>> GOOD MORNING, EVERYBODY.
I HOPE YOU ALL ENJOYED THE DAY
YESTERDAY AND ARE READY TO
FINISH UP WITH A LITTLE BIT OF A
LIGHTER DAY TODAY, SO WE HAVE
TWO MORE SESSIONS, BOTH OF WHICH
WE'LL HAVE A DISCUSSION PANEL,
AN EXTRA DISCUSSION PANEL IN THE
AFTERNOON TO TALK ABOUT
REAGENT AND THOSE KIND OF
THINGS.
FOR THOSE WHO WANT TO ORDER
LUNCH, WE HAVE THE SAME SYSTEM
TODAY AS WE DID YESTERDAY.
SO LUNCH CAN BE ORDERED USING
THE QR CODE ON THE AGENDA OR ON
THE FLYERS ON YOUR TABLES.
THAT'S FROM CHOPT.
PLEASE DO THAT BY 9:30, IF
THAT'S WHERE YOU WANT TO GET
YOUR LUNCH FROM.
TODAY IS A LITTLE BIT SHORTER OF
A DAY TOO.
WE WILL ADJOURN IN EARLY
AFTERNOON, SO BY 3:15 HOPEFULLY,
UNLESS THE DISCUSSION IS VERY
RIVETTING, WE'LL KEEP GOING, B
BUT -- I DON'T THINK THERE'S ANY
ADDITIONAL HOUSEKEEPING ITEMS.
SO TODAY WE'RE GOING TO START
WITH SESSION 3, WHICH IS THE
UTILITY OF ANIMAL MODELS IN
RESPIRATORY VIRUS TRANSMISSION.
WE HAVE OUR MODERATOR IS SABRA
KLEIN, AND OUR FIRST SPEAKER
WILL BE DELIVERING HIS TALK
VIRTUALLY.
AND THE REST WILL BE IN PERSON.
>> GOOD MORNING, EVERYONE.
HOW IS EVERYBODY DOING?
CAN EVERYBODY HEAR ME?
>> WE CAN HEAR YOU WELL.
>> THANK YOU, SABRA.
I APOLOGIZE FOR NOT ATTENDING IN
PERSON.
I SUFFERED FROM A BIT OF A HEAD
COLD AND I DIDN'T WANT TO SHARE
THAT WITH THE ENTIRE CONFERENCE.
BEFORE I START, I WANT TO THANK
MICHELLE AND STEVEN FOR ACTUALLY
PUTTING TOGETHER A FANTASTIC
WORKSHOP.
I LEARNED A LOT YESTERDAY, AND
I'M LOOKING FORWARD TO LEARNING
A LOT MORE TODAY.
FOR THOSE OF YOU WHO DON'T KNOW
ME, MY NAME IS JACCO BOON, AND I
AM IN THE DEPARTMENT OF MEDICINE
AT WASHINGTON UNIVERSITY IN
ST. LOUIS, MISSOURI, IN THE
MIDDLE OF THE COUNTRY.
AND I'M GOING TO TALK A LITTLE
BIT ABOUT THE HAMSTER MODEL AND
HOW IT'S USED IN SARS-COV-2 AND
INFLUENZA.
THIS IS THE LAYOUT OF THE TALK.
A LITTLE BIT OF HISTORY, THEN,
OF COURSE, THE EXPLOSION OF
HAMSTER RESEARCH FOLLOWING THE
EMERGENCE OF SARS-COV-2, SOME OF
THE ADVANTAGES AND CURRENT
NEEDS.
SO WHEN I LOOKED AT INFLUENZA
VIRUS IN PUBMED AND SYRIAN
HAMSTERS, I RAO HELY FOUND ONE
PAPERS.
ONE FROM 1965 WHO USED PRA VIRUS
SHOWING PULMONARY LESIONS IN
SYRIAN GOLDEN HAMSTERS FOLLOWING
INFECTION.
THIS IS REALLY WHAT I WOULD CALL
A SEMINAL STUDY IN 2018, ALMOST
50 YEARS LATER, SHOWING THAT THE
SYRIAN HAMSTER MODEL COULD BE
USED TO STUDY HUMAN INFLUENZA
VIRUS INFECTION.
AND WHAT THEY SHOWED IS THAT
SOME HUMAN INFLUENZA VIRUSES,
H1N1 AND TYPE B VIRUSES -- HIGH
TITERS -- WHAT WAS EVEN MORE
SURPRISING IS THEY FOUND IN THE
CASE OF SOME VIRUSES SUCH AS
THIS H1N1 CALIFORNIA VIRUS THAT
YOU ACTUALLY SAW AIRBORNE
TRANSMISSION FROM THE INFECTED
DONOR ANIMALS TO THE EXPOSED
CONTACTS.
BUT THIS WAS THE EXTENT OF
INFLUENZA VIRUS FIELD IN SYRIAN
HAMSTERS UNTIL THE PANDEMIC,
WHEN GROUPS STARTED LOOKING AT
SIMILARITIES AND DIFFERENCES
BETWEEN THE MODELS AND I'LL TALK
A LITTLE ABOUT THAT LATER.
SO ANOTHER REALLY IMPORTANT
PAPER I THINK THAT STARTED THE
SARS-COV-2 FIELD AS WELL AS THE
HAMSTER FIELD WAS THIS PAPER AND
THIS WAS FOLLOWING THE EMERGENCE
OF THE ORIGINAL SARS COV IN
2003, AND THEY SHOWED UPON
INTRANASAL INOCULATION YOU GOT
VERY TIE TIT.
TERS BOTH IN THE
UPPER AIRWAY SHOWN IN BLACK AS
WELL AS IN THE LUNGS FOLLOWING
INTRANASAL INFECTION.
THEY CONFIRMED THIS USING VERY
NICE CHEMISTRY DETECTING VIRAL
PROTEIN IN THE UPPER AIRWAYS AS
WELL AS IN THE LOWER AIRWAYS.
BUT IMPORTANTLY THERE WAS NO
EVIDENCE FOR CLINICAL SYMPTOMS
CONSISTENT WITH SARS-COV-2.
THIS IS STILL SOMETHING UNTIL
THIS DAY THAT A LOT OF
SCIENTISTS WOULD LIKE TO SEE
WEIGHT LOSS, PERHAPS EVEN DEATH
FOLLOWING VIRUS INFECTION.
HOWEVER, I THINK THESE TWO
PAPERS LED TO THIS TEAM AND
ANOTHER TEAM TO HONE IN ON THE
SYRIAN HAMSTER EARLY DURING THE
SARS-COV-2 PANDEMIC, AND THIS IS
WORK THAT WAS LED BY OUR
COLLEAGUES WITHIN THE NETWORK,
TOGETHER WITH ANOTHER GROUP ALSO
AT HONG KONG UNIVERSITY SHOWING
HOW GOOD THE SYRIAN GOLDEN
HAMSTER WAS FOR SARS-COV-2
STUDIES.
JUST TO SUMMARIZE THEIR WORK AS
WELL AS MANY, MANY OTHER
PEOPLE'S WORK AND I APOLOGIZE IF
YOUR PAPER IS NOT SOMEWHERE IN
MY PRESENTATION, BUT THE OVERALL
CONCLUSION WAS THAT THE SYRIAN
HAMSTER RECAPITULATES MANY
ASPECTS OF HUMAN CLINICAL
DISEASE.
WE GOT SEVERE PATHOLOGY AFTER
INFECTION, THE VIRUS HAS SIMILAR
TROPISM, SO SEVERE PATHOLOGY
SPECIFICALLY BEFORE THE
EMERGENCE OF THE OMICRON
VARIANT, MORE ON THAT LATER.
WE SAW SOMEWHAT SIMILAR TROPISM
COMPARED TO HUMAN INFECTION WITH
PREDOMINANTLY UPPER AIRWAYS, THE
NASAL TURBINATES AND THE LUNGS
WITH SOME DISSEMINATION TO EXTRA
PULMONARY TISSUE SUCH AS THE
OLFACTORY BULB AS PRESENTED
YESTERDAY IN A BEAUTIFUL TALK,
AS WELL AS THE HEART AND THE THE
INTESTINE.
THIS IS A LITTLE BIT MORE
CONTENTIOUS A STATEMENT BUT
LIMITED TO -- APPLICATION IN THE
BRAIN, AT LEAST NOT IN OUR HANDS
AS WELL AS MANY OTHER LABS.
THIS IS NOT TO SAY THERE ARE NO
NEUROLOGICAL PHENOTYPES
FOLLOWING SARS-COV-2 INFECTION,
WHICH IS SHOWN -- AS WELL,
FOLLOWING THE EXCESSIVE
INFLAMMATION CAUSING THE
NEUROLOGICAL SYMPTOMS.
