO WE CAN
LOOK SPECIFICALLY FOR LINES THAT
MAYBE HAVE LOCI THAT WE KNEW
ABOUT PREVIOUSLY SO WE COULD SEE
DOES THIS STILL MATTER FOR
LONG-TERM DEC OR IF WE SAY WE
KEEP HITTING ON THE SAME
CHROMOSOME LINE LOCUST BECAUSE
IT SEEMS TO OVERPOWER EVERYTHING
ELSE, WE CAN SPECIFICALLY
EXCLUDE THAT.
THAT'S IN THE FUTURE.
>> STANLEY?
>> I HAD A QUESTION ABOUT THE
COLLABORATIVE CROSS FIND, SO
ORIGINALLY THE IDEA WAS TO
DEVELOP A THOUSAND DIFFERENT
LINE ANDS I DON'T KNOW IF THEY
DEVELOPED THAT BUT UNC FOLKS
HAVE ACCESS TO THE FACILITY.
HOW DO YOU DECIDE WHICH 1 YOU
USE?
DO YOU JUST PICK 50 AND 25, YOU
GET A RANGE YOU DON'T HAVE TO
WORRY ABOUT THE OTHER THOUSAND?
NI THINK THERE ARE ABOUT SEIVET
LINEUPS THAT ACTUALLY MADE IT
FULL INBREEDING, 1000 IS A
CRAZY, CRAZY IDEA.
ONE HUNDRED WOULD BE NICE.
THEY DIDN'T GET THERE.
THERE WAS A MASSIVE BREEDING
CAPACITY BOTTLENECK THAT BRED IN
GENERATION 8 OR SO WHERE A LOT
OF LINES DIED OUT BUT WHAT IS
LEFT IS STILL INCREDIBLE
LOAMACYY USEABLE.
WE HAVE A STRONG HISTORY OF
HAVING INFECTED THESE MICE WITH
ORIGINAL SARS.
SARS 2, SO WE CAN PRETTY WELL GO
IN AND SAY, THESE LINES ISSUES
THESE LINES, IF YOU ARE COMING
INTO THE CC NOT HAVING USED IT
BUYER, WHAT I WOULD RECOMMEND IS
GET 10 LINES, ANY 10 LINE ANDS
YOU WILL ALMOST CERTAINLY SEE A
LOT OF PHENOTYPIC VARIATION
BETWEEN THOSE AND THEN PICK THE
1S THAT ARE INTERESTED TO YOU
AND DO AN F2 CROSS IS THE
FASTEST WAY TOWARDS MAPPING, IF
YOU WANT TO YOU CAN GET ALL 71
IN AND INFECT ALL OF THEM AND
GET THE FULL RANGE.
WHEN YOU'RE GETTING THOSE 10
LINES DEPENDING ON WHO'S DOING
THE EXPERIMENT IN OUR GROUP,
WHAT THEY'RE EXPERIENCE WITH
MICE IS, WE MIGHT SAY, LIKE GIVE
US THE 1S THAT AREN'T JERKS SO
THAT WE DON'T HAVE TO WORRY
ABOUT ESCAPES IN THE BL3.
OR IF IT'S SOMETHING WE KNOW WE
NEED TO GET THROUGH PRETTY
QUICKLY, WE WILL ASK FOR GOOD
BREEDERS BECAUSE SOME OF THEM
WILL HAVE LITTERS OF 2-3 MICE
ONLY SO GETTING ENOUGH ANIMALS
TO COMPLETE YOUR EXPERIMENT IN A
TIMELY MANNER WITH A ROBUST
NUMBER OF MICE CAN BE
CHALLENGING SO YOU DON'T WANT
THAT TO BE A LIMITATION DOWN THE
ROAD.
SO IF YOU'RE STARTING OFF 10
LINES, GOOD BREEDERS, NICE MICE.
>> ANOTHER QUESTION, OTHERWISE
WE CAN ASK DURING THE PANEL
SESSION AS WELL.
ALL RIGHT.
WE WILL ASK IT DURING THE PANEL
SESSION EMPLOY SO THANK YOU VERY
MUCH.
>> THANK YOU.
>> OUR NEXT SPEAKER IS JUSTIN
FRERE, FROM NEW YORK UNIVERSITY
WHO WILL DISCUSS LEVERAGING THE
GOLDEN HAMSTER MODEL OF
SARS-COV-2 INFECTION TO DEFINE
SHORT AND LONG-TERM CONSEQUENCES
OF DISEASE EMPLOY.
>> SAW AM, THANK YOU SO MUCH FOR
THE INTRODUCTION, I AM JUSTIN
FREER, AND I AM A POST DOC AT
THE LABORATORY AT NYU AND I'M
EXCITED TO TALK TO YOU ABOUT THE
SARS-COV-2 HAMSTER INMODEL
INFECTION.
SO CARS KICKOFF 2 AND OOTS A
RESPIRATORY VIRUS AND REPLICATES
RAPIDLY IN THE RESPIRATORY
TRACT.
THIS IN TURNS CAUSES A MASSIVE
AMOUNT OF INFLALLATION AND ALL
THE PULMONARY DISTRESS--IT SEES
IN EVERY ORGAN FROM HEAD TO TOES
AND HAS BECOME DEFINED BY
NEUROLOGIC SEQUEL I SUCH AS
[INDISCERNIBLE].
SO THE CLINICAL PICTURE IT'S
MORE COMPLICATED THAT A LOT OF
PATIENTS, THE SYMPTOMS DON'T
FADE AFTER THE VIRUS CLEARED AND
THEY STILL HAVE SYMPTOMOLOGY OR
NEW SYMPTOMOLOGY KNOWN AS LONG
COVID.
SO AGAIN THIS AFFECTS EVERY
ORGAN FROM THE HEAD TO THE TOE
ANDS NOTABLY LYE CHARACTERIZED
BY ABNORMALITIES, BRAIN FOG,
DEPRESSION, ANXIETY, CHRONIC
PAIN.
SO IN THE LAB, A LOT OF OUR WORK
OVER THE PAST FEW YEARS HAS BEEN
TRYING TO SEE HOW IT'S MEDIATING
THESE PATHOLOGIES BOTH AT THE
ACUTE INFECTION TIME POINTSAs
WELL AS AFTER VIRAL CLEARANCE
AND HOW IS IT DOING THIS IN
INFECTED TISSUES LIKE THE LUNGS
AS WELL AS UNINFECTED TISSUES
OUTSIDE THE RESPIRATORY TRACT.
AND DOING THIS IS USING THE
GOLDEN HAMSTER MODEL, AND IT'S
BEEN REALLY NICE FOR US BECAUSE
IT HAS INDIG NOSA2, THE I HAVE
ROLL RECEPTOR FOR SARS-COV-2 AND
THAT'S SIMILAR TO HUMAN ACE 2,
AND IT INTERACTING WITH THE
SPIKE PROTEIN.
AND WHAT'S NICE IS THAT HAS ARK
LOWEDITOUS INFECT WILD-TYPE
HAMSTERS WITH NATURALLY
OCCURRING VARIANTS OF SARS-COV-2
WITHOUT HAVING TO GENETIC LIE
MODIFY THE HAMSTER OR THE VIRUS
ITSELF SO WE DON'T REALLY NEED
HAMSTER TRAINS.
SO 1 OF THE UPSIDES OF THIS IS
IT'S PRETTY SIMILAR TO WHAT WE
SEE IN HUMAN ANDS AS A RESULT WE
SEE GOOD TISSUE IN THE HAM
TERHOST, SO WE CAN SEE THE
STAINING HERE IN SARS-COV-2 AND
S, RED AND GREEN, IN THE LUNGS
AND IN THE LUNGS OF INFECTED
HAMSTERS AND HERE ON THE RIGHT
WE SEE 3D IMMUNO HISTOCHEMICAL
LABELING AND 3D IMAGING IN THE
LUNGS OF THESE SAME HAMSTERS IN
ADDITION ALVEOLAR SECTION, WE
CAN USELESS SEE INFECTION OF
SOME OF THESE SMALL AND MEDIUM
SIZED BRONCHIALS WHICH AGAIN IS
VERY SIMILAR TO WHAT WE SEE IN
HUMANS.
SO AGAIN, THE OTHER NICE THING
IS THAT AS VARIANTS HAVE EMERGED
IN HUMANS THROUGHOUT THE
PANDEMIC, THE FACT THAT THE ICE
2 RECEPTOR IN HAMSTERS IS PRETTY
SIMILAR TO THE RECEPTOR IN
HUMANS MEANS THE HAMSTERS CAN BE
INFECKED WITHOUT HAVING TO
GENETICALLY MODIFY THEM OR ADAPT
THEM IN IMI WAY, AND WHAT'S NICE
IS THEY CAUSE PATHOLOGY, SO
WE'RE SHOWING THE PATHOLOGY
CAUSED BY A COUPLE OF DIFFERENT
VARIANTS.
SO IN HEALTHY LUNGS HAVE YOU
WHITE OPEN AIR WAY SPACES, THIS
IS IMPORTANT FOR LUNG
FUNCTIONING AND HOWEVER ONCE WE
GET INFECTED WITH THE WASHINGTON
STRAIN, CAN YOU SEE DARK PURPLE
OCCURRING IN THE LUNGS SO THIS
THE SCIENCE OF THESE INTO THE
LUNGS AND FURTHER MORE WHEN WE
INFECT HAMSTERS, YOU CAN SEE
VERY SIMILAR PATHOLOGIES EVEN
SOMETIMES CHANGING IN THEIR
SEVERITY WITH SPECIFIC VARIANT
STRAINS, THIS HAS BEEN A NICE
TOOL FOR THAT.
BUT EVEN MORE THAN SARS, THE
HAMSTER IS ALSO A MODEL FOR
INFLUENZA VIRUS IN PARTICULAR
H1N1 INFLUENZA WE WERE WITH THE
2009 STRAIN AND WE CAN SEE IN
HAMSTER LUNGS, HERE WE'RE
STAINING FOR IBAC1, AND AS IN
FLU, YOU SEE SIGNS OF FLU
PNEUMONIA, AND YOU GET NICE
INFILTRATION.
SO THIS FLU MODEL HAS BEEN
USEFUL IN OUR STUDY OF SARS AS
WE'VE BEEN ABLE TO COMPLETE SARS
AND KICKOFF 2 AND FLU SIDE BY
SIDE AND LOOK AT DIFFERENCES IN
HOW THEY'RE CAUGHT IN THE
DISEASE BY DOG COMPARISONS
BETWEEN THE VIRUSES.
SO IN GENERAL, I'VE EXCITED TO
TELL YOU THE STORY ABOUT HOW
WE'VE BEEN TRYING TO MODEL THE
LONG ACUTE AND SARS-COV-2
INFECTION.
THIS PROJECT STARTED OUT A FEW
YEARS AGO, RIGHT WITH WHEN WE
WERE FIRST STARTING TO LEARN
ABOUT LONG COVID.
SO OUR FIRST ATTEMPT TO TRY CAN
MODEL THIS IN HAMSTERS WAS WE
REALLY NEED TO DO A SYSTEMIC
SCREEN, LOOKINGA THE ORGANS THAT
WERE IN THE AFFECTED HAMSTER,
AND REDUCING ANY PATHOLOGIES
THAT WERE ACUTE INFECTION TIME
POINTS AND FOLLOWING INFECTION,
THAT MIGHT MEDIATE THESE POST
ACUTE SEQUEL I INFECTION AND THE
WAY WE WANT TO IDENTIFY
SARS-COV-2 DOING IN A UNIQUE
WAY, IS WE WANT TO COMPETE IT
HEAD-TO-HEAD WITH ANOTHER VIRUS,
AND THIS IS WHERE THE FLU MODEL
COMES IN, SO THIS IS IN SOME
CASES BUT IT DOESN'T SEEM TO
OCCUR TO THE SAME EXTENT AT
LEAST TO THE SAME SEVERITY WE
SEE IN LONG COVID.
SO HERE WE FIRST INFECTED
HAMSTERS WITH FLU AGAIN, THIS IS
THE H1N1 FLIEN FLU STRAIN OR
SARS-COV-2, FOR THE WASHINGTON
STRAIN.
WE DID AT TIME LOOKING AT
REPLICATION DYNAMICS OF VIRUSES
IN THE LUNGS OVER TIME AND SEE
THAT THE FLU AND SARS-COV-2 HAVE
SIMILAR INFECTION KINETICS, BOTH
PEAK AT THE SAME TIME, AND THEY
CLEAR BETWEEN DAYS 5 AND 7,
COMPLETE BY 14, AND THIS
PRESENTED A NICE MODEL IN WHICH
WE CAN COMPARE THE 2 VIRUSES SO
OUR FIRST MAINLIOR SPRM IS WE
DID A LARGE LONGITUDINAL COHORT,
SARS-COV-2 INFLECTED, FLU INFECT
TD HAMSTERS AND THEN WE LET THEM
GO OUT TO 3 DAYS POST INFECTION,
AND 31 DAYS POST INFECTION
AGAIN, AT LEAST 3 WEEKS IT SEEMS
BEYOND VIRAL CLEARANCE TO SEE IF
WE CAN SEE WHAT'S GOING ON IN A
BUNCH OF DIFFERENT ORGANS THAT
MIGHT BE RELATED TO PERSISTENT
PATHOLOGY FROM THESE INFECTIONS.
SO AT THESE END POINTS WE TOOK
TISSUES, NERVOUS SYSTEM TISSUES
AND NONNERVOUS SYSTEM TISSUES SO
WE PARTNERED WITH
[INDISCERNIBLE] LAB AND LED BY
ALEX [INDISCERNIBLE].
THEY WERE NEUROTESTIFIES THAT
HELPED US DISSECT OUT FUNCTIONAL
BRAIN AREAS SO WE WERE ABLE TO
TAKE BRAIN AREAS THAT WERE
RELEVANT TO THE SYMPTOMOLOGY OF
LONG COVID AND WE'RE ABLE TO DO
SEQUENCING ON ALL OF THESE
DIFFERENT ORGANS AND TISSUE
AREAS AS WELL AS DO EFTHIMIOS ON
O LOGIC CHARACTERIZATION.
SO THERE WERE A LOT OF RESULTS,
FROM THIS KREEN.
AND I WILL KIND OF BREEZE
THROUGH THEM QUICKLY BECAUSE A
LOT OF THIS IS ALREADY PUBLISHED
BUT 1 OF THE PRIMARY FINDINGS IS
THAT WE SAW SARS-COV-2 AND FLU,
SEEM TO HAVE SARSIMILAR RESPONSE
IN ALL THE ORGANS THAT WOE
LOOKEDDA THE AND THIS WAS
CHARACTERIZED BY A MASSIVE
SYSTEMIC INFLAMMA ARE TOY
RESPONSE AND ANTIVIRAL RESPONS.
SO HERE WE HAVE A HEAT MAP
SHOWING GENES SIMULATED IN
SARS-COV-2 AND FLU INFECTED
HAMSTERS, EVERYTHING IN RED IS
UPREGULATED SO YOU CAN
APPRECIATE THAT BETWEEN
SARS-COV-2 AND FLU, THESE ARE
UPREGULATED ACROSS THE BOARD AND
IT SEEMS LIKE THIS IS A SYSTEM
WIDE INFLAMMA ARE TOY
RESPONSE--AND WHAT WE ENDED UP
SEEING WAS WE SAW SMM NO
DIFFERENCE BETWEEN SARS-COV-2
AND FLU, AND REALLY THE ONLY
MAJOR DIFFERENCE WE SEEM TO SEE
IN THE VIRUSES IN THE OLFACTORY
BULBS OF HAMENTSTERS SO I SAID
THEY HAD MOSTLY RETURNED TO BASE
LINE AND THERE WAS 1 NOTABLE
EXCEPTION TO THAT AND THAT'S
THIS THE LAST COLUMN.
SO YOU'LL APPRECIATE THAT THE
IT'S STILL RED AT 31 DAYS POST
INFECTION.
SO THIS WAS BIZARRE FOR US
BECAUSE WE DIDN'T EXPECT TO FIND
THIS, THIS WAS 1 SINGLE AREA OF
THE BRAIN, WE DIDN'T SEE
PERRISTENT INFLAMMATORY
RESPONSES AND IT WAS A SPECIFIC
PHENOMENA, SO A LOT OF OUR
RECENT WORK HAS BEEN TRYING TO
GO AFTER THIS AND FIGURE OUT
WHAT MIGHT BE DRIVING THIS
PERSISTENT INFLOOMATORY
RESPONSE, SO DR. PERLMAN'S TALK
WAS ON A NICE PRIMER FOR THIS
BUT 1 OF THE WAYS WE WANT TO DO
THIS WAS BY DOING A PRETTY
THOROUGH CHARKTIZATION OF HOW
SARS-COV-2 WAS AFFECTING THE
OLFACTORY SYSTEM, SO HERE BEFORE
JUMPING INTOAUR DATA I WANTED TO
GIVE A BRIEF PRIMER AND COPYING
DR. PERLMAN'S PRESENTATION ON
SIMILAR ANATOMY, SO HERE SHOWING
HISTOLOGY SLIDES, HAMSTER
OLFACTORY SYSTEM, THIS WAS TAKEN
FROM A CORONAL CUT FROM FAR DEEP
INTO THE HAMSTER SAMPLE AND A
BIT INTO THE CRANIAL CAVITY AND
YOU CAN SEE THE NAIS AT PASSAGES
HERE, AIR GOES THROUGH HERE AND
THESE ARE THE OLFACTORY BULBS IN
THE BRAIN.
O IT'S COMPRISED OF THE
OLFACTORY BULB AS WELL AS THE
EPITHELIUM, AND THIS IS IN THE
FRONT, THE CRANIAL CAVITY AND IT
PRIMARILY CONTROLS INITIAL
SENSATION OF SMELL AND IT IS
HEAVILY FUNCTIONING TO THE
OLFACTORY EPITHELIUM, THE
EPITHELIUM ITSELF IS IN THE BACK
AND TOP PART OF THE NOSE AND
IT'S AN EPITHELIAL LAYER WHERE
YOU HAVE THE NEURAL SENSORYS
HOUSED AND SO HERE WE SHOWING A
ZOOMED IN OF THE OLFACTORY AND
EPITHELIUM AND WE HAVE THEM
MARKED FOR SENSORY NEURONS AND
YOU CAN SEE THE SIMILAR
STRUCTURE TO WHAT WE SEE IN THE
CARTOON WHICH IS THE CELL BODIES
STACKED ON TOP OF EACH OTHER AND
THEN THE STRUCTURES PROTRUDING
OUT INTO THE NOSE AND THESE ARE
WHERE THE NEURONS ARE GOING TO
STICK OUT AND AS AIR AND FLEEING
BY, WHAT YOU INHALE, THEY WILL
CATCH THE MOLECULES AND SIGNAL
ACTION PROTENTIALS THAT WILL
INDICATE THE DETECTION OF SMELL.
SO THESE WILL PASS FROM THESE
NEURON CELL BODIES WHICH ARE
LOCATED IN THE EPITHELIUM
THROUGH AXONAL PROYEKSS WHICH
PROJECT OFF OF THE BASAL LATERAL
SIDE OF THE CELLS, THESE WILL
CLUSTER INTO LARGE NEURAL
BUNDLES AND THESE BUNDLES WILL
FLOW INTO THE OLFACTORY BULB AND
THERE THEY WILL SYNAPSE HIGHER
NEURONAL CELLS WITH THE AREAS
HERE CALLED GLUE MAR I HERE
EMPLOY AND LOOKING AT THE BULB
ITSELF, THIS IS PART OF THE
BRAIN, PART OF THE CENTRAL
NERVOUS SYSTEM,OOSE IMPORTANT TO
APPRECIATE THAT THE PORTION OF
THE BULB IS COMPRISED FROM THESE
NEURON ANDS THIS WAS INTERESTING
AND SOMETHING THAT WILL BECOME
IMPORTANT IN A BIT.
SO IT'S BEEN KNOWN THAT THIS
AFFECTS THE EPITHELIUM, AND THE
EFFECTS IS LOSS OF SMELL, SO WE
WANT TO DO THIS IN A HAMSTER.
SO WE INFECKED A BUNCH OF HAM
TERS NAISALLY, AND TOOK OLYMPIC
FACTOR EPITHELIUM AT 1, 3, 5, 7,
10, 30 AND 45 DAYS POST
INFECTION, AND WE DID PC Rs
AND WE LOOKED FOR SARS-COV-2
MATERIALS AND THIS IS LOOKING
FOR SUBGENOMIC AND TRANSCRIPT
PRODUCED WHEN SARS-COV-2 IS
REPLICATING AND AGAIN WE CAN SEE
THAT SARS-COV-2 IS REP
LITICATING NICELY AND WE CAN SEE
THE INCREASE AT 3 DAYS POST
INFECTION PEAK AND AGAIN
FOLLOWING SIMILAR DYNAMICS TO
WHAT WE SAW IN THE LUNG TISSUE.
NOTE ABLE WE WANT TO LACK AT AN
ANTIVIRAL RESPONSE AND DID A
SIMILAR QPC R AND IN THE
INTERFERON STIMULATED YEEN AND
WE SAW THAT STIMULATED YEEN
PRESENCE SEEMED TO MIRROR PRETTY
WELL WITH VIRAL PRESENCE AS WELL
AND PEEKING AT 3 DAYS POST
INTERACTION AND CLEANING AT
10-15.
SO WE ALSO BROUGHT UP A
SECONDARY LONGITUDINAL COHORT WE
USE TO DO HISTOLOGICAL STUDIES
WITH AND WE WANT TO SEE WHAT
THIS LOOKED LIKE IN THE
OLFACTORY EPITHEMUAL ITSELF, SO
FIRST WE PROBE FOR SARS-COV-2
INFECTION AND SHOWED INFECTION,
AND HOPEFULLY CAN YOU APPRECIATE
THAT IN RED AND GREEN, IN THE
INFECTED ANIMALOT RIGHT, TON OF
THE OLFACTORY EPITHELIUM IS
INFECTED BY SARS-COV-2.
SO THIS SEEMS TO BE A RAM PANT
INFECTION AFFECTING SEEMINGLY
THE MAJORITY OF THE TISSUE AT 3
DAYS POST INFECTION AT VIRAL
PEEK.
FURTHER MORE, WE WANT TO SEE
WHAT THIS INFLAMMA ARE TOY
RESPONSE LOOKED LIKE OUTSIDE OF
OUR QPC R AND HERE WE'RE
STAINING FOR [INDISCERNIBLE] AND
WHAT YOU CAN HOPEFULLY
APPRECIATE IS IT'S VERY RED AT 3
DAYS POST INFECTION.
SO THIS ISHT FERON RESPONSE IS
VERY PERSISTENT AND PREVALENT
THROUGHOUT THIS TISSUE.
SO OUR NEXT QUESTION THEN WAS,
SO WE--DURING OUR INFECTION
DIDN'T SEE THE OLFACTORY SENSORY
NEURONS INFECTED THEMSELVES.
THIS HAS BEEN SHOWN CONNIST
SEBTLY THROUGHOUT THE LITERATURE
BUT WITH ALL THIS INFECTION AND
ALL THIS ANTIVIRAL RESPONSE
HAPPENING INSIDE THE TISSUE, WE
ARE CURIOUS WHAT IS HAPPENING TO
THIS TISSUE EMPLOY SO HERE WE
STAIN PER PUNISHING DRMARKER IN
THE EPITHELIUM AND WE KEIN THESE
CELL BODIES, THESE STRUCTURES
BUT AS SOON AS YOU CAN GET TO
THE INFECTION AT 3 DAYS POST
INFECTION, YOU LOSE A LOT OF
THAT STRUCTURE AND THERE'S A
MASSIVE DISRUPTION, SO CAN YOU
SEE OLFACTORY SENSORY NEURONS
THERE, A LOT OF SLUFFING OFF
INTO THE NOSE, THE 1S THAT ARE
LEFT HAVE LOT THEIR SILLIARY
STRUCTURE AND THEY'RE NOT HAVING
FUN.
IS IT INTERESTINGLY WHENY WOO
LOOK LATER WE SEE THEM DISAPPEAR
ALTOGETHER.
SO AT 5 DAYS POST INFEC, A
SIGNIFICANT PORTION OF THE
EPITHELIUM WAS CLEAR OLFACTORY
SENSORY NEURONS AND THE 1S WE
DID SEE WERE OFTEN TIMES
SLUFFING OFF INTO THE NOSE AND
ONCE WE LOOK AT 10 DAYS POST
INFECTION, THE PRESENCE WAS JUST
COMPLETELY ALMOST ELIMINATED AND
WHILE THIS DID SEEM TO RECOVER
OVER TIME, AT 30 DAY POST
INFECTION, THE OLFACTORY
EPITHELIUM WAS SIGNIFICANTLY
THIN COMPARED TO WHAT WE SAW IN
OUR MOK INFECTED ANIMAL ANDS
THERE ARE DEFINITELY LESS
OLFACTORY SENSORY NEURONS.
SO HERE PUTTING SOME
QUANTITATIVE NUMBERS TO THIS,
LOOKS LIKE SOME OF THE DOTS ARE
NOT SHOWING CORRECTLY BUT YOU
CAN SEE IT THERE SO HERE WE
MEASURED THE THICKNESS PUTTING
NUMBERS TO WHAT WE JUST IN
HISTOLOGY AND HOPEFULLY YOU CAN
APPRECIATE THAT.
IT CAN PEEK AT 5 DAYS POST
INFECTION AND RECOVERS A BIT BY
30 DAYS POST INFECTION IT
DOESN'T GET BACK UP TO BASE LINE
LEVELS.
FURTHER MORWHEN WE LOOK AT
OLFACTORY NEURON, WE STARTED
COUNTING THE NUMBER OF FLUORONS
FOR MICROTISSUE, WE CAN SEE
AGAIN AN EVEN MORE DRAMATIC
EFFECT BY 30 DAYS POST INFECTION
EVEN THOUGH THEY STARTED COMING
BACK, THEY DIDN'T REALLY ACHIEVE
SIMILAR LEVELS TO WHAT WE SAW AT
BASE LINE.
SO, AGAIN, OUR NEXT QUESTION
THEN WAS IF THESE OLFACTORY
SENSORY FIEWRONS ARE DYING OR
BEING CLEAR INDEED SOME SENSE AS
A RESULOF SARS OVERLAPPING 2
INFECTION, AND THESE ARE NOT
NECESSARILY INFECTED WHAT'S
HAPPENING IN THE BULB AND
UPSTREAM, WHAT'S HAPPENING
TO--AND WHAT'S HAPPENING TO
THESE UPSTREAM CELLS AND ARE
THEY GOING TO CONTINUE FIRING
WHEN THEY'RE DEFERVE 8ED OF
THEIR SENSORY INPUT SO THE FIRST
THING WE WANT TO GO AFTER AND
WHAT'S HAPPENING UPSTREAM?
SO WE SO IT'S A NEURONAL GENE
AND WHEN JIEWRONS FIRE MORE THEY
PRODUCE MORE PH OS, WHEN THEY
FIRE LESS, THEY PRODUCE LESS
EMPLOY SO IT'S A GENERAL MARKER.
AND HOPEFULLY YOU CAN APPRECIATE
THAT AFTER SARS-COV-2 INFECTION,
IT GOES DOWN AND STAYS DOWN.
NEXT WE WANT TO DO IS PROVIDE
DEFINITION ON THIS SO WE WENT
INTO OUR RNA SEQUENCING DATA CAN
AND TRIED TO GENERATE ANY
OLFACTORY BULB SEQUENCING WE
HAVE ON FILE AND ABLE TO COMPILE
A SEQUENCING SET FOR OLFACTORY
BULBS WHERE WE HAVE THIS TISSUE
AT 1, 2, 3, 4, 14, 16, 31 DAYS
POST INFECTION SO THERE ARE A
COUPLE DIFFERENT INFECTIOUS
PFUs THAT ARE DIFFERENT
THROUGHOUT THIS COHORT BUT THEY
ALL MODEL IS NICELY AND WE'RE
ALL IN A RANGE OF 100 TO 10,000
PFU.
AND SO WHEN WE LOOK FOR FOSTER
NURSED, WE SEE SIMILAR IN QPC R
WHICH IS DOWN REGULATED AND
SEEMS TO STAY DOWN REGULATED
THROUGH 30 DAYS POST INFECTION.
WHATEVER ELSE IS REGULATED WE
SAW A BUNCH OF OTHER IMMEDIATE
EARLY ACTIVITY SO THIS INCLUDES
ARK, AND FOS3, AND THIS GOES UP
AND DOWN WITH NEURONAL ACTIVITY
SO IT DOES SEEM THAT FACTORY
BULB AND SEEMS TO BE
PERSISTENTLY UPREGULATED
FOLLOWING SARS-COV-2 INFEC
EMPLOY HOWEVER, WITH THIS DATA
SET, WE'RE ALSO THEN CURIOUS,
WHAT ELSE IS HAPPENING IN THE
OLFACTORY BALB?
WE HAVE THE DATA SET, AND HERE
AGAIN WE SAW THE MAJOR
ONTOLOGIES THAT POPPED OUT ON
THE ANALYSIS FOR TYPE 1
INTERFERON AND IMMUNE RESPONSES,
SO HERE THIS IS KIND OF SHOWING
GENE ENRICHMENT ANALYSIS ON ALL
OF OUR DIFFERENT DATA SETS,
SHOWING DAYS POST INFECTION AND
OUR X-AXIS AND THE ENRICHMENT
CORE FOR THE ANALYSIS FOR THIS
ON THE Y-AXIS WITH THE BALLS AT
THE TOP HERE, THAT'S MY STUFF,
THAT'S FORTUNATE SHOWING
SNIFFING CANC AND APOLOGIES ON
THAT BUT IN THE TYPE 1
INTERFERENCE WE AWE IT
UPREGULATED DURING ACUTE
INFECTION AND HAVE A BUMP AT 14
DAYS INFECTION AND BEING
SIGNIFICANT AT LEAST THROUGH 1
MONTH POST INFECTION.
FURTHER MORE WHEN WE LOOK AT
OTHER ASSPECS OF THE IMINE
RESPONSE SUCH AS CHEMOKINE
ACTIVITY, WE SAW IT WHICH IS
THAT THESE MARKERS SEEM TO BE
UPREGULATED EVEN LATER FOLLOWING
INFECTION SO THERE'S A LATE
RISING ONTOLOGY THAT NOTABLILY
SEEMED TO START RISING AFTER 3
DAYS POST INFECTION, SO AFTER
THE PEAK OF VIRAL INFECTION, SO
THIS IS FOR CHEMOKINE ACTIVITY
INSIDE THE OLFACTORY.
WHEN WE'RE INTERESTED THEN IN
SEEING WHAT CELL TYPE MIGHT
DRIVE THIS ONTOLOGY, WE LOOK AT
THE GENE ENRICHMENT ANALYSIS FOR
THE CELL TYPES AND WE SAW THAT
MICROGLOIL CELLS WERE 1 OF THE
MAIN ONTOLOGIES THAT POPPED OUT
FOR DATA AND WE SEE ENRICHMENT
SCORES FOR MICROGLIAL PRESENCE
GOING UP, EVEN ACTUALLY
FOLLOWING THE PEAK OF INFECTION
AND REMAINING ELEVATED AND GOING
UP AND FURTHER PEEKING AT 14
DAYS POST INFECTION, SO AGAIN
THIS IS AFTER WE WOULD EXPECT
VIRAL CLEARANCE THIS HAVE GONE A
BIT INTRIGUING TO US.
SO WE NEXT WANTED TO SEE IT SO
WOE TURN TO OUR HISTOLOGICAL
COHORT AND WE STARTED STAINING
FOR MXA, IT A MARKER FOR MA
ROUGH ATOM PHAGE OR MICROGLIAL
CELL POPULATIONS AND APPRECIATE
THAT EVEN THOUGH IT'S UP AT 3
AND 5 DAYS POST INFECTION, IT
REMAINS ELEVATED OUT FOR 30 DAYS
POST INFECTION AND YOU CAN SEE
IT BLOTCHY AND ALMOST
EXCLUSIVELY MAINTAINED IN THIS
OUTSIDE AREA AROUND THE
OLFACTORY BULB, SO THE LOCAL
EYATION OF THIS SIGNAL WAS VERY
IMPORTANT TO US BECAUSE IF YOU
RECALL, THIS IS ALSO WHERE ALL
OF THESE OLFACTORY SENSORY
NEURONS FROM THE PERIPHERY ARE
COMING IN.
SO WHEN WE ALSO LOOKED AT IB1
SIGNATURES FOR THE INFILTRATION,
WE SAW THE SIGNATURE PEAKED AT
14 DAYS POST INFECTION AND
HOPEFULLY APPRECIATE AGAIN THAT
THERE ARE ALET BIT HARDER TO SEE
MORE GREEN DOTS AROUND THE
OUTSIDE HERE SO AGAIN IT LOOKS
LIKE THE MICROGLIAL CELLS OR
WHATEVER IS POSITIVE HERE ARE
MIGRATING TOWARDS THE OUTSIDE OF
THE OLFACTORY BULB AND SEEM TO
BE GOING THROUGH THE SAME PLACES
THAT THIS INTERFERON SIGNAL IS
PRESENT.
SO WE THEN QUANTIFIED IT AND WE
FOUND THAT WHEN YOU LOOK AT THE
OUTSIDE OF THE OLFACTORY BULB,
THE MICROGLIAL CELLS OR THE
POSITIVE CELLS SEEM TO BE THE 1S
THAT ARE PRODUCING THE MXAs,
THEY ARE IRPT FERON RESPONSIVE
AND THEY SEEM TO BE INTERFERON
RESPONSE AND I HAVE HELD THROUGH
30 DAYS POST INFECTION AND MORE
OVERWHELMINGLY WE LOOKED AT THE
INSIDE OF THE BULB AGAIN, AND WE
LOOK AT POSITIVE OR NEGATIVE FOR
THE OLFACTORY SENSORY NEURONS
THERE, WE FOUND THIS RESPONSE
ACTUALLY EXTENDED TO THE INSIDE
OF THE OLFACTORY BULB, AND YOU
SEE IMAGES OF THIS HERE, AND
THIS IS THE OUTSIDE OF THE
OLFACTORY BULB, WE SEE THESE
LARGE PLAQUES OF THE CELLS THAT
ARE POSITIVE FOR MXA, AND WITH
WHEN WE ELECTRIC AT THE INSIDE,
EVEN THOUGH THEY WERE MORE
DISPERGSED YOU CAN SEE THESE
CELLS ARE ALSO POSITIVE FOR MXA,
SO IT SEEMS LIKE THE INTERFERON
REPONS IS PREVALENT THROUGHOUT
THE BULB.
SO NEXT QUESTION, WHY ARE THEY
ACTIVATED IN WHY ARE THEY
PRODUCING TYPE 1 INTERFERON?
IS THIS A PERSISTENT INFECTION,
IS SARS-COV-2 GETTING INTO THE
OLFACTORY BULB, DOES IT AFFECT
THE OLFACTORY BULB IS IT A SIDE
EFFECT OF THE INFECTION IN SO
EVERY WAY WE COULD TELL IT WAS
NO, WE COULDN'T SEE ANY SIGNS OF
THE SARS-COV-2 PRESENCE, HERE
WE'RE STAINING FOR SARS-COV-2 S
AND N AND FOLLOWING ACUTE AND
FOLLOWING INFECTION WE COULDN'T
SEE ANY THERE.
SO OUR NEXT THOUGHT WAS LET'S
LOOK BACK TO THE SEQUENCING DATA
AND SEE IF WE CAN FIND MARKERS
OF MICROGLIAL ACTIVITY AND WE
LOOKED INTO THE LITERATURE TO
FIND OUT WHAT MICROGLIAL
ACTIVITY THERE WERE AND LOOKING
INTO THE ALZHEIMERS LITERATURE
AND WE ENDED UP SEEING THAT A
TEEP UPREGULATION FOR THAT
ENTIRE MONTH POST INFECTION WIND
O THAT WE'RE LOOKING AT OF
MICROGLIAL MARKERS THAT WERE
SPECIFIC FOR PH AGO-6 ACTIVITY
AND WHERE MICROGLIA CAN MEET
PARTS OF NEURONS TO HELP GUIDE
DEVELOPMENT AND INNERVATION.
SO HERE WE SAW UPREGULATION OF
THE COMPLEMENT C1 A AND 3, AND
THEIR REQUISITE RECEPTORS WHICH
ARE ALL USED TO MARK DEBRISS AND
OTHER SYNAPTIC PRUNING SITES IN
THE SYSTEM BY THE MICROGRIA.
AND WE AWE ADDITION, SUCH AS
TRAN 2 AND THE COMPONENT
SIGNALING COMPLEXES AND FURTHER
MOIRE WHEN WE LOOK AT GENE SET
ENRICHMENT ANALYSIS SIMILAR TO
WHAT WE HAD SEEN EARLIER, WE SAW
A SIMILAR LATE RISING PHENOTYPE,
WITH THE BRUINING THAT SEEMS TO
CORRELATE WITH THE MICROGLIAL
SIGNATURE.
SO I AT THIS POINT IT LOOKS LIKE
MICRODPLIA SEEM TO BE FLOCKING
TOWARDS THE OLYMPIC PACTRY BULB
AND UPREGULATING IT ON THE TYPE
1 IRPT FORON RESPONSIVE, HOWEVER
OUR NEXT QUESTION IS WHAT ARE
THEY GOING OUT TO BECAUSE WHAT
ARE THEY GOING TO EAT IN AND
BASED ON THE LOCALIZATION OF THE
SIGNAL LINE WHERE ALL THESE
THINGINGS WERE HAPPENING, OUR
FIRST THOUGHT WAS ARE THEY
ACTUALLY EATING ALL OF THESE
RESIDUALY DEBRIS FROM THE
NEURONS FOLLOWING SARS-COV-2
INFECTION, AND SO, WE WANT--FROM
OUR HISTOLOGY, COHORT, WE
QUANTIFIED THE IN. TENSITY OF
OMP, AGAIN MARKERS FOR OLFACTORY
SENSORY NEURON IN THE PATIENT IN
THIS TISSUE AT LATER TIME POINTS
POST INFECTION AND EVEN THOUGH
OLFACTORY SENSORY NEURONS SEEM
TO HAVE PEAK CLEARANCE AT AROUND
5 DAYS POST INFECTION AND THERE
SEEMED TO BE DELAYED CLEARANCE
OF THE OMP SIGNATURE INSIDE THE
OLFACTORY BULB SO TO US IT LOOKS
LOAMACYIC THESE MIGHT BE EATING
SOME OF THE DEBRIS FROM THE
OLFACTORY SENSORY, FROM ACUTE
INFEC.
SO OUR MODEL FOR WHAT'S GOING
ON, WE THINK THAT THEY'RE NOT
NECESSARILY INFACTED BUT DAMAGE
TO THE EPITHELIUM CAN CAUSE
THESE TO SLUFF OFF INTO THE NOSE
AND YOU GET SENSORY NEURON LOSS.