AND SHORT-TERM MEMORY LOSS.
THERE WAS MORE SEVERE DISEASE IN
ELDERLY ANIMALS AND
IMMUNOCOMPROMISED ANIMALS.
VERY IMPORTANT TO ME AND MANY
OTHERS IS THAT AIRBORNE
TRANSMISSION WAS READILY
DETECTED, WHICH REALLY KIND OF
OPENED UP THE FIELD, I THINK
REALLY HELPED ALSO THE HAMSTER
MODEL BEING SO PROMINENT FOR
SARS-COV-2 RESEARCHERS.
AND THEN ANOTHER PAPER FROM
BEN'S LAB SHOWING THAT IMMUNE
MEMORY FROM PREVIOUS INFECTIONS
OR VACCINATIONS PREVENTED
DISEASE.
AND THIS WORK -- AND MORE ON
THAT LATER.
SO IN TERMS OF AIRBORNE
TRANSMISSION, WHAT HAVE WE
LEARNED SO FAR?
ME, MY LAB AND MANY OTHER LABS
HAVE DONE A LOT OF WORK ON THIS.
THE THINGS WE'VE LEARNED IS THAT
CONTACT TRANSMISSION IS MORE
EFFICIENT IN AIRBORNE
TRANSMISSION.
PROLONGED EXPOSURES ALSO
INCREASED TRANSMISSION
EFFICIENCY.
IN OUR SYSTEM, AIRBORNE
TRANSMISSION WAS OPTIMAL AT 24
HOURS POST INOCULATION OF THE
DONOR ANIMAL BUT IT CAN LAST AS
LONG AS THREE TO FOUR DAYS WITH
SOME OF THE OTHER VARIANTS.
TRANSMISSION CAN BE DETECTED
REALLY EARLY, ALMOST 30 MINUTES
AFTER EXPOSURE.
AND IN OUR VERY SIMPLE SYSTEM
WHERE THE TWO ANIMALS ARE
SEPARATED BY A SMALL SPACE AND
DIRECTIONAL AIR FLOW, WE CAN SEE
100% TRANSMISSION AT EIGHT
HOURS.
TRANSMISSION IS MORE EFFICIENT
WITH HIGHER TITERS AND IS
INFLUENCED BY PARTICLE SIZE.
SO LARGER PARTICLES TEND TO BE
THE ONES THAT ACTUALLY CARRY THE
VIRUS AND TRANSMIT.
THERE'S DIFFERENCES IN ARRAY OF
AIRBORNE TRANSMISSION BETWEEN
STRAINS OF COV2.
OMICRON, 100% TRANSMISSION OF
THE ORIGINAL VIRUS, AS WELL AS
D614G, BETA, DELTA, YOU NAME IT,
AND THIS REALLY FELL OFF A CLIFF
WITH THE EMERGES OF THE
BA1 OMICRON VARIANT.
THIS HAS PICKED UP A LITTLE WITH
THTHESE.
SO YESTERDAY -- THERE'S NO
AIRBORNE TRANSMISSION WITH
THE -- SO THIS SEEMS TO VARY A
LITTLE BIT DEPENDENT ON THE
STRAIN AND I HOPE OTHERS WILL
ACTUALLY -- WHAT IT IS ABOUT
THESE VIRUSES AND WHAT FEATURES
ARE ASSOCIATED WITH EFFICIENT
AIRBORNE TRANSMISSION IN SYRIAN
HAMSTERS.
AND BECAUSE IT'S SO EFFICIENT,
ONWARDS OR SECONDARY
TRANSMISSION FROM THE DONOR TO
THE PRIMARY AND THEN THE PRIMARY
TO THE SECONDARY AND EVEN
TERTIARY ANIMAL CAN BE READILY
DETECTED, WHICH IS SOMETHING
THAT I'LL TOUCH UPON LATER AS
WELL, WHICH IS REALLY HELPFUL
FOR STUDYING COMMUNITY
TRANSMISSION OR -- DAYCARE TYPE
OF TRANSMISSION MODEL.
SO PROBABLY THE MOST EFFICIENT
USE OF THE HAMSTER SO FAR HAS
BEEN TO DEMONSTRATE THE MANY
DIFFERENT VACCINES, PRE-CLINICAL
VACCINES OR EVEN CLINICAL
VACCINES IN THE HAMSTER MODEL.
I LOOKED THIS UP, I THINK THE
FIRST PAPER IN HAMSTER MODEL MAY
HAVE BEEN THIS ONE SHOWING THAT
THE AD26 VACCINE PROTECTED
AGAINST SEVERE DISEASE, BUT
THERE HAVE BEEN MANY, MANY
OTHERS.
-- MOST OF THE PEOPLE IN THE
ROOM HERE.
I WOULD LIKE TO NOTE THAT SOME
OF THESE PAPERS WERE ACTUALLY
STUDYING MUCOSAL VACCINES.
FOR EXAMPLE HERE, YOU'RE LOOKING
AT THE CHADOX VIRUS AND VINCENT
MUNSTER AS WELL AS OUR
CONTRIBUTION AND HERE A
LENTIVIRAL VECTOR WHERE PEOPLE
WERE LOOKING AT INTRANASAL
VACCINES AND SOMETIMES COMPARE
THEM TO INTRAMUSCULAR DELIVERY
SHOWING THAT INTRANASAL VACCINES
WERE ACTUALLY MORE EFFECTIVE IN
PREVENTING UPPER -- LOWERING
UPPER AIRWAY TITERS THAN
INTRAMUSCULAR.
TO COME BACK TO THIS ONWARDS
TRANSMISSION OR COMMUNITY
TRANSMISSION MODEL, AGAIN,
VINCENT'S LAB AS WELL AS MY LAB
HAVE USED THIS TO SHOW HOW
MUCOSAL VACCINES CAN IMPACT THIS
COMMUNITY SPREAD, AND HOW IT MAY
NOT ONLY PROTECT AGAINST THE
INITIAL INFECTION BUT MAY ALSO
LOWER THE RISK OF ONWARDS
TRANSMISSION.
SO THIS IS JUST ONE PANEL IN A
FIGURE IN 2003 SHOWING IN BLACK
THERE'S AN INFECTED ANIMAL, IN
WHITE IS AN UNINFECTED OR AN
ANIMAL THAT NEVER GOT IT.
SO WHAT HE SHOWED IS THAT IF YOU
WERE PREVIOUSLY INFECTED, P.I.,
YOU ACTUALLY HAD A VERY LOW RISK
OF ONWARDS TRANSMISSION, EITHER
GETTING INFECTED OR ACTUALLY
SHEDDING THE VIRUS TO A SECOND
OR THIRD ANIMAL.
THE NEXT BEST THING WAS
INTRANASAL IMMUNIZATION FOLLOWED
BY INTRAMUSCULAR IMMUNIZATION
AND THEN -- AND WE RECENTLY GOT
OUR PAPER ACCEPTED IN SCIENCE
ADVANCES SHOWING VERY SIMILAR.
SO THIS IS LOOKING AT SECONDARY
TRANSMISSION, SO FROM THE
PRIMARY CONTACT TO THE SECONDARY
ANIMAL.
LOOKING AT -- SHOWN HERE ON Y
AXIS IS THE PERCENT SARS-COV-2
POSITIVE CONTACTS.
SO IN THE NAIVE ANIMALS, YOU
HAVE ALMOST AN 80% TRANSMISSION
RATE FROM THE PRIMARY TO THE
SECONDARY ANIMAL.
THIS DEPENDS A LITTLE BIT ON HOW
LONG YOU EXPOSE AND WHAT TIME
YOU EXPOSE.
TO INTRAMUSCULAR IMMUNIZATION
WITH MRNA, YOU ACTUALLY STILL
GET A LOT OF ONWARDS OR
SECONDARY TRANSMISSION.
HOWEVER, IF YOU USE A MUCOSAL
VACCINE, REFERENCED HERE AS
CHAD-S, VECTOR CHIMPANZEE ADD
FLOW VIRUS VECTOR VACCINE, YOU
ACTUALLY BLOCK SECONDARY
TRANSMISSION NOT ONLY TO OTHER
VACCINATED ANIMALS BUT ALSO TO
NAIVE ANIMALS.
SO I THINK THIS KIND OF
HIGHLIGHTS ONE OF THE UNIQUE
FEATURES OF THE SYRIAN HAMSTER
MODELS IN TERMS OF THE ABILITY
TO TRANSMIT THE VIRUS AND TO USE
LARGE ENOUGH ANIMALS AND GROUP
SIZES TO REALLY ASSESS THE
IMPACT OF VACCINE TRANSMISSION
AND -- TRANSMISSION OF THE
VIRUS.