THIS LEADS TO THE FORMATION OF
SOME NEURONAL DEBRIS AS A RESULT
OF THIS INTERACTION AS WELL AS
THE STOP AS WELL AS THE
INHIBITION OF TIGER
DATABASEINALLING GOING TOWARDS
DEEPER AREAS OF THE OLFACTORY
BULB LEADING TO A DECREASE IN
NEURONAL FUNCTION IN THIS AREA,
FURTHER MORE THAT DEBRIS ISN'T
JUST DOING NOTHING, IT'S
RECRUITING WHAT APPEARS TO BE A
MACROPHAGE OR MICRODPLEEL
POPULATION THAT IS TYPE 1
INTERFERON RESPONSIVE AND THIS
EPPEDS UP DOING IS CAUSING THE
OLFACTORY BULBS TO BE
IBT--INTEGRATE FLAMED WITH A
TYPE 1 SIGNATURE FOR GREATER
THAN A MONTH FOLLOWING THE
INFECTION.
SO, EVEN THOUGH THIS IS THE IN
OLFACTORY BULB AND MOST OF THE
SMELL, THE OLFACTORY BULB ITSELF
IT HEAVILY CONNECTED TO THE SUCH
AS THE EMILY CIBOL SYSTEM,
MEMORY, AND BEHAVIOR AND A LOT
OF ACTUALLY HUMAN MRI STUDIES
HAVE SEEN THAT IN POST COVID
INDIVIDUALS WHEN YOU DO--WHEN
YOU LOOK AT THE BRAIN, YOU CAN
SEE GRAY MATTER CHANGES IN A LOT
OF AREAS THAT ARE FUNCTIONALLY
LINKED TO OLFACTORY BULB, THERE
IS A HYPOTH IS THAT 1 MIGHT BE
DRIVING THIS, FOR DEC OR
INFLAMMATORY PATHWAYS OR A LESS
OF SENSORY INPUT DUE TO
INHOSPITALITY, SO WE THINK THERE
IS A MECHANISM THAT MIGHT
PROVIDE AN EXPLANATION FOR WHY
THESE HAPPEN AND SARS-COV-2
INFECTION EVEN WITHOUT CHRONIC
OR PERSISTENT INFECTION.
SO IN CONCLUSION WE HAVE SHOWN A
FEW THINGS ABOUT LONG COVID BUT
KIND OF TAKING A STEP BACK AND
TALKING ABOUT THE HAMSTER MODEL
GENERALLY.
IT'S BEEN A REALLILY GREAT
PLATFORM FOR ACUTE SARS-COV-2
PATHOLOGY AND PARTICULARLY FOR
MODELING PATHOLOGY, HAMSTERS
LOSE A SENSE OF SMELL SO IT'S
BEEN GREAT TO USE THEM FOR
INVESTIGATOR OF ACUTE INFECTION,
FURTHER MORE SYSTEMIC INFLAMMA
ARE TOY RESPONSES HAVE BEEN
INSIGHTFUL FOR LEARNING ABOUT
HOW THE DISEASE REFLEBTS THE
ENTIRE BODY, THE TROAPISM IS
NICE FOR MODELING THE INFECTION,
HOWEVER, HAMSTERS AS HAS BEEN
SAID A FEW TYPES TODAY, THEY'RE
NOT MICE AND WE ARE EMILY THED
BY REEAMENTS SO WE DON'T HAVE
ALL THE ANTIBODY SAYS WE LIKE TO
HAVE, WE HAVE A LOT OF CELL TYPE
SPECIFIC GENETIC MOLL MODELS WE
LIKE TO HAVE IN MOUSE ELEMENTS
AND THEIR CONTINUES TO BE
DEVELOPING BIO-INFORMATICS
PIPELINES SO WE'RE LIMITED TO
OMICS TECHNOLOGIES AND EVEN
THOUGH IT IS A VALUABLE MODEL,
THERE IS ROOM TO GROW AND I
THINK IT'S WORTH IT.
WITH THAT THANKS TO THE LAB, AND
A WHOA LOT OF OTHER PEOPLE WHO
HELPED OUT IN CREATING THIS
PROJECT.
I CAN TAKE ANY QUESTIONS.
>> THANK YOU VERY MUCH EMPLOY WE
HAVE TIME FOR A FEW QUESTIONS.
SEEMA?
>> IN THE HAMSTER MODEL, AFTER
30 DAYS, WHAT IS THE--LIKE
RECURRENT INFECS IN HUMANS?
>> IT'S A IMRAIT QUESTION.
WE ACTUALLY HAVEN'T DONE
RECURRENT INFECTIONS THE CLOSEST
WE'VE DONE IS A POST DOC IN OUR
GROUP A YORE OR 2 AGO DID A
TODAY WHERE HE WAS LOOKING AT
KAN--KANA INFECTION WITH 1
STRAIN OF SARS-COV-2 LEAD TO
ANOTHER INFECTION OF SARS-COV-2
AND WE HAD SEEN YOU DO GET
ADEQUATE PROTECTION FROM PRIOR
VARIANT STRAINS WITHIN THE O
WILL FACTORY--AND WE CONTINUE TO
LOOK FOR DAMAGE AGAIN AT THAT
TIME.
>> I HAVE 1 QUESTION FOR THE
KLEIN GROUP AT WASH-U HAD SHOWN
IN THE CONTENTS OF WEST NILE
VIRUS THAT INTERFERON-GAMMA CAN
LEAD TO MICROFLEEL ACTIVATION
LEADING TO NEURONAL SYNAPSE
PRUNING.
IN YOUR MODEL HAVE BEEN ABLE TO
DIFFERENTIATE THE IMPACT OF TYPE
1 VERSUS TYPE 2 INTERFERON, AND
ESSENTIALLY INDUCE ANTIVIRAL
YEENS, OR THINKING IT'S TIME 1
BUT IT CAN BE TYPE 2 IN A MODEL
SHE ALSO SHOWED A ROLE FOR
T-CELLS AS WELL, SECRETING
INTERFERON-GAMMA EMPLOY.
>> YEAH, GREAT QUESTION AND IT'S
SOMETHING WE LOOKED AT FOR THE
T-CELL AND WE LOOKED FOR T-CELLS
AND WE THOUGHT WE WERE GOING TO
FIND T-CELLS AND WE HAVEN'T
FOUND THEM SOY DO SEE INCREASED
PRESENCE IN OLFACTORY EPITHELIUM
IMNLLY BUT THEY'RE NOT THERE IN
MASSIVE AMOUNTS ESPECIALLY IN
THE BULB ITSELF.
IT'S DRIVEN MORE LOCALLY.
WHEN WE LOOK CLOSELY, WE CAN
UPDATE THE HAMSTER HAN OITATION,
WE CAN SEE SOME TYPES OF TYPE 1
FIRNT FERON BEING EXPRESSED IT'S
LOW LEVELS, BUT WE SEE IT AT 10
AND 14 DAYS POST INFECTION AND
WE THINK ALL OF THAT'S LIKELY
HAPPENING LOCALLY, THERE'S ALSO
SOME TISSUE EXPWRIEWKS PATTERNS
WHICH I DIDN'T SHOW HERE WHICH
HAVE US THINKING THAT THE
INTERFERON ITSELF IS ACTUALLY
COMING FROM INSIDE THE OLFACTORY
BULB AND WITHOUT THE T-CELLS
THERE, WE'RE NOT SURE WE WOULD
BE PRODUCING THE FAMILIES GAMMA
ITSELF BUT WITH THAT WE ALSO
DON'T SEE A LOT OF ISSFERON
GAMMA SEQUENCING EITHER.
>> PETER?
>> SO WHAT ISOLATE OR VARIANT DO
YOU THINK THIS WAS?
AND SO CERTAIN VARIANTS WITH
LOSS OF SMELL?
YOUR THOUGHTS ON THAT?
>> GREAT QUESTION AND 1 I HAVE
BEEN THINKING ABOUT, MYSELF,
UNFORTUNATELY HAVEN'T DONE THOSE
STUDIES YET BUT IT'S ON THE
AGENDA FOR SURE, SO THIS WAS
DONE USING ANCESTRAL VARIANT,
THIS WAS THE WASHINGTON U STRAIN
FOR THE STUDIES BUT WE TARTED
WO,ING WITH,A DITIONAL TRAINS
AND 1 OF OUR FIRST THINGS WE
WANT TO DO IS TEST SMELL LOSS
FOR THOSE VARIANT ANDS DO A
COMPARISON AND SEE IF WE GET
THESE LONGITUDINAL AND
INFLAMMATORY PHENOMENON.
>> A LOT OF NICE DATA.
NYOU SHOWED EARLY STAGE ALMOST
[INDISCERNIBLE] EACH OTHER IN
TERMS OF GENETIC THAN THE
[INDISCERNIBLE] HAVE YOU DO ANY
CO INFECTION YET?
>> I HAVEN'T YET BUT IN THE
[INDISCERNIBLE] LABORATORY BUT
THEY HAD DONE DAY 1 POST
INFECTION FLU FOLLOWED BY SARS
AND VICE VERSA AND SHOWED
PROTECTIVE EFFECTS FROM 1 TO THE
OTHER.
IF I REMEMBER CORRECTLY, PRIOR
EXISTING SARS INFECTION LIMITED
SPREAD OF FLU BUT I WOULD DOUBLE
CHECK THAT, I AM HAPPY TO TALK
AFTER TO GIVE YOU THAT PAPER.
>> THANK YOU VERY MUCH.
GREAT TALK.
>> AND OUR FINAL SPEAKER OF THIS
SESSION IS SABRA KLEIN FROM
JOHNS HOPKINS UNIVERSITY AND SHE
WILL TALK ABOUT SEX DIFFERENCES
IN ANIMAL MODELS OF INFLUENZA
AND SARS-COV-2.
>> THANK YOU.
AND THANK YOU TO THE ORGANIZERS
FOR INCLUDING THIS TOPIC.
TODAY RATHER THAN GIVE A STORY
WITH A SINGLE VIRUS, I WILL GIVE
YOU A LOT OF VIGNETTES,
HOPEFULLY TO GIVE YOU THE MODELS
AVAILABLE AND THAT WE UTILIZED
TO ANSWER QUESTIONS ABOUT
DIFFERENCES BETWEEN MALES AND
FEMALES EMPLOY SO I THOUGHT WE
COULD START WITH WHAT SEX AS A
BIOLOGICAL VARIABLE MEANS.
BECAUSE I KNOW EACH OF YOU
WRITES THIS IN YOUR NIH GRANTS.
SO WHAT THIS IS REFERRING TO ARE
THE BIOLOGICAL DIFFERENCES
BETWEEN FEMALES AND MALES.
FEMALES HAVING 2 X CHROMOSOMES,
MALES HAVE AN X AND A Y, THERE
CAN BE DIFFERENCES IN SEXUAL
DEVELOPMENT THAT CAN LEAD TO
DIVERSITY IN SEX CHROMOSOME
COMPLEMENT AND I WILL TALK ABOUT
THAT SO I WILL HOLD YOU THERE.
BUT 1 THING THAT'S REALLY
IMPORTANT ON THE TINY
Y-CHROMOSOME IS A GENE CALLED
SRY.
AND WHAT SRY ENCODES FOR IS THE
DETERMINING IN FACTOR AND THAT
WILL BE IMPORTANT IN SOME OF OUR
DISCUSSIONS TODAY AS WELL.
SO I WILL ASK YOU TO HOLD ON TO
THIS.
BUT 1 THING THAT TEST ESTIMATE
THAD DETERMINING FACTOR DOES IS
EARLY ON EMBRYONIC DEVELOPMENT
ALL OF US HUMANS AND MICE ALIKE
IS WE ALL START OUT WITH A
BI-POTENTIAL GONAD THAT CAN GO
AND DEVELOP EITHER WAY AND IN
THE PRESENCE OF CERTAIN SIGNALS
INCLUDING THIS TESTIS THAT IT
DEVELOPS INTO A TESTES, BUT IN
THE ABSENCE OF SRY WE WILL GOET
THAT DEVELOPMENT AT TISSUE INTO
THE FETAL COMPARTMENT
MALPRODUCTIVE TRACT.
SO I WILL TALK TODAY ABOUT SEX
CHROME SOAPS, I WILL TALK TODAY
ABOUT SEX CHROMOSOME GENES BUT
I'M ALSO GOING TO TALK A BIT
TODAY ABOUT THE GONEADAL
STEROIDS THAT HAVE THE PRESENCE
OF OVARIES OR THE PRESENCE OF
TESTES.
SO THESE HORMONES THESE ARE
STEROID HORMONES WHEN WE THINK
ABOUT STEROIDS WE THINK ABOUT
THE TBLIEWK O CORTICOIDS AND HOW
THEY FUNCTION AND HOW THEY CAN
DO THINGS LIKE BEAUTIFULLY
DAMPEN INFLAMMATION BUT TODAY I
WILL TALK ABOUT THESE GONADAL
STEROIDS, ANDROGEN ROUGH
ATOMIENS AND ESTROGENS HOW THESE
CAN BE IMMUNOMODULATORY.
AND PART OF HOW THEY'RE
IMMUNOMODULATORY, IS THAT EVERY
SINGLE STYLE WE HAVE LOOKEDDA
THE IN HUMAN AND THEY HAVE
RECEPTORS FOR THESE HORMONES.
SO THEY'RE LIP OPHILIC,
STEROIDS, THEY'RE RAPIDLY
CROSSING THE BILAYER WHERE THEY
COMBINE TO LAYERS LOCATED
INTRACELLULARLY TO HAVE
CANNONICLE GENOMIC EFFECTS.
SO I LIKE TO THINK ABOUT
SPHEROIDS AS TRANSCRIPTIONAL
REGULATES OF IMMUNE CELL
FUNCTION.
SO ULTIMATELY WHEN WE THINK
ABOUT VIRAL DECS WHICH IS WHY
WE'RE ALL HERE, TALKING ABOUT
MODELING THESE VIRAL DECS MY
GROUP AND OTHERS HAVE GENERALLY
SPEAKING THE GOOD, BAD AND UGLY
OF ALL THIS REGULATION EMPLOY SO
WHEN I'M OFTEN ASKED IN A
QUESTION AND ANSWER SESSION TO
DESCRIBE HOW FEMALES ARE BETTER
I WOULD NEVER SAY THAT FEMALES
ARE NECESSARILY BETTER THAN
MALES AND AS WE GO THROUGH OUR
DAT AWE WILL SEE THIS, IT'S THAT
THEY'RE DIFFERENT AND THERE CAN
BE PLUSES AND MINUSES TO AN
ABROAD BRUSH STROKE TYPE OF WAY,
GREATER IMMUNE RESPONSES IN
FEMALES, FEMALES ARE MORE LIKELY
TO DEVELOP IMMUNE MEDIATED
PATHOLOGY, BUT THERE ALSO MAY BE
MORE LIKELY TO CLEAR VIRUSES AND
I WILL SHOW YOU EVIDENCE OF THIS
TODAY, WHEREAS FOR MALES WE HAVE
AN INCREASED LIKELIHOOD OF I
HAVE ROLL PERSISTENCE AND ALSO
REDUCED VACCINE IMMUNITY, AGAIN,
I'M GOING TO REVIEW THAT WITH US
TODAY.
SO AS WE TALK ABOUT
PATHOGENESIS, WHICH IS AGAIN WHY
WE'RE HERE, WE'RE TALKING ABOUT
MODELING PATHOGENESIS, I OFTEN
THINK ABOUT SEX DIFFERENCES IN
TERMS OF THIS DEC RESPONSE
DAMAGE MODEL WHERE I THINK WE'RE
ALL VERY ACCUSTOMED TO THINKING
ABOUT OUR DAMAGE THAT CAN OCCUR
AS A RESULT OF THIS CASE VIRAL
INFECTION AS RANGING FROM A LOW
DEGREE OF DAMAGE TO A HIGH
DEIMREE OF DAMAGE AND IF WE PLOT
THIS, AGAINST AN IMMUNE RESPONSE
WHICH CAN RANGE FROM BEING WEAK
TO STRONG, HERE'S YOUR DISEASE
RESPONSE CURVE.
YOU CAN GET A HIGH DEGREE OF
DISEASE AND HOST TISSUE CULTURE
MEDIA WITH THE WEAK IMMUNE
RESPONSE THAT DOESN'T ALLOW FOR
CONTROL OF VIRUS REPLICATION BUT
IF WE WE FOLLOW THIS ALL THE WY
AROUND WE CAN GET A HIGH DEGREE
OF DISEASE IF WE HAVE AN
OVEREXUBERANT IMMUNE RESPONSE
WHICH CAN LEAD TO IMMUNO
PATHOLOGY.
SO ALL I'VE DONE, I'M NOT
CREATIVE, I DREW A SIMPLE LINE
AND I'M HIGHWAY POGHTICIZING AND
WE WILL DISCUSS DATA FROM MY
GROUP ARE SUPPORTING THIS
HYPOTHESIS BUT THAT WE WOULD SEE
A MALE BIAS, MEANING MALES MAY
SUFFER A WORSE OUTCOME THAN
FEMALES IF DISEASE IS BEING
CAUSED BY AN INABILITY TO
CONTROL VIRUS WHEREAS WE MIGHT
PREDICT WE WOULD SEE A FEMALE
BIAS, MEANING FEMALES MAY SUFFER
A WORSE OUTCOME IN THAT DISEASE
IS BEING CAUSED BY AN
OVEREXUBERANT OR MAYBE A LONGER
LASTING IMMUNE RESPONSE.
SO 1 VIRUS WE HAVE NOT TALKED
ABOUT HERE, I WANT TO PUT THIS
OUT HERE AND AIMING FOR DIVERSIY
WITH BROAD BRUSH STROKES.
SO INFLUENZA B WE HAVE 2 STRAINS
THAT ARE INAUR QUADRIVALENT
VACCINE.
WE HAVE A B-VICTORIOUS STRAIN
AND A B-[INDISCERNIBLE] STRAIN.
WORKING WITH DANIEL PEREZ, HE
WORKED WITH REVERSE GENETIC ANDS
DEVELOPED DIFFERENT STRAINS THAT
HAVE A MUTATION IN PB2 THAT
ALLOW THESE VIRUSES TO
PRODUCTIVELY INFECT MICE.
OKAY, SO CLINICAL ISOLATES DO
NOT INFECT MICE.
SO WHAT I'M SHOWING YOU HERE ARE
DATA FROM THE B-VICTORIA, BUT WE
SEE A SIMILAR PATTERN WITH
B-YAMAGADA.
AND WHAT WE INITIALLY DID AND WE
TOOK OUR ADULT MICE, TYPICALLY
8-10 WEEKSA THE TIME WE BEGIN
THESE INFECTIONS AND WE INTRA
NAISALLY INFECTED MALES IN GREEN
AND FEMALES IN PURPLE AND WE
NOTE THAD MALES GOT SICKER,
FEMALES DIDN'T GET SICK HARDLY
AT ULTIMATELY.
SO WE TOOK MICE THAT WERE
JUVENILE, THEY WERE 23 DAYS AND
JUST AFTER THEY WERE WEANED, WE
INFECTED THEM WITH FLU B, THE
HOLD, MALES GOT SICKER THAN
FEMALES AND WE'RE LOOKING AT
RECTAL TEMPERATURE, WE DO RECTAL
TEMPERATURE, CLINICAL MARKERS OF
DISEASE, WE DO BODY MASS SO YOU
WILL SEE THAT THROUGHOUT.
SO THEN WE TOOK AGED MICE, THESE
MICE WERE 17 MONTH AT THE TIME
WE DID THESE INFECTIONS AND CAN
YOU SEE DEFINITELY THEY GET
SICKER BUT MALES AGAIN GET SICK.
FEMALES REALLY DON'T EVEN AT 17
MONTHS OF AGE.
WHEN WE MEASURE VIRAL TITERS IN
THE LUNGS, AND WAWE FIND
DEPENDING ON THE AGES OF THESE
ANIMALS, WITH YOU'VE NILES,
CLEARING VIRUS FASTER THAN
EITHER ADULT OR OLDER ADULT, BUT
WHAT WE FIND DPREEN MALES,
PURPLE FEMALES, LOG SCALE MALES
HAVE HIGHER VIRAL TITERS THAN
FEMALES DURING PEAK VIRUS
REPLICATION EMPLOY SO AT YOU'VE
NILES WE SEE THIS 3 AND 5 IN
ADULTS AND BY 7 MALE VS NOT
CLEARED AND FOR THE OLDER
ANIMALS MALES STILL HAVEN'T
CLEARED BY 9 DAYS AFTER
INFECTION AND WE'VE SHOWN THAT
THIS GREATER VIRUS REPLICATION
IN MALES DRIVES TYPE 1
INTERFERON RESPONSES SO IF WE
TREAT THEM WITH TAMAVIR, WE CAN
ELIMINATE THE SEX DIFFERENCE
BECAUSE WE CAN MAKE MALES LOOK
LIKE FEMALES.
FEMALES THAT GET OSILTAMAVIR,
DON'T LOOK THAT DIFFERENT THAN
THOSE THAT DON'T GET IT BECAUSE
THEY ARE NOT GETTING SICK IN
THESE MODELS.
FOR BOTH OUR YOUNG ADULTS 8-10
WEEKS OF AGE AS WELL AS OLDER
ADULT AS THE 17 MONTHS OF AGE.
YES, WE MEASURE THESE IN THE
LAB, AND HERE TO SHOW WHAT WE
SEE IF YOU MEASURE TESTOSTERONE,
THEY HAVE SLOW CIRCULATING
CONCENTRATIONS OF TESTOSTERONE
IN ARK DULT HOOD, THAT GOES UP,
THAT INCREDIBLE DIVERSITY, THOSE
ARE THE DOMINANCE HIERARCHIES IN
THE CAGES WHICH I'M CERTAIN ALL
OF YOU HAVE OBSERVED SO HERE ARE
THE SUBORDINATE MALES.
AND THEN AGING WHILE MICE DON'T
GO THROUGH THE SAME MENOPAUSE IN
TERMS OF FEMALES THEY DO
EXPERIENCE REPRODUCTIVE
SENESCENCE, NOT EVEN OUR FEMALES
BUT OUR MALES WE SEE A DECLINE
IN ANDROGENS BUT EACH WITH THESE
CHANGES INSTERIES, I THINK IT'S
REALLY IMPORTANT TO NOTE, WE SEE
THE SEX DIFFERENCE ACROSS
DIVERSE AGES SO THIS STARTED TO
SUGGEST TO US THAT MAYBE WITH MY
ENDOCRINOLOGY BACKGROUND, MAYBE
THESE GONADAL STEROIDS WERE NOT
INVOFFED.
SO WE HAVE LOTS OF DIFFERENT
WAYS THAT WE CAN START TO TEASE
APART IF IT'S NOT HORMONES, THEN
WHAT IS IT?
AND SO, AS SOMEONE WHO'S ROLELY
INTERESTED IN SEX BASED
VIROLOGY, IF IT'S NOT GONADAL
STEROIDS, THEN MY BRAIN GOES TO
SEX HORMONES BECAUSE THAT'S THE
DIFFERENCE.
WE HAVE LOTS OF TRANSGENIC MICE
THAT ARE AVAILABLE THAT ARE BRED
IN MY LAB THAT ARE NOT SMECIALLY
AVAILABLE AND 1 OF THEM IS
CALLED THE FCG, SO IF I CAN
BRING YOU BACK TO MY FIRST SLIDE
AND THAT SRY GENE THAT ENCODES
FOR TESTES DETERMINING FACTOR
WHAT'S DONE IN THE MICE IS THE
SRY YEEN HAS BEEN DELETED TO THE
Y-CHROMOSOME AND TRANSLOCATED TO
CHROMOSOME 3 SO THAT IT CAN BE
AUTOSOMEALLY INHERITED EMPLOY SO
NOW YOU CAN HAVE XY MICE,
THERE'S OUR LITTLE Y, X, CAN YOU
HAVE XY-MICE BUT DON'T HAVE THE
SRY SO THAT BI ON POTENTIAL
GONAD DEVELOPS INTO OVARIES.
WE HAVE OVARIES THAT SECRETING
ESTROGEN, WE ALSO HAVE X-MICE
THAT INHERIT THE SRY, AND SO
DESPITE HAVING 2 X-CHROMOSOMES
THEY HAVE THAT TESTES
DETERMINING FACTOR SO THAT GONAD
DEVELOPS INTO TESTES, SO CAN YOU
GET XXY-MALES OR XY-FEMALES SO
HERE WE FIND HERE FEMALES IN
PURPLE, OUR XX-FEMALES,
XX-ANIMALS WITH OVARIES AND WE
FIND THAT XX-MAILS SO THEY HAVE
2 X-CHROMES BUT THEY HAVE TEEOF
THE EASE THEY LOOK
INDISTINGUISHABLE.
THEY ARE PROTECTED.
MALES THAT HAVE AN X AND A Y
CHROMOSOME COMPLEMENT AS WELL AS
FEMALES MEANING ANIMALS WITH
OVARIES WITH AN XY COMPLEMENT
THAIZ GET AS SICK AS THE
BIOLOGICAL MALE.
OKAY?
SUGGESTING THAT IT'S SEX
CHROMOSOME COMPLEMENT MORE THAN
SEX STEROIDS HORMONES THAT ARE
MODULATING THIS DIFFERENCE AND
YOU CAN SEE THIS HERE, BOTH IN
TERMS OF VIRAL TITERS AND
INTERFERON BETA PRODUCTION.
SO WE ALSO AND I HAVE JEN LU,
SHE'S A POST DOC IN MY GROUP,
SHE'S HERE WITH US, WE'VE DONE
WORK WITH SARS-COV-2 AS WELL.
WHAT I'M SHOWING YOU HERE ON
THIS SLIDE IS USING THE NEW
VARIANT SO THIS IS A CLINICAL
ISOLATE OF THE NEW VARIANT.
IT WAS DISCUSSED STANLEY
BEAUTIFULLY DISCUSSED THIS N501
Y-MUTATION AND WHY IT'S SO
IMPORTANT FOR ALLOWING THE VIRUS
TO BIND TO MOUSE ACE 2, SO IF WE
USE THIS VARIANT, WE'VE ALSO
USED THE MA-10 FROM RALPH BARIC,
AND WE ALSO PUBLISHED USING MALE
AND FEMALE HAMSTERS INFEC WIDE
THAT WUSH WASH 1.
OKAY, SO 3 DIFFERENT MODELS AND
WHAT WE CONSISTENTLY FIND IS
THAT DURING THIS ACUTE PHASE OF
INFECTION, MALES IN GREEN,
FEMALES IN PURPLE, MALES GET
SICKER THAN FEMALES.
BUT HERE WE CONSISTENTLY, WE SAW
THIS WITH HAMSTERS AND WE'RE
SEEING THIS AND I SHOULD STATE
THESE ARE C57 BLACK 6 MICE, WE
HAVE USED BOTHIAC SON AS WELL AS
CHARLES RIVERS SO BOTH Y& C R
IF WE'RE GOING TO GET REAL
TECHNICAL AND THESE ARE WHAT I
WOULD CALL MIDDLE AGE, I WOULD
SAY THESE ARE MATURE MICE.
THEY ARE 30 WEBBINGS OF AGE
WHICH REALLY DOES CORRESPOND
WITH PEOPLE ROUGHLY IN THEIR 30S
TO EARLY 40S ACCORDING TOIAC SON
LABS.
SO CAN YOU SEE WE DON'T SEE A
DIFFERENCE IN VIRAL TITERS SO
THIS IS SAYING TO US, THERE
ISN'T A SEX DIFFERENCE IN THESE
RESISTANCE TYPES OF MECHANISMS,
SO ALREADY A LITTLE BIT
DIFFERENT THAN WHAT I SHOWED YOU
FOR FLU B.
WE DO LOOK IN THE LUNGS AND WE
FIND FEMALES HAVE UPREGULATION
OF CYTOKINES AND CHEMOKINES, WE
ALSO FIND SEVERAL IMMUNE CELLS,
I REALIZE I ONLY PUT MACROPHAGES
ON HERE BUT WE ALSO SEE THIS FOR
CERTAIN T-CELL POPULATIONS WHICH
I WILL COME BACK TO IPT GREATER
CLUING CD4, POSITIVE T-CELLS ARE
GREATER IN FEMALES DURING THIS
ACUTE PHASE OF INFECTION.
IF I SHOWED YOU THAT JEN HAS
CARRIED ANIMALS OUT TO ROUGHLY
21, 28 DAYS, SHE MEASURES
ANTIBODY, FEMALES ALSO HAVE MORE
ANTIBODY THAN MALES.
SO FEMALES, THEY'RE BY AND LARGE
DOING A LITTLE BIT BETTER.
SARS-COV-2 IN THESE MICE, IT
CAUSES A MILD DISEASE NOT LIKE
WHAT WE SEE WITH THINGS LIKE
INFLUENZA A, SO EVEN THOUGH I'M
SHOWING YOU FEMALES HAVING
GREATER CYTOKINE CONCENTRATIONS,
THIS IS NOT TO THE DEGREE OF
CYTOKINE CHANGES THAT WE SEE IN
THE LUNGS OF FLU A INFECTED
MICE.
BUT AGAIN WE CAN TAKE THESE
DIFFERENT TRANSGENIC MICE, I'VE
ALREADY INTRODUCED YOU TO THE 5
CORE TYPE OF MICE, WE USE THOSE
TO EPISAIT SEX CHROMOSOMES FROM
SEX GONADAL HORMONES AND WE FIND
THAT FEMALES ARE THE CLOSED
PURPLE, MALES ARE THE CLOSED
GREEN AND AGAIN X-FEMALE, SO
XEXTINGUISHING-WITH OVARIES, XY
MALES SO XY WITH TESTES AND CAN
YOU SEE THAT XX MALES, IT'S A
BIT INTERMEDIATE BUT THEY DO
MUCH BETTER THAN XY MALES AND XY
FEMALES DO WORSE THAN XX FEMALES
SUGGESTING THAT THERE IS A SEX
CHROME CHROME COMPLEMENT EFFECT.
NOW WHAT WE CAN DO IF WE SEE A
SEX CHROMOSOME COMPLEMENT CENTER
FOR EXCELLENCE ON AGING, MUCH
LIKE WE SAW OBSERVED WITH FLU B,
WE CAN THEN WORK WITH A
DIFFERENT TYPE OF TRANSGENIC
MOUSE MODEL, THESE ARE REKERRED
TO AS STAR Y MICE WHERE THERE'S
BEEN A MUTATION IN THE AUTOSOMAL
REGION OF THE X-ROAMSOME, WHICH
ALLOWS FOR US TO LOOK AT
DIFFERENT DOSAGES OF THE
X CHROMOSOME IN THE PRESENCE OR
ABSENCE OF THE Y CHROMOSOME.
BECAUSE WE HAVE AN INHERENT
DIFFERENCE IF WE'RE TALKING
ABOUT THE POLES OF BIOLOGICAL
SEX, FEMALES HAVING 2 X
CHROMOSOMES, MALES ONLY HAVING
1.
AND WHILE THROUGH A PROCESS OF X
CHROMOSOME INACTIVATION, THAT
SHOULDN'T MATTER.
WE SHOULD ALL HAVE THE SAME
NUMBER OF [INDISCERNIBLE] OR THE
SAME EXPRESSION OF X-LINKED
YEENS BUT ROUGHLY 15% OF THE
GENES ON THE X-CHROMOSOME IN
HUMANS ESCAPE X INACTIVATION.
SO THERE ARE SEVERAL GENES IN
INCLUDING TOLL-LIKE RECEPTOR 7
THAT ARE ON THE CHROMOSOME THAT
ESCAPE X ACTIVATION AND CONNIST
ISENTLY SHOW GREATER EXPRESSION
IN FEMALES AS COMPARED WITH
MALES.
SO NOW WHAT WE CAN DO IS WE CAN
HAVE MICE THAT DO NOT HAVE THE
SRY, THAT HAVE EITHER 2 COPIES
OF AN X-CHROMOSOME OR 1 COPY OF
AN X-CHROMOSOME, SO NOW WE SRO X
X FEMALES, WE CAN HAVE X O
FEMALES SO THIS WOULD BE CONSIST
WENT A DIFFERENT IN SEXUAL
DEVELOPMENT RESULTING IN TURNER
SYNDROME IN HUMANS.
WE CAN HAVE MICE THAT HAVE AND
INHERIT THE SRY ON THEIR Y
CHROMOSOME THAT HAVE EITHER 1
COPY OF THE X CHROMOSOME OR 2
COPIES OF THE X CHROMOSOME SO
YOU CAN HAVE XY MALES OR
EXTINGUISHING XY MALES SO THIS
TERMS OF THINKING ABOUT CLINICAL
RELEVANCE WOULD BE CONSIST WENT
AN INDIVIDUAL WHO MIGHT HAVE
[INDISCERNIBLE] DISORDER AND SO
NOW WHEN WE INFECT THOSE MICE
WITH THE NEW VARIANT OF
SARS-COV-2, YOU CAN SEE THIS
CLEARLY, ANIMALS THAT HAVE 2
COPIES OF THE X CHROMOSOME ARE
PROTECTED AGAINST ACUTE
SARS-COV-2 DISEASE.
MALES HAVING A SINGLE COPY OF
THE X CHROMOSOME, FEMALES HAVING
A SINGLE COPY OF X CHROMOSOME
GET SICKER BUT I THINK IT'S VERY
INTERESTING THAT HAVING THE
SINGLE COPY OF THE X CHROMOSOME
IN THE PRESENCE OF Y MAKES
ANIMALS THE SICKEST SUGGIEST
THRG IS SOMETHING ABOUT THE
Y-CHROMOSOME WHICH IS MANY
REGULATORY GENES EMPLOY SO WHAT
DO YOU DO WHEN WE SEE THIS
EMPLOY WELL, WE CAN THEN GO AND
LOOK AT SOME OF THE YEENS THAT
MAY BE ESCAPING X INACTIVATION,
I MENTIONED TOLL-LIKE RECEPTOR
7, WE PUBLISHED A LOT ON THAT.
BUT THERE ARE ALSO OTHER YEENS
THAT UPGENERATED ETDICALLY
REGULATE CELL ACTIVITY THAT CAN
DIFFER BETWEEN THE SEXES AS A
RESULT OF HAVING 2 COPIES OF THE
X-CHROMOSOME, SO WE CAN LOOK AT
THOSE X-LINKED YEENS THING
LOOSIC KDM6 A, KDM5 C BOTH OF
WHICH ARE SHOWN HERE AND
CDPOSITIVE T-CELLS IN SARS-COV-2
INFECTED MICE, AND XXINDIVIDUALS
HAVE GREATER EXPRESSION OF THESE
EPIGENETIC REGULATING GENES IN
THEIR REGULATING T-CELLS, SO
THEN WE HAVE CONDITIONALLY KNOCK
THESE OUT OF SPECIFIC CELLS,
MAYBE NOSE INFLAMMATORY
MACROPHAGES AS AN EXAMPLE.
MAYBE CD4 POSITIVE T-CELLS TO
LOOK AND SEE ARE THESE
PARTICULAR X-LINKED YEENS
MEDIATING THE SEX DIFFERENCE WE
OBSERVE EMPLOY SOW FLEW A, I
WOULD BE REMISS IF I DIDN'T GO
BACK TO WHERE THIS ALL BEGAN FOR
ME.
WE HAVE SHOWN THAT WHETHER WE
WORK WITH PRA, WHETHER WE WORK
WITH ACOW 09, WHETHER WE WORK
WITH H3 N2 VIRUSES, WHAT WE FIND
WITH FLU A ARE THAT FEMALES GET
SICKER THAN MALES.
THEY LOSE MORE WEIGHT, IF YOU
DON'T LIKE WEIGHT AND YOU WANT
TO TALK ABOUT SOME OF THE MORE
CLINICALLY RELEVANT MEASURES
THAT STANLEY BROUGHT UP AND YOU
LOOK AT THAT DIFFUSION CAPACITY
THAT EXCHANGE OF OXYGEN ACROSS
THE LUNGS, HERE I'M SHOWING YOU
14 DAYS AFTER INFECTION EMPLOY
SO 14 DAYS, MALES IN GREEN,
THEY'VE RETURNED.
THEY'RE BETTER THAN BASE LINE
HEALTH.
FEMALES HAVE NOT YET RETURNED
BUT AGAIN, IF YOU DON'T LIKE
THAT, HERE, CAN YOU SEE THE
STIFFLED NICE, CAN YOU SEE WHAT
IT LOOKS LIKE IN TERMS OF THE
OXYGEN EXCHANGE, MALES ARE BACK
UP TO NORMAL BASE LINE HEALTH,
FEMALES ARE NOT.
THIS IS NOT DUE TO RESISTANCE
MECHANISMS, WE FOUND IN NAIVE
MICE MALE ANDS FEMALES HAVE
SIMILAR TITERS OF REPLICATING
THEM IN THE LUNGS, THEY HAVE A
LOT MORE INFLAMMATION, CYTOKINE,
THAT CYTOKINE STORM CONSISTENTLY
SHOWED MUCH GREATER IN FEMALES
AS COMPARED WITH MALES.
SO WHAT IS IT ABOUT MALES, WELL,
HERE IS WHERE I THINK ANDROGENS
TERN OUT TO BE REALLY GOOD.
I KNOW AGAIN, CLINICALLY WE
ALWAYS TALK ABOUT THESE DPLU CO
CORTICOIDS BUT I WANT TO
CONVINCE THAT YOU ANDROGEN ROUGH
ATOMIENS ALSO DO A BEAUTIFUL JOB
DAMPENING INFLAMMATION.
AND SO SOME OF THE OTHER MODELS
WE HAVE ARE TEASING APART SEX
DIFFERENCES SO WE CAN DO
SURGICAL INTERVENTIONS, WE CAN
TAKE OUT THE TESTES, SO BY
TAKING OUT THE EAST EASE WE'VE
REDUCED CIRCULATING ANDROGEN
CONCENTRATIONS TO NONDETECTIBLE
LEVELS.
WE WANT TO KNOW, IS IT THAT 1
HORMONE?
WELL WE CAN TAKE THOSE MICE AND
WE CAN GIVE AN IMPLANT, A TIME
RELEASED IMPLANT THAT'S GOING TO
SECRETE EXOGENOUS IN THIS CASE
TESTOSTERONE INTO CIRCULATION
AND WE FIND THAT MALES THAT THEN
GET INFECT WIDE AICAL 09, IF
THEY HAVE ENDOGENOUS OR
EXOGENOUS ANDROGEN ROUGH
ATOMIEN, THEY DON'T GET US SICK
AS MALES THAT HAVE NO
CIRCULATING ANDROGENS.
TO SHOW YOU THIS ISN'T SO MUCH
ABOUT RESISTANCE, BUT MAYBE IT'S
A BIT MORE ABOUT HOW WELL THEY
TOLERATE VIRUS, I'M SHOWING YOU
VIRAL LOADS, 7 DAYS, COMPARED
WITH BODY MASS AND SO WHAT I
WANT YOU TO LOOK AT IS THAT ALL
ANIMALS WHETHER THEY HAVE THEIR
TESTES OR NOT, WHETHER THEY HAVE
ANDROGENS OR NOT, THEY HAVE A
VERY SIMILAR VIRAL LOAD, BUT THE
IMPACT OF HAVING THAT VIRAL LOAD
ON DEC REALLY DIFFERS, AND YOU
CAN SEE THAT BY THE SLOPE OF
THESE LINES.