SO FOUR YEARS INTO THE PANDEMIC,
SO WHAT ARE HAMSTERS CURRENTLY
USED FOR?
I'M SURE I'M MISSING SEVERAL
REALLY IMPORTANT SEMINAL STUDIES
HERE, BUT I THINK THE FIELD HAS
SETTLED ON A LOT OF THESE.
AND THEY FALL IN ONE OR MORE OF
THESE CATEGORIES.
SO THERE ARE GOING TO BE MORE
AND MORE VACCINE STUDIES COMING
DOWN, BEING DONE RIGHT NOW IN
MANY DIFFERENT LABS.
PEOPLE WILL IMPROVE MUCOSAL
VACCINES, THEY WILL IMPROVE
VECTOR VACCINES, MRNA VACCINES,
RECOMBINATION VACCINES.
PEOPLE WILL START TO STUDY
VACCINE EFFICACY AGAINST
PRE-EXISTING IMMUNITY AS WE AND
OTHERS HAVE DONE ALREADY.
PEOPLE ARE LOOKING AT VARYING
VACCINES AND THE EFFICACY
AGAINST NEWLY EMERGING
SARS-COV-2 VIRUSES, WHICH IS
SOMETHING MY LAB DOES QUITE A
BIT OF, AND THEN UNDERSTANDING
CORRELATES OF IMMUNE PROTECTION.
I THINK THE HAMSTER WILL START
TO BE USED MORE FOR ANTIVIRAL --
MEASURE STUDIES SUCH AS SMALL
MOLECULES.
THIS WILL REQUIRE -- THIS IS NOT
MY AREA OF EXPERTISE, I THINK IT
WILL REQUIRE A SIGNIFICANT
INVESTMENT IN UNDERSTANDING THE
PHARMACOKINETICS AND DYNAMICS
AND SMALL MOLECULE CLEARANCE IN
THE HAMSTER COMPARED TO SOME OF
THE OTHER MORE ESTABLISHED SMALL
ANIMAL MODELS.
SO I THINK MORE AND MORE PEOPLE
WILL DO FUNDAMENTAL BIOLOGY OF
SARS-COV-2 INFECTION,
PATHOGENESIS AND TRANSMISSION.
THIS IS A PAPER FROM MY LAB
LOOKING AT THE SIGNIFICANCE OF A
RNA MOTIF CALLED S2M IN THE
3 PRIME REGION OF SARS-COV-2,
AND WE SHOWED IT ACTUALLY WAS
DISPENSABLE FOR SARS-COV-2, AND
THEN WHILE WE WERE WRITING THIS
PAPER, THE VIRUS ITSELF ACTUALLY
EVOLVED AND DELETED A SLIGHTLY
DIFFERENT S2M REGION, A
DIFFERENT SEQUENCE BUT STILL IT
DELETED THE S2M IN ITS GENOME.
SO THE CURRENT VARIANTS OF SARS
CHSARS-COV-2 NO LONGER CARRY IT.
SEVERAL PAPERS, AND MORE WILL
COME, ARE LOOKING AT MUTATIONS,
THE ROLE OF ORF AND DELETION
MUTANT VIRUSES ON PATHOGENESIS
REPLICATION AND HOPEFULLY
TRANSMISSION.
HE SEVERAL GSEVERAL GROUPS ARE T
THE ROLE OF CO-INFECTION ON
SARS-COV-2 DISEASE, REPLICATION
AND TRANSMISSION.
AND THIS IS NOT JUST INFLUENZA
VIRUS BUT ALSO OTHER
CORONAVIRUS.
I HOPE THE FIELD WILL EVOLVE
FURTHER TO INCLUDE MANY, MANY
MORE PATHOGENS SUCH AS RSV,
OTHER CORONAVIRUS, ET CETERA.
AND THEN BARCODED SARS-COV-2,
SOMETHING THAT MY LAB HAS BEEN
WORKING ON EXTENSIVELY IN THE
LAST COUPLE OF YEARS.
SO WE RECENTLY POSTED A
PRE-PRINT ON BIOARCHIVES.
THIS IS WORK DONE BY A VERY
TALENTED GRADUATE STUDENT,
POSTDOC IN MY LAB, TRENDE.
WHAT WE DID IS ESSENT ESSENTIALK
AT SOME OF THE LITERATURE ON
BARCODED VIRUSES.
WE DECIDED TO INTRODUCE,
TOGETHER WITH ANOTHER LAB, DAVE
WANG AT WASHINGTON UNIVERSITY,
WE DECIDED TO INTRODUCE A NINE
NUCLEOTIDE BARCODE IN ORF10 IN
SARS-COV-2.
GNNGNNGNN.
WE INDIVIDUALLY GENERATED OVER
200 UNIQUE VIRUSES THROUGH THIS
METHOD.
WE PULLED THESE AT EQUAL AMOUNTS
OF INFECTIOUS TITER AND THEN
MEASURED THE RELATIVE BARCODE
FREQUENCY IN THIS POOL OF OURS.
IT'S SHOWN HERE IN THE TOP BAR
IN THIS KIND OF RAINBOW.
ONE PICTURE ACTUALLY MATCHES THE
COLORS THROUGHOUT THE
PRESENTATION.
WE HAD TO DO A LOT OF UP-FRONT
INVESTIGATIVE STUDIES TO MAKE
SURE THAT THE VIRUSES WERE GOOD,
AND WE DIDN'T HAVE EITHER GROSS
ATTENUATION OR INCREASED
REPLICATION.
SO IN VITRO, ALL THE VIRUSES
REPLICATED TO THE SAME TITER AND
IN VIVO, COMPARING THE BARCODE
FREQUENCY BETWEEN THE INNOCULUM
AND 32 AND 72 HOURS POST
INOCULATION IN HAMSTERS IN
VARIOUS TISSUES BUT WE DIDN'T
REALLY SEE A SIGNIFICANT
INCREASE IN ONE BARCODE OVER
ANOTHER OVER MANY, MANY
DIFFERENT EXPERIMENTS.
SO I'M NOT GOING TO TALK ABOUT
THE ENTIRE PAPER BUT I KIND OF
WANT TO SHOW A VIGNETTE OF SOME
THINGS YOU CAN DO WITH THIS
REALLY UNIQUE RESOURCE.
SO THE FIRST THING THAT WE DID
IS TO SHOW THAT THESE BARCODED
VIRUSES ACTUALLY TRANSMIT AND
HOW EFFICIENT IS SARS-COV-2
TRANSMISSION.
SO IN OUR MODEL SYSTEM, AGAIN
IT'S RELATIVELY STRAIGHTFORWARD,
SO WE TAKE A DONOR ANIMAL, WE
INOCULATE IT WITH 100,000
INFECTIOUS UNITS, BARCODED
VIRUSES.
WE WAIT 24 HOUR, THEN EXPOSE OUR
CONTACT ANIMAL FOR A BRIEF
AMOUNT OF TIME, EIGHT HOURS, AND
WE CAN DO EITHER DIRECT CONTACT
OR AIRBORNE.
THIS IS DATA FROM AIRBORNE
EXPOSURES.
AND THEN WE WAIT A TOTAL OF
72 HOURS BEFORE WE SACRIFICE THE
CONTACT ANIMAL.
WE COLLECT THE TISSUES, HERE I
JUST AM SHOWING YOU THE NASAL
TURBINATES AND SEE THE TRACHEA
AND THE LUNGS, AND WE JUST
MEASURE HOW MANY BARCODES CAN WE
DETECT.
AND THERE'S A COUPLE THINGS THAT
I WANT YOU TO TAKE AWAY FROM
THIS SLIDE.
SO FIRST OF ALL, YOU DETECT A
LOT OF DIFFERENT BARCODES.
IN THIS PARTICULAR ANIMAL, WE
HAD 13 DIFFERENT BARCODES.
THIS ONE WE HAVE 20 UNIQUE
BARCODES.
SO AS I INDICATED EARLIER, SARS
TRSARS-COV-2 AIRBORNE TRANSMISSN
IN HAMSTERS IT REALLY VERY
EFFICIENT.
THERE ARE MULTIPLE BARCODES
THROUGHOUT THE VARIOUS TISSUES
AND MANY ARE SHOWN HERE.
SO EACH CIRCLE IN THE VENN
DIAGRAM IS A DIFFERENT TISSUE.
WE'VE ACTUALLY ALSO DONE THIS AT
DAY 1 AND DAY 2 POST EXPOSURE
AND WE ACTUALLY SEE MORE
TISSUE-SPECIFIC BARCODES.
FOR EXAMPLE, THIS BLUE ONE HERE
IS REALLY SPECIFIC TO THE LUNGS
BUT NOT THE NASAL TURBINATES,
SUGGESTING THAT THE VIRUS CAN
TEUMLY BYPASS THE UPPER AIRWAYS
AND DIRECTLY INFECT THE LOWER
AIRWAYS.
SO THIS IS ONE THING THAT WE
DID, WE LOOKED AT EXPOSURE
TIMES.
WE CAN SEE THAT THERE IS A
SIGNIFICANT DIFFERENCE IN THE
AMOUNT OF BARCODES DETECTED IF
YOU EXPOSE FOR ONE HOUR, FOUR
HOURS OR EIGHT HOURS.
AND I INCLUDED THIS SLIDE FOR
SABRA, WE COMPARED THE NUMBER OF
BARCODES OR AIRBORNE
TRANSMISSION BETWEEN MALE AND
FEMALE HAMSTERS AND THERE ARE NO
CLEAR DIFFERENCES IN THE AMOUNT
OF BARCODES DETECTED, SUGGESTING
THAT THERE'S NO LARGE DIFFERENCE
IN TRANSMISSION OR
SUSCEPTIBILITY BETWEEN BOTH
SEXES.
SO WHAT ARE THE ADVANTAGES OF
THE HAMSTER?
THEY'RE CLEAR LEHIGHLY
SUSCEPTIBLE TO SARS-COV-2 AND
THEY'RE PERMISSIVE TO OTHER --
AND INFLAWN IS A VIRUS.
I THINK THIS OPENS UP A WHOLE
NEW FIELD THAT -- TO STUDY THE
ROLE OF COINFECTIONS, PRIOR
INFECTIONS ON TRANSMISSION,
SUSCEPTIBILITY OF THE VIRUS, ET
CETERA.
HAMSTER ARE RELATIVELY CHEAP,
EASY TO HOUSE AND HANDLE.
I SEE TOO MANY PAPERS LOOKING AT
REALLY SMALL NUMBERS, AND I
DON'T THINK THAT IS JUSTIFIED IN
THE CONTEXT OF SYRIAN HAMSTERS.
WE REALLY SHOULD USE LARGER
GROUP SIZES AND REPEATING
EXPERIMENTS.
AND WE SHOULD INCLUDE MALE AND
FEMALE HAMSTERS WHEN REQUIRED.
THE HAMSTERS, THE GENOME IS
AVAILABLE, THE ANIMALS ARE
INBRED, AND THIS WAS ALLUDED TO
YESTERDAY BUT WE HAVE EVIDENCE
TO STRONGLY SUGGEST THAT, REALLY
SIMPLIFYING SOME OF THE
TRANSCRIPTIONAL STUDIES
INCLUDING SINGLE CELL RNA SEQ
THAT ARE AVAILABLE.
THERE ARE A HUGE GAP IN
IMMUNOLOGICAL REAGENTS AND
ASSAYS THAT ARE AVAILABLE.
I THINK MY LAB AS WELL AS OTHERS
HAVE REALLY PUSHED THIS ENVELOPE
AND THE ASSAYS THAT ARE NOW
ONLINE REALLY IMPROVE THE MODEL
AS A WHOLE.
JUST TO SHOW YOU TWO EXAMPLES OF
THIS, THIS IS A PAPER CURRENTLY
UNDER REVISION FROM MY LAB
SHOWING OR DEVELOPING A B CELL
ELISPOT ASSAY FOR SYRIAN
HAMSTERS MEASURING BOTH THE IGG
AS WELL AS THE IGA ANTIBODY
RESPONSE AT THE CELLULAR LEVEL
IN THESE ANIMALS.
SO THIS IS JUST HOW THIS LACKS.
AND WE CAN MEASURE, AND BEEF
DONE THIWE'VEDONE THIS NOW IN DT
VACCINE CONTEXTS.
THIS IS AN EXAMPLE OF A PROTEIN
VACCINE, NOVAVAX, IGG AND IGA
LEVEL IN BOTH THE SPLEEN AND D
DLN.
AND WE CAN DETECT EITHER
INTRANASAL VECTOR OR MRNA, WE
CAN DETECT IT IN MULTIPLE
DIFFERENT TISSUES.
WE ALSO IMPLEMENTED THE T-CELL
ELISPOT.
THIS IS A COMMERCIAL ASSAY,
AVAILABLE FROM MED TECH, AND WE
MADE SURE THAT IT WORKED AND IT
WAS ROBUST, CONFIRMING SOME OF
THE STUFF THAT PEOPLE HAVE DONE
IN MICE SHOWING THE OPTIMAL TIME
POINT FOR TISSUE COLLECTION,
OPTIMIZING THE ASSAYS FOR
MEASURING T-CELL RESPONSES IN
BOTH THE SPREEN, DRAINING LYMPH
NODE AS WELL AS IN HERE.
I'M SHOW SHOWING YOU T-CELL RES
THROUGH -- USING EX VIVO
DEPREETDEPLETION ASSAYS, FOLLOWG
IMMUNIZATION, TAKE THE TISSUES,
THE SPLEEN, DRAINING LYMPH NODE
OR EVEN THE LUNGS, SHOWING THAT
IN THIS PARTICULAR SITUATION
FOLLOWING PROTEIN-BASED
NANOPARTICLE VACCINE THAT THE
T-CELL RESPONSE IS PREDOMINANTLY
CD4 MEDIATED.
SO IF YOU DEPLETE THE CD4s,
YOU HAVE A TENFOLD REDUCTION IN
THE AMOUNT OF SPOTS DETECTED
WHEREAS IF YOU DEPLETE THE
CD8s, YOU DON'T REALLY GET ANY
DIFFERENCE.
SO BEFORE I OPEN THE FLOOR TO
QUESTIONS, BUT THERE'S STILL
MANY NEEDS FOR THE HAMSTER
MODELS.
SO WE NEED MORE VALIDATED
REAGENTS.
I WANT TO EMPHASIZE THE
VALIDATED PART.
AND THIS NEEDS TO BE VALIDATED
IN MANY WAYS, PREFERABLY USING
MASS SPECTROMETRY SHOWING
WHATEVER YOU PULL DOWN OR WHAT
THE ANTIBODY DETECTS IS REALLY
WHAT YOU WANT IT TO BE, AND NOT
CROSS-REACTIVE TO SOMETHING
ELSE.
AND THIS IS IN MY CASE REALLY
IMPORTANT FOR FLOW CYTOMETRY,
B220, CD19, IFN GAMMA, CD103, ET
CETERA.
IN MY LAB, IDENTIFYING MHC CLASS
I AND II ALLELES AND DISCOVER
ROW OF T-CELL EPITOPES, ALLOWING
US TO DEVELOP TRE MA TETRAMERS.
WE WANT TO KNOW MORE ABOUT B
CELL AND T-CELL UTILIZATION.
I THINK THIS IS ANOTHER EXCITING
AREA OF RESEARCH, THE DWARF
HAMSTER, WHICH CONTINUES TO BE
MORE SUSCEPTIBLE TO EVEN THE
CURRENT VARIANCE OF SARS-COV-2,
I THINK WE NEED TO NOT LOSE
SIGHT OF THE DEVELOPMENT OF
TOOLS AND AGENTS FOR THIS.
SO WITH THAT I'D LIKE TO THANK A
LOT OF DIFFERENT PEOPLE AND
FUNDING SOURCES.
SO IN MY LAB, A LOT OF PEOPLE,
SPECIFICALLY TRACI, TAMARAND AND
REED, THE WANG LAB, KLEIN LAB,
ELLEBEDY LAB, DIAMOND LAB.
I WANT TO MAKE A PLUG FOR THE
SAVE COMMUNITY THAT WAS STARTED
VERY EARLY AFTER THE PANDEMIC
AND IT'S BEEN A FANTASTIC
COMMUNITY THAT WAS STARTED VERY
EARLY AFTER THE PANDEMIC,
SHARING DATA, RESOURCES AND
WHATNOT.
SINCE THE START, THIS COMMUNITY
ACTUALLY CONTINUES TO EXIST AND
IF YOU WANT TO KNOW MORE ABOUT
IT, PLEASE REACH OUT TO ME.
WITH THAT, I'LL TAKE ANY
QUESTIONS.
>> YES, LET'S START WITH SOME
QUESTIONS.
>> HI, I REALLY LIKED YOUR TALK.
I ALSO REALLY LIKE YOUR PUSH
FOR -- SPECIFIC REAGENTS.
I THINK WE ALL REALLY PEN FIT BT
FROM THAT.