MALES HAVE TAKEN--THEY ANDROGEN
ROUGH ATOMIENS, THE SLOPE
DOESN'T JUST STEEP, THEY'RE NOT
GETTING AS SICK AT THAT VIRAL
LOAD LEVEL AS COMPARED WITH
MALES THAT DON'T HAVE ANDROGEN.
AND WE CAN COMPARE MALES AND
FEMALES WHICH I'VE SHOWN YOU AND
HERE THE FEMALES, OPEN ORANGE,
MALES CLOSED BLUE, AND WE CAN
NOW GET FEMALES IN ANDROGEN,
THAL CASE WE CHOSE TO GIVE THEM
HIHYDROTESTOSTERONE, THE DIRTY
LITTLE SECRET ABOUT TESTOSTERONE
IS THAT IT CAN BE CONFIRMED BY A
ENZYME INTO AROPA TAISS--SYNTH
INTO ESTRADISCIPLINARY OLDER
PEOPLE SO A LOT OF THESE EFFECTS
THAT ARE OCCURRING IN MALES ARE
REALLY THROUGH BINDING AT TIMES
TO THE ESTRO GENRE SEPTORSOR,
AND BEING AROMAITIZED SO WE
DIDN'T WANT THAT SO WE TOOK A
NONAROMAITIZABLE ANDROGEN, CAVE
THAT TO FEMALES AND NOW YOU CAN
SEE WE CAN MAKE FEMALES LOOK
INDISTINGUISHABLE FROM MALES IF
WE GIVE THEM ANDROGENS.
THEY DON'T GET US THERE.
AND WE'VE GONE ON TO SHOW THAT
IT'S PARTIALLY BY DAMPENING A
LOT OF INFLAMMATORY IMMUNE
RESPONSES IN THE LUNGS IS HOW
THEY'RE PROTECTING VERY SIMILAR
TO HOW WE KNOW THAT THE
CORTICOIDS PROTECT.
SO IF EVERYBODY LIVES BECAUSE
WITH ACALO9 EVERYBODY LIVES AND
WE LOOK AT ADAPTIVE TIME POINTS
WE FIND THAT FEMALES HAVE
GREATER ADAPTIVE IMMUNITY, WE
LOOK AT ANTIBODY IN THEIR SERUM
OR LOOK LOCALLY, I DON'T EVEN
KNOW HOW TO GET BACK OR WE LOOK
LOCALLY IN THE BRONCO ALVEOLAR
LAVAGE, WE CAN LOOK IN THE LUNGS
FOR ANTIBODY SECRETING CELLS,
ALL DIFFERENT KINDS OF FLAVORS
OF T-CELLS AND WHAT WE FIND
FEMALES THE OPEN SYMBOLS, THE
MALES THE CLOSED, I WAS CHARGED
FOR COLOR.
FEMALES HAVE MORE OF THESE
CELLS, MORE OF THIS ANTIBODY.
SO WE CAN--THIS GOT US THINKING
ABOUT VACCINES AND SO IF WE WORK
WITH ENACTIVATED AICAL 09 DOING
A PRIME BOOST AND THEN LOOK AT
THE RESPONSES AND NOW I WANT TO
START TO BRING IN AGING.
STANLEY SPOKE ABOUT AGING WE
THINK ABOUT AGING BUT I THINK
ABOUT ANALLING DIFFERENTLY.
I THINK ABOUT IT AS A TIME AS
OUR HORMONES ARE STARTING TO
SERIOUSLY DEMINNISH.
AND SOPHISTICATED TO ME IT'S
ALMOST A NATURAL MODEL STUDYING
HOW CHANGES IN GONADAL HORMONES
MIGHT IMPACT VACCINE DOSED
IMMUNITY.
SO HERE ARE ARK DULLS 2-3 MOCKS
OF AGE, AT THE TIME WE'RE
VACCINATING AND COLLECTING
SAMPLES AND AGAIN OUR ADULTS ARE
17-18 BECAUSE OF THE DURATION OF
TIME IT TAKES WITH THESE VACCINE
MODELS SO THE YOUNG ADULTS ARE
DARKER GREEN AND PURPLE AS
COMPARED WITH THE AGE.
SO IF YOU LOOK IN THE DRAINING
EMILY NODES, ADULT FEMALES LET'S
JUST TART OFF, THEY JUST HAVE
MORE B-CELLS THAN THEIR MALE
COUNTERPARTS.
WE LOSE THAT WITH AGING.
WE CAN MEASURE GERMINAL TRUSTEES
B-CELLS, WE CAN MEASURE PLASMA
BLAST, ASULTAN MALS HAVE MORE OF
THESE CELLS THAN ADULT MALES AND
WE LOSE THAT WITH AGING.
IN FACT, CAN YOU SEE IT LOOKS
PRETTY BAD FOR BOTH AGED MALES
AND FEMALES.
NEUTRALIZING ANTIBODIES AND WE
LOSE THAT SEX DIFFERENCE IN THE
AGING WE'VE DON CYSTENTLY SHOWN
IS GRITTER FOR FEMALES AS
COMPARED WITH MALES SO IT'S
CORRELATIONAL BUT I HOPE CAN YOU
SEE IF WE MEASURE
ESTRADISCIPLINARY OLDER PEOPLE
IN ALL THESE ANIMALS YOU CAN SEE
THAT THE PEAK OF
ESTRADISCIPLINARY OLDER PEOPLE
CORRELATES WITH THE INCREASES IN
ALL OF THESE DIFFERENT CELL
TYPES IN THESE ANTIBODY
RESPONSES, IT'S NOT WHAT WE SEE
WHEN WE MEASURE ANDROGENS IN
THESE ANIMALS.
SO SOME OF THE WAYS WE CAN USE
HORMONES THERAPEUTICALLY, WE CAN
TAKE AGED ANIMALS LIKE AGED
FEMALES AND MAYBE IT'S MY AGE
THAT I'M STARTING TO REALLY
THINK ABOUT THESE THINGS AND CAN
JUST INSERT A LITTLE
ESTRADISCIPLINARY OLDER PEOPLE,
EXOGENOUSLY IN THOSE ANIMALS AND
ESTRADISCIPLINARY OLDER PEOPLE
IS ABBREVIATED E2, SO WHEN WE
COMPARE THAT WITH ARK DULT
FEMALES IN AGED FEMALES, AFTER
WE DO OUR PRIME BOOST
VACCINATION, AGED FEMALES HAVE
LOWER ANTIBODY RESPONSES, I'VE
ALREADY SHOWN YOU THIS, BUT IF
YOU GIVE AN AGED FEMALE,
ESTRADISCIPLINARY OLDER PEOPLE,
THE ANTIBODY RESPONSE LOOKS
INDISTINGUISHABLE FROM EITHER
ADULT FEMALES THAT HAVE
EXOGENOUS ESTRADIOL, OR FROM
FEMALES THAT HAVE CLINICAL
GENETICS DODGE NOWS ESTRADIOL,
AND AGAIN WE GET TO WORK WITH
ANDY PECOS, WITH ALL HIS GENETIC
REVERSE SYSTEMS, AND HE MONITORS
WHAT'S GOING ON OVER 32 THROUGH
THE SURVEILLANCE EFFORTS OF SEER
AND INSERTS MUTATIONS INTO
VIRUSES SO THAT WE'RE DRIFTING
AICAL 09, SO WE CAN WORK WITH
THE VIRUSES TO THE MATCH, BUT
THEY DON'T EVEN GET SICK IF
THEY'VE BEEN VACCINATED, THE WAY
AGED FEMALES DO, THEY GET SICK,
BUT THEY DON'T DIE, AND THEY DO
GET SICK.
BUT IN AN AGED FEMALE, WHO GOT
THE ESTRADIOL, SHE LOOKS
INDISTINGUISHABLE FROM THE ADULT
FEMALES.
SO I THINK I WILL HAVE THIS BE
MY LAST AND WE CAN END AFTER
THIS BUT AGAIN GOING BACK TO
THOSE FCGMICE AS 4 CORE GENOTYPE
MICE, WHAT I WANT TO SHOW YOU IS
WE SEE SOMETHING VERY DIFFERENT
WITH VACCINE INDUCED IMMUNITY WE
DO WITH VIRAL PATHOGENESIS, IN
WHICH CASE IF WE TAKE THOSE 4
GENOTYPES WHAT WE FIND IS THAT
FEMALES, MEANING ANIMALS THAT
HAVE OVARIES REGARDLESS OF
WHETHER THEY HAVE AN XX OR XY
CHROMOSOME COMPLEMENT THEY MAKE
MORE ANTIBODY TO OUR VACCINE
THAT IS ANIMALS THAT HAVE TESTES
REGARDLESS OF THE SEX CHROMOSOME
COMPLEMENT AND THIS IS REFLECT
INDEED GERMINAL CENTER B-CELLS,
PLASMA BLASTS AND CHALLENGE WITH
THE DRIFTED VIRUS, THOSE ANIMALS
THAT HAVE MORE ANTIBODY, MORE
CELLS THEY CLEAR VIRUS BETTER
THAN ANIMALS THAT HAVE TESTES,
SO WHAT I THINK I'VE DONE, I'VE
SHOWN WE HAVE LOTS OF MODELS
THAT I THINK ARE CRITICAL IN
HIGHLIGHTING THE ROLE BIOLOGICAL
MODELS PLAYS, I THINK SEX IMPACS
OUR LIFE STRATEGIES, I THINK
IT'S EVOLUTIONARY AND REALLY
IMPACTS THIS NOTION OF
RESISTANCE VERSUS TOLERANCE, WE
HAVE TRANSGENIC MICE, WE HAVE
SURGICAL MODELS, AGING TO SHOW
YOU ROLE OF COMPLEMENT VERSUS
SEX STEROIDS OTHERS BOTH FOR
VIRAL PATHOGENESIS BUT ALSO IN
THE CONTEXT OF VACCINATION AND
SO WITH THAT I WILL END.
THANK YOU.
>> SO FOR THE PANEL DISCUSSION,
WE WILL LIKE TO ASK THE SPEAKERS
TO JOIN UP FRONT AND SO WE WILL
HAVE OPEN QUESTIONS FOR ALL
PANEL MEMBERS OR ANY TYPE OF
GENERALIZED QUESTION THAT WOULD
BE DIRECTED IN GENTLEMEN TO THE
PANEL.
>> GO FOR IT.
>> I HAVE 1 FOR SABRA, I LOVE
IT, I LOVE ALL OF IT.
BUT I AM SO CURIOUS ABOUT SOME
OF THE OBSERVATIONS THAT MOVE
MADE IN ALL OF THESE VARIOUS
MODELS THAT YOU'VE USED, HOW IT
RELATES TO, YOU KNOW THINGS THAT
HAPPEN IN HUMANS ESPECIALLY WHAT
I BELIEVE LIKE THE END WITH THE
ADDITIONAL ESTRO GENERATED IN
THE AGED FEMALE, A LOT OF PEOPLE
TALKING ABOUT MENOPAUSE RIGHT
NOW WITH THE NEW HEALTH
INITIATIVES AND THINGS.
WHAT DOES THAT MEAN AND HOW
CAN--YOU KNOW HOW IS IT
RELATING?
NSO IF I START AT THE END WITH
VACCINES, MENOPAUSE, HORMONE
REPLACE AM THERAPY, THERE'S
REALLY, THERE HAS BEEN
ABSOLUTELY NOTHING DONE WITH
VACCINATION, SO THAT'S SOMETHING
THE DIRECTION AS I AGE THAT I'M
TAKING THE LAB BECAUSE I WANT TO
KNOW BUT I DO THINK, YOU KNOW
THESE MOUSE STUDIES ARE REALLY
SUGGESTIVE THAT THERE GOING TO
BE A DIFFERENCE.
THE OTHER THING, YOU KNOW THAT
WE'VE GOTTEN INTERESTED IN, YOU
KNOW WHEN WE REPLACE HORMONE IN
MICE THESE ARE TIME RELEASED
CAPSULES SO IT'S LIKE PUTTING
THEM ON HORMONE REPLACEMENT
THERAPY BUT SOMETHING WE'RE ALSO
VERY INTERESTED IN IS THINKING
ABOUT HORMONES AS ADJUVANTS.
LIKE DO YOU NEED A BOLUS AT THE
TIME OF VACCINATION AND COULD
THAT BE SUFFICIENT OR DO YOU
NEED THIS CONSTANT RELEASE OF
THE HORMONES?
SO THOSE ARE SOME OF THE THINGS
WE'RE THINKING.
>> OTHER QUESTIONS?
MICHELLE?
>> I WOULD LIKE TO ASK A GENERAL
QUESTION OF THE PANEL.
SO WHEN YOU ARE PREPARING
VIRUSES, YOU'RE GROWING UP THEM
UP IN THE LAB OR SUBCULTURE OR
ANIMALS, AND YOU ARE INFECTING
DIFFERENT ANIMAL MODELS, IS THIS
STANDARDIZED AMONG THE VIRUSES
THAT YOU USE ACROSS LABS OR IS
EVERY LAB DOING THEIR OWN THING
AND ACTUALLY USING LOTS OF YOU
KNOW DIFFERENT LEVELS OF VIRUS
AND SO ON?
>> I CAN DEFINITELY SAY THAT IN
CORONA VIROLOGY IT VARIES FROM
LAB TO LAB.
THERE WERE MASSIVE CONVERSATIONS
KIND OF ACROSS, ESTABLISHED NEW
INVESTIGATORS WHEN SARS 2 HIT
BECAUSE OF THE ISSUES OF THE
CLEAVAGE SITE AND HAVING A
STABLE VIRUS STOCK, SO THERE
WERE KIND OF YOU KNOW COMMON
CONDITIONS THAT WE WOULD ALSO
USE BEFORE LIKE GROW INVIRO
CELLS AND TAKE BEFORE YOU HAVE
COMPLETE CPC AND SERUM AND ALL
OF THAT BUT I DON'T THINK
THERE'S ANY SORT OF COMPLETE
TANNED ARDIZEATION ACROSS THE
FIELD.
YOU GET BETTER AT IT OVER TIME
AND PASS ON TIPS AND TRICKS BUT
ABSOLUTELY NOTHING.
>> I THINK THAT'S EXACTLY THE
CASE ALSO WITH INFLUENZA, FOR
THOSE OF US WORKING IN THESE
AREAS, I DO THINK THAT WE
SEQUENCE WHAT WE'RE GROWING.
THERE IS--SO YOU'VE GOT SOME
INTERNAL STANDARDIZATION, SOME
OF THE WAYS THAT WE GO ABOUT
CREATE YOU KNOW THE SEED STOCKS,
VERSUS THE WORKING STOCKS ARE
VERY STANDARDIZED BUT NO, I
THINK WE TRY TO SEEK CONSISTENCY
WITHIN OUR LABS AND GOING BEYOND
AND AND GOING TO WHAT EVERYONE
ELSE IS CORKING WITH IS PROBABLY
A NEAR IMPOSSIBLE TASK.
NAT THE END THE DAY DOES THAT
AFFECT THE RESULTS AND THE WAY
YOU ARE ABLE TO COMPARE RESULTS
BETWEEN LABS.
>> YEAH, AS YOU KNOW ABOUT THE
SAVE PROJECT WHICH KEY DID AS A
COMMUNITY FOR STUDYING
SARS-COV-2, THERE ARE STOCKS
SENT OUT TO EVERYBODY SO THAT IN
THEORY WE COULD ALL JUST GET 10
OR 15 DOSES OF VIRUS AND USE ALL
THAT FOR OUR PRPLTS AND I THINK
THAT WE SAW PRETTY GOOD
CONFORMITY OF RESULTS BECAUSE OF
THAT, BUT THERE'S ALSO OTHER
VARIABLES SUCH AS--EVEN IF 1
MOVED FROM 1ITATE TO
ANOTHERITATE, THERE'S OFTEN
CHANGES IN MICROBIOMES AND
ANIMAL FACILITIES AND THAT
CHANGES HOW INFECTIONS OCCUR
EVEN APPROXIMATE 1 USES THE
STOCKS IN MICE, SO I AGREE WITH
THE ANSWER, IT'S PRETTY
DIFFICULT TO DO IT, IT'S IDEAL
IF WE COULD BUT IT'S PRETTY
DIFFICULT.
>> BUT I THINK WE SAW THAT AT
LEAST FOR THE SARS-COV-2 WORK
THAT SOMETIMES ACROSS VERY
DIFFERENT SPECIES, DIFFERENT
LABS ISSUES EVEN DIFFERENT
STRAINS OF VIRUS, WE START TO
SEE--IF WE TART TO SEE
CONVERGING EVIDENCE THAT WE CAN
SEE SOMETHING LIKE THE ANASMIA,
LIKE THE INFLAMMATION OF THE
OLFACTORY BULB, ACROSS, THEN
THIS I THINK TO ME, THAT'S
ALMOST MORE EXCITING CONVERGING
EVIDENT BECAUSE IT DOESN'T MEAN
THAT YOU KNOW YOU HAVE TO STAND
ON 1 FOOT, GROW THE VIRUS THIS
WAY, INFECT THE ANIMAL WITH ITS
HEAD TILTED ONLY THIS WAY IN
ORDER TO REPLICATE WHAT SOMEBODY
HAS DONE.
>> I THINK WITH THE DEEP
SEQUENCING WE'VE DONE IN THE
LABORATORY THAT'S BECOME MUCH
MORE PROMINENT THAN IT HAS IN
THE PAST WITH VIRUS STOCKS.
I THINK WE KNOW WHAT FREQUENCY
OF THAT MUTATION IS PRESENT
WITHIN OUR VIRUS STOCKS,
PRECOVID, I'M NOT SURE HOW MANY
OF US ACTUALLY DEEP SEQUENCED
OUR VIRUS STOCKS--
>> ONLY IF WE WERE SHIPPING IT--
>> --IT PROVIDES VARIATION TO
WHAT WE'RE SEEING IN THE VIRAL
STOCKS BETWEEN LAB RAARE TOYS AS
WELL.
--LABORATORIES AS WELL.
>> I WAS WONDERING HOW MUCH ALL
OF YOU GUYS LOOK AT DIFFERENT
INOCULATION ROUTES AND
[INDISCERNIBLE] BECAUSE I THINK
EVERY SINGLE 1 OF YOU IS
ANYTHING IN THE INTRA NAISAL
ROUTE, DO YOU SEE TRY FULL
[INDISCERNIBLE], DO YOU SEE
DIFFERENT PATHOGENESIS AFTER
THAT DO YOU TRY TO INOCULATEAR O
SOLS OR IS THAT SOMETHING YOU'RE
NOT CAPABLE OF DOING.
>> WE'VE TRIED ESPECIALLY WITH
THE HAMSTER FOR SARS OVERLAPPING
2 TRIED A BUNCH OF DIFFERENT
INOCULATION METHODS, EVERYBODY
TRIED EVERYTHING PRACTICES INTRA
NAISAL TO IV OR OCULAR
DRIPPINGS, NAIS AT PASSAGES WITH
DIFFERENT DOSES RANGING FROM 100
UP TO MILLIONS OF PFU AND IT WAS
SURPRISING HOW INCONSISTENT THE
RESULTS WERE SO IN THE LUNGS THE
VIRUS IS REP LITICATED GENERALLY
THE SAME LEVELS ACROSS ALL THESE
DIFFERENT CONTRACT TRANSMISSION,
GET SIMILAR, NOT ENTIRELY SURE
BECAUSE IT'S CONTRACT
TRANSMISSION AND HOW MUCH IS
COMMUNICATED WEAN THE DIFFERENT
ANIMALS BUT YOU SEE SIMILAR
VIRAL DYNAMICS.
THE ONLY THING THAT CHANGED IS
THE SUBTLE SHIFT IN THE KINETICS
WITH HIGHER DOSES, IT SEEMS LIKE
HIGHER ANTIGEN IN THE FRONT, BUT
SO YOU SEE A QUICKER TRIGGER SO
IT'S BEEN INTERESTING HOW
CONSISTENT THE INFECS ARE FOR
SARS.
INITIATIVES THE ONLY OTHER THING
I MIGHT ADD IN ARK DITION TO
INOCULATION ROUTES AND I
COMPLETELY AGREE AND WE'VE DONE
THAT WITH SEVERAL DIFFERENT
VIRUSES, I THINK ALL OF US WITH
WE'RE FIRST STARTING WITH A NEW
VIRUS, WE DEFINITELY DO LD50S
WITH OUR MICE AND ANYTHING BACK
TO THE QUESTION ABOUT, YOU KNOW
CONSISTENCY ACROSS LABS, 1
OBSERVATION THAT I'VE MADE AT
LEAST WITH FLU IS THAT LOOKING
AT THE MOUSE LD50 DOSE ALLOWS US
TO THEN DEAL WITH THAT MAYBE
THAT PR8 VIRUS IS A LITTLE MORE
PATHOGENIC IN THIS GROUP THAN
THIS GROUP BUT YOU CAN MATCH IT
ON THE MLD 50 AND WHEN YOU DO
THAT YOU SEE THAT YOUR DATA IS
CONSISTENT WITH DATA FROM A
COMPLETELY DIFFERENT LAB WHO
GREW THEIR PR8 STRAIN FROM
ANOTHER COMPLETELY DIFFERENT LAB
EMPLOY SO THOSE KINDS OF THINGS
I DO THINK HELP.
I THINK ALL FIELDS STILL DIFFER
IN HOW MUCH THAT'S A NORM, I
THINK FOR FLU, THAT'S DEFINITELY
A NORM.
YOU ASKED ABOUT VOLUME, I DON'T
THINK--I KNOW WE DO NOT PLAY
MUCH WITH DIFFERENT VOLUMES, FOR
THE INNOCULUM WE KEEP THAT
CONSISTENT BUT MAYBE CHANGE HOW
MUCH VIRUS IS GOING IN, BUT
AGAIN THAT ALSO DIFFERS
DEPENDING ON THE ROUTE.
NYEAH, I THINK A LOT OF IT IS
ALMOST PREDICTABLE SO IF YOU USE
A SMALLER VOLUME AND YOU DO AN
INTRA NAISAL INOCULATION YOU
WON'T COVER AS MUCH OF THE LUNGS
SO YOU GET A DIFFERENT RESULT.
IF YOU USE AEROSOL VIRUS, THE
CHALLENGE IS MAKING SURE YOU
HAVE THE SAME DOSE AS YOU WOULD
DELIVER DIRECTLY TO THE LUNGS
BUT INTRA NAISAL INOCULATION SO
WE'VE DONE DIFFERENCES IN
VOLUMES MOSTLY NOT FOR GOOD
REASONS BUT WE MADE MISTAKES AND
WE SAW DIFFERENCES THERE.
AEROSOL WE HAVEN'T TRIED BUT I'M
PRETTY SURE THAT IF YOU GET
ENOUGH VIRUS INTO THE LUNGS, OF
AN ANIMAL IT'S SUSCEPTIBLE,
BECAUSE THING WITH THE HAMSTER
SYSTEM THEY DON'T GET VERY SICK
RIGHT NETWORK THEY HAVE MODERATE
WEIGHT LOSS, THEY GET PEL
MONITORARY INFILTRATE AND
PNEUMONIA YOU ABOUT THEY ALWAYS
RECOVER.
>> WE HAVE GREAT HISTOLOGY, I
REMEMBER FROM THE
BEGENERATEDDING WE HAD GREAT
HISTOLOGICAL CHANGES THAT WERE
USEFUL FOR FOLLOWING BUT THERE
YOU HAVE A BIGGER CHANGE THAN
WHAT YOU CAN GIVE BECAUSE YOU'RE
NOT GOING TO KILL THE ANIMALS.
>> I HAVE A QUESTION FOR LISA
AND SORRY TO PUT YOU ON THE
SPOT, THE COLLABORATIVE CROSS
MOUSE MODEL HAS HELPED US BETTER
UNDERSTAND THE IMPACT OF MOUSE
GENETICS ON OUR PHENOTYPIC RATES
THAT 1 LOOKS AT BUT 1 OF THE
CHALLENGING PARTS OF THE
COLLABORATIVE PROCESS IS JUMPING
INTO USING THE COLLABORATIVE
CROSS AND THAT SEEMS TO BE A
BARRIER TO HAVE MUCH MORE WIDE
EXPRESS AND UNDERSTAND AND
UNDERSTANDING WHAT THE
PHENOTYPIC LOCI HOW THEY
CORRELATE CAN YOU SORT OF
PROVIDE MORE INSIGHT INTO HOW 1
CAN UTILIZE THAT MODEL,
WHATEVER, PATHOGEN WE MAY BE
INTERESTED IN?
>> YEAH, I WILL SAY MESSIAH
EXPERIENCE WITH THE CC HAS BEEN
TARTING ON SECOND BASE ALREADY
BECAUSE I'M LOCATED AT UNC WHERE
MOST OF THE MICE ARE AND A LOT
OF THE INVESTIGATORS WHO ARE
INVOLVED IN CREATING THIS
RESOURCE SO I DEFINITELY GAME IN
WITH A MASSIVE ADVANTAGE IN THAT
SENSE.
BUT THERE ARE A NUMBER OF MOUSE
GENETICISTS AT UNC, AND OCRANGE,
AND JACKSON HAS A LOT OF PEOPLE
WORKING WITH THEM AS WELL, AND
THEY ARE REALLY GOOD TO
COLLABORATE IN TERMS OF GENETICS
AND THE MODELING ASPECTS OF THAT
WHICH, YOU KNOW PEOPLE HAVE
TRIED TO TEACH ME HOW TO USE R
AND I CAN'T SAY IT'S BEEN
PARTICULARLY SUCCESSFUL, I HAVE
SPENT MY TIME LEARNING OTHER
THINGS, I WILLAD MITT, SO DOING
THE SCREENING INFECTING A LOT OF
MICE IS HONESTLY THE EASY PART.
CAN YOU GET GREAT PHENOTYPES AND
UNTIL YOU CAN CORRELATE WITH
WITH THE GENETICS AND DO THE
MODELING, YOU HAVE A PILE OF
DATA THAT CAN'T REALLY GO
ANYWHERE, SO, HAVING A PIPELINE
AND THERE'S FAIRLY STANDARDIZED
SCRIPTS IF YOU HAVE YOUR DATA
ORGANIZED PROPERLY THAT AREN'T
COMPLETELY PLUG AND PLAY BUT CAN
GET YOU TO QTLs PRETTY EASILY,
IN MY EXPERIENCE, THE BIGGEST
CHALLENGE HAS BEEN YOU HAVE A
QTR L, THIS REGION FROM
CHROMOSOME SENCH IS TIGHTLY
ASSOCIATED WITH THE PHENOTYPE OF
INTEREST AND HOW DO YOU GET TO A
CAUSATIVE YEEN AND HOW DO YOU
VALIDATE THAT SO A REAL
CHALLENGE FOR US HAS BEEN BOTH
IN GRANTSMANSHIP AND IN WRITING
PAPER ANDS HAVING PEOPLE WHO
WILL SUPPORT THE GENETIC SIDE
AND SUPPORT THE VIROLOGY SIDE,
AND RECOGNIZE THAT YOU MIGHT
HAVE AIAN LOT OF PRELIMINARY
DATA AND YOU MIGHT NOT HAVE A
COMPLETE STORY BUT IF A REVIEWER
COMPLETELY DISCOUNTS GENETICS
HALF OF IT, OR THE VIROLOGY HALF
OF IT AND BEING ABLE TO PUSH
THAT ACROSS THE FINISH LINE AND
GETTING PAPERS AND MORE MONEY TO
DO MORE WORK IS THE MOST
IMPORTANT PART.
>> UNT HAS DONE BASE LINE
MEASUREMENTS AS WELL AS
VALIDATED ANTIBODIES THAT 1 USES
FOR IMMUNE ANALYSIS.
IT'S A LOT OF THAT STUFF
PUBLICLY AVAILABLE, AND IF SO,
WHERE COULD 1 FIND IT IN.
>> A LOT OF IS SUPPOSED TO BE
AVAILABLE ON JACK'S MOUSE FETAL
COMPARTMENT NOAM BUT I SHOULD
DOUBLE CHECK ON THAT.
ALTERNATIVELY AS YOU KNOW E-MAIL
MATTER FERRIS.
NYEAH.
--MARTY FERRIS.
>> WE HAVE A VIRTUAL ATTENDEE
WITH THEIR HAND RAISED SO I WANT
TO CALL ON THEM.
KAY BARNS?
>> YEAH, HI, JUST WONDERING WHAT
WAS YOUR VIEW ON THE USE OF
THOSE ANIMAL MODELS KNOWING THE
SEX DIFFERENCES IS QUITE EXTREME
OR THE USE OF THEM FOR
EVALUATING MEDICAL
COUNTERMEASURES, WOULD YOU
CONSIDER USING BOTH SEXES BUT
MAYBE HAVING BIGGER GROUPS TO
COUNT FOR THAT DIFFERENCE?
>> OKAY SO IF I'M UNDERSTANDING
YOU CORRECTLY, YOU WANT TO KNOW
WHAT YOU OR ANYBODY AS AN
INVESTIGATOR SHOULD DO IF THERE
ARE THESE DIFFERENCES AND YOU'RE
BEING EXPECTED TO INCLUDE BOTH
BUT NOT THE KINDS OF MEMBERS
OBVIOUSLY THEY AM, I DO THINK
THAT'S A CHALLENGE AND I DO
THINK IT'S BEEN A PROBLEM WITH
THE 6 AS A BIOLOGICAL VARIABLE
REQUIREMENT.
BECAUSE I THINK IN SOME CASES IT
INCREASES HETEROGENEITY WHEN
YOU'RE LUMPING EVERYBODY
TOGETHER WHICH CAN NOT BE A GOOD
THING.
I THINK IN MANY CASES, WHAT I AM
SEEING IS JUST USING DATA SUCH
AS MINE THEN AS JUSTIFICATION
THAT IF YOU'RE STUDYING THE
PATHOGENESIS, FOR EXAMPLE, 3 B,
MAYBE IT IS BETTER TO GO AHEAD
AND USE MALE MICE BECAUSE THEY
GET SICK, AND YOU COULD STUDY IS
SEE BEAUTIFUL EFFECTS OF
DEFINITE ANTIVIRALS, IF YOU WERE
STUDYING ANTIVIRALS ON
EFFECTIVENESS ESPECIALLY NOVEL
ANTIVIRALS, I DON'T KNOW WHY YOU
WOULD GIVE THAT TO FEMALE MICE
WHEN THEY JUST DON'T GET SICK.
SO, YOU KNOW I THINK THERE ARE
RATIONAL WAYS TO UTILIZE WHAT WE
KNOW ABOUT SEX DIFFERENCES TO
THEN ASK WHAT QUESTION ARE YOU
ASKING AND COULD THAT BE
JUSTIFICATION FOR A SINGLE SEX
STUDY.
>> THANK YOU SO MUCH, YEAH, I
AGREE.
IT'S A VERY BIG CHALLENGE, ISN'T
IT?
>> YEAH, IT IS.
>> SO TO FOLLOW UP ON THAT, I
MAY HAVE MISSED THIS FROM YOUR
PRESENTATION, HOW MUCH HAS BEEN
DONE AT BASE LINE BETWEEN MALES
AND FEMALES IN TERMS OF INNATE
ANALYSIS, CYTOKINE ANALYSIS,
WHAT ARE DIFFERENCES INITIALLY
NNO, THAT'S SUCH A GOOD
QUESTION.
WE ABSOLUTELY ALWAYS DO THAT.
WE TEND TO NOT FIND A LOT OF
BASE LINE DIFFERENCES.
, YOU KNOW EITHER IN TERMS
OF'MUNE CELLS FROM FLOW
CYTOMETRY, OR PROTEIN CONTENT IN
THE LUNGS, CYTOKINES,
CHEMOKINES, THEY REALLY DO NOT
SEE A LOT OF BASE LINE
DIFFERENCES, SO, YOU KNOW I
THINK MUCH OF WHAT WE'RE SEEING
NOT SO DIFFERENT, I KNOW WE'RE
TALKING ABOUT RESPIRATORY
VIRUSES, BUT I MEAN, I COULD
REPLACE WHAT WE DO WITH
RESPIRATORY VIRUSES AND SHOW YOU
BEAUTIFUL WORK FROM OTHER
INVESTIGATORS WHO DO ALLERGY AND
ASTHMA MODELS AND I CAN SHOW YOU
THEY DON'T SEE BASE LINE
DIFFERENCES BUT THEY WILL GIVE
THEIR MICE THE HOUSE DUST MIGHTS
ALL OF A SUDDEN EVERYTHING THEY
SEE WITH FLU A, THEY SEE WHERE
FEMALES HAVE THIS OVER EXUBERANT
CYTOKINE RESPONSE, MORE TISSUE
DAMAGE, ET CETERA, SO IT'S
REALLY IN RESPONSE TO THE
STIMULUS, IT DOESN'T EVEN HAVE
TO BE A VIRUS.
>> STACEY?
>> SO THERE WAS AN INTERESTING
PAPER IN OBESITY AND SEX BIAS
AND LOOKINGA THE ENDOTHELIAL
CELLS AND LAKE LOOKING AT BASE
LINE, THE DIFFERENCES BETWEEN
ENDOTHELIAL MALE AND FEMALE, IT
WAS IN THE RACE, IN THE CELLS
THEMSELVES THAT MALES JUST TEND
TOWARDS MORE OF A
PROINFLAMMATORY RESPONSE EVEN AT
THAT ENDOTHELIAL CELL LEVEL SO
THOUGHT THAT WAS FASCINATING
NSEEMA?
>> I WANTED TO--MY
INTERPRETATION OF THE QUESTION
WAS FOR LISA ABOUT THE
COLLABORATIVE CROSS AND THE
PHENOTYPES YOU SEE ARE YOU DOING
THEM IN BOTH GENDERS--BOTH
SEXES, RIGHT, I'M SORRY, NEED TO
BE BETTER WITH THAT, IN BOTH
SEXES IN ORDER TO ENSURE THEY
ARE REPRODUCIBLE AND SEX BIAS
THERE.
NYEAH, SO PAINFULLY WE ARE DOING
IT IN BOTH SEXES AND WE DO
SOMETIMES SEE SOME DIFFERENCES.
I WOULD RECOMMEND AGAINST SOME
OF THE MALE CC LINES IF
POSSIBLE, BUT WE ARE STUDYING
THEM FOR OUR APPROPRIATE
STUDIES.
>> AND NOT TO MAKE THIS'
REFERENDUM I SEE ON THING AS THE
STUDY SECTION BUT THIS IS MORE
OF A PROGRAM OFFICE IS THERE ANY
APPETITE AT NIH TO, I DON'T WANT
TO SAY LOOSEN SEX AS A
BIOLOGICAL VARIABLE BECAUSE I
THINK IT'S IMPORTANT WE CONSIDER
IT BUT I THINK SEEMA MADE POINTS
WHERE IT MIGHT BE APPROPRIATE TO
SELECT 1 OVER THE OTHER
BIOLOGICALLY THAT CAN I TELL YOU
BECAUSE OF THE WAY THE RULES ARE
WRITTEN, BECAUSE IT'S SO RIGID
EVEN IF YOU HAVE AN ABSOLUTELY
GOOD SCIENTIFIC RATIONAL, CAN
YOU GET DESTROYED BY THE STUDY.
IT'S HAM STRUNG, SOME STUDY OF
SEX AS A BIOLOGICAL BARRIER.
>> I FULLY AGREE WITH YOU
BECAUSE TOO QUICKLY I HAVE
PEOPLE GOING, OH IT DOESN'T
MATTER.
BUT YOU HAD ENDS OF 2, YOU KNOW?
SO I FULLY GET THAT, I THINK
WHAT'S HAPPENED IS IT'S GETTING
APPLIED AND THIS IS MUCH MORE
LESS ABOUT PROGRAM OFFICERS,
MORE ABOUT THE CENTER FOR
SCIENTIFIC REVIEW.
IT GETS APPLIED IN A VERY LAZY
WAY WHERE YOU KNOW, REALLY IF
YOU DO PROVIDE THE RATIONAL, YOU
KNOW I WILL JUST KEEP GOING BACK
TO FLU B, YOU KNOW THAT MALES
GET SO MUCH SICKER, YOU HAVE
DESIGNED ALL THESE REALLY COOL
NOVEL ANTIVIRALS THAT YOU WANT
TO TEST THAT WOULD BE FANTASTIC
RATIONAL, YOU KNOW IN I WILL
SAY, IT'S THE PROBLEM IS, YOU
JUST HAVE REVIEWERS SAYING, BOTH
THERE, YES, NO, IF THEY'RE BOTH
THERE, CHECK THE BOX, IF THEY'RE
NOT.
SO I DO AGREE WITH YOU, BUT I
DON'T SEE HOW IT'S CHANGING.
>> I WOULD AGREE, WE GOT REAMED
ON A PREGNANCY GRANT BECAUSE
WE'RE NOT USING MALES, AND I'M
LOOK OKAY, BECAUSE THERE WAS A
LAZY REVIEWER AND NOBODY ON THE
STUDY SECTION CALLED THEM OUT
AND SAID HOW ARE YOU GOING TO DO
THAT?
SO I THINK THERE'S SOME GENERAL
REEDUCATION OF STUDY SECTIONS
THAT NEED TO HAPPEN AND IT'S
TRUE WHICH WE WON'T GET INTO,
WHERE GAIN OF FUNCTION, FOR A
REVIEWSHER, EVERYTHING'S GAIN OF
FUNCTION.
NOTHER QUESTIONS?
I HAVE 1 FOR JUSTIN.
IN YOUR WORK, LOOKING AT
MICRODPLEEL ACTIVATION,
OLFACTORY BULB IMPACS, THE
OMICRON VARIANTS IN HAMSTERS
DIFFERED IN THE WAY THEY
REPLICATE IN THE UPPER VERSUS
LOWER RESPIRATORY TRACT, HAVE
YOU SORT OF COMPARED DIFFERENT
VARIANTS IN THEIR IMPACT ON THE
UPPER RESPIRATORY TRACT.
>> WE HAVEN'T OURSELVES, I
THINK, [INDISCERNIBLE] LAB HAS
DONE A TON OF WORK I THINK ON
SPERKS LEECIALY LOWER
RESPIRATORY TRACT IN OMICRON
VARIANT WORK, SO I THINK I REF
FERENCED A LOT OF THEIR WORK IN
TERMS OF THE PERSISTENT
OLFACTORY PATHOLOGY, DEFINITELY
IS SOMETHING WE'RE INTERESTED
AND HAVEN'T DONE YET, ESPECIALLY
BECAUSE WE WOULD EXPECT BASED ON
AT LEAST SOME OF THE PRELIMINARY
DATA AND THE DATA THAT'S
AVAILABLE NOW WE MIGHT HAVE
HIGHER REPLICATION THAT THAN
THAT REAP YOJ WHICH MIGHT DRIVE
THIS PHENOMENON.
SO IT'S SOMETHING THAT WE'RE
DEFINITELY THINKING ABOUT AND
HOPING TO EXPAND INTO THAT AREA
FOR, FOR SURE.
AND JUST TO TIE IT TO THE
PREVIOUS DISCUSSION, HAVE YOU
LOOKED AT MALE-FEMALE IN HAM
TERS.
>> SO IF FEMALE HAMSTERS THEY
GET TERRITORIAL SO WE CAN ONLY
SINGLE HOUSE THEM AT A TIME.