MY QUESTION IS ABOUT THE
TRANSMISSION OF OMICRON IN THE
HAMSTER MODEL, BECAUSE IT JUST
DOESN'T TRANSMIT THAT WELL IN
HAMSTERS AND IT'S
COUNTERINTUITIVE WHAT WE'RE
SEEING IN A HUMAN POPULATION.
SO WHAT DO YOU THINK THAT MEANS,
HOW MUCH DO YOU THINK THE
HAMSTER MODEL REALLY RE REPRESES
TRANSMISSION IN HUMANS?
>> THAT'S A VERY FAIR POINT.
WE AND OTHERS AS WELL AS YOUR
LAB IS STILL SEEING DIRECT
CONTACT TRANSMISSION SO I THINK
IT IS DOWN TO EFFICIENCY OF
INFECTION.
IF I HAD INFINITE AMOUNT OF
RESOURCES IN THE LAB I WOULD
LOVE TO DO -- TITER DOWN HOW
MUCH VIRUS IS NEEDED TO ZERO
CONVERT AN ANIMAL.
MY PREDICTION IS THAT SOME OF
THE MORE RECENT OMICRON
VARIANTS, THIS NUMBER GOES UP BY
LET'S SAY TENFOLD COMPARED TO
SOME OF THE ORIGINAL SARS-COV-2
VIRUSES.
THIS IS ENOUGH, GIVEN NOW WE
KNOW WHAT THE BOTTLENECK IS,
IT'S SOMEWHERE AROUND 10 TO 20
DIFFERENT PARTICLES IN A
TRANSMISSION SETUP THAT EVEN
THAT TENFOLD INCREASE IN
INFECTIOUSNESS OR INFECTABILITY
FOR LACK OF A BET TERM BETTER TN
THESE OMICRON VARIANTS IN A
HAMSTER MODEL WILL REDUCE
TRANSMISSION EFFICIENCY.
SO I WOULD LOVE FOR SOMEONE TO
DO THAT.
IT COULD ALSO BE THAT THE DONOR
ANIMAL, THERE'S NOT AS MUCH
VIRUS IN THE UPPER AIRWAYS OF
THE DONOR ANIMAL.
MAYBE THERE'S A CHANGE IN HOST
RESPONSE, CREATING DIFFERENT
PARTICLE SIZES.
SO IT COULD BE A MULTITUDE OF
THINGS BUT I THINK IT'S AN
IMPORTANT QUESTION.
NOT TO LOSE SIGHT OF THE FACT
THAT THE HAMSTER IS STILL NOT AN
EXACT REPLICA OF THE HUMAN FOR
SURE.
STEPHANIE.
>> HI.
YES, GOOD MORNING.
REALLY GREAT TALK.
I LOVE THE ELISPOT DEVELOPMENT.
THAT'S GOING TO BE REALLY
HELPFUL FOR THE FIELD.
I'M CURIOUS, YOUR FEMALE HAMSTER
STUDIES, WERE THOSE ONLY SINGLY
HOUSED?
HAVE YOU GROUP-HOUSED FEMALES
AND DO YOU HAVE ANY TIPS AND
TRICKS BECAUSE THEY DO GET QUITE
TERRITORIAL.
>> SO NO.
SO ALL -- SO WE WERE VERY
FORTUNATE WITH OUR ANIMAL
STUDIES COMMITTEE IN THAT WE
COULD USE OUR MOUSE CAGING, OUR
MOUSE BIOCONTAINMENT CAGING FOR
ALL OUR HAMSTER STUDIES WITH
SARS-COV-2.
FOR THOSE OF YOU WHO'VE WORKED
WITH THESE CAGED SYSTEMS,
THEY'RE NOT VERY LARGE.
SO WE'VE NEVER DONE GROUP
COMMUNITY TRANSMISSION.
WE ONLY LOOK FROM ONE DONOR
ANIMAL TO ONE CONTACT ANIMAL IN
EVERY SINGLE SITUATION.
SO ALL THE DATA AND ALL THE DATA
POINTS THAT I SHOWED YOU ARE ONE
TO ONE PAIRINGS.
AND THEN FOR THE ONWARDS
TRANSMISSION, FOLLOWING OUR
EIGHT-HOUR EXPO SEU W EXPOSURE,S
SOMEWHERE BETWEEN LET'S SAY THE
SLEEPOVER THAT STACEY WAS
TALKING ABOUT AND MAYBE THE
SHOPPING, THE KROGER EVENT, MORE
OF A NIGHT OUT, I WOULD SAY, IS
THEN USING THAT CONTACT ANIMAL,
PUT IT SEPARATE FOR TWO TO THREE
DAYS, AND THEN USE THAT AGAIN IN
A ONE TO ONE TRANSMISSION SETUP.
>> GREAT, THANK YOU.
>> HI, IT'S SEEMA.
I FIRST HAD A COMMENT AND THEN I
HAD A QUESTION.
MY COMMENT WAS THAT I THINK YOU
MAYBE MISSPOKE A LITTLE BIT AND
I'M JUST GOING TO CORRECT YOU A
SMIDGE IN THAT YOU SAID THAT IT
WAS A NUMBER OF PARTICLES THAT
INITIATED AN INFECTIOUS BASED ON
YOUR BARCODE DATA, AND THAT'S
ACTUALLY NOT RIGHT.
THE WAY TO INTERPRET THAT IS
IT'S THE NUMBER OF UNIQUE
GENOTYPES THAT INITIATED AN
INFECTION, NOT CORRELATE TO THE
NUMBER OF PARTICLES THAT
ACTUALLY WENT ACROSS.
I THINK THAT'S SOMETHING THAT'S
MISINTERPRETED A LOT, AND I
WANTED TO SORT OF LIKE CALL THAT
OUT BECAUSE I THINK WE SORT OF
AS WE DO THESE SORT OF
BOTTLENECK EXPERIMENTS, IT'S
REALLY IMPORTANT TO MAKE THAT
CLEAR.
AND MY -- SO MY QUESTION WAS
THEN ABOUT EXPULSION IN YOUR
SYSTEM.
SO OBVIOUSLY CLOSE CONTACT
TRANSMISSION, ALL MODES OF
TRANSMISSION ARE FEASIBLE, AND
WHAT DO WE KNOW ABOUT THE
AEROSOL DISTRIBUTION THAT'S
EXPELLED FROM HAMSTERS AND WHAT
DO YOU KNOW ABOUT THE DIFFERENT
PERHAPS EXPULSION AMOUNTS FROM
HAMSTERS INFECTED WITH
SARS-COV-2 VARIANTS?
>> THOSE ARE -- THANK YOU,
SEEMA.
THOSE ARE EXCELLENT QUESTIONS
AND YEAH, THANKS FOR CORRECTING
ME ON THAT.
SO IT'S TRUE, THIS IS GENOTYPES
DETECTED.
I MUST SAY THAT I THINK THERE
WILL BE DIFFERENCES BETWEEN THE
INFLUENZA SYSTEM AND THE
SARS-COV-2 SYSTEM.
THAT I THINK THE VARIOUS LABS
WILL HAVE TO FIGURE OUT.
GIVEN THE SENSITIVITY OF THE
HAMSTER TO SARS-COV-2 AND THE
FACT THAT IT'S NOT A SEGMENTED
VIRUS, I DO THINK THE RATIO OF
TRANSMITTED AND INFECTION IS
CLOSER TO WHATEVER ONE TO ONE IT
IS THAN FOR INFLUENZA VIRUS, BUT
YOU'RE RIGHT, I SHOULD REFER TO
IT AS GENOTYPES.
IN TERMS OF THE PARTICLE SIZE,
THIS IS SOMETHING THAT I'M
INTERESTED IN BUT I WILL REFER
YOU TO WORK FROM WAYLAND YEN
WHO'S DONE REALLY INTERESTING
WORK, SHE MEASURED FROM HER LAB
INFECTED -- DIFFERENT SIZES HOW
IT IS ASSOCIATED WITH RNA VIRAL
LOAD.
THEN SHE USED HER IMPACTER THAT
ALLOWS PARTICLES OF DIFFERENT
SIZES TO PASS THROUGH FROM THE
DONOR TO THE RECIPIENT ANIMAL,
AND SHE SHOWED THAT FOR SARS
KROSARS-COV-2, YOU NEED LARGER
PARTICLE SIZES, WHEREAS YOU GET
SMALLER BOTTLENECK WITH SMALLER
PARTICLES AND IF YOU LOWER THE
PARTICLE SIZE BEYOND THE CERTAIN
SET POINT, THERE'S ACTUALLY NO
TRANSMISSION DETECTED.
>> GREAT TALK.
THANK YOU SO MUCH.
I HAVE A COMMENT AND A QUESTION
AS WELL.
THE FIRST ONE IS SHARING
INFORMATION ABOUT THE HOUSING
WHICH WAS A QUESTION THAT CAME
UP YESTERDAY.