WE HAVE TRIED ON WORKING WITH
FEMALE HAMSTERS AND WE HELD
THEM, DOUBLE CAGED BECAUSE WE
THOUGHT IT WAS A TRIAL AND WE
ENDED UP HAVING TO POLIT THEM
FOR LIKE A WEEK.
SO WHAT WE HAVE DONE IS MOSTLY
FOCUS ON THE RESULTS OF HAMSTERS
ON THAT PILOT VALIDATION STUDY
AND WE'VE BEEN ABLE TO SEE THIS
PERSISTENT PHENOMENON IN THE
OLFACTORY BULBS BUT WE DO AT
LEAST FROM OUR PRELIMINARY DATA
SEE IT.
>> I WILL ASK A NONSTUDY
SECTION QUESTION.
YOU KNOW, I STRUGGLE WITH THIS
TOO, HOW DO PEOPLE THINK ABOUT
TRYING TO MODEL A DISEASE THAT'S
ONLY OBSERVED IN A SMALL
FRACTION OF HUMANS THAT MAY HAVE
AN IDIO PATHIC COMPONENT BUT
HAPPENS IN A HUNDRED% OF THE
ANIMAL MODEL BUT IT'S IMPORTANT
TO FIND MODELS FOR THIS THING
BUT WHAT DOES IT MEAN WHEN 5% OF
HUMANS GET ANASMEI AND BUT A
HUNDRED PERCENT IN IN HAMSTERS
AND HOW DO WE THINK ABOUT THAT?
>> THAT'S SOMETHING WE STRUGGLE
WITH BECAUSE WE WANT OUR ANIMAL
MODEL TO BE AS REPRESENTATIVE AS
POSSIBLE BUT PEOPLE WILL SAY I
FOUND THIS INTERESTING ASPECT OF
WHERE A VIRUS WAS LOCATED IN AN
AUTOPSIES, WHY CAN'T YOU SHOW
THIS OR LIKE YOU SAID WHY DO ALL
YOUR HAN ANIMAL VS THIS WHEN ALL
NOT ALL PEOPLE DO I THINK PART
OF THAT STEMS FROM LIKE WHEN
WE'RE DEVELOPING THESE MODELS
WE'RE USUALLY GOING FOR PRETTY
ROBUST DISEASE BECAUSE WE WANT
IT TO BE RELIABLE AND WE WANT IT
TO BE EASILY MEASURED AND THE
LIKE ARDS DEVELOPMENT VERSION OF
COVID IS SOMEWHAT MORE
INTERESTING TO STUDY JUST
BIOLOGICALLY THAN THE COMMON
COLD VERSION OF COVID IS.
SO I THINK IF WE BACKED OFF ON
DOSES AND MUCH LESS IN THE WAY
OF SIM TOMS WE PROBABLY WOULD
HAVE TAKEN--THEY TYPE OF
VARIABLE RESPONSE WHICH IS
HORRIBLE FOR GENERATING FIGURES
AND SAYING SOMETHING IS
STATISTICALLY SIGNIFICANT BUT
MUCH MORE REPRESENTATIVE OF WHAT
HAPPENS IN PEOPLE.
>> I WAS JUST ANYTHING TO ADD TO
THAT, IN THE FIRST TALK,
STACEY'S TALK, ALL THE VARIABLES
THAT 1 COULD THINK ABOUT,
WHETHER IT'S MALE, FEMALE,
OBESITY, OTHER UNDERLYING
CONDITIONS HOW DOES 1 ACTUALLY
CHOOSE AN ANIMAL MODEL THAT
COMES NEAR FOCUSED ON MIMICKING
WHAT HAPPENS IN PEOPLE BECAUSE
ALL THE SINGLE VARIABLES, EVEN
THE COLLABORATIVE, OOGHTS A
SINGLE VARIABLE MICE OR HOWED IN
CONDITIONS MOST OF THE TIME WHEN
THEY DON'T GET EXPOSED TO OTHER
PATHOGENS SO THIS IS REALLY--I
COULDN'T THINK OF A WAY TO DO
IT, IT SEEMS OVERWHELMING IN YOU
TRY TO INCLUDE EVERYTHING.
>> I HAVE ANOTHER QUESTION ON
THE SEX DIFFERENCE
[INDISCERNIBLE] FOR YOU.
YEAH, SO THERE IS MOW BASE LINE
DIFFERENCE BETWEEN MALES OR
FEMALES BUT AFTER INFECTION OR
CHALLENGE, MIGHT BE SOMETHING
DIFFERENT.
SO THEN DO WE KNOW WHETHER THE
IMMUNITY, WHICH TARGET BECOME
DIFFERENT?
WE HAVE SO MANY DIFFERENT MOUSE,
TRAITS HAVE YOU ANY OF THEM
[INDISCERNIBLE]?
>> YEAH, I THINK IT DEPENDS ON
THE QUESTION YOU'RE ASKING AND
THE TIME POINTS THAT YOU LOOK AT
AS TO WHETHER THESE DIFFERENCES
AND YOU KNOW YOU COULD SAY, THE
SEX DIFFERENCE BUT I THINK IT'S
EVEN TRUE WITH AGE DIFFERENCE
CAN IMPACT BOTH THE INNATE AND
THE ASBESTOS DATA PROTECTIONIVE
IMMUNE RESPONSES.
AND THERE'S AMPLE DATA ON THE
LITERATURE ON THAT.
THANK YOU.
NANY OTHER QUESTIONS?
ONLINE?
NONE?
ALL RIGHT, I THINK WE WILL END
THERE.
THANK OUR PANELISTS FOR YOUR
TIME.
I THINK NOW IS LUNCH.
>> WE'LL GET STARTED IN THE NEXT
SESSION, DEFINING IMMUNE
RESPONSES IN ANIMAL MODELS OF
RESPIRATORY VIRUS INFECTION.
PARTICULARLY I'M DELIGHTED TO
INVITE RYAN LANGLOIS TO SPEAK,
AT THE UNIVERSITY OF IOWA I WAS
HIS FIRST MENTOR, I DON'T KNOW
IN THE LAST CENTURY TIME.
>> CHAIRED MY THESIS COMMITTEE
AS WELL.
THIS IS HIS FAULT IF IT GOES
WRONG.
>> THE TITLE IS DIRTY MICE,
BETTER REKA PIP LATE VACCINE
RESPONSES OBSERVED IN HUMANS.
>> THANK YOU VERY MUCH.
THANK YOU FOR THE INTRODUCTION.
WE'LL TALK ABOUT A MODEL THAT
MAKES ONE ATTEMPT TO NORMALIZE
ONE CRITIQUE OF SMALL ANIMAL
STUDIES, THAT THEY ARE LIVING IN
CLEAN HOUSING, DON'T EXPERIENCE
A NUMBER OF INFECTIONS THAT WE
ALL EXPERIENCE.
THIS IS JUST A MODEL.
I'M TRYING TO TARGET ONE OF THE
POTENTIAL CAVEATS THIS IS THE
MODEL.
HE WENT TO THE PET STORE, GET
MICE, FILLS WITH PATHOGENS,
VIRUSES, FUNGI, BACTERIA, A
WEALTH OF INFECTIONS THAT OTHER
ANIMALS EXPERIENCE FREE LIVING
IN THE WILD.
YOU CO-HOUSE PET STORE MICE.
I'LL TALK ABOUT A SLIGHT TWIST
ON THE SYSTEM.
CO-HOUSE THESE PET STORE MICE,
GENETICALLY DIVERSE, WITH
LABORATORY MICE, GENETICALLY
IDENTICAL, YOU CAN SORT OF
RETAIN MOUSE GENETICS, A LOT OF
MOUSE GENETIC TOOLS.
A COLLABORATIVE CROSS,
COMPOUNDING DIVERSITY WOULD BE
FUN BUT THIS IS GENETICALLY
IDENTICAL MICE.
WE CO-HOUSE THEM FOR 60 DAYS.
THIS GIVES ENOUGH TIME FOR THESE
MICE TO GET ACUTE AND CHRONIC
INFECTIONS, THEY WILL RESOLVE
THE ACUTE INFECTIONS BY THIS
TIME POINT.
THEY WILL HAVE RETURNED TO WHAT
THE NEW NORMAL IS, NEW BASELINE,
WITH A COMPLEX IMMUNE HISTORY
AND INFECTION EXPERIENCE
HISTORY.
THIS IS SOME DATA WHERE I'M
GOING TO TRY TO CONVINCE YOU WE
LIKE TO EMBRACE THE
HETEROGENEITY OF THE MODEL
SYSTEM, OTHERWISE YOU'LL GO
INSANE.
THIS IS OLDER DATA NOW.
WE'VE BEEN CONSTANTLY UPDATES, I
SHOULD MAKE SURE I GET THAT DATA
TABULATED SOMETIME.
THIS IS LOOKING AT CO-HOUSING,
WE MEASURE CDTA CELL ACTIVATION
IN THE BLOOD, IF THE MICE
WRITTEN EXPECTED, IF THERE WERE
CHANGES BASEALLY TO THE IMMUNE
SYSTEM.
AFTER 60 DAYS OF CO-HOUSING THIS
IS WHAT IT LOOKS LIKE, FROM EACH
STACK AT DIFFERENT DATE,
DIFFERENT EXPERIMENT, DIFFERENT
MOUSE FROM THE PET STORE.
WE HAD TO TAKE A BREAK DUE TO
ANOTHER VIRUS CIRCULATING AROUND
THIS TIME.
WE CAN SEE WE COME BACK, THE
PHENOTYPES ARE STILL CONSISTENT.
THIS MULTI-DIMENSIONAL SCALING
ANALYSIS ON THE RIGHT I'M
SHOWING YOU DIFFERENT
COMBINATIONS OF PATHOGENS THEY
HAVE BY SEROLOGY.
EACH CLUSTER IS A UNIQUE
PATHOGEN COMBINATION.
AND OVERLAID ON TOP OF THAT IS
THE PERCENT OF HEAT MAP OF THE
CD8 T-CELL ACTIVATION.
THERE AREN'T ANY REALLY MAJOR
HOT SPOTS, RIGHT?
A PARTICULAR PATHOGEN
COMBINATION IS THE MAJOR DRIVER
OF T CELL T-CELL ACTIVATION.
THIS IS OVER 700 MICE WHICH
YIELDED 150 UNIQUE PATHOGEN
COMBINATIONS.
WE NOW HAVE OVER 1600 MICE, MUCH
LARGER SET OF PATHOGENS, DOESN'T
MATTER WHAT THE INDIVIDUAL
PATHOGENS ARE, HAVING A COMPLEX
PATHOGEN EXPOSURE HISTORY
BASEALLY CHANGES YOUR IMMUNE
SYSTEM.
WE DID THIS FACTOR ANALYSIS OF
MIXED DATA.
THIS IS BASICALLY TAKING EACH OF
THESE POTENTIAL PATHOGENS
INDIVIDUALLY AND ASKING HOW THEY
IMPACT THE PHENOTYPE WE'RE
INTERESTED IN, WHICH WAS THE
CD44 EXPRESSION.
IF IT HAD ZERO IMPACT, THIS
WOULD LIVE ON THE ORDINAL HERE.
IF IT HAD A LARGE IMPACT WOULD
EXIST WAY OUT HERE.
WHAT YOU CAN APPRECIATE PATHOGEN
COMBINATION IS CLOSE TO THE
BASELINE HERE, SO THE PATHOGEN
COMBINATIONS DON'T REALLY DRIVE
CHANGES, IT'S JUST HAVING SOME
PATHOGEN EXPOSURE THAT MATTERS.
BECAUSE SEX AS A BIOLOGICAL
VARIABLE HAS HEAVILY DISCUSSED
SO FAR I WANT TO THROW THIS
PIECE OF DATA IN HERE TOO.
SO, ONE CAVEAT TO THIS MODEL
SYSTEM BECAUSE WE'RE CO-HOUSING
ANIMALS WE HAVE TO ONLY CO-HOUSE
FEMALES.
OTHERWISE THE MALES WILL FIGHT
AND FOR OBVIOUS REASONS WE CAN'T
TAKE A MALE PET STORE WITH
FEMALE MICE.
SO TO GET AROUND THAT POTENTIAL
CAVEAT WHAT WE CAN DO IS TAKE
FOMITES FROM PET STORE MICE,
BEDDING IS FILLED WITH PET STORE
MOUSE FECES, URINE, FUR, OTHER
THINGS, TRANSFER THAT OVER.
THIS IS THE FOMITE TRANSMISSION.
THERE ARE SOME DIFFERENCES,
DIVERSITY ISN'T QUITE AS LARGE.
BUT WHAT I'M NOT SHOWING HERE IS
IF WE LOOK AT T-CELL ACTIVATION
IT'S STILL THE CAME.
IN TERMS OF ACTIVATING T-CELL
RESPONSE AND BASAL CHANGES THERE
WAS NO DIFFERENCE IN FOMITES.
FOR SIMPLICITIES WE'RE SWITCHING
TO FOMITES FOR A LARGE NUMBER OF
OUR EXPERIMENTS.
IT HELPS ADDRESS SEX AS A
BIOLOGICAL VARIABLE WHICH STUDY
SECTION CARES A LOT ABOUT.
OKAY.
THE OTHER THING I WANTED TO
POINT OUT FROM THE MODEL SYSTEM
AS A VIROLOGIST, I CARE ABOUT
THE VIRUSES HERE, THERE'S A LOT
OF NOVEL AND UNDER OR
UNAPPRECIATED VIRUSES FROM MICE.
THERE'S A LOT OF OBVIOUSLY
PATHOGENS THAT ARE NATURAL IN
MICE THAT WE DON'T SEE IN OUR
TRADITIONAL SPF PANELS DEVELOPED
BY JACKSON AND CHARLES RIVER.
I'M SHOWING YOU SOME VIRUSES
THAT YOU DON'T NORMALLY SORT OF
SEE.
THE NOROVIRUS, NHV, COMMON,
CLASSIC.
IN THIS ANALYSIS, THIS IS A
DIFFERENT WAY TO LOOK AT DATA,
SO IF THE PET STORE MOUSE HAD
THAT PATHOGEN, THIS IS ALSO FROM
ABOUT A THOUSAND MICE, IF THE
PET STORE MOUSE HAD THE PATHOGEN
EVERY TIME IT WOULD BE 100%.
THE HEAT MAP IS HOW FREQUENT DID
IT TRANSMIT WHEN PRESENT.
WHAT YOU CAN SEE ASTRO VIRUS 2
IS OUR WINNER HERE, IN EVERY
SINGLE PET STORE MOUSE,
TRANSMISSION EFFICIENCY IS
ALMOST 100%.
SOME OTHER VIRUSES HAVE VARYING
DEGREES.
BUT SOMETHING I WANT TO POINT
OUT THAT'S IMPORTANT, FOR
EXAMPLE, IN THIS ORIGINAL
ANALYSIS IN THIS PAPER, WE AT
THE FAMILY LEVEL MISSED SOME
VIRUSES, LOOKING DEEPER WE FIND
A ENVELOPE ALPHA CORONAVIRUS
QUITE FREQUENTLY IN THE PET
STORE MOUSE AND NOVEL
CORONAVIRUS DISTANTLY RELATED TO
THIS COVID REQUIRES HERE.
EVEN MORE IMPORTANTLY EVERYTHING
HERE WITH A STAR IS NOT SCREENED
BY ANY COMMERCIAL AVAILABLE KIT
THAT IS USED TO LOOK AT
SPF-NESS, FOR LACK OF BETTER
TERM.
A LOT OF THESE THINGS ARE
PRESENT IN OUR DIRTY MOUSE
MODEL.
I CAN TELL YOU, I WON'T NAME THE
VENDOR HERE, BUT VERY FREQUENTLY
IN SPF MICE WE PURCHASED WE FIND
ASTRO VIRUS 1 QUITE FREQUENTLY.
THERE'S A LOT OF THINGS STILL
GOING ON THAT COULD BE MISSED
BECAUSE THEY ARE NOT BEING
LOOKED FOR.
SO OUR LAB HAS DEVELOPED
SCREENING SYSTEMS, PCR PRIMERS
AND ELISAS FOR ALL OF THESE.
EXAMPLE OF HETEROGENEITY, WE GET
THE PET STORE MICE, WE DON'T
KNOW WHAT STAGE OF THE INFECTION
THEY ARE IN, ARE THEY IN ACUTE
INFECTION, CHRONIC STAGE.
THIS IS THE MARINE KO BU VIRUS.
WE GET PET STORE MICE, RESTED
THEM, BLED THEM EVERY WEEK.
AT WEEK ZERO THIS ANIMAL HERE
THAT WAS POSITIVE, SUSPECT
RATHER, ALL THESE WERE NEGATIVE.
WE LOOK AT 14 DAYS LATER SOME
ANIMALS DID SEROCONVERT,
SUGGESTING THEY WERE IN THE
ACUTE PHASE OF INFECTION
PRE-CONVALESCENS.
MANY MORE CONVERTED.
WE'RE PROVIDING OUR SPF MICE
WITH BOTH ACUTE AND CHRONIC
INFECTION.
OUR GOAL IS TO USE THIS MODEL TO
BETTER RECAPITULATE HUMAN IMMUNE
RESPONSES FROM HUMANS WHO SEE
LOTS OF INFECTIONS OVER THE
COURSE OF THEIR LIFE.
THIS IS WHAT THIS LOOKS LIKE.
I SHOWED YOU THIS IN THE FIRST
SLIDE.
WE CAN GIVE THEM A VACCINE.
WHEN WE'VE DONE THIS WE GAVE AN
INFLUENZA VACCINE, AND THIS IS A
SEASONAL FLU VACCINE THAT WE
BOUGHT FROM A COMPANY THAT IS
USED IN HUMANS.
WE GAVE IT TO MICE.
WE TOOK OUT THE PBMCs AND
MEASURED VACCINE RESPONSE OF
GENES, DID A GENE SET ENRICHMENT
ANALYSIS TO COMPARE RESPONSES IN
MICE, RESPONSES OBSERVED IN
CLINICAL TRIALS FROM HUMANS
WHERE PBMCs WERE ALSO TAKEN
AND DEEP SEQUENCED.
I HOPE YOU CAN APPRECIATE IF
YOU'RE FAMILIAR WITH THE GRAPHS
THIS CURVE IS SHOWING YOU A HIGH
SET OF ENRICHMENTS TO VACCINE
RESPONSE IN HUMANS IN THE DIRTY
MICE, SO THIS IS REALLY GREAT,
RIGHT?
THIS IS PRETTY PHENOMENAL
ENRICHMENT IN FACT.
AND THIS IS SORT OF
ANTI-ENRICHMENT, VERY LITTLE TO
NO ENRICHMENT IN THE SPF MICE.
SPF MICE ARE NOT RECAPITULATING
THE VACCINE RESPONSIVE GENES IN
HUMANS BUT THE DIRTY MICE ARE.
RIGHT AT THE BASELINE YOU'RE
HAVING APPLES TO ORANGES
COMPARISONS IF YOU'RE MODELING
VACCINE RESPONSES AND SPF MICE,
BECAUSE YOU'RE NOT MODELING
TRANSCRIPTIONAL CHANGES THAT
HAPPEN IN A HUMAN.
SO THAT IMPACTS THE END RESULTS
WHICH YOU CARE ABOUT, CORRELATE
OF PROTECTION.
IN THIS CASE I'LL SHOW ANTIBODY
RESPONSES.
HERE WE'RE LOOKING AT THE
ANTIBODY RESPONSE AGAINST THE
VACCINE.
AND SO WHAT YOU CAN APPRECIATE
IS THAT THERE ARE SOME
DIFFERENCES THAT YOU OBSERVED
HERE, RIGHT?
SORRY.
WHERE IF YOU LOOK AT SPF MICE,
YOU HAVE -- SORRY, I'LL COME UP
HERE, LOOKING AT SPF MICE HAVE A
STRONGER ANTIBODY RESPONSE.
AND THAT CORRELATES TO BETTER
PROTECTION, AND THE PROTECTION
IS WORSE IN THE DIRTY MICE.
SO IT IS EASIER TO PROTECT AN
SPF MICE WITH INFLUENZA VACCINE
THAN TO PROTECT A DIRTY MOUSE.
SO YOU GET THIS SORT OF
UNNATURAL LEVEL OF IMMUNITY WHEN
YOU'RE VACCINATING AN SPF ANIMAL
COMPARED TO DIRTY ANIMAL.
AND SO WITH MY ESTEEM COLLEAGUE
OVER HERE, WE WANTED TO
RECAPITULATE THIS WITH ANOTHER
VACCINE MODALITY SO THE PREVIOUS
VACCINE WAS THE KILLED INFLUENZA
VACCINE, SPLIT KILLED VACCINE,
THIS IS THE SARS-COV-2 VACCINE.
SO WE VACCINATED ANIMALS, MEHUL
AND JENS ANALYZED SERUM
RESPONSE.
YOU GET A MUCH STRONGER RESPONSE
EVEN INITIALLY TO THE PRIME, A
WHOPPING RESPONSE COMPARED TO
THE PRIME.
AND YOU EVEN GET A BETTER BOOST
WHEN YOU BOOST.
HERE, THE BOOST IS REALLY
REQUIRED IN DIRTY MICE.
I THINK THIS REALLY NICELY
DEMONSTRATES WHAT WE OBSERVED IN
HUMANS WHERE HUMANS DID NEED
THAT BOOST FOR mRNA VACCINES
TO REACH PEAK OPTIMAL LEVEL.
YOU GET QUITE A GOOD RESPONSE TO
THE PRIMARY IN SPF MICE.
SO DIRTY MICE NEED THIS BOOST
JUST LIKE THE HUMANS DO.
SO I THINK THIS IS REALLY A
HELPFUL MODEL AND I HOPE WOULD
IMPROVE mRNA VACCINES, NOW YOU
CAN MAKE MODIFICATIONS TO TURN
THIS TO LOOK MORE LIKE THIS, TO
IMPROVE THE IMMUNOGENICITY, THE
STRENGTH OF THE VACCINE
RESPONSE.
THAT'S ONE HOPE OF HAVING A
MODEL SYSTEM THAT BETTER
RECAPITULATES PEOPLE.
ANOTHER COMPONENT BETTER
RECAPITULATED IN DIRTY MICE IS
DURABILITY.
OFTEN ESPECIALLY IN THE FLU
WORLD YOU VACCINATE MICE WITH
FLU MODALITIES ALMOST SEE NO
DIMINUTION OF VACCINE RESPONSE.
IT'S STABLE, EXTREMELY ROBUST,
HIGHLY DURABLE.
THIS IS NOT TRUE IN HUMANS,
RIGHT?
FOR INFLUENZA VACCINE, FOR A LOT
OF VACCINES, IT'S TRUE FOR THE
SMALLPOX VACCINE BUT MANY
VACCINES SUFFER FROM DURABILITY
ISSUES.
SO THERE'S NO OBSERVABLE OR
OBVIOUS DURABILITY ISSUE IN THE
SPF MICE BUT YOU DO SEE SOME
DURABILITY ISSUES IN THE mRNA
VACCINE IN THE DIRTY MICE.
THIS IS ANOTHER AVENUE WHERE
THIS IS BETTER RECAPITULATING
THE RESPONSE IN HUMANS, WE ARGUE
IT'S UNNATURAL AND NOT
REPRESENTATIVE OF WHAT'S SEEN IN
HUMANS.
DOES THIS MATTER?
OF COURSE IT DOES.
SO MEHUL AND JENS ASKED ARE
ANTIBODIES ABLE TO NEUTRALIZE TO
THE SAME DEGREE.
THEY ARE NOT.
THIS WORKS BETTER IN SPF MICE
THAN DIRTY MICE.
AND IN AN EXPERIMENT THAT I WAS
GLAD MEHUL DECIDED TO DO, WHAT
DOES THE BREATH LOOK LIKE, WHEN
LOTS OF VARIANTS ARE ARISING,
WHAT'S THE BREADTH OF THESE
RESPONSES, IT'S WORSE IN DIRTY
MICE, REPERTOIRE IS NARROWER,
WORSE ON EVERY FRONT.
THIS RECAPITULATES IN HUMANS,
THE BREADTH OF THE RESPONSE IS
QUITE WEAK.
THERE ARE A LOT OF WAYS TO
CREATE THESE DIRTY MOUSE MODELS,
AND I THINK ALL OF THEM HAVE
VALUE.
I THINK ALL OF THEM CAN TEACH US
SOMETHING, AND WE CAN LEARN
SOMETHING INTERESTING FROM THEM.
I WANT TO GIVE ONE QUICK EXAMPLE
OF A RECENT EXPERIMENT THAT WE
DID WITH COLLABORATORS FROM
NORWAY.
SO, THIS IS OUR PET STORE
CO-HOUSING.
HIS MODEL SYSTEM, HE STERILIZES
MICE, TAKES SPF MICE, HOUSED IN
A BARN-LIKE ENVIRONMENT EXPOSED
TO COWS, PIGS, THERE ARE OTHER
FREE-LIVING MICE THAT CAN COME
IN, THEY DON'T EXCLUDE THOSE
NECESSARILY.
AND SO THEY ARE EXPOSED TO LOTS
OF THINGS THEY WOULD SEE IN
NATURE.
THEY TAKE ONE SMALL STEP FURTHER
AND WILL DIRECTLY CO-HOUSE THEM
WITH WILD MICE THEY CAPTURE IN
NORWAY.
NOW THERE'S WILD MICE WITH THE
SPF MICE IN THE BARN-LIKE
ENVIRONMENT.
OKAY.
WE JUST LOOKED AT THE POTENTIAL
PATHOGEN EXPOSURES.
I'M SHOWING SEROLOGY.
WHAT I HOPE YOU CAN APPRECIATE
IS THAT ACTUALLY PET STORE MICE
WITH SIGNIFICANTLY DIRTIER.
NOT NECESSARILY SURPRISING.
SO PET STORE MICE GET AD LBITUM
ACCESS TO FOOD AND WATER.
THEY DON'T HAVE TO FIND FOOD AND
WATER.
IF THEY GET SICK THEY DON'T HAVE
TO BE ABLE TO RUN FAST FROM A
HAWK.
THEY ARE QUITE PROTECTED, SO
IT'S EASIER TO KEEP THEM ALIVE.
WILD MICE, IF THEY GET SICK
THAT'S A SIGNIFICANT DETRIMENT
TO THEIR THEIR SURVIVAL.
WE FOUND NO NATURAL MASS
PATHOGENS.
WE DID DEEP SEQUENCING ANALYSES
TO SEE WHAT VIRUSES WERE
PRESENT, ARE THERE NOVEL VIRUSES
PRESENT.
I WAS A BIT SURPRISED, I'LL JUMP
HERE, TO FIND VERY FEW VIRUSES
THAT WERE PRESENT, AS WE LOOKED
AT SIX ANIMALS, A TON OF TISSUE,
ONLY FOUND TWO CORONAVIRUSES
PRESENT.
TYLER'S MIRIAM AND THIS VIRUS.
WE COMPARED THE SPF PANEL OF ALL
OF OUR ANIMAL MODELS, AGAIN
ABOUT A THOUSAND, TO ABOUT 40
FROM PREBIN AND LOOKED AT
PATHOGEN COMBINATIONS.
YOU CAN SEE THERE ARE SOME
DIFFERENCES.
HERE ARE PET STORE CO-HOUSING,
SOME EXIST IN THIS CLUSTER, IF
YOU WOULD OVERLAY SPF MICE A LOT
EXIST OVER HERE, SO DIRTINESS
KINDS OF CORRELATES WITH WHERE
THE CO-HOUSING OR HOW THE
CO-HOUSING OCCURRED.
DOES THAT MATTER?
DOES THE DEGREE OF DIRTINESS IN
THIS SYSTEM MATTER BECAUSE HIS
ARE SUBSTANTIALLY LESS DIRTY.
MAYBE, AND MAYBE JUST A LITTLE
BIT.
I WON'T OVERINTERPRET DATA.
THIS IS THE ANTIBODY RESPONSE WE
SENT HIM, EXACT SAME VACCINE, HE
VACCINATED HIS ANIMALS, WE
VACCINATED A SET OF ANIMALS,
DIRECTLY COMPARED THE RESPONSE
TO THE SAME VACCINE.
THIS IS, AGAIN, THE SPLIT KILLED
FLU VACCINE.
MOSTLY THE SAME BUT THERE ARE
SOME SMALL DIFFERENCENS.
Ns ARE REASONABLY LARGE, SOLID
DATA, SMALL CHANGES, VERY SMALL
IF YOU SQUINT, MOSTLY
OVERLAPPING.
BY AREA UNDER THE CURVE YOU SEE
A FEW PLACE WAS STATISTICAL
DIFFERENCES, FALLOUT BETWEEN OUR
MICE AND THEIR MICE.
BUT, AGAIN, THESE ARE VERY SMALL
CHANGES, JUST BEING DIRTY MAKES
A MAJOR DIFFERENCE AND IS THE
MAJOR SORT OF IMPACT HERE.
SO QUICKLY IN CONCLUSION, I
THINK THERE'S LOTS OF DIFFERENT
MICROBES, A LOT OF DIVERSITY AND
HETEROGENEITY IN THE INFECTIONS
WE AS HUMAN EXPERIENCE
THROUGHOUT OUR LIFETIME.
WE HAVE CONVINCING DATA THAT
SUGGESTS THE DIRTY MICE DO
BETTER RECAPITULATE THE VACCINE
RESPONSES SEEN TO MULTIPLE
DIFFERENT VACCINE MODALITIES.
AND DIRTINESS DOES IMPACT OR
DOES VARY BASED ON MODEL YOU
USE, THERE CAN BE SMALL CHANGES
IN RESULTS YOU WOULD SEE BASED
ON MODEL SYSTEMS, YOU KNOW,
WHICH SHOULDN'T BE TERRIBLY
SURPRISING.
SO WITH THAT I WILL STOP.
I'LL THANK ALL THE FOLKS HERE
WHO DO THIS WORK.
SO FRANCES SHEPHERD, THE WORK IN
PREBIN BY AUTUMN SANDERS.
I'LL STOP AND TAKE QUESTIONS.
>> BEAUTIFUL TALK.
THE DIRTY MOUSE MODEL, A CLEAR
ARGUMENT TO MAKE FOR USING
THESE.
THEY DON'T GET TERRIBLY EXCITED
WHEN I BRING IT TO OUR BRANCH.
HOW DO YOU SUGGEST WE DEAL WITH
THAT?
ARE THERE THINGS WE CAN DO?
CAN WE LIKE INFECT MICE WITH
LIKE, I DON'T KNOW, INFLUENZA
AND SARS, ANY OF THE VIRUSES YOU
FIND HERE TO DO THAT INSTEAD?
>> YEAH, THAT'S A GREAT POINT.
I FAIL TO BRING THIS UP AND NEED
TO REMEMBER TO INTRODUCE THIS AT
THE BEGINNING.
WE DO ALL OF OUR EXPERIMENTS IN
A MODIFIED BSL3 FACILITY.
MIGHT SEEM LIKE OVERKILL BUT I'M
HAPPY BECAUSE WE DO A LOT OF
DEEP SEQUENCING OF OUR SPF MICE
HOUSED IN OUR FACILITY AND NEVER
FIND PATHOGENS WITH THE
EXCEPTION OF ASTRO-1, SOME
PATHOGENS ARE EXTREMELY
ENVIRONMENTALLY STABLE.
I THINK THE OVERKILL IS
WARRANTED.
THAT BEING SAID, THERE ARE OTHER
WAYS TO ACHIEVE THE SAME GOAL.
SO SKIP AND TIFFANY HAVE THE
SEQUENTIAL INFECTION EXPERIMENT,
DONE SEVERAL TIMES AT SEVERAL
ITERATIONS, MIKE DIAMOND IS
WORKING ON A SIMILAR SYSTEM
USING TRACTABLE NATURAL MOUSE
PATHOGENS FROM DIFFERENT
KINGDOMS WHERE YOU CAN DELIVER
THEM WITHOUT BIOSAFETY CONCERNS.
DAVE AND SKIP COMPARED
TRANSCRIPTIONAL RESPONSES AND
THEY WERE QUITE OVERLAPPING,
AGAIN FITTING WITH THE GENERAL
THESIS THAT SOME IMMUNE
EXPERIENCE IS BETTER THAN NONE
SO I THINK DIRECT INFECTION
SYSTEM COULD WORK.
YES?
>> SO, BEAUTIFUL DATA, AS
ALWAYS.
ONE THING I STARTED TO THINK
ABOUT THIS TIME IS WHETHER THERE
ARE MICROBIOME CHANGES, AND IS
THE MICROBIOME WHICH MAYBE IS
MODIFYING THOSE DE NOVO ANTIBODY
IMMUNE RESPONSES, YEAH,
MICROBIOME.
>> THERE ARE MICROBIOME CHANGES.
A BEAUTIFUL QUESTION.
WE'RE WORKING ON A PROJECT NOW
NOW TRANSFERRING FECES, WE
FILTER, WE DON'T FILTER, TO DO
THIS.
WE'RE WORKING TO DEWORM THEM.
I WILL TELL YOU ACTUALLY 100% OF
THESE ANIMALS HAVE WORMS.
THEY ARE GROSS.
SO, YES, WE DON'T KNOW.
THERE ARE MICROBIOME CHANGES.
THOSE CHANGES ARE ALSO IMPACTED
BY VIRUS INFECTION, I DON'T KNOW
THAT I LOVE THIS WORD DYSBIOSIS
BUT YOU SEE GUT MICROBIOME
CHANGES AFTER FLU INFECTION,
QUANTITATIVELY AND QUALITATIVEY
DIFFERENT.
A PAPER WAS RECENTLY REJECTED,
ONE OF THE REASONS BECAUSE WE
USED PRAs, TO BRING THAT BACK,
IF YOU WERE HERE COME TALK TO
ME.
>> I REJECTED THAT BECAUSE --
NO, I WANTED TO ACTUALLY ASK
ABOUT THAT BECAUSE I HAVEN'T
SEEN THE DATA.
MY QUESTION WAS IF YOU SEE THE
SAME CHANGES TRANSCRIPTIONALLY
AS WELL AS BREATH, LONGEVITY OF
ANTIBODY RESPONSES AGAINST
NATURAL INFECTION THAT YOU SEE
AGAINST VACCINATION.
>> BEAUTIFUL QUESTION.
WE HAVEN'T DONE AS MUCH AS WE
SHOULD.
WE DID FOCUS ON VACCINES BUT DID
SOME OF THOSE, AGAIN, PR 8, YOU
DID SEE THE CHANGES.
THEY WEREN'T AS PRONOUNCED AS
WITH THE VACCINE.
VACCINE RESPONSES WITH QUITE
DRAMATIC, NOT AS DRAMATIC WITH
ACUTE INFECTION.
>> RYAN, FANTASTIC.
HOW MUCH OF THE CHANGES THAT
YOU'RE SEEING ARE ACTIVATION
BASED, BASED ON CO-INFECTION, OR
TRAINED IMMUNITY?
>> GREAT QUESTION.
I WILL SAY WE'VE DONE SOME
EXPERIMENTS WHERE WE GO OUT WAY
FURTHER THAN 60 DAYS OF
CO-HOUSING AND SEE VERY, VERY,
VERY SIMILAR RESULTS.
I WOULD SORT OF SUGGEST THAT
TRAINED IMMUNITY MIGHT LIKELY IS
NOT INVOLVED.
THERE AREN'T A LOT OF CHRONIC
INFECTIONS HERE BUT THERE ARE
SOME.
SO IT GETS A LITTLE BIT
DIFFICULT TO DISENTANGLE.
WITH SOME EXPERIMENTS THAT SABRA
WAS SUGGESTING MIGHT GET AT THAT
EXACT QUESTION.
IT'S TRICKY, YEAH.
>> I HAD A QUESTION.
WHEN YOU LOOK AT THE MICE THAT
YOU GET FROM THE DIRTY VENDOR AS
OPPOSED TO CLEAN VENDOR DO YOU
SEE SIMILAR DIFFERENCES, HAVE
YOU EVER DONE THIS?
>> YEAH, WE'VE DONE A COUPLE
EXPERIMENTS WHERE WE GOT MICE
FROM THE PET STORE, WHERE THE
MICE HAD BEEN SITTING IN THE PET
STORE FOR A LONG TIME.
THEY CLEARED A LOT OF THEIR
ACUTE INFECTION.
AND THOSE WERE SUBSTANTIALLY
LESS DIRTY.
IN THOSE SITUATIONS WE DID NOT
SEE MAJOR IMPACT WHICH COULD
SUGGEST THAT IT'S NOT
MICROBIOME, ACTUALLY SOME ACUTE
INFECTIONS ARE NECESSARY BUT OUR
N IS WAY TOO LOW.
>> WHEN I SAID DIRTY VENDOR I
DIDN'T MEAN PET STORE.
>> OH, I SEE.
WE'VE RECENTLY STARTED SCREENING
OUR MICE FOR ASTRO-1.
YES, YOU'RE ASKING ABOUT THE
VENDOR I SHOULDN'T NAME, YEAH, I
DON'T KNOW.
>> OKAY.
>> PEOPLE SHOULD PCR THEIR MICE,
I'M GUESSING A LOT OF YOU HAVE
ASTRO.
>> ONE QUESTION ABOUT THE WORMS
IN PARTICULAR, WHEN YOU CO-HOUSE
DO YOU SEE THE WORMS CROSSING
BETWEEN THEM, AND I GUESS
CURIOSITY WOULD THAT BE MORE
PERSISTENT INFECTION IF THEY DO
CROSS OVER A AND DO YOU SEE
CYTOKINES THAT WOULD MORE
EOSINOPHILIA IN THE BLOOD.
>> IT'S NOT LIKE AT THE 60 OR
PLUS TIME POINT.
WE HAVE DRAMATIC Th2-LIKE
RESPONSE, SO I DON'T THINK
THERE'S A MAJOR IMPACT.
ARE THE WORMS THERE?
YES.
AND YOU CAN SEE THEM, YOU KNOW,
WITHOUT A MICROSCOPE WHEN YOU
CUT THE GI TRACT OPEN.
THEY ARE GROSS.
>> HAS ANYONE USED THIS DIRTY
MOUSE MODEL IN THE CONCEPT OF
ALLERGY AND ASTHMA TO TEST
HYGIENE HYPOTHESIS?
>> YES, IT RECAPITULATES WHAT
YOU MIGHT EXPECT.
A PAPER FROM STEVE JAMISON IN
NATURE IMMUNOLOGY USING HOUSE
DUST MITES, AND UNPUBLISHED DATA
FROM MINNESOTA, THEY GO IN THE
OPPOSITE DIRECTION LOOKING AT
AUTOIMMUNE DISEASE, IT ALSO FITS
WITH WHAT YOU MIGHT EXPECT FROM
HYGIENE HYPOTHESIS, A FUN DIRECT
TEST OF THAT HYPOTHESIS.
YEAH, GREAT QUESTION.
ALL RIGHT.
THANK YOU.
>> NEXT SPEAKER IS TALKING ABOUT
RESPONSES TO RESPIRATORY
INFECTION WITH FOCUS ON
INFLUENZA.
>> THANK YOU, I CAN PARTICIPATE
IN THIS AMAZING WORKSHOP.
YES, I KEPT THE TITLE LIKE IT
WAS SUGGESTED, AND I'LL DIVE
RIGHT IN.
YEAH, SO THE INFLUENZA VIRUS
CHALLENGE MODEL WE DISCOVERED
THIS ALREADY A LITTLE BIT TODAY
BUT SO IT'S TYPICALLY INTRANASAL
INFECTION THAT WE USE AT MOUNT
SINAI, WE DON'T DO AEROSOL FOR
THE FLU.
THIS MEANS DEEP LUNG INJECTION,
USING 50 MICRO LITERS OF VOLUME,
IT GOES STRAIGHT INTO THE LUNG.
THE MOUSE MODEL IS NOT A GOOD
TRANSMISSION MODEL ESPECIALLY
ADULT MICE SO WE CANNOT USE IT
FOR THAT.
SOME LIMITATIONS OF THE MOUSE
MODEL ALSO FOR FLU WE'RE STUCK
TO SOME MORE DISRUPTED STRAINS
LIKE PURATE AND NEED MORE
VIRUSES MORE RECENT FOR
VACCINATION STUDIES FOR EXAMPLE
IF YOU WANT SOME ANTIBODY
RESPONSES AND PROTECTION
STUDIES.
THIS IS ESPECIALLY TRUE FOR H3N2
AND INFLUENZA B VIRUSES.
AND ALSO FOR THE MOUSE INFLUENZA
MODEL, AS OTHER MODELS, WE CAN
STUDY IMMUNE ANIMALS VERSUS
PREEXISTING IMMUNITY,
STERILIZING IMMUNITY THAT
PREVENTS VIRUS IMPLICATION A
LOT, THERE'S NOT A LOT OF CHANCE
FOR VIRUS TO REPLICATE IN THE
LUNG.
INFECTION PERMISSIVE IMMUNITY
WHICH BASICALLY IS A
BREAKTHROUGH INFECTION.
INFECTION PERMISSIVE IMMUNITY IS
SOMETHING THAT'S THE LAB HAS
BEEN FOCUSING ON A LOT BECAUSE
IT CAN PROVIDE PARTIAL
PROTECTION, SOMETIMES BROADLY
PROTECTIVE, VERY BROADLY
REACTIVE PROTECTION.
BUT ALSO CAN BE ASSOCIATED WITH
ENHANCED DISEASE BECAUSE YOU MAY
HAVE IMMUNE RESPONSES
CROSS-REACTIVE THAT NOT PROTECT
HOST RESPONSE TO INFECTION.
YOU MAY HAVE MODULATION OF
DISEASE, WHEN THERE'S INFECTION
PERMISSIVE IMMUNITY.
INFECTIONIVE PERMISSIVE IMMUNITY
CAN BE ADAPTIVE, ALSO WE HAVE TO
THINK OF INNATE IMMUNITY WHICH
BY DEFAULT IS ALSO INFECTION
PERMISSIVE.
SO, THIS IS INFECTION PERMISSIVE
IMMUNITY, SOMETHING WE FOCUS ON
FOR A LONG TIME IN THE LAB.
I WON'T BRING VACCINATION STUDY
BUT WE USE VACCINES AS A KIND OF
WAY TO INDUCE PREEXISTING
IMMUNITY THAT IS INFECTION
PERMISSIVE.
BECAUSE I DON'T WANT TO SAY THAT
FLU VACCINES MAY NOT WORK, I USE
VINTAGE VACCINE, 2005 TO 2006,
BASICALLY VACCINE 20 YEARS IN
THE FRIDGE, IN OUR FRIDGE.
DEFINITELY EXPIRES, OBVIOUSLY,
PRE-PANDEMIC, 2009, ANOTHER
REASON WE USE THIS VACCINE.
TYPICALLY FLU VACCINES AIM AT
INDUCTION OF NEUTRALIZING
ANTIBODIES TARGETING H-A.
A SINGLE VACCINATION OF 3 MICRO
GAPS OF H-A WILL RESULT IN LOW
HI TITERS GIVING INFECTION
PERMISSIVE IMMUNITY.
STUDY I'M GOING TO BRING, THE
FIRST PART, WE VACCINATE MICE
THREE TIMES, OR ONE TIME, OR NOT
AT ALL WHICH IS PBS I HAVE A
INTRAMUSCULAR ROUTE, FIRST IS
FEMALE MICE.
THREE WEEKS AFTER WE CHALLENGE
FIRST TIME WITH H1N1, A NEW
CALEDONIA FROM 99, NC 99, INFECT
WITH H1N1 OR GIVE MOCK
INFECTION.
THIS NC99 IS HUMAN ISOLATE, THIS
WORKS WELL IN MICE.
WE WAIT FOR FOUR WEEKS, WE
RECHALLENGE THESE MICE FOUR
WEEKS LATER WITH X31, ACTUALLY A
PURATE-BASED BACKBONE VIRUS WITH
THE HA AND NA FROM THE STRAIN
FROM 68.
THIS IS ANOTHER PROBLEM FOR THE
MOUSE MODEL.
WE NEED GOOD H3N2 VIRUSES, STILL
ON A BACKBONE FROM A LONG TIME
AGO.
I'LL BRING THE STORY ABOUT HOST
RESPONSE, NOT THE VACCINE STUDY
BUT HOST RESPONSE TO INFECTION.
I WILL CHOP IN THREE PARTS.
FIRST WE HAVE VACCINE RESPONSE
WHICH WE WILL CORRELATE WITH
PROTECTION AND THE OUTCOME OF
THE HOST RESPONSE AFTER THE
FIRST INFECTION AND THEN THE
HOST RESPONSE AFTER SECOND
INFECTION.
VACCINE RESPONSE AS I MENTIONED
IS SINGLE TIV VACCINATION GIVES
HARDLY ANY HA ANTIBODIES, YOU
HAVE TO VACCINATE THREE TIMES TO
DETECT ELISA VIRUS.
A SINGLE VACCINATION GIVES YOU
HI TITER DETECTABLE.
WE DIDN'T CHECK HA ELISA TITERS
AGAINST H3, IT'S ALSO VERY LOW,
YOU START TO FIND IN
TRIPLE-VACCINATED ANIMAL.
SPLIT VACCINE, IT HAS NP, IF YOU
VACCINATE THREE TIMES YOU GET
SOME NP ANTIBODIES.
I'M MENTIONING BECAUSE NP
ANTIBODIES MAY PROVIDE SOME
PROTECTION DURING INFECTION.
SO THEN WE CHECK, WE'RE GOING TO
FOCUS ON THE HOST RESPONSE AND
PROTECTION DURING THE FIRST
INFECTION WITH H1N1.
WHAT WE CAN SEE IS ANIMALS WITH
THIS DOSE OF VIRUS THAT ARE NOT
VACCINATED, SO ANIMALS NOT
VACCINATED, WILL THEY START TO
LOSE 20% OF BODY WEIGHT AND
RECOVER.
IN THE MOUSE MODEL YOU LOSE
WEIGHT UNTIL DAY 7, 8, MICE WILL
SUCCUMB OR START TO RECOVER.
ANIMALS VACCINATED WITH TIV ONCE
OR THREE TIMES, THEY STILL SHOW
THIS KIND OF 5% WEIGHT LOSS
AFTER INFECTION, THEN THEY
RECOVER.
THEY ARE PROTECTED BUT IT'S NOT
AS MUCH AS A MOUSE FOR EXAMPLE
THAT GETS THE MOCK CHALLENGE.
WHEN WE LOOK AT VIRUS TITERS IN
THE LUNGS THE NON-VACCINATED
ANIMAL GETS A NICE VIRUS TITER,
WHICH PEAKS IN THIS CASE AROUND
DAY 2.
IT GOES UP, SO REPLICATING
BASICALLY.
BUT THEN IT STARTS TO GO DOWN BY
DAY 3.
A SINGLE VACCINATED MOUSE, THEY
STILL HAVE VIRUS TITER IN THE
LUNGS, DETECTABLE DAY 2 AND DAY
3, NOT DAY 1, STILL REPLICATING
BUT NOT THERE NOT DETECTABLE DAY
1.
TRIPLE VACCINATED HAS
STERILIZING IMMUNITY BASICALLY
YOU CANNOT DETECT VIRUS TITERS
AT THE EARLY TIME POINTS
POST-INFECTION.
SO NOW WE COME TO THE HOST
RESPONSE.
I PLOT AGAIN THIS VACCINE,
SORRY, PLOT AGAIN THE VIRUS
TITER ON THE RIGHT.
EVERY ROW IN THIS HEAT MAP IS A
SINGLE MOUSE.
IN THE SIDEBAR YOU CAN SEE THE
ONE DAY, TWO DAY, THREE DAY
MARKS FOR EVERY GROUP.
SO WE HAVE THE FIVE VACCINATION
GROUPS HERE.
WHAT YOU CAN SEE IS IS ONLY MICE
THAT DID NOT GET THE VACCINE
THAT DID GET INFECTED SHOW
DIFFERENT CYTOKINES, DIFFERENT
TIME POINTS POST-INFECTION,
CORRELATES TO SOME EXTENT WITH
VIRUS TITERS.
SINGLE VACCINATION MOUSE, MICE,
THAT GOT A BREAKTHROUGH
INFECTION YOU CAN STILL SEE
VIRUS TITERS IN THE LUNGS, LUNG
ARE SILENT IN TERMS OF
CYTOKINES.
THE VACCINE ALLOWS REPLICATION
BUT THE CYTOKINE PROFILE DOES
NOT REFLECT THE VIRUS TITERS
THAT WE DETECT.
SO THEN WE CHECK MYELOID CELLS
DAY 7 POST INFECTION, BASICALLY
HERE.
THAT'S WHERE IT BECOMES
INTERESTING.
UNVACCINATED MICE THAT GET SICK
AFTER THE FIRST INFECTION, THEY
WILL LOSE ALL OF THEIR ALVEOLAR
MACROPHAGES.
THEY WILL HAVE INFLAMMATORY
MONOCYTES COMING INTO THE LUNGS.
A SINGLE OR TRIPLE VACCINATION
PREVENTS LOSS OF ALVEOLAR
MACROPHAGES TO SOME EXTENT BUT
ALSO PREVENTS INFLAMMATORY
MONOCYTES TO COME IN.
AND SO WHEN WE THEN DO FLOW
CYTOMETRY, YOU'RE GOING TO OF
COURSE FOLLOW THE FATE OF THE
ALVEOLAR MACROPHAGES BY FLOW
CYTOMETRY.
THESE ARE BASELINE, PBS VACCINE
AND MOCK CHALLENGE.
SO WHEN YOU VACCINATE ANIMALS
YOU SEE THE SAME, NOTHING IS
HAPPENING AT THE ALVEOLAR
MACROPHAGE STAGE WHICH WE
EXPECTED.
IF YOU INFECT ANIMALS WITHOUT
VACCINATION YOU LOSE ALVEOLAR
MACROPHAGES.
WHEN YOU INFECT ANIMALS
VACCINATED YOU SEE ALL OF A
SUDDEN THIS BLOCK COMING UP
WHICH ARE CYCLIC POSITIVE CELLS
CD 11 NEGATIVE.
ALL MACROPHAGES ARE POSITIVE,
THIS IS NOT THEORETICALLY AN
ALVEOLAR MACROPHAGE BASED ON
SURFACE MARKERS.
WE THOUGHT MAYBE THESE ARE
MONOCYTES INFILTRATING AND
THEREFORE UPREGULATE CYCLIC F,
MAYBE STILL IN THE PROCESS PROF
UPREGULATING CD11C.
WE CHECKED SURFACE MARKERS, IN
THE LUNGS, BASICALLY
EOSINOPHILS.
SO WHAT WE SAW IS HERE YOU SEE
FOLLOWING THE CELL POPULATIONS
IN THE LUNGS AT ONCE.
IN SALMON ARE THE MACROPHAGES, I
PUT A BOX AROUND THE EXPECTED
EOSINOPHILS.
IN ANIMALS THAT GET NO VACCINE
AND NO CHALLENGE YOU GET THE
ALVEOLAR MACROPHAGE BUT NO
EOSINOPHILS.
SAME WITHOUT CHALLENGE.
IF YOU CHALLENGE WITHOUT VACCINE
THEY LOSE ALVEOLAR MACROPHAGES,
YOU DON'T SEE MUCH CHANGE IN THE
EOSINOPHIL COMPARTMENT, YOU SEE
DARK BLUE INFLAMMATORY MONOCYTES
COMING IN.
IF YOU VACCINATE AND CHALLENGE,
YOU SEE A LOT OF EOSINOPHILS,
MACROPHAGES ARE STILL SAVED.
IF YOU VACCINATE THREE TIMES,
WHICH MEANS THEY GOT THE
STERILIZING IMMUNITY, YOU GET
EVEN MORE EOSINOPHILS.
VACCINATION PERIPHERY RESULTS IN
EOSINOPHIL IN THE LUNGS, IF YOU
VACCINATE MORE YOU GET MORE
EOSINOPHILS.
PREVIOUS INFECTION DOES NOT
RESULT IN EOSINOPHIL INFLUX,
VACCINATION IN THE MUSCLE THAT
TRIGGERS THIS EOSINOPHIL INFLUX
IN THE LUNGS.
SO THEN WE CONFIRMED THIS IN
ANOTHER MOUSE BACKGROUND, THIS
IS SIX MICE, WE VACCINATE MICE
IN THE PREVIOUS VACCINE STUDY WE
USED ADJUVANT, IN THIS CASE DID
NOT USAGE -- USE THE ADJUVANT.
VACCINE PLUS CHALLENGE HAVE MORE
EOSINOPHILS COMING INTO THE
LUNGS, ANIMALS WITHOUT VACCINE
THEY HAVE NO EOSINOPHILS COMING
IN THE LUNGS.
THIS INFLUX, EOSINOPHIL INFLUX
IS ALSO ASSOCIATED WITH TYPE 2
CYTOKINE RESPONSE IN THE LUNGS
OF VACCINATED MICE SO IT GETS
CLASSICAL SUSPECTS LIKE IL-4, 5,
13, CYTOKINES THAT ARE TYPICAL
FOR TYPE 2 IMMUNE RESPONSE.
BASICALLY WE SAW THIS
INTRAMUSCULAR VACCINATION
RESULTS IN EOSINOPHIL INFLUX IN
BREAKTHROUGH AND STERILIZING
IMMUNITY, TWO GENETIC
BACKGROUNDS, BLACK 6 AND BALB/C,
CORRELATES WITH PROTECTION, NOT
ENHANCED DISEASE, BUT IMPACT
DRIVERS OF EOSINOPHIL INFLUX IS
NOT KNOWN.
EOSINOPHILS IN THE LUNGS ARE
INTERESTING AND IN THE FIELD OF
RESPIRATORY VIRUSES TYPICALLY
ASSOCIATED DOGMATICALLY CLASS
FIGHT AS PATHOLOGICAL SYMPTOMS
LIKE FROM DURING RSV INFECTION,
CONSIDERED SOMETHING THAT THEY
DON'T WANT, VACCINE-ASSOCIATED
ENHANCEMENT OF RESPIRATORY
DISEASE IS ASSOCIATED WITH
EOSINOPHIL INFLUKES AND ALLERGIC
ASTHMA THERE'S A LOT OF
EOSINOPHILS COMING IN.
HOWEVER, IF WE THINK OUTSIDE OF
THIS FIELD OF RESPIRATORY
VIRUSES AND RESPIRATORY
DISEASES, EOSINOPHILS ARE ALSO
INVOLVED IN PROTECTIVE RESPONSES
AGAINST HELMINTHS AND IMPORTANT
FOR TISSUE REPAIR, SOMETHING
THAT WOULD ALSO OCCUR DURING
RESPIRATORY INFECTION IF YOU
HAVE A LOT OF DAMAGE IN THE
LUNGS.
SO BECAUSE WE THOUGHT THIS WAS
INTERESTING, ONE OF MY STUDENTS
WHO USED TO WORK IN RSV GOT
FASCINATED AND WROTE A REVIEW ON
THE FUNCTIONS.
YOU'LL FIND SOME OVERVIEW OF THE
FUNCTIONS OF EOSINOPHILS AND
IT'S NOT ALWAYS ASSOCIATED WITH
PATHOLOGY.
WE ALSO SAW THIS ACTUALLY IN
OTHER RESPIRATORY VIRUS MODELS.
FOR EXAMPLE WHEN WE DID
SARS-COV-2 IN HAMSTERS DID
RNAseq ON LUNGS OF HAMSTERS
THAT GOT OPTIMAL DOSE OF PFIZER
mRNA VACCINE AND CHALLENGED
WITH VIRUSES AND CAUSED
BREAKTHROUGH INFECTIONS, AND
COULD SEE USING THE ALGORITHM,
TRANSCRIPTOME, EOSINOPHILS WERE
SEEN WITH BREAKTHROUGH INFECTION
IN VACCINATED ANIMALS WHICH ARE
DARK BOX PLOTS.
IN SARS-COV-2 CHALLENGE AND
MEHUL WILL TELL YOU MORE WE ALSO
SAW WITH TYPE 2 RESPONSE IN
VACCINATED ANIMALS WHEN THEY
RECEIVED RECOMBINANT SPIKE
PROTEIN WITH ADJUVANT GIVEN IN
THE MUSCLE BASICALLY IL-4, 5,
13, AND EOTAX IN.
WE DIDN'T SEE THIS PROFILE IN
LUNGS ININFECTED ANIMALS WHEN
GIVEN INTRANASAL THROUGH MUCOSAL
SITES WITH MUCOSAL ADJUVANTS.
I WON'T GO INTO THAT.
THIS IS SOON TO BE PUBLISHED AS
WELL AND AVAILABLE AS PRE-PRINT.
WE FOCUS ON OTHER HOST RESPONSES
TO INFECTION THAT ARE OCCURRING
LONGER AFTER INFECTION, AND
FIRST ONE IS GERMINAL CENTER B
CELLS IN THE LUNGS, AND
PERIBELONGAL LIMP NOTES -- LYMH
NODES 24 DAYS POST INFECTION.
WE TOOK THE LYMPH NODES 24 DAYS
AND MEASURED GERMINAL CENTER B
CELLS IN THE LUNGS MEASURED
OUTSIDE LYMPH NODES, WHAT YOU
CAN SEE HERE IS GL 7 WE CAN
STAIN ON B CELLS IN THE LUNGS
CLOSE TO THE BRONCHUS.
WHY ARE THEY INTERESTING?
THEY CORRELATE WITH IMMUNITY AND
CAN ALSO BE SOURCES OF DE NOVO
ANTIBODY RESPONSES IN THE LUNGS.
AND SO WHEN WE CHECK THESE
LEVELS OF GERMINAL CENTERS IN
THE LUNGS IN VACCINATED AN
UNVACCINATED ANIMALS YOU CAN SEE
TIV VACCINATION INTERFERENCE,
THEY ARE NICELY INDUCED IN
INFECTS THE ANIMALS WITHOUT
VACCINATION.
WHEN YOU LOOK IN THE GERMINAL
CENTERS IN THE LYMPH NODES YOU
CAN SEE VACCINATION IS
DEFINITELY NOT INHIBITING THIS,
CONTRARY IF YOU VACCINATE THREE
TIMES YOU GET MORE GERMINAL
CENTERS AFTER INFECTION IN THE
PERIBRONCHIAL LYMPH NODES.
WE LOOKED AT TISSUE RESIDENT
MEMORY T CELLS, THEY CAN
RECOGNIZE COULD BE SERVED
EPITOPES, PROVIDE IMMUNITY AND
WE KNOW THEY ARE TISSUE
RESIDENTS BECAUSE WE FOCUS ON
CELL SURFACE MARKERS, YOU CAN
EXCLUDE BY IV LABELING AND ALSO
HERE YOU CAN SEE TIV VACCINATION
INTERFERES WITH INDUCTION AFTER
INFECTION, YOU GET VERY NICELY
IN MICE NOT VACCINATED.
SO NOW WE CAN LOOK AT HOST
RESPONSE DURING INFECTION WITH X
31 VIRUS, THE PROTECTION YOU CAN
SEE.
BASICALLY ANIMALS THAT GOT VERY
SICK THE FIRST TIME ARE FULLY
PROTECTED DURING REINFLECTION.
ANIMALS NOT VACCINATED THE FIRST
TIME COME TO THE H INFECTION BY
DAY 7.
ANIMALS WITH VACCINE WITHOUT
CHALLENGE SOME ARE PROTECTED AND
THEY RECOVER BY DAY 6, 7.
SOME WE LOSE.
THEN ANIMALS THAT RECEIVE ONE OR
THREE VACCINATIONS PLUS
INFECTION, THEY STILL LOSE MORE
BODY WEIGHT THAN ANIMALS THAT
GOT VERY SICK THE FIRST TIME BUT
START TO RECOVER EARLY ON BY DAY
4.
THIS IS ALSO REFLECTED IN THE
LUNG HOMOGENOUS WHEN WE CHECK
VIRUS TITERS, ANIMALS THAT GOT
SICK FIRST TIME AFTER INFECTION,
WELL THEY CONTROL THE H3N2 VIRUS
DURING INFECTION COMPLETELY, ALL
THE OTHER GROUPS INCLUDING TIV
AND TRIPLE TIV MICE THAT
RECEIVED INFECTION THEY STILL
HAVE VIRUS IN THEIR LUNGS.
AND ALSO I HAVE TO SAY THAT
THESE ARE THE MICE THAT HAVE THE
TRMs AND GERMINAL CENTER B
CELLS, THESE ARE MICE THAT DO
NOT HAVE THESE BECAUSE OF
VACCINATION.
SO THEN WE TAKE THE BLOOD FROM
THESE MICE AND WE'RE GOING TO
CHECK THAT IT'S DIFFERENCES IN
CROSS-REACTIVITY IN SERUM OF
THESE MICE.
TO DO THIS WE DID A KIND OF IN
VITRO/IN VIVO MICRO
NEUTRALIZATION ASSAY.
WE USED THESE IMMUNE COMPLEXES
TO KIND OF INFECT NAIVE MICE AND
SEE HOW MUCH THEY WERE PROTECTED
AGAINST DISEASE FROM H3N2
CHALLENGE.
WE SEE THE OPPOSITE PROTECTION
THAT WE SAW IN THE PREVIOUS
SLIDES.
SO MICE THAT GOT VIRUS THAT WAS
INCUBATED WITH SERUM FROM NAIVE
MICE OR SERUM FROM MICE WITHOUT
CHALLENGE OR RECEIVED SERUM FROM
MILES THAT RECEIVED H1N1 WITH NO
VACCINATION DIED, ALL THE MICE
THAT RECEIVED VIRUS INCUBATED
WITH TIV PLUS NEW CALEDONIA
CHALLENGE ALL WERE PROTECTED
FROM SEVERE DISEASE DURING THIS
H3N2 CHALLENGE.
BASICALLY WHAT WE SAW WAS
COMBINATION OF TIV VACCINATION
FOLLOWED BY NC 99 CHALLENGE WAS
REQUIRED TO HAVE PROTECTED
SERUM.
THIS MEANS BECAUSE THE MICE THAT
RECEIVED PBS WITHOUT, SO NO
VACCINE PLUS NEW CALEDONIA
CHALLENGE WAS NOT PROTECTED,
THIS MEANS PROTECTION FROM TIV
PLUS NEW CALEDONIA CHALLENGE
RELYING MORE ON ANTIBODIES
RATHER THAN B CELLS OR MUCOSAL.
WHAT MAKES THIS HYBRID IMMUNE
SERA, FROM INFECTION AND
VACCINATION TOGETHER, BETTER
COMPARED TO CONVALESCENT SERUM?
MORE CROSS-REACTIVE ANTIBODIES,
HOW CAN WE INDUCE BY VACCINATION
OR HOW CAN WE INDUCE AND TARGET
IBALT AND TRMs WITH MUCOSAL
VACCINES.
THEN WE STILL HAVE THE INNATE
IMMUNITY THAT I DIDN'T ADDRESS
YET.
SO, EVERYBODY KNOWS INFECTION
WILL RESULT IN INNATE IMMUNE
RESPONSES SHORT-LIVED.
HOWEVER, THERE'S MORE EVIDENCE
FOR INNATE RESPONSES, SO WHAT WE
WANTED TO CHECK IF THERE WAS
STILL KIND OF A CHANGE INNATE
RESPONSIVENESS BY 28 DAYS POST
FIRST INFECTION.
THIS IS FOUR WEEKS AFTER THE
H1N1 INFECTION.
WE DID THIS EXPERIMENT WHERE WE
INFECTED MICE OR GIVE THEM A
MOCK CHALLENGE.
FOUR WEEKS LATER TOOK THE
BRONCHIAL ALVEOLAR LAVAGE AND
CHECKED ESPECIALLY THE ALVEOLAR
MACROPHAGE COMPARTMENT, I'VE
SHOWN INFECTION CAN DEPLETE THE
MACROPHAGES AND THEY COME BACK.
WE SAW PHENOTYPIC CHANGES ON
MACROPHAGES OF MICE THAT WENT
THROUGH INFECTION.
THEY HAVE HIGH EXPRESSION BUT
ALSO THEY HAVE LOWER CYCLIC F
EXPRESSION, AT LEAST POPULATION
LIKE THIS.
THIS IS SOMETHING THAT HAS BEEN
DESCRIBED BEFORE IN OTHER
STUDIES.
FOR EXAMPLE THIS STUDY FROM
MISHARIN, SORRY IF I SAY IT
WRONG, TYPICALLY ALVEOLAR
MACROPHAGES THAT COME INTO THE
LUNGS AND THEN BECOME RESIDENTS,
THIS WAS FIRST DESCRIBED IN
FIBROSIS MODEL WHERE THEY INDUCE
FIBROSIS.
THIS IS ALSO SUGGESTED FOR
TRAINED IMMUNITY WHERE PEOPLE
CLAIM ALVEOLAR MACRO FEIGNS CAN
BE TRAINED BY FIRST FLU
INFECTION AND HAVE DIFFERENT
MARKERS OF EXPRESSION, HOWEVER
THERE'S A GOOD DOCUMENTARY
RECENTLY WHERE THEY SAY WE HAVE
TO BE VERY CAREFUL WHEN WE STUDY
TRAINED IMMUNITY INDUCED BY FLU
IN THE LUNGS BECAUSE HAVE YOU
ALVEOLAR MACROPHAGES THAT MIGHT
GET TRAINED, INFLAMMATORY
MONOCYTES BECOME RESIDENT AND
THEREFORE HAVE DIFFERENT
PHENOTYPE, NOT PERCEIVED OR
TRAINED BY THE FIRST FLU
INFECTION.
SINGLE CELL RNA SYNC ON BALT
FLUIDS IN THESE MICE TO SEE
COMPOSITION AND REMODELING IF
FIRST FLU INFECTION.
BY SINGLE CELL RNA-SEQ CAN
DISCRIMINATE IMMUNE CELLS IN THE
BALF FLUID, WE DEFINE UP TO SIX
MACROPHAGES CLUSTERS AS WELL AS
B CELLS AND T CELLS THAT BECAME
RESIDENT AFTER FIRST INFECTION.
YOU CAN SEE INFECTION OR MOCK
CHALLENGE WILL GIVE DIFFERENT
PROPORTIONS OF DIFFERENT
MACROPHAGES SUBSETS AS WELL AS T
CELLS.
WE WONDERED IF THIS LUNG
REMODELING BY INFECTION WHICH
H1N1 CAN BE PROTECTIVE OR NOT,
CAN IT AFFECT OUTCOME WITH FLU
B, AND WHY WE TAKE FLU B?
INFLUENZA A AND B ARE
ANTIGENICALLY UNRELATED.
WE INFECTED ANIMALS WITH H1N1 OR
GIVE THEM MOCK CHALLENGE, WAIT
FOUR WEEKS, REINFECTED WITH
B/MAR FROM 2004 CHALLENGED WITH
THREE DOSES, IF YOU GIVE 520 PFU
THERE'S SOME PROTECTIVE EFFECT
FROM PREVIOUS H1N1 SO ANIMALS
THAT RECEIVED H1N1 FIRST THEY
LOSE TYPICALLY 10% LESS BODY
WEIGHT COMPARED TO NAIVE
ANIMALS.
WHEN WE GIVE 2500 PFU IT'S
BIGGER.
10,000 PFU YOU STILL CAN KILL
MOST OF THE ANIMALS ONLY ONE
MOUSE WILL SURVIVE DESPITE
HAVING H1N1 FIRST.
THERE'S SOME PROTECTION FROM
FIRST INFECTION, YOU CAN STILL
OVERRULE IF YOU GIVE ENOUGH
VIRUS OF THE WE WANTED TO KNOW
IF CD 11+ CELLS ARE PROTECTED.
WE TRIED TO TRANSFER BY
TRANSFERRING CELLS FROM ANIMALS
THAT GOT NC 99 CHALLENGE OR MOCK
CHALLENGE.
SO THEREFORE WE COLLECTED
POSITIVE CELLS FROM BALF FLUIDS,
TRANSFERRED INTO NAIVE MICE,
CHALLENGED WITH B VIRUS 24 HOURS
LATER, FOLLOWED 14 DAYS TO
MONITOR FOR MORBIDITY AND
MORTALITY.
WE CAN SEE THAT CD11+ CELLS CAN
PROVIDE FULL PROTECTION AGAINST
THE B VIRUS INFECTION.
ANIMALS THAT RECEIVED CD11 CELLS
IN MICE MOCK CHALLENGED, SO THEY
LOSE 50% OF THE MICE AFTER B
VIRUS CHALLENGE.
BUT IF FOUR TIMES MORE CELLS
FROM ANIMALS THAT RECEIVED H1N1
FIRST, WE LOSE 100% PROTECTION
BUT GET 100% MORTALITY.
YOU CAN TRANSFER PROTECTION BY
H1N1-EXPOSED CD11+ CELLS BUT TOO
MUCH AND YOU LOSE PROTECTION AD
TURN TO MAR AT THAT TIME, THIN
LINE BETWEEN PROTECTION AND
DISEASE.
WHY IS THIS?
WE DON'T KNOW.
CAN BE DUE TO THE FACT CONTEXT
DEPENDENT BECAUSE YOU TAKE VIRUS
EXPOSED MACROPHAGES, PUT THEM
INTO NAIVE ANIMAL, FOR EXAMPLE
THOSE EPITHELIA HAVE NOT BEEN
EXPOSED TO H1N1 OR COMPONENTS
HAVE NOT BEEN EXPOSED TO VIRUS
BEFORE.
SO WE DON'T FLOW IF THERE'S A
FEED -- KNOW IF THERE'S A
FEEDBACK MECHANIC , STILL FIELDS
WE CAN EXPLORE.
WE WERE INTRIGUED BY LINE
REMODEL, THIN LINE BETWEEN
PROTECTION AND DISEASE.
WE DID SECOND EXPERIMENT
INFECTED WITH H1N1, ALSO
INCLUDED A SECOND GROUP INFECTED
WITH SARS-COV-2, THE OMICRON
STRAIN OR HAD A MOCK CHALLENGE
GROUP.
TWO DIFFERENT GENETIC
BACKGROUNDS.
THE MICE WITH THE HUMAN H2 WE
ALREADY DISCUSSED TODAY, 129
MICE ARE WILDTYPE MICE, MORE
SUSCEPTIBLE TO SARS-COV-2.
WE USED A BETA STRAIN TO
REINFECT, THIS STRAIN CAN ALSO
INFECT WILDTYPE MICE AND CAUSE
ABOUT 10 TO 15% BODY WEIGHT LOSS
IN 129 MICE, LETHAL IN K18 MICE.
WE SEE THIS IS THE MOCK
CHALLENGED ANIMAL, THEY DON'T
LOSE WEIGHT, DEPENDING ON
GENETIC BACKGROUND WE CAN SEE
K18 MICE ARE FULLY PROTECTED
FROM BETA INFECTION, WHEN FIRST
INFECTED WITH BA.1 VIRUS.
FIRST INFECTION WITH NEW
CALEDONIA THE 9 DID NOT PROTECT
FROM BODY WEIGHT LOSS, BASICALLY
GAVE THE SAME AS ANIMALS NOT
PRIOR INFECTED.
WE CAN SEE IN 129 MICE PRIOR
INFECTION WITH FLU KIND OF
PROTECTS THEM TO SOME EXTENT AND
GIVES FIRST RECOVERY, HOWEVER
THE BA.1 PRIOR INFECTION RESULTS
IN NO RECOVERY BY DAY 4.
WHEREAS BETA INFECTION GIVES
SOME RECOVERY BY DAY 3 ALREADY.
THERE'S A DIFFERENCE IN AGAINST
BACKGROUNDS REFLECTED IN
PROTECTION AFTER SECONDARY
INFECTION WITH UNRELATED VIRUS.
WE CHECKED CYTOKINE PROFILES,
VERY DIFFERENT CYTOKINE PROFILES
DEPENDING ON FIRST TREATMENTS
BEFORE THE BETA INFECTION, MOCK
INFECTED BA OR 101 INFECTION,
DIFFERENT BETWEEN K18 MICE, TOP
PANEL, VERSUS 129 MICE, BOTTOM
PANEL.
WE SEE BA.1 PRIOR INFECTION WILL
RESULT IN IL-2 AND IL-9 IN K18
MOUSE, DIFFERENT IF YOU
PREVIOUSLY INFECT WITH FLU.
WE TAKE 129 MICE, BA.1 PRIOR
INFECTION WILL ACTUALLY RESULT
IN VERY STRONG TYPE 2 RESPONSE,
IL-15, 4, 5, SO THIS IS
DIFFERENT AGAIN WHEN YOU
PREVIOUSLY INFECT ANIMALS WITH
FLU.
PRIOR INFECTION DEPENDING ON
VIRUS GIVERS DIFFERENT CYTOKINE
RESPONSE, ALSO DIFFERENT BETWEEN
DIFFERENCE MOUSE STRAINS.
WE TOOK CD11+ CELLS AND
STIMULATED BECAUSE IT'S GOING TO
STIMULATE ALVEOLAR MACROPHAGES
AROUND SEE BIG DIFFERENCE
BETWEEN BA.1 AND DIFFERENT MOUSE
STRAINS, SO K18s UPREGULATE
IL-19, 17, 22, AGAIN THIS TYPE 2
PROFILE IL-4 REDUCTION.
THIS IS MY LAST DATA SLIDE.
I SHOWED THAT ACTUALLY TYPE 2
RESPONSE CAN CORRELATE
PROTECTION IN VACCINATED MICE
AFTER FLU BUT ALSO CORRELATES
WITH LACK OF PROTECTION IN
SARS-COV-2 IN REINFLECTION.
WE'VE EXPANDED TO OTHER MOUSE
MODELS THAT ARE KNOWN TO HAVE
TYPE 2 RESPONSE THAT IS NOT
CORRELATING WITH PROTECTION AND
SO WE LOOK AT ALLERGY RESPONSES
TO COMPARE WHAT WE SEE IN FLU
AND COVID MODELS, WITH SOME
ALLERGY MODELS.
WE SENSITIZE MICE AGAINST A
STANDARDIZED PROTOCOL, FOLLOWED
BY SOME INTRANASAL CHALLENGES
AND LET THESE MICE FULLY RECOVER
AND AGAIN WITH 2,000 PFU OF H1N1
NEW CALEDONIA VIRUS.
ALLERGIC MICE LOSE MORE BODY
WEIGHT AFTER FLU CHALLENGE, IN
BLACK 6 MICE IT'S NOT BASED ON
BODY WEIGHT BUT WE CAN SEE IN
BOTH MOUSE STRAINS THAT THE
VIRUS TITER IS HIGHER OTHER NOT
LOWER IN ALLERGIC MICE COMPARED
TO NONSENSE NON-SENSITIZED.
BASICALLY TO SUMMARIZE THIS,
POINTS FOR FUTURE ATTENTION WE
SEE HOST IMMUNE RESPONSES IN
MICE WITH MORE COMPLEX EXPOSURE
HISTORY ARE DIFFERENT SO THERE'S
DISEASE MODULATION AND THEN
ESPECIALLY TYPE 2 RESPONSE WE
CAN SEE CORRELATES WITH
PROTECTION.
AND WE SEE DIFFERENCE BETWEEN
TRAINED AND POTENTIALLY
PERSISTENT IMMUNE IMMUNITY,
ASSOCIATED WITH LUNG REMODELING.
WE NEED BETTER MODELS FOR
ENHANCED RESPIRATORY DISEASE,
BUT WE NEED SOME IN MOUSE MODELS
BECAUSE THAT'S NOT ESTABLISHED.
THIS WILL BE LINKED TO
PREEXISTING IMMUNITY.
WE HAVE TO CONSIDER HOST
CONDITION WHEN WE LOOK AT OUR
MOUSE MODELS.
AGAINST BACKGROUNDS PLAYS
IMPORTANT ROLE, CO-MORBIDITIES,
WE HAVE ADVANCED AGE, OBESITY,
DIABETES, ENVIRONMENTAL FACTORS.
CHALLENGED STRAINS ARE NEEDED
THAT MATCH MOST RECENT VACCINE
STRAINS TO SEE VACCINE MEDIATED,
ESPECIALLY FOR H1N 2 AND
INFLUENZA B VIRUS.
THESE ARE THE PEOPLE THAT DO TE
WORK AND MY COLLABORATORS AT
MOUNT SINAI.
THANK YOU.
>> WE HAVE TIME FOR ONE
QUESTION.
>> NICE WORK.
I HAVE A QUESTION.
WOULD IT BE GOOD TO LOOK AT
PATHOLOGY IN YOUR INFLUENZA B
INFECTION OR CD, POSITIVE, YOU
GIVE MORE DOSE THE LUNG TISSUE
IS MORE DAMAGED FROM IMMUNE
SYSTEM OR THE VIRUS, WHAT CAN
YOU SAY ABOUT THE PATHOLOGY OF
THE TISSUE OR CAN YOU SPECULATE?
>> SO WE SEE LESS PATHOLOGY, BUT
WE SEE LUNG REMODEL.
SO WE SEE IBALT, LESS PATHOLOGY,
SO THEY ARE PROTECTED.
>> WE'LL SAVE THE OTHER
QUESTIONS FOR THE PANEL.
>> LAST SPEAKER BEFORE THE BREAK
IS NEELTJE VAN DOREMALEN.
>> CLOSE?
>> ALMOST.
>> THANK YOU.
>> MUCOSAL IMMUNE RESPONSES IN
ANIMAL MODELS OF SARS-COV-2,
CURRENT RESEARCH AND FUTURE
DIRECTIONS.
>> ALL RIGHT.
SO, THANKS FOR THE INVITATION,
I'M VERY EXCITED TO BE HERE,
REALLY ENJOYED TODAY ALREADY.
AND I'M GOING TO TALK ABOUT
MUCOSAL IMMUNE RESPONSES IN
ANIMAL MODELS OF SARS-COV-2.
IF I CAN MOVE MY SLIDES, THAT
IS.
OKAY.
SORRY ABOUT THAT.
SO I GOT FIRST INTERESTED IN
THIS WHEN WE DID OUR VACCINE
STUDY IN NON-HUMAN PRIMATE, IN
EARLY 2020.