OUR EXPERIENCE HANDLING HAMSTERS
AND HOUSING THEM SOCIALLY,
MALE/FEMALE, DID NOT HAVE ANY
ISSUE.
I WOULD SAY IT'S ALSO SOMETIMES
DEPENDENT ON THE VENDOR.
THE ONLY ONE ISSUE WE HAD WAS
WITH MALES.
WE JUST SWITCHED THE VENDOR AND
IT WAS NEVER AN ISSUE FOR THE
PAST FOUR YEARS.
AND THE OTHER QUESTION IS BASED
ON ANTIVIRALS AND HAMSTERS, AND
YOU BRIEFLY MENTIONED ABOUT IT.
WITH ALL THE STUDIES WE HAVE
DONE, IT SEEMS TO BE
CHALLENGING, SO I KNOW YOU
MENTIONED THAT MORE WORK IS
IMPORTANT SO WHAT ARE YOUR
THOUGHTS ON THAT?
IS IT THE TYPICAL CLASS OF THE
SMALL MOLECULES ANTIVIRALS OR
IT'S JUST THE SYSTEM METABOLISM
IN HAMSTERS?
THANK YOU SO MUCH.
>> YEAH, I DON'T HAVE ANY
FURTHER COMMENTS ON THE FIRST
ONE ABOUT SOCIAL BEHAVIOR.
WE TRI TRY TO SINGLE-HOUSE THEM
WHENEVER POSSIBLE.
SIZE AND AGE MAKES A HUGE
DIFFERENCE, SO IF YOU PUT A
6 MONTH OLD MALE WITH A 6 WEEK
OLD MALE, ONE WILL LOSE AND YOU
CAN GUESS WHICH ONE.
AND IT WILL KILL IT, IN FACT.
SO THERE ARE -- YOU'VE GOT TO BE
CAREFUL AND I THINK SAME WITH
FERRETS, I'VE NEVER WORKED WITH
THEM BUT I'VE HEARD THAT IF YOU
WORK WITH THEM A LOT, YOU HANDLE
THEM A LOT, THEY ARE ACTUALLY
MUCH MORE -- MUCH KINDER THAN IF
YOU JUST GET THEM STRAIGHT FROM
THE VENDOR AND THEY'RE NOT --
THEY'RE NEVER HANDLE PERSONALLY.
SO I SUSPECT THAT HAMSTERS MAY
HAVE A SIMILAR BEHAVIOR THERE.
IN TERMS OF ANTIVIRAL MOLECULES
AND SMALL MOLECULES, THIS IS NOT
MY AREA OF EXPERTISE AT ALL.
IT COULD BE METABOLISM, THERE
COULD BE SOMETHING ELSE GOING
ON, BUT I HOPE SOMEONE WILL
INVESTIGATE THIS MORE, PERHAPS
VODEVELOP AKNOCKOUT MOUSE, KNOCR
TO DEVELOP ONE OF THE PUMPS FOR
EXAMPLE OR BREAKDOWN MOLECULES
THI WWHICH WAS DONE FOR MICE AS,
BUT CERTAINLY IT'S IMPORTANT.
>> JUST A COMMENT ON THE
BEHAVIOR OF HAMSTERS.
IN THE PAST 10 YEARS, WE USUALLY
PRODUCED MORE THAN SEVERAL
THOUSAND OF HAMSTERS AND THIS --
BEHAVIOR, WE OBSERVED THEM.
THEY ARE VERY FRIENDLIMENT I FR.
I HAVE BEEN WORKING WITH A
SOCIAL BEHAVIORAL SCIENTIST.
IN ORDER TO ENCOURAGE THE FEMALE
TO BE MORE AGGRESSIVE, THEY HAVE
TO TEMPORARILY SEPARATE THEM FOR
A WHILE.
IF THEY ARE TOGETHER, YOU KEEP
THEM, I KEEP THEM FOR MONTHS,
THEY ARE VERY FRIENDLY, THEY
HUDDLE WITH EACH OTHER.
SO I DON'T THINK THAT THE
FIGHTING IS A BIG ISSUE.
>> OKAY.
THANK YOU.
, JACCO, THANK YOU SO
MUCH FOR JOINING US EVEN
REMOTELY.
[APPLAUSE]
I'D LIKE TO INVITE DIEGO DIEL
FROM CORNELL UNIVERSITY TO THE
PODIUM.
TO GIVE HIS TALK ENTITLED "THE
FERRET AND FELINE MODELS OF SARS
SARS-COV-2 INFECTION AND
PATHOGENESIS.
THANK YOU.
>> THANK YOU, SABRA, FOR THE
INTRODUCTION AND THANKS,
MICHELLE AND STEVEN FOR THE
OPPORTUNITY TO BE HERE TO TALK A
LITTLE BIT ABOUT THE WORK WE
HAVE BEEN DOING ON MODELS THAT
ARE NOT AS TRADITIONAL MODELS OF
SARS-COV-2 INFECTION AND
PATHOGENIC.
SO JUST A BRIEF OUTLINE OF THE
TALK.
I'M GOING TO GIVE A BRIEF
BACKGROUND AND THEN WE'LL TALK A
LITTLE BIT ABOUT THE WORK THAT
WE HAVE DONE ON FERRET MODEL
INCLUDING STUDIES ON AGE-RELATED
SUSCEPTIBILITY TO SARS-COV-2 AND
THEN COMPARATIVE PATHOGENESIS OF
A FEW SARS-COV-2 COV-2 VARIANTS
ON THAT MODEL, AND THEN WE'LL
TALK MORE ON THE FELINE MODEL ON
COMPARATIVE PATHOGENESIS OF VAIR
YENTS, CHARACTERIZATION OF
VIRULENCE AND VIRUS-HOST
INTERACTIONS WITH FOCUS ON THE
ROLE OF SPIKE ON PATHOGENESIS
AND R3A.
SO MY LAB ESTABLISHED A PIPELINE
FOR STUDYING THE EVOLUTION OR
THE PHENOTYPES, GENOTYPE TO
PHENOTYPE ON SARS-COV-2.
I LED THE CORNELL TESTING LAB,
ESTABLISHMENT OF THAT COVID
TESTING LAB THE A CORNELL AND
OVER THE PANDEMIC, WE WERE ABLE
TO TEST OVER 2.1 MILLION
SAMPLES, SO THAT PROVIDED A
PERFECT PIPELINE OF SAMPLES AND
ISOLATES THAT WE LEMPLED TO
ACTUALLY STUDY SOME OF THE
DIFFERENCES BETWEEN THE VIRUSES.
SO OVER THE COURSE OF THE
PANDEMIC, WE SEQUENCED OVER 3500
SAMPLES AND OBTAINED CLOSE TO A
THOUSAND ISOLATES OF THE VIRUS.
THE GOAL OF THAT PIPELINE WAS
REALLY TO LOOK AT THOSE VIRUSES,
CHARACTERIZE THEM IN VITRO AND
THEN ASSESS THEIR PATHOGENESIS
IN ANIMAL MODELS.
WE REALLY TRIED TO FOCUS OUR
ANIMAL MODEL WORK ON SPECIES
THAT WERE NATURALLY SUSCEPTIBLE
TO THE VIRUS AND WE ALL KNOW
THAT THE INTERACTION BETWEEN THE
SPIKE OF THE VIRUS WITH THE
RECEPTOR, IT A KEY DETERMINANT
OF THE TROPISM AND HOST RANGE OF
THIS VIRUS.
SO AGAIN, OUR FOCUS WAS ON
IDENTIFYING AND WORKING SPECIES
THAT WERE NATURALLY SUSCEPTIBLE
TO THE VIRUS.
INITIALLY, WE LOOKED AT THE
LITERATURE TO SEE WHAT WAS KNOWN
ABOUT U. SUSCEPTIBILITY OF THESE
ANIMALS TO CORONAVIRUS, AND
THERE WAS A PAPER PUBLISHED IN
2008 THAT ACTUALLY CHARACTERIZED
THE SUSCEPTIBILITY OF BOTH
FERRET AND CATS TO SARS 1, AND
THEY KE DEMONSTRATED REPLICATIOF
THE VIRUS IN THE LUNG.
FERRETS IN THE UPPER AND LOWER
RESPIRATORY TRACT OF CATS, THEY
ALSO CHARACTERIZE EXPRESSION OF
THE ACE 2 RECEPTOR THROUGHOUT
THE RESPIRATORY TRACT.
AND THE DISTRIBUTION OF ACE
2 EXPRESSION IN THOSE SPECIES IS
VERY SIMILAR TO THAT WHAT IS
FOUND IN HUMANS.