SO WE LOOKED AT CHADOX1, THE
ASTRAZENECA VACCINE, ENCODED TO
THE GENOME OF ADENOVIRUS VECTOR
VACCINE WHICH IS REPLICATION
INCONSEQUENT, THE SPIKE PROTEIN
IS NOT ACTUALLY EXPRESSED ON THE
OUTSIDE OF THE ADENOVIRUS.
WE DID A STUDY IN NON-HUMAN
PRIMATE, NOT THE SMALL ANIMAL
MODEL OF COURSE.
WE GAVE SINGLE OR DOUBLE
VACCINATION, FOUR WEEKS APART
AND CHALLENGED FOUR WEEKS AFTER
THAT.
AS A CONTROL USED CHADOX1.
AFTER CHALLENGE WE TOOK MULTIPLE
NASAL SWABS AND DAY 1, 3, 5, 7,
NECROPSY ON DAY 7 TO SEE IF LUNG
TISSUE WAS PROTECTED.
YOU CAN CLEARLY SEE IN SERUM WE
GET INDUCTION OF VIRUS
NEUTRALIZE BE ANTIBODIES.
WHEN WE LOOK AT T-CELL
RESPONSES, WHEN WE SIMULATED
PBMCs YOU CAN SEE INCREASE OF
SPOT FORMING UNITS, THEY WERE
PRODUCING INTERFERON GAMMA AFTER
STIMULATION.
MOST IMPORTANTLY WHAT WE NOTICED
WHEN WE LOOKED AT LUNG TISSUE
THESE VACCINATED ANIMALS WERE
PROTECTED.
THAT WAS GREAT.
THIS VACCINE WAS DEVELOPED.
IT HAS BEEN APPROVED IN
180-SOMETHING COUNTRIES, FIRST
YEAR WAS SUPPOSED TO HAVE
PROTECTED 6 MILLION PEOPLE,
SAVED THAT MANY LIVES.
ONE OF THE THINGS I NOTICED IN
THE STUDY, THIS IS A SURPRISE TO
ME, PROBABLY SHOULDN'T HAVE BEEN
BUT WAS, WHEN WE LOOKED AT NASAL
SWABS THERE WASN'T A DIFFERENCE,
WE WERE NOT THE ONLY ONES TO
SHOW THIS.
OTHER NON-HUMAN PRIMATE STUDIES
SHOWED THE SAME THING.
MORE IMPORTANTLY WE NOW KNOW
VACCINATED INDIVIDUALS WE SEE
THIS TOO.
SO WHEN YOU'RE VACCINATED, YOU
CAN STILL GET INFECTED, YOU CAN
STILL SHED INFECTIOUS VIRUS.
WHEN WE LOOK AT VIRAL RNA IN THE
BEGINNING IT'S VERY LITTLE
DIFFERENT BETWEEN VACCINATED
INDIVIDUALS AND UNVACCINATED
INDIVIDUALS.
SO, THAT'S OF COURSE AN ISSUE
BECAUSE IF YOU'RE VACCINATED,
YOU STILL SHED VIRUS, YOU'RE
TRANSMITTING THE VIRUS TO OTHER
PEOPLE.
AND SO THAT REALLY HAS TWO MAJOR
CONSEQUENCES.
FIRST ONE IS THAT
IMMUNOCOMPROMISED INDIVIDUALS
ARE STILL SUSCEPTIBLE TO SEVERE
DISEASE AND WE WOULD RATHER
PROTECT THEM FROM SEVERE
DISEASE.
THE SECOND ONE CONSTANTLY
DEALING WITH, VARIANTS OF
CONCERN, SARS-COV-2 HAS BEEN
ABLE TO ADAPT TO HUMAN HOST AND
ALSO MOVE AWAY FROM THE IMMUNE
RESPONSE.
TO UNDERSTAND THAT BETTER, I
THINK WE NEED TO UNDERSTAND
SYSTEMIC VERSUS MUCOSAL
IMMUNITY.
SYSTEMIC COMMUNITY IS
CIRCULATING THROUGHOUT THE BODY.
MUCOSAL IS MORE LOCALIZED.
THE KEY PLAYERS ARE TISSUE
RESIDENT T- AND B-CELLS AS WELL
AS DIFFERENT ANTIBODIES
INCLUDING SECRETORY IgA.
WE KNOW FROM STUDIES IN HUMANS
MUCOSAL IMMUNITY IS IMPORTANT.
IN THIS STUDY IN 2022, YOU CAN
SEE HUMANS HAD HIGH AMOUNT OF
MUCOSAL IgA, WERE PROTECTED
AGAINST OR HAD LOWER RISK OF
OMICRON BREAKTHROUGH INFECTIONS.
WE DIDN'T FIND THIS IN HUMANS
THAT HAD HIGH IgG IN THEIR
MUCOSA.
WE LOOKED AT VACCINATED
INDIVIDUALS, THIS IS A STUDY
DONE IN 2022 AS WELL, LOOKED AT
MUCOSAL IgA IN HUMANS THAT
WERE VACCINATED THAT EITHER DID
OR DID NOT HAVE PREVIOUS
SARS-COV-2 INFECTION.
THEY WERE REALLY ONLY ABLE TO
DETECT MUCOSAL IgA IN PEOPLE
THAT HAD A SARS 2 INFECTION
SUGGESTING CURRENT VACCINATIONS
WE'RE GIVING ARE NOT GIVING US A
GOOD MUCOSAL IMMUNE RESPONSE.
I STARTED CARING ABOUT IT, WHY
SHOULD YOU CARE ABOUT MUCOSAL
IMMUNITY?
MOST VIRUSES WE WORK WITH AFFECT
THE MUCOSAL ROUTE.
INFLUENZA A, SARS-COV-2, RSV,
ALL THESE VIRUSES.
REALLY THE MAJORITY OF VIRUSES
INFECT BY MUCOSAL ROUTE, WE
SHOULD CARE ABOUT MUCOSAL
IMMUNOLOGY, IT'S NOT THAT WELL
UNDERSTOOD RIGHT NOW.
KEY PLAYERS WITH MUCOSAL
IMMUNITY IN THE UPPER
RESPIRATORY TRACT ARE TONSILS,
THEY FORM A THE WALDEYER'S RING,
SECONDARY ORGANS.
WITHIN THESE TISSUES, YOU GET
IMMUNE RESPONSE THAT'S SIMILAR
TO WHAT WE'RE FAMILIAR WITH IN
LYMPH NODES, YOU HAVE NAIVE T
CELLS, IF THEY ARE EXPOSED TO
ANTIGEN THEY GET ACTIVATED, CD6
IS DOWNREGULATED, UPREGULATION
OF CD44, T CELLS CAN MIGRATE.
THEY START UPREGULATING, CD69
AND CD1 ALLOWING CELLS TO STAY
IN THE RESPIRATORY TRACT, THEY
CAN STAY FOR MONTHS OR YEARS
AFTER INFECTION.
SO STRUCTURE OF RESPIRATORY
MUCOSA, YOU'RE LOOKING AT UPPER
RESPIRATORY TRACT, LOWER
RESPIRATORY TRACT REGARDING
EPITHELIUM LOOKS DIFFERENT BUT
OVERALL WHAT I WANT YOU TO LOOK
AT IS MUCUS LAYER ON TOP, MUCOSA
LAMB NAH ON THE BOTTOM.
THE LAYER IS IN PLACE TO CATCH
DIFFERENT THINGS YOU BREATHE IN
ALL THE TIME, BACTERIA, VIRUSES,
ALSO OTHER ALLERGENS.
WITHIN THAT MUSOUS LAYER YOU CAN
FIND PEPTIDES AND ANTIBODIES
SUCH AS SECRETORY IgA.
LAMINA COPA, DIFFERENT IMMUNE
CELLS, REALLY NICE RED D C
PROBING THROUGH, TRYING TO FIND
ANTIGENS.
WE'LL TAKE THROUGH LYMPH NODES,
TONSILS AND STARTS IMMUNE
RESPONSE.
YOU CAN SEE OTHER IMMUNE CELLS
THERE.
AND THIS RESPONSE FUNCTIONS AS
FIRST LINE OF DEFENSE AGAINST
PATHOGENS, PLASMA CELLS
PRODUCING, FOR EXAMPLE, IgA,
THAT GET ACTIVELY TRANSPORTED
ACROSS EPITHELIUM AND CAN
DIRECTLY NEUTRALIZE VIRUSES WITH
OTHER FUNCTIONS, TISSUE RESIDENT
T CELLS CAN START PRODUCING
CYTOKINES AND CHEMOKINES, THERE
BY PRODUCING ANTI-VIRAL STATES
BY INDUCTION OF ISG, CAN ALSO
RESULT IN ACTIVATION OF OTHER
IMMUNE CELLS, RECRUIT RECRUITME
LAMINA PROPRIA WITHIN THREE
HOURS AFTER.
ONCE THEY ARE SITTING IN A LUNG
THEY CAN ACT VERY QUICKLY.
THIS IS AN INFLUENZA STUDY DONE
THAT SHOWED IN IMMUNE CELLS THEY
ONLY HAVE TISSUE RESIDENT T
CELLS, DON'T HAVE ANTIBODIES AT
ALL, WHEN THESE MICE WERE
INFECTED WITH INFLUENZA LUNGS
WERE FULLY PROTECTED SHOWING T
CELLS ARE INCREDIBLY IMPORTANT
IN PROTECTING LUNG TISSUE.
SO WE'VE BEEN TALKING ABOUT
ANIMAL MODELS THROUGHOUT HERE.
THE MAIN MODELS THAT I LIKE TO
LOOK AT ARE THE MOUSE, HAMSTER,
FERRET, A SMALL ANIMAL MODELS,
EVEN A FERRET IS QUESTIONABLE
WHETHER IT'S A SMALL ANIMAL
MODEL, IT NEEDS VERY LARGE CAGES
TO WORK WITH.
MY FAVORITE MODEL RIGHT NOW IS
THE MOUSE.
THAT ISN'T BECAUSE I THINK THE
MOUSE HAS A RESPIRATORY TRACT
THAT LOOKS LIKE THE HUMAN
RESPIRATORY TRACT, IT'S PURELY
BECAUSE OF THE REAGENTS WE HAVE
AVAILABLE.
IF YOU WANT TO DO DEEP DIVE INTO
IMMUNOLOGY, IT'S GOING TO BE
PRETTY DIFFICULT TO THAT WITH
HAMSTERS AND FERRETS, IMPORTANT
TO BE RESOLVED BECAUSE MUCOSAL
IMMUNOLOGY CAN BE STUDIED ON ANY
ANIMAL MODEL, INCLUDING HUMANS.
SO WE HAVE DIFFERENT TOOLS TO
INVESTIGATE MUCOSAL IMMUNE
RESPONSE IN MICE.
I REALLY LIKE LOOKING AT
LYMPHOID TISSUE.
YOU CAN SEE TWO TRIANGLES,
LOCATED RIGHT ABOVE OF THE SOFT
PALLIATE, YOU CANNOT SEE THESE
WITH THE NAKED EYE, YOU NEED
GOOD PATHOLOGISTS TO HELP
COLLECT TISSUES, LUCKILY WE HAVE
THOSE.
NOW THEY ARE FUNCTIONING LIKE
THE TONSILS IN HUMANS, DIFFERENT
LOCATION IN UPPER RESPIRATORY
TRACT, IT'S REALLY GOOD AT
MODELING.
HERE WE BASICALLY STAIN FOR
DIFFERENT IMMUNE POPULATIONS
WITHIN THE MOUSE.
WE'RE LOOKING AT T- AND B-CELLS
ON TOP.
REALLY NICE
COMPARTMENTALIZATION, NAIVE
MOUSE, DOING STUDY WE CAN ALSO
TAKE THAT MOUSE, PUT IT IN
MEDIA, INTUBATE, COLLECT AND ASK
WHAT ANTIBODIES DID THOSE B
CELLS PRODUCE IN THAT TIME.
YOU CAN TRUE TO UNDERSTAND THAT
ANTIBODIES WE GET IN UPPER
RESPIRATORY WE GET.
WE CAN UNDERSTAND THE LOWER
RESPIRATORY TRACT AND LOOK AT
SERUM TO LOOK AT RESPONSE.
WE NOW HAVE T- AND B-CELL
ELISPOTS.
WE CAN LOOK AT SPIKE SPECIFIC
RESPONSES IN MICE AND ONE OF MY
FAVORITE TOOLS, SOMEBODY ELSE
MENTIONED EARLIER, DOING AN IV
INFECTION WITH TD 45 BEFORE WE
DO FLOW CYTOMETRY.
WE USE ANTI-CD45, INJECT RETRO
ORBITICALLY, CIRCULATES THROUGH
THE BODY OF THE MOUSE BUT NOT
PENETRATING THE TISSUES.
WE THEN HARVEST THE TISSUES AND
DO FLOW CYTOMETRY B CELLS ARE
CD4 POSITIVE, C CELLS ARE NOT.
DOES INTRANASAL VACCINATION
PROTECT?
WE WENT TO THE MOUSE MODEL,
VACCINATED ANIMALS VIA
INTRAMUSCULAR ROUTE WITH mRNA,
THREE WEEKS APART.
WE WAITED SIX WEEKS.
THIS IS WHERE WE DIVIDED MICE IN
TWO GROUPS AND EITHER GAVE
mRNA VACCINATION
INTRAMUSCULARLY AGAIN OR GAVE
INTRANASAL VACCINATION CHADOX,
THAT I WAS TALKING ABOUT BEFORE.
HARVESTED TISSUES 3 AND 14 DAYS
AFTER VACCINATION.
WE'RE LOOKING AT THREE DAYS POST
VACCINATION, ONLY LOOKING AT THE
CELLS THAT WERE CD45 NEGATIVE,
RESIDENTIAL CELLS IN THE LUNGS.
WE THEN USED UNSUPERVISED
ANALYSIS TO LOOK AT WHAT CELL
POPULATIONS WE CAN FIND IN THE
LUNG TISSUE.
WE PULLED THAT APART LOOKING AT
TWO VACCINE GROUPS, SO YOU CAN
IMMEDIATELY SEE IN THE mRNA
GROUP YOU HAVE CERTAIN
POPULATIONS THAT ARE
OVERREPRESENTED, COMPARED TO THE
CHADOX GROUP.
YOU CAN LOOK AT WHAT MARKERS ARE
UPREGULATED.
PAY ATTENTION TO 1 AND 5, BROWN
AND ORANGE, IN THE TIFFANY PLOT,
UPREGULATED CD11C, MAY BE
DENDRITIC CELLS.
IF WE GO BACK TO TRADITIONAL
GATING, ONLY LOOK AT CD11C, WE
SEE NO DIFFERENCE IN VACCINE
GROUPS.
IF WE ADD CD11B TO TRADITIONAL
GATING NOW WE'RE STARTING TO SEE
DIFFERENCES BETWEEN VACCINE
GROUPS.
THIS IS A REALLY NICE WAY OF
GOING IN LIKE WITHOUT AN IDEA
WHAT SHOULD CHANGE BUT SEEING
WHAT ARE THE DIFFERENCES WE'RE
FINDING.
WE THEN LOOKED AT TISSUE
RESIDENT CD8 T CELLS WHICH MAY
BE MY FAVORITE CELL POPULATION
AT THE MOMENT.
WE'RE LOOKING AT LUNG TISSUE,
AND THE QUESTION WE ASK IS WHAT
PERCENTAGE OF CD8 T CELLS IN THE
LUNG IS RESIDENTIAL.
WE TACK ALL -- TOOK ALL THE
CELLS IN THE LUNG.
YOU CAN SEE IN DAY 3 WE GET
INFLUX OF CD8 T CELLS IN THE
LUNG TISSUE AND STILL THERE AT
DAY 14.
WE CAN ASK HAVE THESE CELLS SEEN
ANTIGEN, AND INDEED CELLS THAT
ARE MOVING INTO THE LUNG WITH
ANTIGEN EXPERIENCE.
SO ARE THESE CELLS SPECIFIC FOR
SPIKE?
THE NEXT QUESTION WE LOOK BY
USING TETRAMER, YOU CAN SEE DAY
3 EVEN THOUGH IT WAS MOSTLY
ANTIGEN EXPERIENCED CELLS THEY
ARE NOT SPIKE SPECIFIC.
SOME ARE BUT THERE'S NOT
SIGNIFICANT INCREASE COMPARED
TO mRNA GROUP.
ON DAY 14 WE ARE ASKING
SIGNIFICANT DIFFERENCE BETWEEN
mRNA AND CHADO X-1 GROUPS THESE
CELLS ARE LIKELY TO STAY IN THE
LUNG.
IF YOU LOOK AT THE PURPLE
POPULATION YOU SEE
CO-EXPRESSION, WE SEE INCREASE
IN ANIMALS WITH CHADOX1 AT DAY
14 BUT NOT DAY 3.
WHAT ABOUT THE NOSE?
THE TISSUE I'M CURRENTLY
INTERESTED, A LOT OF PEOPLE HAVE
LOOKED AT THE LUNG BEFORE,-DAY
DATA IN UPPER RESPIRATORY TRACT,
STILL LACKING.
SO HERE WE'RE LOOKING AT ANTIGEN
EXPERIENCE CELLS, A NUMBER OF
THEM.
ON DAY 3 INFLUX OF CELLS IN THE
NOSE WITH THE CHADOX1 ON DAY 14
HAS EQUALIZED.
WHEN WE LOOK AT B CELLS SPIKE
SPECIFIC WE SEE DIFFERENCE
BETWEEN ANIMALS WITH CHADOX1
AND mRNA VACCINATION.
IN PARTICULAR ON DAY 14.
AND B CELLS ALSO MUCH MORE
LIKELY TO EXPRESS CD 69 AND
CD103.
WE ISOLATED CELLS FROM LYMPH
NODES, CERVICAL LYMPH NODE IS
DRAINING UPPER RESPIRATORY
TRACT, INGUINAL IS CLOSE TO
MUSCULAR VACCINATION.
WE SEE CELLS POSITIVE FOR CD8
TETRAMER SPIKE SPECIFIC, ANIMALS
WITH A CHADOX1 VACCINATION MUCH
HIGHER POPULATION IN CERVICAL
LYMPH NODES, SUGGESTING A LOCAL
RESPONSE WE'RE MEASURING.
WE CAN LOOK AT B CELLS, CD19
EXPRESSING ON THE SURFACE AS
WELL AS IgA, ON THE NOSE TOP
PANEL DAY 3 INFLUX OF CELLS IN
THE UPPER RESPIRATORY TRACT, IN
THE LUNGS THAT'S NOT HAPPENING
YET.
DAY 14 WE SEE THIS POPULATION
INCREASING UPPER AND LOWER
RESPIRATORY TRACT.
THIS IS EXPRESSION ON THE
SURFACE OF B CELLS, CAN ALSO
LOOK AT ACTUAL PRODUCTION OF
IgA USING B CELL.
WE'RE COMPARING CELLS ISOLATED
FROM THE LUNGS, LOOKING AT THE
SPLEEN THERE ISN'T A DIFFERENCE,
RESPONSE SIMILAR TO mRNA AND
CHADOX1 VACCINATED, CONTROL
ANIMALS DIDN'T HAVE IgA B
CELLS SITE SPECIFIC.
WE'RE ONLY SEEING IgA
PRODUCTION IN ANIMALS THAT GOT A
CHADOX1 VACCINATION.
WE USED NASOPLEX TO LOOK AT
ANTIBODY RESPONSES, YOU'RE
SEEING NINE ANTIGENS IN ONE
WELL.
AND FURTHER OUT FROM THE MIDDLE,
THE HIGHER THE BINDING RESPONSE
TO DIFFERENT ANTIGENS.
YOU CAN SEE mRNA GROUP IN
BLUE, CHADOX1 IN PURPLE, IN
SERUM 14 DAYS AFTER VACCINATION
NOT SEEING DIFFERENCE BETWEEN
THE TWO GROUPS.
WHEN WE LOOK IN RESPIRATORY
TRACT WE SEE STRIKING
DIFFERENCE.
NASAL ASSOCIATED LYMPHOID TISSUE
IS REPRESENTATIVE OF THE UPPER
RESPIRATORY TRACT, BRONCHIAL
ALVEOLAR LAVAGE IN LOWER,
BEAUTIFUL EXPRESSION OF IgA
AND IN THE CHADOX1 BUT NOT THE
INTRAMUSCULAR VACCINATION.
SO, IS THIS VACCINATION ABLE TO
REDUCE SHEDDING?
WE DID A STUDY IN HAMSTERS.
WE CHALLENGED ANIMALS AND TAKE
ORAL SWABS, YOU CAN SEE UPON
INTRANASAL VACCINATION WE SEE
REDUCTION IN THE AMOUNT OF
INFECTIOUS VIRUS SHED.
AND ON TOP OF THAT WE ALSO SEE A
REDUCTION IN AMOUNT OF
INFECTIOUS VIRUS IN LUNG TISSUE,
IMPORTANT BECAUSE WE DON'T
NECESSARILY WANT TO SACRIFICE
IMMUNITY IN LOWER RESPIRATORY
TRACT FOR IMMUNITY IN UPPER
BECAUSE THAT'S ASSOCIATED WITH
SEVERE DISEASE.
IN NON-HUMAN PRIMATE WE COMPARED
CHADOX 1 AND 19, GAVE TWO
VACCINATIONS FOUR WEEKS APART,
CHALLENGED FOUR WEEKS LATER,
SHEDDING IS REDUCED COMPARED TO
OUR CONTROL ANIMALS WHICH IS IN
CONTRAST TO WHAT WE SAW WITH
INTRAMUSCULAR VACCINATION.
LIKEWISE WHEN WE LOOK AT
BRONCHIAL A ALVEOLAR LAVAGE WE
SUE VERY LITTLE VIRUS.
LOWER RESPIRATORY TRACT WAS
NICELY PROTECTED IN VACCINATED
ANIMALS.
WE WANTED TO UNDERSTAND WHAT
FUNCTIONALITY IS OF THESE
ANTIBODIES.
THIS IS IN NON-HUMAN PRIMATE.
THIS WAS WORK DONE IN OUR LAB,
SHE SHOWED ANTIBODIES IN SERUM
WERE ABLE TO NEUTRALIZE VIRUS
RESULTING IN COMPLEMENT
ACTIVATION, OXYGENATION AND
PHAGOCYTOSIS AND N CASE CELLULAR
CYTOTOXICITY.
IgA WAS INDUCED, WE LOOKED AT
RESIDENTIAL T CELLS, TOOK A
LAVAGE SEVEN DAYS AFTER
VACCINATION, YOU CAN SEE CDAND
CD8 T CELLS START PRODUCING
INTERFERON GAMMA TNS ALPHA AND
SPIKE AT THAT TIME.
I THINK MUCOSAL IMMUNOLOGY WE
HAVE A LOT OF GAPS IN OUR
KNOWLEDGE STILL.
I ONLY MENTIONED A FEW HERE.
I KNOW THERE ARE MORE.
THESE ARE THE MOST IMPORTANT
ONES THAT WE SHOULD TRY TO
ANSWER RIGHT NOW.
I'VE SHOWN INDUCTION OF IMMUNE
PARAMETERS IN RESPIRATORY TRACT.
I'VE SHOWN SOME PROTECTION.
THOSE ARE NOT CORRELATEIVE
PROTECTION.
WE REALLY NEED TO UNDERSTAND
CORRELATE OF PROTECTION.
WHAT DO WE NEED TO PROTECT UPPER
RESPIRATORY TRACT?
SECONDLY, WHAT MUCOSAL IMMUNITY
IS INDUCED BY INFECTION WITH
VIRUSES, HOW DOES IT DIFFER WITH
VACCINES?
I'M ONLY SHOWING VACCINATION
DATA, VERY LITTLE WORK IN
SARS-COV-2 FIELD LOOKING AT
MUCOSAL IMMUNOLOGY, DIFFERENT
STORY WITH INFLUENZA.
BUT I THINK IT'S IMPORTANT TO
UNDERSTAND DIFFERENCE.
WE KNOW WE CAN GET REINFECTED
WITH SARS-COV-2 SO HOW IS THAT
IMMUNITY INDUCED BY INFECTION.
ARE DIFFERENCES BETWEEN MUCOSAL
IMMUNOLOGY AND UPPER AND LOWER
TRACTS, THE ANSWER IS YES BUT WE
NEED TO UNDERSTAND THIS BETTER.
AND FINALLY ARE CERTAIN VACCINE
PLATFORMS BETTER AT INDUCING
MUCOSAL IMMUNE RESPONSE, I THINK
THE ANSWER IS YES, SUBUNIT
VACCINE IS NOT AS GOOD, FOR
EXAMPLE, AS A LIVE ATTENUATED
VIRUS BUT WE NEED TO SHOW THIS.
OKAY.
WHAT IS NEEDED?
AS I MENTIONED BEFORE
HOST-SPECIFIC REAGENTS ARE
IMPORTANT HERE.
HAMSTERS AND FERRETS, CURRENTLY
DIFFICULT.
WE WANT TO UNDERSTAND WHAT'S
HAPPENING IN HUMANS AS WELL.
SO CAN RESULTS BE OBTAINED IN
ANIMAL STUDIES BE TRANSLATED TO
HUMANS?
I THINK EXPERIMENTAL INFECTION
STUDIES ARE FASCINATING.
WE NEED TO DO MORE OF THOSE.
WE CAN ALSO GO TO PEOPLE THAT
HAVE INFECTION AND START ASKING
WHAT IS HAPPENING IN UPPER
RESPIRATORY TRACT.
WE NEED TO OPTIMIZE A TOOL.
THINK ABOUT HOW YOU COLLECT
THOSE, RETRO ORBITAL INFECTION
OF CD45, NASAL TURBINATE DURING
INJECTION OF CD45 IN 9 NOSE
BECAUSE YOU GET MORE
CONTAMINATION OF CELLS COMING
FROM THE BONE MARROW.
I ALREADY TOLD RYAN I LOVE THE
DIRTY MICE, WE SHOULD BE USING
THEM.
OVERALL WE NEED TO INCREASE
UNDERSTANDING OF MUCOSAL
IMMUNOLOGY ESPECIALLY IN
RESPIRATORY TRACT.
WITH THAT I WOULD LIKE TO THANK
EVERYBODY THAT'S INVOLVED IN
THESE STUDIES, IN PARTICULAR
VINCENT FOR ALLOWING ME TO DO
STUDIES IN HIS LAB AND THE
PEOPLE THAT HELPED, AND FINALLY
MY LONGSTANDING COLLABORATORS
TERESA LAMBE AT SARAH GILBERT AT
OXFORD UNIVERSITY.
>> WE HAVE TIME FOR A FEW
QUESTIONS.
>> THAT WAS NICE.
I'M CURIOUS, USING THE mRNA
VACCINE, YOU USED ADENOVIRUS
VACCINE.
ADENOVIRUS VACCINE WILL
INTERNALIZE AND REPLICATE, AT
LEAST YOU'LL GET INFECTION IN
NEIGHBORING CELLS, OR INTO
CELLS.
HAVE YOU TRIED JUST LIKE
INACTIVATED VIRUS?
LOOKING EITHER GIVEN
INTRAMUSCULARLY VERSUS
INTRANASALLY TO THE RESPONSE IN
THE NALT?
>> WE HAVEN'T HAD TIME YET.
WHAT VACCINE PLATFORM WE USE IS
GOING TO BE REALLY, REALLY
IMPORTANT IN WHAT IMMUNE
RESPONSE YOU GET.
SUBUNIT WE NEED TO USAGE
ACKNOWLEDGE ADVANCE.
>> IN FERRETS ACTIVE REPLICATION
RATHER THAN ACTIVATED WHOLE
VIRUS PROVIDES SOME LEVEL OF
PROTECTION FROM TRANSMISSION IN
OUR CASE, THAT'S WHERE THE
MUCOSAL IMMUNITY MIGHT BE
IMPORTANT FOR A VIRUS, AT LEAST
LOW LEVEL, LIVE ATTENUATED
VACCINES, POTENTIAL PLATFORMS
MIGHT BE INTERESTING.
>> I HAVE A STUDENT IN SEPTEMBER
LOOKING AT TRANSCRIPTOMICS OF
THE VACCINATION, TRYING TO
UNDERSTAND WHAT THE IMMUNE
RESPONSE ACTUALLY LOOKS LIKE,
WHAT INFLAMMATION WE'RE SEEING
AFTER VACCINATION.
I THINK THAT'S GOING TO GET AT
THOSE QUESTIONS AS WELL.
>> A SIMILAR FOLLOW-UP QUESTION,
BECAUSE YOU'RE FOLLOWING WITH
THE LIVE VIRUS DO YOU HAVE ANY
ANTI-VECTOR IMMUNITY OR DO YOU
NOT HAVE ANTI-VECTOR COMMUNITY
BECAUSE mRNA PROVIDES PURE
MUCOSAL IMMUNITY, IS THIS
SOMETHING TOO NEEDS TO BE
CONSIDERED.
>> WITH THE CHADOX 1 IT DOESN'T
HAVE SPIKES ON THE SURFACE.
SOME STUDIES NOW, A LOT OF
PEOPLE ASK ABOUT VECTOR
IMMUNITY, A VERY IMPORTANT
QUESTION TO ASK.
SHE'S VACCINATED WITH ONE
CHADOX1, COMPARED TO A SINGLE
VACCINATION, SEEING FAIRLY
SIMILAR RESPONSES.
WE HAVE ALWAYS SHOWED WHEN WE DO
A BOOST WITH THE SAME VACCINE WE
DO GET A BOOST IN RESPONSE AS
WELL.
I THINK VECTOR IMMUNE RESPONSES
WILL LIKELY PLAY A ROLE.
I DON'T KNOW HOW MUCH OF A ROLE
THEY WILL PLAY.
>> TYPE 3 INTERFERENCE VERY
IMPORTANT FOR MUCOSAL IMMUNITY.
DID YOU TEST WHETHER -- THE
RESPONSE?
>> NO, I FULLY AGREE, THOSE ARE
STUDIES THAT WE HAVEN'T HAD THE
TIME YET TO DO.
YEAH, THOSE ARE ALL THINGS.
WE DO TRANSCRIPTOMICS IN THESE
ANIMALS, WE JUST FINISHED A
STUDY WHERE WE CHALLENGED THESE
MICE THAT HAD THESE
VACCINATIONS, AND WE HAVE
TISSUES, WHICH WE'RE GOING TO
LOOK EXACTLY AT THAT.
>> THE REASON I ASK, RECEPTOR
AND KNOCKOUT, HAVE YOU TRIED TO
CONVINCE THE WORLD TO USE THEM,
UNTIL RECENTLY -- YEAH, I NEED
MORE.
THANK YOU.
>> ONE QUICK QUESTION, IN A
WORLD IN WHICH WE AGENT
LIMITATIONS ARE NOT AN ISSUE,
MAYBE COST LIMITATIONS WHAT IS
YOUR PREFERRED MODEL FOR THESE
THINGS?
>> THE HUMAN.
>> WHAT IS YOUR PREFERRED SMALL
ANIMAL MODEL?
>> I THINK FOR SARS 2, JUST A
PRACTICALITY THING, THE HAMSTER
IS REALLY GOOD AT LOOKING AT
TRANSMISSION STUDIES.
IF YOU WANT TO TAKE IT FURTHER
AND LOOK AT TRANSMISSION OF THE
VIRUS, WHEN YOU START LOOKING AT
THE FERRET, THE PHYSIOLOGY OF
THE LUNG IS MUCH MORE LIKE A
HUMAN.
>> THANK YOU.
WE'LL TAKE A 15-MINUTE BREAK NOW
AND COME BACK AT 3:30.
WE'LL HAVE THE LAST TALK OF
THE SESSION, AND WE'LL HAVE A
PANEL DISCUSSION.
SO THE SPEAKER NOW IS MEHUL
SUTHAR, WHO WILL TALK ABOUT
EXAMINING THE INNATE AND
ADAPTIVE IMMUNES RESPONSES TO
SARS-COV-2 INFECTION.
>> THANK YOU SO MUCH.
I WANT TO THANK THE ORGANIZERS
ESPECIALLY MICHELLE FOR GIVING
ME OPPORTUNITY TO PRESENT WORK
IN MY LAB ABOUT MOUSE MODELS OF
SARS-COV-2.
WE ANCHOR ON WHAT OBSERVATIONS
WE'RE MAKING IN HUMANS TO
DEVELOP SOME LEVEL OF
UNDERSTANDING IN MICE AND FEED
THAT BACK INTO WHAT MIGHT BE
HAPPENING IN HUMANS BASED ON
OBSERVATIONS OF MICE.
IF THE SWISS CHEESE MODEL THAT
DEMONSTRATES THE MANY LAYERS OF
IMMUNITY THAT ARE PRESENT, THE
SARS-COV-2 VARIANTS OVERCOMING
THROUGH HOLES IN OUR IMMUNE
SYSTEM.
ELEMENTS OF INNATE RESPONSE, B
CELL RESPONSE, C TELL, RESIDENTS
CIRCULATING PLAYING A ROLE.
WE THINK ABOUT THIS WE WOULD
LIKE TO MODEL DIFFERENT ASPECTS
IN MICE.
ONE OF THE FIRST THINGS IN EARLY
2020 WAS WORK CLOSELY TO HELP
DEVELOP SARS-COV-2 MOUSE MODELS
IN NATURE AND DEVELOP MODELS AND
UNDERSTAND INNATE AND ADAPTIVE
IMMUNE RESPONSE, SOME WORK WE
RECENTLY -- WORK RECENTLY
ACCEPTED TO UNDERSTAND
CONTRIBUTION OF T CELLS IN THE
RESPIRATORY TRACT, PRIMARY
INFECTION.
LASTLY I'LL TOUCH ON WORK THAT
IS IN PREPARATION TO UNDERSTAND
AND MODEL VACCINE BREAKTHROUGH,
AS MICHAEL ALLUDED TO, POTENTIAL
PROTECTIVE ROLE OF EOSINOPHILS
IN THIS CONTEXT.
WE'VE DISCUSSED THIS BUT N 501Y
OCCURRED PRIOR TO THAT,
INVESTIGATORS IN BEIJING HAD
PASSAGED THEIR STRAIN OF I
BELIEVE B1 IN MICE RESULTING IN
A MUTATION AT N 51Y ALLOWS FOR
MOUSE INFECTION, HELPED
INTERACTION WITH MOUSE ACE-2
WITH HISTIDINE AT 353.
THAT WAS FURTHER PASSAGED IN
BALB-C MICE, MUTATIONS IN THE
SPIKE PROTEIN, N 501Y WAS
RETAINED, THIS VIRUS PICKED UP
NK 417N NOW NATURALLY OCCURRING
IN VARIANTS, AGAIN PRIOR TO
EMERGENCE OF VARIANTS.
WE SAW MUTATION AT POSITION 655Y
THAT OCCURRED WITHIN THE SPIKE
PROTEIN AND COLLECTIVELY THIS
ALLOWED FOR THIS VIRUS TO CAUSE
WEIGHT LOSS IN MICE, NICE LUNG
VIRUS REPLICATION, LIMITED
EXTRA-PULMONARY VIRUS
REPLICATION, SOME IN THE HEART
HE OBSERVED, THIS REPLICATED IN
THE LARGE AIRWAY WITH LESS
REPLICATION IN THE SMALLER
AIRWAYS BUT THERE WAS
INFLAMMATION THAT OCCURRED, I'LL
SLOW SOME DATA WE PRODUCED USING
THIS VIRUS WE STARTED TO WORK
WITH CMA 3 P20 BUT THE BETA
VARIANT IS A WORK HORSE, AFFECTS
MICE, DOSE DEPENDENT, AGE
DEPENDENT, WEIGHT LOSS 8 TO 20
WEEKS, SIMILAR TO SARS-COV-1,
YOU SEE INCREASE IN WEIGHT LOSS
WITH INCREASE IN AGE, 5 TO 10%
INTRANASALLY RESULTING IN 30%
MORTALITY IN THESE MICE.
MOST STUDIES WE TYPICALLY USED 1
TIMES 10 TO THE 6 AND COMPARED
SIDE BY SIDE WITH MOUSE ADAPTED
STRAINS IN DATA NOT SHOWING BUT
VIRUS REPLICATES WELL IN THE
LUNGS, CLEARED INFECTIOUS
VIRUS-WISE BY DAY 10 POST
INFECTION, INDUCED NICE
SPIKE-SPECIFIC ANTIBODIES,
NUCLEOCAPSID ANTIBODIES,
NEUTRALIZING, AND YOU SEE VERY
NICE INNATE IMMUNE RESPONSES,
SEVERAL SPEAKERS HAVE SHOWN
PRIOR TO THIS PRESENTATION.
IN MY LAB WE HAVE A BASIC
QUESTION, WHAT IS HAPPENING IN
THE LUNGS AND HOW DOES THIS LOOK
SIMILAR TO WHAT WE SEE IN
HUMANS.
ACROSS MULTIPLE MODELS WE USE A
LABELING TECHNIQUE TO TAG TISSUE
RESIDENT CELLS VERSUS
CIRCULATING CELLS, ISOLATED BY
FLOW CYTOMETRY, AND FOLLOWING
SARS-COV-2 INFECTION THIS IS
WORK PUBLISHED IN 2020, 2021, WE
SEE INFLUX OF NEUTROPHILS,
EOSINOPHILS, MONOCYTES COMING
IN, INFLAMMATORY MONOCYTES,
DIFFERENTIATED BASED ON LY 6
HIGH AND LOW, AND USE LABELING,
ISOLATE CELLS, WE USE MAGNETIC
SORT COLUMN OR MAGNETIC
DEISOLATION TO ISOLATE, TAKE
NEGATIVE CELLS WHICH ARE TISSUE
RESIDENT AND PERFORM SINGLE CELL
RNAseq, LOOKING AT TISSUE
RESIDENCE, INNATE IMMUNE CELLS,
PERFORM CLUSTERING ANALYSIS,
SHOWING SIMILAR TO FLOW
CYTOMETRY, ALL THESE POPULATIONS
OF CELLS, SIMILAR TO WHAT
MICHAEL MENTIONED, ALVEOLAR
MACROPHAGE POPULATION DECREASE
FOLLOWING INFECTION, REBOUNDING
30 DAYS AFTER INFECTION.
ONE OF THE THINGS WE OBSERVED IN
THIS SINGLE CELL RNAseq IS
UPREGULATION, WHAT ROLE DOES IT
PLAY?
WE GOT OUR HANDS ON KNOCKOUT
MICE, WE SEE GREATER WEIGHT LOSS
BY DAY 3, DAY 4, POST-INFECTION,
SOME MICE DO RECOVER.
60% MORTALITY DEMONSTRATING
IMPORTANCE THAT CCR 2 PLAYS AND
INNATE IMMUNE RESPONSE
PROTECTING AGAINST INFECTION.
WE SEE SLIGHT INCREASE IN VIRAL
LOAD.
IN WILDTYPE MICE WE SEE MUCH
MORE RESTRICTED VIRUS
REPLICATION IN LARGE AIRWAYS.
CCR 2 KNOCKOUTS MORE EXPANDED
REPLICATION BEYOND JUST LARGE
AIRWAYS, WE SEE IT IN THE SMALL
AIRWAYS, INNATE RESPONSE IS VERY
IMPORTANT FOR RESTRICTING WHERE
THIS VIRUS REPLICATES WITHIN THE
LUNGS.