THERE'S A MORE RECENT PAPER THAT
PUBLISHED AND DEMONSTRATED
EXPRESSION ALSO IN THE
RESPIRATORY MUCOSA AND THE
OLFACTORY MUCOSA OF BOTH
SPECIES.
SO VERY SIMILAR ACE DISTRIBUTION
TO THAT WHAT IS FOUND IN HUMANS.
THE OTHER ASPECT THAT WE LOOKED
AT IS THE PRECE THE RECEPTOR DIS
HIGHLIGHTED HERE.
WHEN COMPARED TO THE HUMAN, TH
THE -- THE CAT HAS FOUR AMINO
ACID DIFFERENCES, THE ONES
HIGHLIGHTED IN YELLOW ARE SHARED
BY BOTH SPECIES.
IT'S NOT THE PERFECT MATCH WHEN
COMPARED TO THE HAMSTER, GOLDEN
HAMSTER HAS ONLY TWO MUTATION --
TWO DIFFERENCES.
WHEN COMPARED TO
THE HUMAN.
SO ONE OF THE FIRST STUDIES THAT
WE DID WAS TO BASICALLY
ESTABLISH THE FERRET MODEL IN
THE LAB.
WE DID AN INFECTIOUS DOSE STUDY
AND ALSO COMPARED SUSCEPTIBILITY
OF TWO DIFFERENT AGE GROUPS OF
FERRETS.
WE USED 6 MONTH OLD ANIMALS
WHICH WE CALLED YOUNG FERRETS
AND THEN WE ALSO HAD AGED
ANIMALS THAT WERE BETWEEN 18 AND
36 MONTHS OF AGE.
DIVIDED THESE ANIMALS IN FIVE
GROUPS, CONTROL ANIMALS, AND
THEN ANIMALS THAT WERE
INOCULATED WITH DIFFERENT
INFECTIOUS DOSES FROM 10 TO 1 TO
10 TO THE 6.
WE MONITORED THESE ANIMALS BY
COLLECTING CLINICALLY AND
COLLECTING SAMPLES FOR
VIROLOGICAL ASSESSMENTS AS WELL.
AS SHOWN HERE, WE ONLY DETECTED
REPLICATION AND SHEDDING OF
INFECTIOUS VIRUS OF THOSE 10 TO
THE 2.
WITH AGED ANIMALS PRESENTING
SLIGHT LEHIGLY HIGHER REPLICATIN
THE UPPER RESPIRATORY TRACT, AND
THAT WAS CONSISTENT THROUGHOUT
THE DOSES THAT WE USED, AND THAT
WAS ALSO CONFIRMED BY
SEROCONVERSION SO ONLY ANIMALS
THAT WERE INOCULATED TENDED TO
BE -- HIGHER FREQUENCY OF AGED
ANIMALS ACTUALLY CONVERTING TO
INFECTION.
SO WITH THOSE RESULTS, WE
DETERMINED THE INFECTIOUS DOSE
IN BOTH AGE GROUPS.
AND AS SHOWN HERE, THE YOUNGER
ANIMALS REQUIRED THREE FOLD
HIGHER DOSE TO ACTUALLY BECOME
INFECTED WITH SARS-COV-2.
THAN THE AGED ANIMALS.
WE LOOKED AT POTENTIAL CAUSES
FOR THIS AGE DIFFERENCE AND ONE
THING THAT WE OBSERVED IN THIS
STUDY IS THAT AGED ANIMALS
EXPRESSED USING QPCR, AGED
ANIMALS EXPRESSED HIGHER LEVELS
OF ACE 2 AND TMPRSS2 IN THE
NASAL MUCOSA.
THERE WERE NO DIFFERENCES IN
EXPRESSION OF THOSE TWO
MOLECULES IN THE LUNG.
SO WE THINK THAT THIS COULD
POTENTIALLY AT LEAST PARTIALLY
EXPLAIN THIS AGE DIFFERENCE.
OBVIOUSLY WE HAVE NOT ASSESSED
IMMUNOLOGICAL FACTORS THAT ARE
ALSO LIKELY DIFFERENT BETWEEN
AGED AND YOUNG FERRETS.
SO THE NEXT STUDY WE CONDUCTED
WAS TO COMPARE THE PATHOGENESIS
OF B1 OR D614G VARIANTS WITH THE
ALPHA VARIANT AS SOON AS THE
VARIANT ACTUALLY EMERGED.
IN THE
U.S., SO WE DESIGNED AN
EXPERIMENT WITH THREE GROUPS,
CONTROL ANIMALS, ANIMALS THAT
WERE INOCULATED WITH B1 AND THEN
WITH THE ALPHA VARIANT.
WE EUTHANIZED ANIMALS O ON DAYS
3 AND 5 TO MONITOR VIRUS
REPLICATION AND TISSUES AND
AGAIN COLLECTED RESPIRATORY
SAMPLES TO MONITOR VIRUS
REPLICATION AND SHEDDING.
WE ALSO MONITORED THESE ANIMALS
CLINICALLY AND AS YOU CAN SEE
HERE, WE HAVE NOT OBSERVED ANY
CLINICAL SIGNS AND I THINK THAT
THIS IS A FEATURE OF THE FERRET
MODEL.
THEY DON'T DEVELOP FEVER, THEY
ALSO DON'T LOSE WEIGHT AFTER
INFECTION, SO IT'S A SUBCLINICAL
INFECTION IN THE FERRET MODEL.
WE LOOKED AT REPLICATION OF THE
VIRUS IN THE UPPER RESPIRATORY
TRACT PCR RESULTS SHOWING ON THE
LEFT HERE.
VIRUS TITRATIONS ON THE RIGHT.
WE DID NOT SEE DIFFERENCES IN
THE LEVELS OF RNA THAT WE
DETECTED THROUGHOUT THE COURSE
OF THE EXPERIMENT.
THERE WAS A SLI SLIGHTLY HIGHERS
OF INFECTIOUS VIRUS IN THE NASAL
SWABS FOR THE D614G VIRUS BUT WE
HAVE NOT OBSERVED THE SAME
DIFFERENCE, THIS WAS NOT
SIGNIFICANT, WE DID NOT OBSERVE
DIFFERENCES IN VIRUS REPLICATION
IN THE OROPHARYNGEAL SWABS AS
WELL.
WHEN WE LOOKED AT TISSUES, WE
DID DETECT RNA IN THE UPPER AND
ALSO LOWER RESPIRATORY TRACT,
BUT WHEN WE PERFORMED VIRAL
TITRATIONS TO LOOK FOR
INFECTIOUS VIRUS, THAT WAS
BASICALLY RESTRIBTED TO THE
UPPER RESPIRATORY TRACT WHERE WE
OBSERVED HIGH LEVELS OF
REPLICATION IN THE NASAL
TURBINATES FOR BOTH VIRUSES AND
IN THE TONSIL WITH NO DETECTION
OF VIRUS ON DAY 3 OR DAY 5 IN
THE LOWER RESPIRATORY TRACT.
ANOTHER INTERESTING OUTCOME OF
THIS STUDY IS WHEN WE LOOKED AT
CHANGES IN THE VIRUS FOLLOWING
INOCULATION, WE OBSERVED THAT
THE VIRUS ACTUALLY CHANGED VERY
QUICKLY FOLLOWING INOCULATION OF
FERRETS AND THOSE CHANGES WERE
LOCATED IN THE RECEPTOR-BINDING
DOMAIN OF THIS SPIKE.
THIS HAS BEEN DESCRIBED BY OTHER
GROUPS, OUR FINDINGS BASICALLY
REINFORCED WHAT WAS SEEN, AND
ONE PARTICULAR MUTATION, THE
Y453 F MUTATION OCCURRED IN A
HIGHER FREQUENCY OF ANIMALS AS
EARLY AS DAY 3 POST INOCULATION.
SO INDICATING THAT THERE'S A
PRETTY HIGH PRESSURE ON THE
RECEPTOR-BINDING DOMAIN, MOST
LIKELY BECAUSE OF THAT
INCOMPATIBILITY OF THE SPIKE AND
THE RECEPTOR.
RECEPTOR-BINDING THE DOMAIN.
GIVEN THESE FINDINGS, WE DECIDED
THAT WE PROBABLY NEEDED A BETTER
MODEL TO STUDY PATHOGENESIS OF
SARS-COV-2 THAN THE FERRET
MODEL.
BECAUSE WE REALLY WANTED TO
ASSESS POTENTIAL TRANSMISSION
AND INFECT -- IDENTIFY VIRULENCE
DETERMINANTS AND DETERMINE THEIR
ROLE IN PATHOGENESIS.