WE'VE DONE SOME WORK LOOKING AT
TYPE I INTERFERON GROUP, RALPH'S
GROUP LOOKED, KNOCKOUT MICE LIVE
IN SARS-COV-2 INFECTION, ONE OF
THE FEW VIRUSES I KNOW OF THAT
DOESN'T KILL IT IN FRONT OF A
KNOCKOUT MOUSE.
TYPE 3 INTERFERONS ARE VERY
IMPORTANT, MAY BE OVERLAPPING
FUNCTION BETWEEN TYPE I AND TYPE
II INTERFERON.
ONE OF THE MORE IMPORTANT
QUESTIONS WE'RE NOW STARTING TO
ASK WHAT IS THE IMPORTANCE OF
THE UPPER VERSUS LOWER
RESPIRATORY TRACT, I'LL SO DATA
FROM T CELLS, GOING BACK TO
SYSTEMATIC ACROSS TIME POINTS
FROM ACUTE INFECTION TO RECOVERY
IN THE UPPER AND LOWER
RESPIRATORY TRACT OF INFECTION.
I'LL SO DATA ABOUT VACCINE
BREAKTHROUGH, WHAT IS THE ROLE
OF THE INNATE IMMUNE RESPONSE?
AND SO WE'RE FAMILIAR THAT T
CELLS ARE VERY IMPORTANT IN
SARS-COV-2 INFECTION IN HUMANS.
T CELLS ARE INDUCED, YOU SEE IN
PERIPHERAL BLOOD.
MANY HAVE SHOWN T CELLS ARE
INDUCED IN PERIPHERAL BLOOD,
MAINTAINED, QUITE STABLE IN
DURABILITY, AFTER INFECTION OR
VACCINATION.
ONE OF THE MORE RECENT FINDINGS
PEOPLE HAD LOOKING IN HUMANS
BREAKTHROUGH INFECTIONS DO
INDUCE NASAL CD4 AND CD8 T-CELL
RESPONSES.
WE WANTED TO ASK IN MOUSE
MODELS, BLACK 6 MOUSE MODELS,
WHAT IS THE ROLE OF T CELLS?
USING LABELING, HARVESTING
TISSUE FROM LUNG AND NASAL
COMPARTMENT, ALSO THE SPLEEN,
I'M NOT SHOWING THAT DATA TODAY.
FOLLOWING INFECTION TO REDUCE
RECRUITMENT OF CD4 AND CD8 T
CELLS IN THE LUNGS AS WELL AS
WITHIN THE NASAL COMPARTMENT,
THIS IS LOOKING AT DAY 8
POST-INFECTION.
WE SEE 3 1/2-FOLD INCREASE IN
CD8 T CELLS IN THE LUNG, 1st
INCREASE OF CD4 T CELLS,
STRONGER INCREASE IN THE NASAL
COMPARTMENT, SUGGESTING CD8 T
CELLS MAY PLAY IMPORTANT ROLE IN
UPPER RESPIRATORY TRACT.
WE PERFORMED PEPTIDE
RESTIMULATION, ANTIGEN SPECIFIC
RESPONSES.
LOOKING AT GRANZYME B,
UPREGULATION IN NASAL AND LUNG
COMPARTMENT FOLLOWING INFECTION.
CD 4 CELLS WE SEE BOTH SINGLE
POSITIVE, INTERFERON GAMMA, TNF,
DOUBLE POSITIVE CELLS.
THERE ARE SLIGHT DIFFERENCES IN
WHAT WE'RE SEEING BETWEEN THE
UPPER AND LOWER RESPIRATORY
TRACT.
FOR INSTANCE IN NASAL
COMPARTMENT, IT'S PREDOMINANTLY
SINGLE POSITIVE TNF ALPHA AND
INTERFERON GAMMA POSITIVE CELLS.
IN THE LUNGS WE DO SEE
POPULATION OF DOUBLE POSITIVE
CELLS FOR CD4.
GO TO CD8 T CELLS WE SEE BETWEEN
THE LUNGS AND NASAL COMPARTMENT
HIGHER LEVELS OF TNF ALPHA
PRODUCING CD8 T CELLS IN THE UP
HE WERE.
IN THE LUNGS WE SEE LOWER LEVELS
OF TNF ALPHA RELATIVE TO
INTERFERON GAMMA SO THERE ARE
ANATOMIC DIFFERENCES BEHIND HOW
THESE CD8 T CELLS FUNCTION
WITHIN THE UPPER AND LOWER
COMPARTMENT.
WE LOOKED AT DURABILITY THROUGH
28 DAYS WITHIN THESE
COMPARTMENTS AS SIMPLY DO THESE
T CELLS -- ARE THEY MAINTAINED
IN COMPARTMENTS DIFFERENTLY.
WE LOOKED IN THE LUNGS.
SORRY, NASAL COMPARTMENT.
WE LOOKED AT ANTIGEN SPECIFIC,
WHETHER TNF ALPHA OR INTERFERON
GAMMA.
WE SEE A SINGLE POSITIVE, SEEMED
TO BE STABLE FOR CD4 CELLS,
DOUBLE POSITIVE CELLS MORE
STABLE WITHIN THE LUNGS COMPARED
TO NASAL COMPARTMENT.
CD8 T CELLS WE SEE DIFFERENT
PROFILE, MORE STABLE IN THE
NASAL COMPARTMENT COMPARED TO
LUNGS.
INTERFERE GAMMA HIGHER IN THE
LUNGS, DIMINISHES, IF YOU LOOK
AT DOUBLE POSITIVE CELLS THESE
CELLS SHOW HIGHER LEVEL AT DAY
7, DIMINISH THROUGH DAY 28.
WE LOOKED AT THE CONTRIBUTION OF
CD4 AND CD8 T CELLS OR BOTH IN
TANDEM, FOLLOWING SARS-COV-2 SO
WE TOOK A VERY SLEDGEHAMMER
APPROACH TO MAKE SURE T CELLS
HAVE BEEN DEPLETED, INJECT MICE
THREE TIMES BEFORE WE INSPECT
AND EVERY WEEK PROVIDE
ADDITIONAL ANTIBODIES THROUGH
DAY 28 POST-INFECTION.
CD8 T CELLS AND CD4 T CELLS
DEPLETED QUITE WELL WITHIN THE
LUNGS AND NASAL COMPARTMENT.
FOR THE MOST PART WE SEE THESE
DEPLETED IN THE BLOOD AS WELL
THROUGH DAY 28, NOT JUST BEFORE
INFECTION, WE FOLLOW THESE
THROUGH INFECTION AS WELL.
SURPRISINGLY, THE LACK OF T
CELLS DOES NOT KILL A MOUSE,
WHICH FURTHER SUPPORTS INNATE
IMMUNE RESPONSE IS QUITE
IMPORTANT FOR PROTECTION.
WE DON'T SEE MUCH OF A
DIFFERENCE IN WEIGHT LOSS WITH
SINGLE DEPLETION OR TANDEM
DEPLETED MICE, MIRRORS WHAT
RALPH'S LAB HAD SEEN MANY YEARS
AGO WITH SARS-COV-1, IF YOU
DON'T HAVE THE CELLS THE MICE
LIVE, QUITE IMPORTANT FOR
DRIVING PROTECTION.
THAT'S ONE OF THE SIMILAR
ASPECTS BETWEEN BOTH VIRUSES.
IN THE LUNGS DAY 3, DAY 10 AND
DAY 28, TYPICALLY WHERE YOU HAVE
VERY LOW LEVELS OF VIRAL RNA.
IN EITHER THE SINGLE OR DOUBLE
DEPLETED MICE BY DAY 28
POST-INFECTION THIS VIRUS IS
PRETTY WELL CONTROLLED.
DAY 10 THERE MAY BE SLIGHT
DIFFERENCE IN VIRAL CONTROL AS
TANDEM DEPLETED MICE CD4 AND CD8
DEPLETED SHOW HIGHER LEVELS OF
VIRUS REPLICATION, IMPACT OF
VIRAL KINETICS.
WITHOUT T CELLS THESE MICE
COMPLETELY CONTROL INFECTION.
LOOKING IN NASAL COMPARTMENT
DEPLETION OF CD4 T CELLS,
SLIGHTLY HIGHER ELEVATED BUT
TANDEM DEPLETED ARE NOT
SIGNIFICANTLY HIGHER.
DAY 28 POST-INFECTION TANDEM
DEPLETED MICE SHOW HIGHER LEVEL
OF VIRUS REPLICATION COMPARED TO
EITHER SINGLE DEPLETED CD4 CD8
OR CONTROL, SUGGESTING
IMPORTANCE FOR CLEARING VIRUS
INFECTION.
WE WANTED TO ASK WHAT ROLE DO
ANTIBODIES PLAY?
IF YOU LOOK AT SPIKE BINDING,
RBD OR NUCLEOCAPSID BINDING, YOU
LOSE ANTIBODY RESPONSES SO IT'S
A T CELL-DEPENDENT RESPONSE,
VERY SIMILAR TO WHAT YOU SEE IN
HUMANS, AS WELL AS OTHER ANIMAL
MODELS.
IF WE GET RID OF CDs 8, NO
IMPACT.
WHEN YOU GET RID OF CD4, LOSE
NEUTRALIZATION, CD8 DON'T REALLY
HAVE EFFECT, IT'S NOT DUE TO
ANTIBODIES OR LACK OF
ANTIBODIES, IT HAS TO DO WITH T
CELLS THAT ARE PRESENT IN THE
UPPER RESPIRATORY TRACT.
WE WANTED TO ASK CAN WE ISOLATE
INFECTIOUS VIRUS IN THE UPPER
RESPIRATORY TRACT, VIRAL RNA
THAT'S PERSISTING OR IS IT
INFECTIOUS VIRUS.
FROM ALL THE MICE WE HAD TANDEM
DEPLETED, ONLY IN THOSE MICE CAN
WE RECOVER INFECTIOUS VIRUS FROM
THESE UPPER RESPIRATORY TRACT,
CAN'T RECOVER THE VIRUS FROM
LOWER RESPIRATORY TRACT.
WE WANTED TO ASK WHERE IS THE
VIRUS REPLICATING.
THIS IS DONE IN COLLABORATION
WITH ROBIN KLEIN'S LAB, MY
FORMER POSTDOC ABIGAIL, THERE'S
ABSOLUTELY NO VIRAL RNA OR VIRAL
PROTEIN STAINING WITHIN THE
BRAIN.
DESPITE HAVING PERSISTENT VIRUS
REPLICATION IN UPPER RESPIRATORY
TRACT WE'RE NOT SEEING THE VIRUS
GO INTO THE BRAIN.
SO THAT I THINK WAS VERY
IMPORTANT OBSERVATION GIVEN SOME
CONFLICTING DATA WE'VE SEEN WITH
THE K18 MOUSE MODEL WHERE YOU
GET INFECTION OF THE BRAIN, IN
THIS MODEL WE DON'T SEE THAT.
THERE'S VARIATION BETWEEN MICE
WHERE IT REPLICATION AND HOW
MUCH BUT REGARDLY WE CONTINUALLY
SEE PERSISTENT VIRAL INFECTION.
WE NEXT WORKED WITH THE NIH TO
ASK DOES THAT PERSISTENT VIRUS
CHANGE OVER TIME.
SO WE DID A COMPLICATED
ANALYSIS, WE HAVE OUR INNOCULUM
STOCK, DEEP SEQUENCE THAT, DEEP
SEQUENCE VIRUS PASSAGE TO SEE IF
THERE'S CHANGES.
WE ALSO DEEP SEQUENCE VIRUS
FOUND AT DAY 3 POST-INFECTION TO
SEE IF THE FIRST THREE DAYS IS
THERE ANY CHANGE THAT OCCURS.
WE DEEP SEQUENCE THE VIRUS WE
GET AT DAY 28 POST-INFECTION.
AND WHAT WE SEE IS THAT THROUGH
EITHER INNOCULUM STOCK,
INNOCULUM WE PASSAGE ONCE, WHAT
WE DO WHEN WE ISOLATE FROM THE
UPPER RESPIRATORY TRACT OR THRE
DAYS POST-INFECTION VIRTUALLY NO
CHANGES.
REALLY NO CHANGES IN VIRUS
SEQUENCES THROUGH DAY 3
POST-INFECTION.
MOST CHANGES OCCUR BY DAY 28
POST-INFECTION.
THESE ARE SOMEWHAT SPORADIC,
THEY OCCUR AT VARIOUS PLACES.
I'LL SHOW THAT IN THE NEXT
SLIDE.
BY DAY 28 YOU DO START TO SEE
QUASI-SPECIES OCCURRING IN THIS
INNOCULUM STOCK.
WHERE ARE THESE OCCURRING?
THEY SEEM TO BE OCCURRING
PRIMARILY WITHIN THE
NON-STRUCTURAL REGION.
AGAIN, DIFFERENT BETWEEN
DIFFERENT MICE.
VERTICALLY ARE DIFFERENT MICE
THAT WERE SEQUENCED.
AND WHERE THESE MUTATIONS FALL.
WHAT'S INTERESTING IS WE
OBSERVED SOME AT LEAST FOR THE
MICE HAD DELETIONS IN ORF6, ORF
6 POTENTIALLY LINKED TO
REGULATION OF THE INNATE IMMUNE
RESPONSE IN THE CONTEXT OF
SARS-COV-2.
SO THERE MAY BE SOME PRESSURE
THAT'S PLACED ON THIS VIRUS AS
IT PERSISTS TO PERSISTENTLY LIVE
IN THE NOSE. THERE MAY BE SOME
CHANGES OCCURRING.
WHAT WAS SURPRISING WE DIDN'T
SEE MANY CHANGES IF ANY WITHIN
THE SPIKE PROTEIN DURING THESE
FIRST 28 DAYS.
THIS IS SOMETHING WE'RE NOW
FOLLOWING UP ON, WHAT THE IMPACT
OF THESE MUTATIONS ACTUALLY ARE
ON PERSISTENCE.
ANOTHER ANGLE WE ALSO WANTED TO
LOOK AT WAS TO ASK NOW THAT
WE'VE DEVELOPED A PERSISTENT
INFECTION MODEL, WE WANTED TO
ASK HOW DO ANTI-VIRALS WE USE
WORK IN THIS MODEL.
WE HANDED THIS OVER TO GEORGIA
STATE UNIVERSITY, THE PAPER IS
IN bioRxiv FOR REVIEW, HIS
LABORATORY RECAPITULATED
OBSERVATIONS WE MADE IN TERMS OF
VIRAL PERSISTENCE AND TESTED TWO
ANTI-VIRAL DRUGS PAXLOVID AND
MALNOPIRAVIR, ARE THEY CAPABLE
OF REDUCING.
IF YOU LOOK AT PANEL E,
MALNOPIRAVIR THROUGH DAY 7 OF
TREATMENT, AND 14 DAYS AFTER THE
INITIAL TREATMENT, MALNOPIRAVIR
WAS ABLE TO ELIMINATE VIRUS IN
UPPER RESPIRATORY TRACT.
PAXLOVID WAS NOT ABLE TO FULLY
REDUCE REPLICATION BUT THIS DOES
ALLOW ONE TO TAKE A MODEL OF
PERSISTENT VIRAL INFECTION AND
ASK QUESTIONS ABOUT HOW
ANTI-VIRALS CAN WORK IN THE
RESPIRATORY TRACT, ESPECIALLY
UPPER, NOW LOOKING AT INNATE
IMMUNE RESPONSE, OTHER ASPECTS
OF I ADAPTIVE IMMUNE RESPONSE TO
UNDERSTAND WHAT IS CONTRIBUTING
TO VIRAL PERSISTENCE.
THE LAST BIT I'LL TALK ABOUT ONE
STORY WE'RE WORKING ON, TO
CONNECT SOME WORK WE'RE DOING
WITH SEROLOGY ASSAYS IN THE
LABORATORY LOOKING AT WANING
IMMUNITY IN THE CONTEXT OF
HUMANS THAT GET VACCINES OR
INFECTION.
WHAT DOES IT MEAN IN VIVO WHEN
ONE GETS A BREAKTHROUGH
INFECTION?
AND SO THERE'S SEVERAL WAYS FOR
US TO MODEL THIS.
AND SO ONE OF THE THINGS WE
WANTED TO BETTER UNDERSTAND IS
THAT WHEN YOU HAVE LOWER LEVELS
OF PROTECTION, YOU HAVE HIGHER
LEVELS OF VIRUS REPLICATION, AS
YOU INCREASE PROTECTION, AND
VACCINE RESPONSES YOU CAN
CERTAINLY DRIVE PROTECTION, AS
WE'VE SEEN IN THE LAST
PRESENTATION.
WE WANTED TO KNOW WHAT DOES THIS
MEAN IN VIVO AND HOW DOES THE
NATURE IMMUNE RESPONSE HELP
CONTROL INFECTION?
THIS IS ANOTHER SCHEMATIC OF THE
APPROACH WE'RE TAKING.
IN THE NAIVE INFECTION I'VE
SHOWN MONOCYTES PLAY A VERY
IMPORTANT ROLE, T CELLS PLAY AN
IMPORTANT ROLE IN THE UPPER AND
POSSIBLY KINETIC OF VIRAL
CONTROL IN THE LOWER RESPIRATORY
TRACT.
INNATE MOLECULES AND CELLS PLAY
AN IMPORTANT ROLE, WHAT DOES IT
MEAN IN THE CONTEXT OF
VACCINATION?
WE'VE SEEN FROM WORK FROM MIKE
DIAMONDS GROUP AND OTHERS
ANTIBODY EFFECTOR MECHANISMS
PLAY A CRITICAL ROLE IN DRIVING
PROTECTION.
T CELLS ALSO PLAY QUITE AN
IMPORTANT ROLE IN WORK DAN
BAROUCH AND OTHERS HAVE DONE AS
WELL.
ONE WAY TO MODEL IN THE
LABORATORY IS BY USING PRIMER
SPIKE PROTEIN WITH ADDAVAX,
VACCINATE MICE AND BOOST THEM 30
DAYS LATER, CHALLENGE THEM.
AND WE USE TWO DOSES TO BE ABLE
TO MODEL WANING IMMUNITY, IN THE
ABSENCE OF BEING ABLE TO
ACTUALLY HOLD THESE MICE FOR SIX
MONTHS OR A YEAR, WE TRY TO GIVE
TWO DOSES SPREAD OUT BY ABOUT
TEN-FOLD, TO BE ABLE TO AT LEAST
GET INFECTION OF THE LUNGS.
THAT'S OUR GOAL.
AND SO WHEN WE NOW VACCINATE
THESE MICE, AT FIVE MICROGRAMS,
.5 MICRO GAPS, SAME LEVEL, WE
CHALLENGE THEM WITH BETA
VARIANT, IN THIS CASE WA 1 SPIKE
PROTEIN, A BIT OF DIVERGENCE IN
SPIKE VERSUS VIRUS.
NAIVE 15 TO 20% WEIGHT LOSS.
FIVE MICROGRAM DOSES DON'T SEE
WEIGHT LOSS, MICE ARE WELL
PROTECTED.
.5 MICROGRAMS INTERMEDIATE
WEIGHT LOSS, FOR THE MOST PART
PROTECTED.
WE SEE IN THE LUNGS WE DON'T SEE
VIRUS REPLICATION, INFECTIOUS
VIRUS AT FIVE.
AT .5 LOG, LOG AND A HALF
REDUCTION.
WE WANTED TO UNDERSTAND INNATE
IMMUNE RESPONSE CORRECTING TO
PROTECTION.
WE LOOKED AT STAINED SECTIONS,
IN SITU HYBRIDIZATION.
THIS VIRUS REPLICATES IN THE
LARGE AIRWAYS, YOU DO START TO
SEE INFLAMMATION AROUND THESE
AREAS OF VIRUS REPLICATION.
.5 5 MICROGRAMS LOTS OF INFECTN
IN THE LUNGS, LOTS OF DEBRIS IN
THE LARGE AIR WAYS WHERE VIRAL
NUCLEOCAPSID STAINING IS
OCCURRING.
AT FIVE MICROGRAMS DON'T SEE
VIRUS REPLICATION IN MICE,
SIMILAR TO WHAT WE'VE SEEN WITH
INFECTIOUS VIRUS REPLICATION, WE
I.D. LABEL, HARVEST IMMUNE
CELLS, PERFORM FLOW CYTOMETRY OR
DO SINGLE CELL RNA SEQUENCING.
WE SEE AT LOW DOSE VACCINATION
WE SEE HIGHER LEVELS OF
INFLAMMATION, SIMILAR TORE H&E,
MUCH HIGHER THAN NAIVE
INFECTION.
ONE OF THE MORE STRIKING
OBSERVATIONS THAT WE MADE IS
THAT IN THE LOW DOSE VACCINATED
MICE YOU SEE HIGHER PROPORTION
OF EOSINOPHILS IN THE LUNGS
COMPARED TO NAIVE INFECTION.
IT IN NAIVE MORE DOMINATED BY
NEUTROPHILS RATHER THAN
EOSINOPHILS.
GRANULOCYTE POPULATION, IT'S
STRIKING, YOU SEE A HIGH
PERCENTAGE OF EOSINOPHILS COMING
IN, COMPARED TO JUST INFECTION
ALONE, THIS CORRESPONDS TO
FREQUENCY OR TOTAL COUNT, TWO
LOG INCREASE IN LEVEL OF
EOSINOPHILS IN LOW DOSE
VACCINATED MICE INFECTED
COMPARED TO NAIVE INFECTED MICE.
THESE EOSINOPHILS INVOLVED IN
TYPE 2 IMMUNITY, KNOWN FOR
DRIVING LUNG PATHOLOGY DURING
ASTHMA AND ASSOCIATED DISEASE.
THERE'S GROWING EVIDENCE
EOSINOPHILS CAN HAVE A
PROTECTIVE ROLE AS WELL.
WE WANTED TO LOOK AT THIS A BIT
MORE CLOSELY.
WE PERFORMED SINGLE CELL RNA
SEQUENCING WHERE IN THIS CASE
UNLIKE WHAT WE DID BEFORE WE
PAIRED THIS WITH CITE-SEQ DATA,
IDENTIFIED CLUSTERS OF IMMUNE
CELLS, ABLE TO PRECISELY
IDENTIFY EOSINOPHILS AND
NEUTROPHILS HERE, AND WE ALSO
MAPPED THIS TO SARS-COV-2.
WE WANTED TO SEE WHAT IMMUNE
CELLS WERE MAPPING OR
ASSOCIATING WITH VIRAL RNA.
WE SEE EOSINOPHIL POPULATION HAS
LOTS OF VIRAL RNA ASSOCIATED,
MAY BE COLLECTING DEB RIB OR
ASSOCIATING WITH DEAD CELL
DEBRIS OR PHAGOCYTOSING VIRAL
RNA.
WE TRIED TO DO IN VITRO
INFECTIONS, DOESN'T LOOK LIKE
THEY ARE INFECTIBLE AS PLAYING
DEVIL'S ADVOCATE THERE, WE DO
SEE LOTS OF VIRAL RNA.
THERE'S BEEN QUITE A BIT OF
LITERATURE WITH MONOCYTES AND
MACROPHAGES, ASSOCIATING WITH
VIRAL RNAs QUITE CONTROVERSIAL
AS TO WHETHER THEY ARE INFECTED
OR NOT.
BUT I THINK IT'S RATHER ACCEPTED
THERE IS VIRAL RNA ASSOCIATED
WITH THESE IMMUNE CELLS, WE
CERTAINLY SEE THAT AS WELL, IN
OUR NAIVE POPULATION AND WITHIN
OUR VACCINE BREAKTHROUGH
INFECTED GROUP HERE.
WE WANTED TO LOOK CLOSELY AT
THIS, AND ISOLATE THESE
COMPUTATIONALLY AND RECLUSTER
THEM BASED ON THIS POPULATION.
MOCK IS IN GRAY.
INFECTED GROUP IN BLUE.
VACCINE BREAKTHROUGH INFECTED
GROUP IS IN RED.
YOU CAN SEE EOSINOPHILS ARE
DIFFERENT TRANSCRIPTIONALLY
BETWEEN NAIVE INFECTED AND
VACCINE BREAKTHROUGH GROUPS.
AND YOU CAN FURTHER SUBCLUSTER
INTO DIFFERENT POPULATIONS AND
THESE ARE AGAIN VACCINE
BREAKTHROUGH GROUPS ARE BOTTOM
RIGHT-HAND GROUP HERE, WHEREAS
THE NAIVE ARE PART OF THE
CLUSTER 3, AND -- SORRY MOCK IN
CLUSTER 3, NAIVE INFECTED MICE
IN CLUSTERS 1 AND 2 HERE.
WE NEXT WANTED TO ASK WHAT KIND
OF EOSINOPHILS MIGHT BE, THERE'S
QUITE A BIT OF LITERATURE IN
TERMS OF AT LEAST IDENTIFYING
SIGNATURES, WHETHER TYPE 1 OR
TYPE 2.
AND SO THERE'S ONE GROUP HERE
HAD ISOLATED MOUSE EOSINOPHILS
BUT THEN EX VIVO TREATED WITH
INTERFERON GAMMA OR IL-4.
WE TOOK TRANSCRIPTIONAL MODULES
AND ASK HOW DO OUR DATA COMPARE,
WITHIN THEIR ASTHMA MODULE IT
DOESN'T LOOK LIKE ENRICHMENT IN
TERMS OF DIFFERENCE BETWEEN
NAIVE AND VACCINE BREAKTHROUGH,
IL-4 DOESN'T SEEM TO STRONGLY
INFLUENCE.
WE DO INSTEAD SEE MORE
INTERFERON GAMMA-RELATED
PHENOTYPE, EOSINOPHILS FROM OUR
VACCINE BREAKTHROUGH INFECTIONS
SEEM TO BE MORE ENRICHED FOR
INTERFERON GAMMA SIGNATURES
COMPARED TO NAIVE INFECTION,
SUGGESTING IN THE LUNGS AT LEAST
THERE MAY BE MORE OF INTERFERON
GAMMA MILIEU, SOMETHING WE'RE
LOOKING AT IN DETAIL.
WE WANTED TO ASK THE MORE
IMPORTANT QUESTION ARE
EOSINOPHILS PROTECTED?
WE TOOK ANTIBODY-DEPLETING
TECHNIQUE, WHICH WE GAVE MICE
CCR3 ANTIBODY, WHICH IS SPECIFIC
TO EOSINOPHILS, DEPLETE
EOSINOPHILS.
WHEN WE DEPLETE THESE MICE WE
ENDED UP HAVING TWO GROUPS OF
DEPLETED MICE, YOU DON'T EVER
GET PERFECT DEPLETION IN MICE,
ESPECIALLY WITH EOSINOPHILS.
WE TRY DIFFERENT TECHNIQUES,
DIFFERENT DOSES, DIFFERENT
REGIMENS.
WE SEEM TO GET TWO GROUPS, ONE
THAT DOESN'T DEPLETE WELL, ONE
THAT DEPLETES VERY WELL.
HERE WE CAN SEE A TWO LOG
REDUCTION IN EOSINOPHILS, IN THE
BREAKTHROUGH GROUP.
WHAT'S INTERESTING IS THAT THE
GROUP IN WHICH WE SAW GREATER
THAN FIVE-FOLD DEPLETION OF
EOSINOPHILS, CONTINUESED TO SHOW
WEIGHT LOSS, ISOTYPE OR POORLY
DEPLETED SHOWED BETTER
PROTECTION IN TERMS OF WEIGHT
LOSS.
NEXT ASKED ABOUT VIRAL LOAD,
INFECTIOUS VIRUS HIGHER LEVELS
IN MICE THAT WERE EOSINOPHIL
DEPLETED, CORRELATES COMPARING
SAMPLES, LOWER -- SORRY, THE
LOWER NUMBERS OF EOSINOPHILS,
HIGHER LEVELS OF VIRUS
REPLICATION, SEEMS TO BE A NICE
CORRELATION BETWEEN THAT.
MORE IMPORTANTLY, IF YOU NOW GO
WITHIN THE HISTOLOGY SAMPLE BETA
INFECTION IN LARGE AIRWAYS,
ISOTYPE CONTROLS WE SEE LOTS OF
VIRUS IN THE LARGE AIRWAY SPACES
WITH LOTS OF INFLAMMATION
OCCURRING.
NOW WHEN YOU GET RID OF
EOSINOPHILS WHAT YOU SEE IS
VIRUS SPREADING TO SMALLER
AIRWAYS, LESS INFLAMMATION
OVERALL, DON'T SEE LARGE AIRWAYS
FILLING UP WITH DEBRIS OR CELLS
SUGGESTING EOSINOPHILS ARE
HELPING RESTRICT VIRUS
REPLICATION AND MAY BE CLOGGING
UP AIRWAYS TO HELP GET RID OF
CELLS OR GET RID OF INFECTED
CELLS AND DEBRIS.
NONETHELESS IT APPEARS TO
EOSINOPHILS ARE IN FACT HAVING
AN ANTI-VIRAL ROLE IN THIS
VACCINE BREAKTHROUGH INFECTION.
WE'VE DONE THIS WITH SCP BUT
but
with mRNA VACCINE, WE FOCUSED
ON MONOCYTES AND MACROPHAGES,
SIMILAR TO EOSINOPHILS WE GET
DIFFERENT CLUSTERS, FROM
INFECTED, NAIVE INFECTED, OR
VACCINE BREAKTHROUGH INFECTION.
ONE IMPORTANT MOLECULE THAT
SEEMS TO BE DIFFERENT BETWEEN
THESE GROUPS IS EOHTAX IN2,
IMPORTANT FOR DRIVING EOSINOPHIL
INTO THE LUNGS.
TOTAL MONOCYTES IN DEPLETED MICE
WE SEE SLIGHTLY INCREASED LEVEL
OF MONOCYTES WITHIN THE LUNGS
COMPARED TO ISOTYPE CONTROL,
SUGGESTING THERE MAY BE FEEDBACK
BETWEEN EOSINOPHILS BEING
RECRUITED, MONOCYTES DEPLETING
CCL 24, MAYBE A FEEDBACK ROPE
BETWEEN THE TWO POPULATIONS,
SOMETHING WE'RE LOOKING AT MORE
CLOSELY NOW.
THIS IS A SUMMARY OF THE
DIFFERENCE BETWEEN NAIVE
INFECTION AND VACCINATED
BREAKTHROUGH INFECTION.
WE THINK THERE'S CERTAINLY A
POSITIVE ROLE IN TERMS OF
EOSINOPHILS DRIVING PROTECTION.
THESE ARE RECRUITED POSSIBLY BY
MONOCYTES, SEEM TO HELP CONTROL
VIRUS REPLICATION.
I HOPE I'VE SHOWN AT LEAST THREE
VERSIONS WHETHER WE LOOK AT THE
INNATE IMMUNE RESPONSE IN THE
CONTEXT OF NAIVE INFECTION
TRYING TO DISSECT WHAT'S GOING
ON, WE'RE NOW LOOKING AT UPPER
VERSUS LOWER RESPIRATORY TRACT,
WHAT ROLE T CELLS PLAY AND
CONTEXT OF BREAKTHROUGH
INFECTION.
SEVERAL PEOPLE I'D LIKE TO
THANK, I'LL POINT OUT A FEW THAT
HAVE WORKED HARD IN THE
LABORATORY, WORKING ON THE
VACCINE BREAKTHROUGH MODEL.
AND MEENAKSHI KAR, ABIGAIL WHEN
WE FIRST STARTED THE MOUSE WORK
IN THE LABORATORY HAD UNDERTAKEN
SOME WORK WITH CCR 2 AND SEVERAL
COLLABORATORS ACROSS EMORY
UNIVERSITY AND OTHER PLACES,
VINEET IS NOW AT EMORY.
I HAVE HIM MISLABELED AT
GALVESTON.
>> I HAVE A QUESTION ABOUT
EOSINOPHIL AND VACCINE
BREAKTHROUGH.
DO YOU THINK THAT THE EOSINOPHIL
THAT YOU'RE SEEING IS AT ALL DUE
TO SLIGHTLY HETEROLOGOUS
CHALLENGE VIRUS SKEWING THINGS
MORE Th2 OR DO YOU THINK THAT
IT'S JUST AN INSUFFICIENT
ANTIBODY, LIKE A BINDING
NON-NEUTRALIZING OR NOT
SUFFICIENT NEUTRALIZING?
>> I THINK IT HAS MORE TO DO
WITH VIRUS INFECTION OF THE
LUNG.
IF THE VIRUS CAN GET INTO THE
LUNGS YOU'RE GOING TO GET
INFECTION OF CELLS, IMMEDIATE
ANTIBODY.
THIS IS MY -- WE HAVE NOT
TESTED, TESTED AGAINST GAMMA
RECEPTOR KNOCKOUT MICE, GETTING
ANTIBODY BINDING.
WHEN YOU LOOK AT BINDING VERSUS
NUTES, THERE'S A LOT OF BINDING
COMPARED TO ANTIBODIES.
THE MOUSE ADAPTIVE STRAIN IS
SIMILAR TO TRIMERIC, THE
PROTEIN, WE SEE THE SAME THING.
AS LONG AS YOU OVERCOME WHATEVER
PROTECTIVE IMMUNITY THERE IS TO
NOT GET THE VIRUS IN THE LUNGS,
IF THE VIRUS GETS IN THE LUNGS
YOU'RE GETTING INFLAMMATORY
RESPONSE.
TIME 1 VERSUS TYPE 2 I DON'T
THINK THERE'S STRONG EVIDENCE IN
HUMANS THERE'S STRONG TYPE 2
RESPONSE.
I THINK THIS TYPE 1 VERSUS TYPE
2, I'M NOT QUITE SURE THAT THIS
NECESSARILY FITS THAT BILL
COMPLETELY.
>> CURIOUS ABOUT THE SARS 2 IN
PHAGOCYTIC CELLS, DIFFICULT TO
DISENTANGLE.
DO YOU FIND LOW LEVEL READS FOR
EPITHELIAL-SPECIFIC MARKERS
SUGGESTING THEY DID PHAGOCYTOSE.
>> WE HAVEN'T GONE TO THAT DEEP
LEVEL.
THERE'S BEEN SOME STUDIES THAT
TRIED TO FLOW SOURCE, MACROPHAGE
POPULATIONS IN CULTURE FOR
HOURS, SHOWING POSITIVE VIRUS
REPLICATION, IT'S STUFF TO STATE
THERE'S VIRUS REPLICATION.
EVEN IF YOU TRY TO IDENTIFY I
STILL DON'T THINK ONE CAN SAY
IT'S VIRUS REPLICATION VERSUS
EATING DEAD CELL DEBRIS.
>> YES, SUPER NICE WORK.
I WAS WONDERING DO YOU THINK THE
VIRUS RESTRICTION BY EOSINOPHILS
IS DIRECT RESPONSE OR SOMETHING
OTHER, ANOTHER FUNCTION.
>> WE HAVE A COUPLE CANDIDATE
GENES WE'RE LOOKING AT.
ONE THAT SEEMS TO BE QUITE
INTERESTING THAT EOSINOPHILS MAY
BE SECRETING.
THERE'S NOT A LOT OF EOSINOPHIL
REAGENTS.
THAT'S THE PAUSE I HAVE IN TERMS
OF BEING ABLE TO LOOK AT
CONDITIONAL KNOCKOUTS, TAKING
OUT CERTAIN MOLECULES.
JUST SHORT OF BEING ABLE TO
ISOLATE EOSINOPHILS FROM MICE,
WHICH IS STILL VERY TOUGH TO BE
ABLE TO SORT THOSE WITHOUT
KILLING THEM.
AND TRANSFERRING THEM.
WE'RE NOT ABLE TO SPECIFICALLY
DISSECT DIRECT ANTI-VIRAL
SECRETION OF ENZYME OR CYTOKINE,
FROM BY STANDARD EFFECT.
>> WERE YOU ABLE TO STAIN FOR
VIRAL PROTEIN IN EOSINOPHILS?
>> WE'VE NOT DONE THAT.
IT'S ONLY BY SORT OF EYE IN
TERMS OF WHEN WE DEPLETE
EOSINOPHILS WE NO LONGER GET
THIS MASSIVE POPULATION THAT'S
WITHIN THAT AIRWAY SPACE,
WHETHER OUR COMPUTATIONAL STUFF
FOR RNAseq SHOWING A LOT OF
VIRAL RNA ASSOCIATED WITH THOSE
CELLS.
>> I HAD A QUESTION.
IN PATIENTS WHO DIED EARLY IN
THE PANDEMIC, WITH LOTS OF
EOSINOPHILS IN THE LUNGS --
>> THEY WERE NEUTROPHIL
DOMINATED IN TERMS OF NAIVE
INFECTION.
FROM VACCINE BREAKTHROUGH
INFECTIONS, WHICH INDIVIDUALS
MAY HAVE GONE TO THE HOSPITAL,
HAD SEVERE DISEASE, WE'VE TRIED
TO LOOK IN THE LITERATURE.
THERE'S NOT A LOT OF DATA TO GET
THIS TRYING TO GET FLUID FROM
INFECTED INDIVIDUALS IS TOUGH,
ESPECIALLY WITH SHORT-LIVED
CELLS LIKE EOSINOPHILS, TIMING
IS IMPORTANT WHEN YOU COLLECT
THOSE SAMPLES AS WELL.
>> THIS QUESTION WAS SUBMITTED
AT THE BEGINNING OF THE TALK.
WOULD IT BE IMPORTANT TO SHOW
THE STRENGTH OF RESPONSE IN
SHOWING MEDIAN FLUORESCENT
INTENSITY OF PERCENT POSITIVE
ANTIGEN SPECIFIC T CELL?
>> WE HAVE LOOKED AT IN TERMS OF
INTERFERON GAMMA STAGE, TNF
ALPHA STAINING, WE DON'T SEE
DIFFERENCE IN INTENSITY.
NO MAJOR SHIFT EITHER POSITIVE
OR NEGATIVE.
>> EOSINOPHIL HAPPEN IN
BREAKTHROUGH INFECTION, LOOKS
LIKE ANIMAL GET PRIME BY
INFECTION OR VACCINE, IN THAT
CASE, SO WHY ONLY THE
BREAKTHROUGH INFECTION YOU SEE
--
>> I THINK IN TERMS OF
BREAKTHROUGH INFECTION THE LUNG
IS BATHED IN ANTIBODIES.
WHEREAS NAIVE MICE DON'T HAVE
THOSE.
I THINK IT HAS A LOT TO DO WITH
ANTIBODIES.
I THINK IN THE SHORT WINDOW OF
DAY 3 POST-INFECTION, WE HAVE
HAVEN'T FORMALLY TESTED ROLE OF
T CELLS BUT CERTAINLY THE LUNG
AND LOTS OF BINDING ANTIBODIES.
>> IF YOU USE KNOCKOUT MICE,
WHAT WOULD YOU EXPECT?
>> B CELL KNOCKOUTS ARE MORE
COMPLEX, IMMUNOLOGICALLY NOT THE
SAME AS NAIVE MICE.
IT'S NOT JUST SIMPLY YOU REMOVE
B CELLS FROM THEM.
I THINK THE BETTER EXPERIMENT
WHICH I'VE ASKED MY FOLKS TO DO
IS DEPLETE T CELLS AND ASK WHAT
HAPPENS IN THAT CONTEXT.
>> I GUESS ON A SIMILAR NOTE
HAVE YOU BEEN ABLE TO GIVE
IMMUNE SERA FROM RECOVERED
HAMSTERS, SORRY, FOR RECOVERED
MICE, IS THAT SIGNIFICANT ENOUGH
TO INDUCE EOSINOPHILS?
>> WE'VE BEEN WORKING TRYING TO
GET THE RIGHT DOSE, RIGHT AMOUNT
OF ANTIBODIES, RIGHT TIMING OF
INFECTION, HASN'T BEEN
SUCCESSFUL YET.
IT HAS MORE TO DO WITH TECHNICAL
THAN ACTUAL PHENOTYPE.
ONE OF THE THINGS WE WANTED TO
DO WAS TAKE THAT PASSIVE IMMUNE
TRANSFER AND PERFORM THAT
RECEPTOR KNOCKOUT MICE TO
DEMONSTRATE FC GAMMA DEPENDENT
SIGNALING THAT'S DRIVING THAT.
>> IF NO MORE QUESTIONS WE CAN
HAVE OUR PANEL DISCUSSION.
>> ONE MORE QUESTION.
IN TERMS OF THE VIRAL PERSISTENT
STORY, I'M NOT CLEAR ON THE
EVIDENCE FOR HOW STRONG IT IS
FOR THE IDEA OF PERSISTENCE IN
HUMANS.
IS THAT STRONGER DATA NOW?
>> YEAH, SO IMMUNOCOMPROMISED
INDIVIDUALS I THINK IT'S QUITE
COMPELLING THAT INDIVIDUALS THAT
ARE EITHER ON B CELL THERAPY,
T-CELL THERAPY, THEY DO HAVE
PERSISTENT VIRAL INFECTION.
IMMUNE COMPROMISED INDIVIDUALS,
ESPECIALLY CANCER INDIVIDUALS,
THERE'S LOTS OF EVIDENCE OUT
THERE SHOWING VIRUS CAN PERSIST
FOR MONTHS ON END IN THEIR UPPER
RESPIRATORY TRACT, AS A MATTER
OF FACT I KNOW ONE OF OUR
CLINICIANS WORKING AT THE CANCER
CENTER AS PART OF OUR U54 GRANT
WOULD COMPLAIN HIS PATIENTS
WEREN'T COMING TO THE CLINIC
EARLY DURING THE PANDEMIC
BECAUSE THEY WOULD TEST POSITIVE
IN THE UPPER RESPIRATORY TRACT,
MANY WOULD MISTREATMENT MISS -E
TREATMENT THEY ARE SUPPOSED TO
GET.
THANK YOU.
>> OKAY.
SO NOW WE HAVE OUR PANEL
DISCUSSION.
>> DOES ANYONE HAVE QUESTIONS
FOR THE PANEL?
>> I HAVE A QUESTION.
WE'RE DISCUSSING DURING THE
BREAK, IN TERMS OF MUCOSAL
IMMUNITY IN THE SMALL ANIMAL
MODEL, ARE WE GOING TO NEAT
ADJUVANTS, ARE THERE POTENTIAL
WITHDRAWAL EFFECTS OF USING
ADJUVANTS IN ANIMAL MODELS AND
IN TRYING TO TRANSLATE THAT TO
HUMANS, SO DO WE NEED ADJUVANTS?
ARE THERE MUTUAL SIDE EFFECTS
WHEN WE MOVE TO HUMANS?
WHAT ARE THE PANELIST FEELINGS?
>> I CAN TAKE THE QUESTION.
WE HAVE TO START SEVERAL
ADJUVANTS FOR MUCOSAL AND
MUSCULAR VACCINATION, AND IN
MICE WE HAVE SEEN IF WE GIVE
RECOMBINANT PROTEIN NOT MUCH
HAPPENS IN TERMS OF SERUM,
ANTIBODY INDUCTION, MEASURING
SERUM OR MUCOSA.
ON THE HAMSTER WE SEE GOOD IgA
AND SEER INDUCTION, SEEMS TO BE
A DIFFERENCE IN MICE OR BIG OR
SMALL ANIMALS.
THAT BEING SAID, IF WE USE MICE
FOR MUCOSAL VACCINATION WE ALSO
HAVE TO PUT SOME GOOD ADJUVANTS,
BUT WE HAVE TO BE CAREFUL
BECAUSE THE RANGE OF TOXICITY IS
VERY SMALL.
SO, FOR EXAMPLE, AGONISTS WE'VE
TESTED WAS TOXICS, SMALLER WERE
ACTUALLY VERY SAFE.
>> A QUICK COMMENT.
AS FAR AS I'M AWARE, CORRECT ME
IF I'M WRONG, NO SMALL ANIMAL
MODEL RECAPITULATES
REACTIGENICITY, FOR SOME HUMANS
HAVE IT BUT NOT ALL.
IT'S A HUGE DRAWBACK IN THE
VACCINE FIELD BECAUSE PEOPLE
THINK REACTIGENICITY, PREDICTIG
WHO MAY GET REACTIGENICITY,
PREVENTING REACTIGENICITY WOULD
BE A HUGE DEAL.
WE'VE TRIED, WITH A FEW
ADJUVANTS WE HAVE YET TO SEE IT.
WE WOULD LIKE TO FIND A MODEL
FOR REACTIGENICITY.
>> YOUR DIRTY MICE MODEL DOES
YOUR ADJUVANT STEAL ADJUVANT.
>> YOU NEED IT.
THE ADJUVANT IS QUITE CRITICAL
IN DIRTY MICE JUST LIKE HUMANS
WHERE IT'S QUITE CRITICAL.
>> ANOTHER PROBLEM IN MICE IS
THE VOLUME FOR MUCOSAL
ADJUVANTS, VERY SMALL VOLUMES IF
YOU WANT TO PREVENT FROM GOING
TO THE LUNGS.
>> I HAVE A QUESTION, MORE
GENERAL QUESTION.
NATURAL INFECTION DOESN'T SEEM
TO INDUCE MUCOSAL IMMUNITY, AT
LEAST WITH PROTECTIVE EXTENT.
WHAT DO YOU THINK WE'RE DOING
DIFFERENTLY THAT WILL HELP GET
AROUND THAT PROBLEM?
>> I MEAN, I THINK WE DON'T
FULLY UNDERSTAND WHAT IS BEING
INDUCED WHEN PEOPLE ARE INFECTED
WITH SARS-COV-2, WHETHER THAT
COMPARES TO WHAT WE INDUCE WITH
INTRANASAL VACCINATION, MUCOSAL
VACCINATION.
SO THAT'S AN INCREDIBLY
IMPORTANT QUESTION TO TRY AND
UNDERSTAND BETTER BECAUSE
CLEARLY WE'RE GETTING THE
INFECTED AND WOULD IDEALLY HAVE
VACCINE THAT LIFELONG PROTECTION
WOULD BE GREAT BUT I DON'T WE'RE
GOING TO GET THERE.
EITHER WITH VIRUS INFECTION, NOT
BEING INDUCED OR OVER TIME GO
AWAY AND YOU'RE SUSCEPTIBLE
AGAIN.
>> ARE YOU SPECIFICALLY ASKING
PROTECTION IN THE UPPER?
>> YEAH, I THINK IT'S VERY HARD
TO DRIVE STERILIZING IMMUNITY IN
THE UPPER RESPIRATORY TRACT.
EVEN IF YOU HAVE A HOMOLOGOUS
INFECTION YOU CAN STILL GET --
HOMOLOGOUS VIRUS, COULD STILL
GET GOOD REPLICATION OF THE
UPPER TRACT BUT DO GOT STRONG
PROTECTION IN THE LOWER TRACT,
WE NEED A DIVERGENT VARIANT,
THAT'S WHERE YOU GET INFECTION
OF THE LUNGS.
>> IT'S ONE OF THE KEY QUESTIONS
BECAUSE WE DO GET REINFECTED,
PEOPLE ARE NOT GOING TO THE
HOSPITAL, TO SUBSTANTIAL EFFECT,
UNLESS THEY ARE
IMMUNOCOMPROMISED OR QUITE AGED
IN NURSING HOMES.
THE GOAL OF MUCOSAL IMMUNITY IS
PREVENT INFECTION, YOU'RE SAYING
THAT MAY BE VERY TYPICAL.
IS IT WORTH FOCUSING ON MUCOSAL
IMMUNITY AS OPPOSED TO OTHER
METHODS OF VACCINATION.
>> SO I THINK FROM WHAT WE KNOW
IN SARS-COV-2 HYBRID IMMUNITY, A
COMBINATION OF VACCINATION,
COMBINATION OF INFECTION OF
MUCOSA, GIVING SUPERIOR
PROTECTION BUT ALSO IT SEEMS
THAT MEMORY AT MUCOSAL SITES IS
NOT AS STRONG AS PERIPHERY.
WE THEY FOCUS IF RECALL OR
MEMORY IS NOT IN THE PERIPHERY,
HAS TO BE RECRUITED TO THE
MUCOSA, HAVE TO CHECK IF WE CAN
FACILITATE OR MAKE THIS GO
FASTER.
FOCUSING ON INDUCING MEMORY AT
MUCOSAL SITES THEMSELVES.
PERIPHERY MIGHT BE THE MEMORY
COMPARTMENT FOR THE MUCOSAL
SITE.
>> WE DON'T KNOW ENOUGH ABOUT
THE UPPER RESPIRATORY TRACT
COMPARED TO THE LUNG.
>> SORRY, YEAH, I WANT TO SORT
OF MORE BROADLY TALK ABOUT WE'VE
BEEN FOCUSED ON VACCINATION,
LETHALITY OF DISEASE OR
REPLICATION, HOPEFULLY
ADMINISTERED OR SYSTEMICALLY
ADMINISTERED BUT WHAT ABOUT
TRANSMISSION AND NATURAL
INFECTION, CAN WE TALK ABOUT
WHAT BARRIERS ARE, DOVETAILING
WITH TOMORROW'S SESSION, WHERE
DO WE NEED TO GO, RIGHT, AS A
FIELD TO REALLY THINK ABOUT HOW
VACCINATION CAN COMBAT NATURAL
INFECTIONS AND BE SORT OF WHAT
WE BILL IT TO BE?
>> YEAH, I'M GOING TO ACTUALLY
GO BACK AND SAY I DO THINK
MUCOSAL IMMUNITY IS SOMETHING WE
SHOULD FOCUS ON BECAUSE STRAIGHT
AFTER INFECTION PEOPLE OFTEN ARE
PROTECTED, I THINK THAT ALSO
GOES INTO TRANSMISSION AS WELL,
WHAT WE LIKELY NEED IN UPPER
RESPIRATORY TRACT IS ANTIBODY
PROTECTION BY CELLS, SO IT'S NOT
EVEN NECESSARILY JUST THAT YOU
GET STABLE PROTECTION OF UPPER
TRACT BUT ANTIBODIES,
POTENTIALLY REDUCING ACTUAL
EFFECTIVENESS OF TRANSMISSION.
SO I THINK WE NEED A LITTLE BIT
MORE THAN JUST WE NEED TO
UNDERSTAND WHETHER ANTIBODIES IN
THE UPPER TRACT ARE ABLE TO DO
THAT.
WHEN WE DO TRANSMISSION STUDIES
WE ONLY VACCINATE SENTINEL, WE
STILL GET TRANSMISSION OF THE
VIRUS, FROM THE DONOR ANIMALS,
TO THE SENTINELS EVEN THOUGH
THEY ARE PROTECTED, YOU STILL
GET INFECTION OF UPPER
RESPIRATORY TRACT.
DONOR ANIMALS VACCINATED,
INTRAMUSCULAR, WE GET LESS
TRANSMISSION OR ABSOLUTELY NO
TRANSMISSION AT ALL, THAT HAS TO
DO WITH COATING OF THE VIRUS BY
ANTIBODIES.
SO I THINK THAT TO LOOK AT
TRANSMISSION, PREVENTION
TRANSMISSION, KEY QUESTION, HOW
CAN WE INDUCE BY VACCINATION.
>> A SMALL ANIMAL MODEL
WORKSHOP, I'LL PLUG WE NEED MORE
MODELS TO STUDY THIS EXACT
THING, NEED MORE TRANSMISSION
MODELS.
A MODEL ABOUT ANOTHER
TRANSMISSION MODEL, THE MORE WE
HAVE WITH DIFFERENT MODELS,
STRENGTHS, WEAKNESSES, TOOLS,
THE BETTER TO ADDRESS THE
QUESTIONS.
IT'S HARD, TRANSMISSION MODELS
ARE NOT EASY.
WE CLEARLY NEED MORE.
>> I WAS GOING TO CALL ON DAVID
MARTINEZ ON ZOOM TO UNMUTE.
>> YEAH, I HAVE A QUESTION FOR
ANNELTJE IN PARTICULAR, ABOUT
YOUR CHADOX DATA.
THE VECTOR IS REMARKABLE AT
ELICITING TISSUE RESIDENT T-CELL
RESPONSES AND IgA RESPONSES IN
ANIMAL MODELS.
HOWEVER, THAT CHADOX VACCINE IN
HUMANS IN A PAPER PUBLISHED
MAYBE OVER A YEAR AGO SHOWED IT
WAS LACKLUSTER.
SO I WAS WONDERING IF YOU COULD
COMMENT ON WHY PERHAPS THERE'S
SUCH A DISCONNECT IN THE ANIMAL
MODEL THAT YOU'RE USING COMPARED
TO WHAT THE DATA THAT WAS SHOWN
IN HUMANS, I THINK BY
ASTRAZENECA.
>> YES, A FANTASTIC QUESTION.
IT GOES BACK TO THE MODELS WE'RE
USING IN THE MOUSE, NOT THE
HUMAN, NEITHER THE HAMSTER OR
FERRET.
HOW THEY ADMINISTER, THEY USE
THE SAME NEBULIZER AS WE USED IN
NON-HUMAN PRIMATES, WE CAN'T USE
THAT IN A HAMSTER BECAUSE IT'S
TOO LARGE BUT BASICALLY CREATING
A CLOUD OF AEROSOLS, VACCINES
THE ANIMALS BREATHE IN.
IT DIDN'T INUSE THE SAME IMMUNE
RESPONSE.
OUR COMPANIES USING DIFFERENT
WAYS OF VACCINATION AND IN CHINA
THEY ARE ACTUALLY DOING A CORE
THING CREATING A CLOUD OF
AEROSOLS, SORT OF INHALE THAT,
IT GOES MORE INTO THE LUNGS.
I THINK THAT MAY BE BETTER WAY
OF GETTING MUCOSAL IMMUNITY, IN
THE NOSE WE DON'T ACTUALLY
INDUCE IT IN THE SAME WAY.
THIS IS WHAT I TRIED TO SAY
DURING MY PRESENTATION, MODELING
IN ANIMALS IS INCREDIBLY
IMPORTANT TO UNDERSTAND
MECHANISMS LIKE HOW DO WE INDUCE
IMMUNITY BUT THEN I THINK WE
NEED TO TAKE IT TO HUMANS AND
SEE WHAT WE SEE IN ANIMALS IS
THAT ACTUALLY HAPPENING IN
HUMANS AS WELL.
>> OTHER QUESTIONS?
>> YEAH, I HAVE ANOTHER FROM THE
Q&A.
BRIAN SAYS AGREE WITH THE PANEL,
UPPER RESPIRATORY TRACT, MUCOSAL
IMMUNITY IS MEMORY.
WOULD THE GOAL BE PROPHYLACTIC
ATTACKS AT SEASONAL COMMISSION,
EPIKINETICS OF SEASONAL DISEASE
AND POPULATION?
IT'S EASIER WHEN SOMEONE COMES
OFF MUTE AND ASKS.
>> DO YOU WANT THE QUESTION
REPEATED?
COULD YOU REPEAT IT?
>> BRIAN?
WE CAN GO ON TO STEPHANIE.
>> YES, THANK YOU.
THIS IS FOLLOW-UP.
FOLLOWING UP WITH WHAT DAVID WAS
SAYING CURIOUS ABOUT DIFFERENCES
IN OUR SERONEGATIVE ANIMAL
MODELS TO HUMAN, EVEN THOUGH
SCREENED SERONEGATIVE FOR
SARS-COV-2 MAYBE OTHER
CORONAVIRUSES, THEY COULD BE
ASYMPTOMATICALLY INFECTED WITH
OTHER RESPIRATORY VIRUSES, COULD
BE SMOKERS, COULD HAVE
POLLUTANTS IN THE HOME SHOWN TO
REDUCE IgA RESPONSES, CURIOUS
YOUR THOUGHTS ABOUT TRANSLATING
TO HUMANS WHICH FOR MUCOSAL
IMMUNITY MAY BE MORE
COMPLICATED.
>> THERE ARE SO MANY VARIABLES
WE NEED TO THINK ABOUT.
I HONESTLY THING THE MOUSE MODEL
WOULD BE WHERE I WOULD LIKE TO
GO FIRST BECAUSE I THINK ABOUT
ACCESSING SO MANY OTHER THINGS.
>> FOR SURE.
>> I THINK IT'S SO DIFFICULT TO
MODEL ALL THEY THINGS, AT WHAT
POINT ARE YOU ASKING TOO MUCH
FOR ONE STUDY?
>> YEAH, FOR SURE.
>> MY APOLOGIES FOR MISSING IT
BUT HAS ANYONE DONE THE DIRTY
EXPERIMENT FOR FERRETS?
MAKING FERRETS DIRTY?
>> YOU DON'T THINK THEY ARE
SPECIFIC PATHOGEN FREE?
THEY ARE NOT OUTBRED DIRTY BUT
WILD DIRTY.
>> THE FACILITIES ARE FAIRLY
CLEAN.
THEY TRY KEEP FLU-FREE COLONIES
BECAUSE FLU RESEARCHERS ARE THE
HIGHEST CONSUMERS I THINK, IF
YOU WOULD AGREE WITH ME, STACY,
HERE, OF THE FERRETS.
WITH THAT, THEY DEFINITELY HAVE
A LOT OF PATHOGENS.
ENTERIC VIRUSES, CORONAVIRUSES
THAT ARE ENTERIC, SOME HAVE
DIARRHEA SOMETIMES, THEY
DEFINITELY HAVE LOTS OF OTHER --
YEAH, WE HAVEN'T DONE THE SAME
SORT OF VIRUM SEQUENCES, I DON'T
KNOW IF THEY ARE KEPT CLEAN.
>> I THINK CLEANER THAN A PET
STORE FERRET FOR SURE.
THE SAME BLEEDER, TRIPLE F AS
WELL BUT GETTING INTO A BL 2
FACILITY WHERE WE GOWN UP AND GO
IN IS DIFFERENT THAN A PET STORE
WITH A HIGH SCHOOL KID CLEANING
THE BIRD CAGES, HAVING POP ON
THEIR HANDS, CLEANING THE FERRET
CAGES.
I THINK THEY ARE CLEANER THAN
YOU THINK THEY ARE BUT THEY ARE
DEFINITELY NOT CLEAN-CLEAN.
OURS COME IN, THEY ARE PRETTY
CLEAN.
WE'VE DONE VIRUM INFORMANT.
SOMEBODY RON USED TO TALK ABOUT
ALL THE TIME, YOU HAVE TO BE
CAREFUL, FERRETS DON'T HAVE
CORONAVIRUSES, BECAUSE IT CAN
INFLUENCE WHAT WE DO WITH THE
FLU BUT WE'VE SEEN WORMS
OCCASIONALLY, THINGS TO GET
TREATED WITH ANTIBIOTICS BUT
NOTHING DRAMATIC.
>> WE'VE LOOKED AT LIKE -- WE'VE
HAD ENTERIC CORONAVIRUSES, IN
THE FERRETS, THEY HAVE COMMON
COLD CAUSING CORONAVIRUS
ANTIBODIES.
I DON'T KNOW.
WE HAVEN'T DONE A DEEPER DIVE ON
ALL THE ANIMALS, YEAH, I'D SAY
SEMI DIRTY.
CAN I SAY SEMI DIRTY?
>> WITH BANK THE SERUM --
>> WE'VE TALKED ABOUT THIS.
>> IT WOULD BE AMAZING, REALLY
COOL.
WE SHOULD DO IT.
WE SHOULD COLLABORATE, ALL YOU
FERRET PEOPLE.
>> YOU COULD DO THIS STUDY THAT
PET STORE FERRETS, COMPARE IT.
>> MICHELLE'S POINT THERE ARE
IMMUNE EXPERIENCED, NOT BRED TO
BE PATHOGEN FREE ALTHOUGH
HAMSTERS ARE, IS THAT RIGHT?
HAMSTERS ARE SPF?
>> THEY ARE SUPPOSED TO BE.
I DON'T KNOW IF ANYBODY HAS TO
TRIPLE F.
THEY ARE NEVER COMPLETELY CLEAN,
I'LL LEAVE IT AT THAT.
>> IF YOU CONSIDER FERRET VERSUS
HAMSTERS, THE CARETAKERS ARE WAY
MORE INTERACTING WITH THE
FERRETS, VERY CLOSED, IT'S LIKE
PETS.
HAMSTERS YOU CAN STILL HOLD, THE
FERRETS THE INTERACTION IS MORE
INTENSE.
THE CHANCE THEY ARE GOING TO GET
MICROBIOMES FROM THE CARETAKERS
IS BIGGER.
>> SPF ANIMALS AREN'T COMPLETELY
FREE OF ANY INFECTION, HOW MUCH
DO THEY NEED TO BE EXPOSED, TO
BE LIKE DIRTY MICE?
>> IT'S A GREAT QUESTION.
PROBABLY NOT A LOT, TO BE
HONEST.
THE PET STORE CO-HOUSING IS NOT
NECESSARY BUT THERE'S OTHER
THINGS THAT CAN BE LEARNED.
THERE'S A LOT OF WAYS TO ATTACK
THE SAME QUESTION.
SOME IMMUNE EXPERIENCE IS BETTER
THAN NONE.
BASED ON WHAT SEEMA IS SAYING,
SUFFICIENT AMOUNT OF IMMUNE
EXPERIENCE IN FERRETS TO AT
LEAST MAKE THEM MORE HUMAN-LIKE.
>> ASTRO-1, YOU DO FIND IT IN
SOME OF YOUR MICE.
BUT YOU DON'T KNOW YET --
>> I DON'T WANT TO SPEAK, BUT I
AGREE ASTRO-1 MAY BE COMMENSAL,
MAYBE NOT DOING THE SAME THINGS
AN ACUTE CORONAVIRUS WOULD DO SO
ASTRO IS A UNIQUE SITUATION.
I DON'T THINK IF THAT'S GOING TO
END UP BEING TRUE BUT THAT'S
INTERESTING.
MAYBE THE MOUSE AND YELLOW
VIRUS, I DON'T KNOW.
MAYBE STACY HAS THOUGHTS ON
THAT.
>> BE CAREFUL WHEN THEY GET
IMMUNOCOMPROMISED, IT WILL GO
STRAIGHT TO THE BRAINS.
WE'VE LOOKED FOR ASTRO IN
FERRETS BECAUSE WE WANTED TO
FIND IT SINCE WE WORK ON ASTRO
VIRUS, NOTHING TO DATE
UNFORTUNATELY.
>> I WANT TO RETURN TO THE NEED
FOR BETTER MOUSE ADAPTED STRAINS
FOR FLU.
HOW DO WE GO ABOUT IDENTIFYING
THESE?
IS THIS JUST LIKE A MASSIVE
SCREENING -- OKAY.
>> DO YOU WANT TO TAKE IT?
WHAT WE DID IN THE PAST, YOU CAN
DO SERIAL PASSAGING LIKE MICE,
IT STARTS WITH MICE START TO
LOSE WEIGHT.
YEAH, YOU MAY HAVE TO TRY THAT
WITH A LOT OF VIRUSES, A LOT OF
MONEY AND A LOT OF WORK.
MAYBE IF WE COORDINATE THAT
WOULD BE VERY GOOD.
>> IS THERE A COMPUTATIONAL WAY
TO SORT OF STRATEGIZE ABOUT
THIS?
LIKE BEST -- I DON'T KNOW.
>> MULTIPLE FACTORS WITH MOUSE
ADAPTIVE VIRUS, THERE'S RECEPTOR
BINDING, INTERFERON ANTAGONISM,
CAN YOU MODEL FOR SURE BUT IN
THE END MANY MODELS, FOR EXAMPLE
PLEA TICKETED HIGHER AFFINITY
FOR RECEPTOR BINDING BUT NOT
THAT'S NOT THE CRUCIAL POINT.
SO MANY FACTORS YOU CAN MODEL
BUT IN THE ENDS YOU HAVE TO TEST
SEVERAL VIRUSES, WHICH MEANS A
LOT OF ANIMALS, A LOT OF MONEY
BUT WORTH IT TO COORDINATE SUCH
AN EFFORT.
>> THE COMPUTATIONAL IDEA IS
FUN, THE IDEA OF PUTTING IN
FEATURES YOU KNOW CAUSING
PATHOGENESIS IN MICE, FEEDING
SOME A.I., WHAT WOULD THE NIH
SAY ABOUT MAKING THOSE UNKNOWN
MUTATIONS WHICH MAY POTENTIALLY
GAIN A FUNCTION AND HOW WE GO
ABOUT DOING THAT.
I THINK THIS IS A SIGNIFICANCE
ISSUE, TAKING ON ANY HUMAN VIRUS
DIRECTLY, DOING PATHOGEN
EXPERIMENTS, I THINK A LOT OF US
ARE QUITE GUN SHY IN THE SPACE.
EXPERIMENTS ARE INCREDIBLY
IMPORTANT AND EXPERIENCES FOLK
ON THE STAGE DID EARLY IN THE
PANDEMIC WITH SARS WERE
ESSENTIAL TO GETTING US OUT OF
THE DEEPEST DARKEST PLACE.
AND WHAT HAPPENED IN THE NEXT
PANDEMIC WHEN THE RULES ARE
UNCLEAR, WHEN THE FUNDING SPACE
FOR THIS IS UNCLEAR.
I THINK THIS ISSUE OF HOW TO
MAKE BETTER MODELS FOR HUMAN
VIRUSES HAS BEEN SIGNIFICANTLY
MUDDIED IN THE CURRENT POLITICAL
CLIMATE.
>> I AGREE.
THAT'S GOING TO GET A LITTLE BIT
DICEY, I THINK.
THAT'S ACTUALLY ONE THING THAT
THE GENETICALLY OBESE ARE REALLY
GOOD FOR, IS PASSAGING FOR MASS
ADAPTATION BECAUSE TYPICALLY YOU
HAVE TO GO TEN OR MORE PASSAGES
TO GET A VIRUS MOUSE ADAPTED,
BACK AND FORTH, SPOSHLY GENETIC
SPLICE, YOU CAN DO TWO OR THREE
PASSAGES, BACK IN THE WILDTYPE
MICE IT IS HOT.
>> IF YOU TAKE TEN MICE, DO TEN
INDEPENDENT TRIALS, YOU GET TEN
DIFFERENT MOUSE ADAPTED VIRUSES.
WHICH HAS VIRTUE BECAUSE IT
GIVES -- SHOWS SHOWS POSSIBLE
ALTERNATIVES TO GETTING TO THE
SAME ENDPOINT.
>> I'LL POINT OUT TOO IT'S NOT
NECESSARILY JUST VIRUS STRAIN
SPECIFIC BUT MATCH BETWEEN VIRUS
AND HOST, DB AEROBICS MICE ARE
MORE SENSITIVE TO FLU THAT DON'T
MAKE A BLACK 6 OR BALD MOUSE SIX
THAT WILL KILL A DDA.
>> OTHER QUESTIONS?
>> YEAH, FROM THE CHAT, WORKING
WITH DIRTY MICE.
I HAVE A QUESTION ABOUT DIRTY
MOUSE MODEL, IS THAT CONTROLLED
LEVEL, DO YOU SCREEN TO AVOID
RISK OF OUTBREAK IN THE MOUSE
FACILITY, HARD TO BALANCE RISK
FOR SAFETY AND FURTHER HOUSE FOR
GENERAL LABS ALTHOUGH IT'S AN
IMPORTANT MODEL FOR INFECTION
STUDY.
YOU MAY HAVE TOUCHED ON THIS AT
THE VERY END BUT MAYBE
CLARIFYING HOW YOU ACTUALLY DO
THIS WORK.
>> YEAH, WE DO DO THIS IN A
MODIFIED BSL3 FACILITY.
I'VE BEEN DOING THIS NOW FOR
SEVEN YEARS.
WE SCREEN EVERY PET STORE
ANIMAL, EVERY CO-HOUSED ANIMAL,
EVERY SPF ANIMAL WE WORK WITH
FOR THESE EXPERIMENTS, WE'VE
NEVER SEEN CONTAMINATION OF SPF
ANIMALS.
I THINK THIS WORK CAN BE DONE IN
TERMS OF PROTECTING ANIMAL
FACILITIES ON CAMPUS.
WE HAVE IMMUNOCOMPROMISED MICE
IN OUR LABORATORY AND FACILITY
AND THEY ARE OKAY.
I THINK IT CAN BE DONE.
IT DOES REQUIRE EXTRA TIME,
MONEY, EFFORT, BUT PERSONALLY
IT'S WORTH IT BECAUSE I'M
WORRIED ABOUT HARMING THE ANIMAL
QUOLL
COLONIES OF COLLEAGUES.
WE DON'T NEED TO SCREEN EVERY
TIME NOW WE KNOW PHENOTYPES ARE
STABLE FOR ALL THE VIRUSES BUT
WE DO IT ANYWAY BECAUSE WE'RE
INTERESTED IN UNDERSTANDING THAT
BIOLOGY, BUT THAT'S ANOTHER
COMPONENT THAT'S QUITE LABOR AND
COST EXPENSIVE.
>> IF THERE ARE NO FURTHER
QUESTIONS, NO MORE FROM ZOOM
>> WE HAD ONE FROM MICHAEL THAT
CAME AT THE END, DID NOT HAVE
TIME TO ANSWER BACK THEN.
>> I HAVE A QUESTION.
OVER HERE.
WE'VE TALKED ABOUT DIFFERENT
ANIMAL MODELS, ARE THERE -- ARE
WE HUNTING FOR NEW SMALL ANIMAL
MODELS, OTHER SPECIES, OR ARE WE
LOCKED INTO MICE, FERRET,
HAMSTERS, HUMANS?
ARE WE SEARCHING FOR NEW ANIMAL
MODELS?
>> I CAN QUICKLY MENTION,
SOMEONE IS HERE DEVELOPING
REALLY LESS TRADITIONAL ANIMAL
MODELS, MAYBE I'LL PHRASE IT.
>> YEAH, A LOT OF ANIMAL MODELS,
TOMORROW I WILL PRESENT THEM.
>> WE HAVE OTHER MODELS.
I DON'T KNOW IF PETER IS GOING
TO TALK ABOUT THE ROBO HAMSTERS,
BUT IT'S A PERFECT EXAMPLE
BECAUSE IT'S A USEFUL ANIMAL
MODEL BUT THEY DON'T BREED VERY
MUCH.
MAKES IT A REAL CHALLENGE.
WE DO HAVE TO NOT ONLY HAVE A
SUSCEPTIBLE ANIMAL THAT GIVES
CERTAIN PROPERTIES, WE HAVE TO
MAKE SURE THEY ARE USABLE.
PETER'S LAB HAS BEEN ABLE TO USE
THEM.
I DON'T THINK TOO MANY LABS
WOULD WANT TO DO IT.
>> IT WILL ESPECIALLY BE
IMPORTANT WHEN YOU WANT TO GO TO
OTHER VIRUSES, BEYOND FLU AND
SARS-COV-2, BECAUSE NOT ALL THE
VIRUSES ARE AS GOOD IN MICE.
>> WE ACTUALLY HAVE A BUNCH OF
ANIMAL MODELS THAT WE USE.
THIS ISN'T NECESSARILY WORK I
DO, THE MUNSTER LAB HAS A LOT.
WE WORKED WITH BATS, PRAIRIE
DOGS, FOR EXAMPLE, A FEW
INTERESTING ANIMAL MODELS.
I THINK THAT WORK IS REALLY
IMPORTANT, IT'S VERY INTERESTING
TO DO.
I THINK YOU RUN INTO THE SAME
PROBLEM AGAIN THAT THERE'S NOT
ENOUGH REAGENTS AND NOW OFTEN
SHIPPING IS REALLY EXPENSIVE TO
GET THEM TO YOUR FACILITY AND
THEN WITH DIFFERENT CAGES AND
DIFFERENT BRANCH LEADERS
LEARNING HOW TO DEAL WITH
ANIMALS, OTHER ANIMAL MODELS ARE
REALLY IMPORTANT, INVESTMENT YOU
NEED TO MAKE AS A LAB IS
SIGNIFICANT.
>> I ALSO THINK IT'S IMPORTANT
TO KIND OF LOOK FOR OTHER ANIMAL
MODELS BECAUSE AT SOME POINT
WHEN NEW VIRUSES EMERGE, SO
THERE WILL BE SUGGESTIONS FROM
WHAT IS THE RESERVOIR, YOU MAY
WANT TO KIND OF BE READY TO HAVE
OTHER ANIMAL MODELS MAYBE
POTENTIAL RESERVOIR, ALREADY
BEEN GOING.
>> MAYBE FOR CORONAVIRUSES AND
OTHER VIRUSES TOO PEOPLE ARE
INTERESTED IN THE BATS.
RECENTLY IN THE NEWS, DISCUSSION
OF THIS BUILDING A BAT FACILITY
AT COLORADO STATE, IT'S RUN
TOO -- NIMBY PROBLEMS OR
BIOSAFETY PROBLEMS, BUT IT'S
GOING TO BE BUILT, BUT THERE'S
SOME PROTESTS ABOUT IT.
IT'S A ALL NOT STRAIGHT FORWARD.
THERE'S SO MANY TYPES OF BATS.
A BAT IS NOT A BAT IS NOT A BAT.
WE HAVE A BAT COLONY, DIFFERENT
VIRUS, ENTIRELY DIFFERENT BATS
AGAIN.
IT GETS COMPLEX.
>> OKAY.
IF THERE'S NO MORE QUESTIONS I
THINK WE CAN THANK THE PANEL FOR
A GREAT DISCUSSION.
>> JUST BEFORE WE CLOSE OUT THE
FIRST DAY OF OUR MEETING, I HAVE
COMPILED A FEW SLIDES OF THE
ANSWERS THAT WE'VE GOTTEN SO FAR
TO SOME QUESTIONS.
SO IF WE COULD BRING UP THAT
SLIDE SET THAT WOULD BE HELPFUL.
OKAY.
SO, YOU KNOW, DON'T GET EXCITED
IF YOUR ANSWER CAME IN LATE AND
IT'S NOT HERE.
WE'LL TALK ABOUT THIS TOMORROW,
RIGHT?
TO SORT OF CONTINUE TO THINK
ABOUT, YOU KNOW, SOME OF THE
REAGENTS NEEDED, SO ONES THAT
CAME IN FIRST ON THE TOP, AN
IMPORTANT POINT, WHICH IS THERE
MAYBE NEEDS TO BE A DATABASE OF
VALIDATED REAGENTS, FAR MORE
IMPORTANT THAN CREATING MORE AND
MORE UNVALIDATED ANTIBODIES,
RIGHT?
AND SO THIS IS SOMETHING THAT
WE'D LIKE TO ENTERTAIN, BUT ALSO
MAYBE SPEND SOME TIME DISCUSSING
TOMORROW BEST WAYS TO APPROACH
SUCH A DATABASE, RIGHT?
IT'S MORE THAN JUST JUST HAVING
A SPREADSHEET BUT GETTING
APPROPRIATE INPUT FROM ANYBODY
DOING RESEARCH IN THE AREAS.
THERE'S A COUPLE ADDITIONAL
SUGGESTIONS FOR VALIDATED HIGH
AFFINITY ANTIBODIES, T CELL
ANALYSIS, INTRACELLULAR CYTOKINE
STAINING, B CELLS BY FLOW ARE
HELPFUL, ANTIBODIES TO ASSESS
NUMBER AND FUNCTIONALITY OF
INNATE IMMUNE CELLS, SO WE'VE
HEARD THIS A FEW TIMES, THE
MEASUREMENTS OF IMMUNE CELLS
WILL BE REALLY IMPORTANT.
SO SECOND QUESTION THAT WHEE IS
NEED FOR USE OF CONSISTENT
VALIDATED CHALLENGE STRAINS OF
VIRUS.
SO SOMEBODY HAS SUGGESTED THAT
IT'S MORE IMPORTANT THAT VIRUS
STRAINS ARE DEEP SEQUENCED, THE
DATA IS AVAILABLE TO UNDERSTAND
THE EFFECTS ON PHENOTYPIC
OUTCOME, THERE COULD BE
QUASI-SPECIES WITHIN THE VIRAL
SAMPLES PEOPLE ARE USING.
PR 8 BASED STRAINS OR MOUSE
ADAPTED STRAINS TO COMPARE IS
ONE IDEA THAT MERITS DISCUSSION.
EARLIER HAVING ANOTHER
DISCUSSION ABOUT USING REVERSE
GENETICS, YOU KNOW, BASED -- --
YOU KNOW, VIRUSES, RIGHT?
I THINK THAT ALSO IS A
MERITORIOUS IDEA THAT SHOULD BE
DISCUSSED AS WELL.
SO, THE LAST QUESTION IS A
LITTLE BIT MORE PHILOSOPHICAL IN
NATURE, WHEN WE'RE TALKING ABOUT
HOW TO COMPARE ANIMAL MODELS TO
THE HUMAN DATA THAT'S AVAILABLE
TO, YOU KNOW, CONTINUE TO ENSURE
SOME CLINICAL RELEVANCY, RIGHT?
INTERESTING SUGGESTION TO STUDY
FAILED CLINICAL TRIALS, WORK
BACKWARDS, TO UNDERSTAND WHY
ANIMAL MODELS WEREN'T
PREDICTIVE.
THAT'S A VERY BIG IDEA BUT ALSO
SOMETHING THAT IS A GOOD
THOUGHT.
AND OF COURSE IT DEPENDS ON THE
QUESTION THAT YOU'RE TRYING TO
ANSWER, RIGHT?
IF YOU'RE LOOKING AT
TRANSMISSIBILITY SOME MODELS ARE
BETTER THAN OTHERS.
LOOKING AT PATHOGENESIS SOME
MODELS ARE BETTER THAN OTHERS.
WHAT I CHALLENGE YOU TO THINK
ABOUT IN YOUR SPECIFIC SITUATION
IF YOU'RE STUDYING TRANSMISSION,
RIGHT, YOU KNOW, HOW DO YOU SORT
OF ENTER THAT CLINICAL RELEVANCY
VERSUS STUDYING THE IMMUNE
RESPONSE, TRANSLATABILITY TO
HUMANS AS WELL.
WE'LL DISCUSS IN MORE DETAIL IN
SESSION 4 TOMORROW.
SO I'D LIKE TO THANK EVERYBODY
WHO GAVE WONDERFUL TALKS TODAY
AND FOR VERY ENGAGING PANEL
DISCUSSIONS, VERY MUCH
APPRECIATE EVERYBODY
PARTICIPATING.