IF THE VIRUS WOULD BE CHANGING
VERY EARLY ON AFTER INOCULATION
ON THESE ANIMALS, THAT COULD
POTENTIALLY MASK AND REALLY
COMPLICATE INTERPRETATION OF THE
RESULTS.
WHEN WE WERE FINISHING OUT THAT
PATHOGENESIS STUDY, I RECEIVED A
CASE OF A CAT WITH VERY SEVERE
SARS-COV-2 INFECTION IN THE
DIAGNOSTIC LAB AT CORNELL, WHERE
I AM DIRECTOR OF THE VIROLOGY
LAB.
AS YOU CAN SEE HERE, THIS IS THE
LUNG OF THE CAT REALLY SHOWING
PRETTY SEVERE COMPROMISE OR
PNEUMONIA ON THAT LUNG.
WHEN WE LOOKED AT THE HISTOLOGY
OF THE LUNG, IT WAS VERY
CHARACTERISTIC SARS-COV-2
INFECTION, RESEMBLING THE
PNEUMONIA THAT IS DESCRIBED IN
HUMANS WITH A DIFFUSE ALVEOLAR
DAMAGE, AND INFLAMMATORY
INFILTRATE, IN THIS CASE, MAINLY
CHARACTERIZED BY MACROPHAGES.
WE ALSO OBSERVED VERY ROBUST
REPLICATION OF THE VIRUS IN THE
LUNG IN THIS PARTICULAR CASE.
AS SHOWN HERE BY IN SITU
HYBRIDIZATION AND
HISTOCHEMISTRY, MAINLY IN
BRONCHIOLAR EPITHELIAL CELLS.
ONE INTERESTING FEATURE ABOUT
THIS PARTICULAR CAT, THIS CAT
WAS DIAGNOSED WITH A
HYPERTROPHIC -- WHICH IS A
CONDITION THAT HAS BEEN
DESCRIBED AS -- IN HUMANS
LEADING TO SEVERE DISEASE.
AND I THINK THERE'S A FEW OTHER
CASES IN CATS THAT DIED FROM
NATURAL INFECTION THAT ALSO
PRESENTED THE SAME HIRE TROPHIC
CARDIOMYOPATHY.
SO WITH THAT, WE DECIDED TO
INITIATE THE ESTABLISHMENT OF
THE FELINE MODEL IN THE LAB.
THE FIRST STUDY WE CONDUCTED WAS
A COMPARISON BETWEEN THREE
VARIANTS, D614G DELTA AND
OMICRON BA1.
AND IN THESE STUDIES, IN ALL
PARAMETERS THAT WE EVALUATED
WHAT WE OBSERVED IS THAT DELTA
IN D614G BEHAVED VERY SIMILAR
ALSO AS THE ANIMALS INOCULATED
WITH THOSE VIRUSES DEVELOPED
FEVER, THEY ALSO HAD A SLIGHT
DECREASE IN BODY WEIGHT, THEY
PRESENTED VERY HIGH VIRUS
REPLICATION IN THE UPPER
RESPIRATORY TRACT AND IN THE
LUNG, AND HIGH LEVELS OF VIRUS
SHEDDING.
AND BOTH OF THESE VARIANTS HAVE
MUCH HIGHER VIRUS REPLICATION
THAN THE OMICRON BA1 VARIANT.
THEY ALSO CAUSED MORE SEVERE
PATHOLOGY AND ALSO HIGHER
ANTIBODY RESPONSES.
SO THE QUESTION THAT CAME UP OF
THIS STUDY IS WHAT WAS THE ROLE
OF THE SPIKE ON THIS, THE
OMICRON BA1 HAD AT LEAST 35
MUTATIONS IN THE SPIKE PROTEIN
ALONE, SO WE COLLABORATED WITH
DR. MARTINEZ SOBRITO AND HE
PROVIDED US WITH THE INFECTIOUS
CLONE BASED ON THE WASHINGTON
D614G -- CONDUCTED THE STUDY IN
THE FELINE MODEL TO ASSESS THE
ROLE OF THE SPIKE ON VIRAL
PATHOGENESIS.
SO AT THREE -- TWO GROUPS WERE
INOCULATED WITH THE D614G OR
WITH THE OMICRON, AND THEN WE
EUTHANIZED ANIMALS ON DAYS 3,
5 OR 14, AND AGAIN MONITORED
CLINICAL SIGNS BY VIRAL
REPLICATION AND IN THE UPPER AND
LOWER RESPIRATORY TRACT AND --
TISSUES.
YOU CAN SEE HERE, WE DID OBSERVE
CLINICAL DIFFERENCES BETWEEN THE
D614G AND THE OMICRON SPIKE
VIRUS WHERE THE ANIMALS
PRESENTED INCREASED TEMPERATURE
BETWEEN DAYS 1 AND 3 AND THEY
ALSO PRESENTED A SLIGHT DECREASE
IN BODY WEIGHT WHILE ALL THE
OMICRON INOCULATED CATS ACTUALLY
GAINED WEIGHT AND THIS IS
CONSISTENT WITH OUR FIRST STUDY
WHERE THESE ANIMALS DON'T
PRESENT CLINICAL SIGNS.
THIS WAS CONFIRMED WHEN WE
LOOKED AT VIRUS REPLICATION.
AS SHOWN HERE, WE DETECTED
SIGNIFICANTLY LOWER LEVELS OF
VARIABLE RNA AND INFECTIOUS
VIRUS IN NASAL SWABS AND OUR
PHARYNGEAL SWABS AND ALSO IN THE
LOWER RESPIRATORY TRACT AND
BRONCHIOLAR LAVAGE FLUIDS IN THE
CURRENT SPIKE INOCULATED
ANIMALS.
WHEN WE LOOKED AT REPLICATION IN
TISSUES, WE AGAIN OBSERVED MUCH
LOWER LEVELS OF REPLICATION OF
THE OMICRON SPIKE WHEN COMPARED
TO THE D614G.
IN BOTH THE UPPER AND LOWER
RESPIRATORY TRACT, SHOWING YOU
HERE REPLICATION IN THE NASAL
TURBINATES.
PRETTY STRIKING PHENOTYPIC
DIFFERENCE BETWEEN THE TWO
VIRUSES.
SO THIS IS A SUMMARY SLIDE AND
FOR SAKE OF TIME I DIDN'T HAVE
TIME TO SHOW YOU THE WORK THAT
WE DID IN VITRO, BUT WHAT WE
OBSERVED IS THAT THE SPIKE, THE
CHIMERA WITH THE OMICRON SPIKE
VIRUS IS ABLE TO INFECT CELLS
JUST AS EFFICIENTLY AS THE D614G
VIRUS, BUT THIS VIRUS IS REALLY
IMPAIRED IN ITS ABILITY TO
SPREAD FROM CELL-TO-CELL.
SO THIS SPIKE OF THE OMICRON HAS
A MUCH LOWER -- ABILITY WHEN
COMPARED TO THE D614G VIRUS.
WE KIND OF OBSERVED TH THE SAMEN
VIVO WHERE THERE WERE A FEW
CELLS INSPECTED WITH THE OMICRON
SPIKE BUT THE VIRUS DIDN'T SEEM
TO BE ABLE TO SPREAD
CELL-TO-CELL AS EFFICIENTLY AS
THE D614F VIRUS.
614G VIRUS.WE BELIEVE THIS IS OE
POTENTIAL MECHANISMS THAT
EXPLAIN THE DIFFERENCE IN
PATHOGENICITY WHEN COMPARED TO
THE D614G VIRUS.
SO THE LAST STORY THAT I WANT TO
TELL YOU, AND I NEED TO RUN
BECAUSE I'M RUNNING OUT OF TIME,
IS ON THE COMPARISON IN THE
EFFECT OF ORF3 ON THE
PATHOGENESIS OF THE VIRUS AGAIN
USING THE FELINE MODEL, WE
COMPARED A VIRUS --
CONTAINING -- VIRUS IN WHICH THE
3A ACCESS REA GEN ACCESSORY GENN
DELETED.
WE DID NOT OBSERVE DIFFERENCES
IN BODY TEMPERATURE BETWEEN THE
TWO VIRUSES, HOWEVER, THE DELTA
RF3 VIRUS INOCULATED ANIMALS DID
NOT PRESENT REDUCTION IN THE
BODY WEIGHT WHILE THE WASHINGTON
1 INOCULATED ANIMALS PRESENTED
DECREASE IN BODY WEIGHT.
WHEN WE LACKED AT REPLICATION IN
THE UPPER AND LOWER RESPIRATORY
TRACT, AGAIN WE OBSERVED
SIGNIFICANT REDUCTION IN
REPLICATION OR SHEDDING LEVELS
OF THE DELTA R3 VIRUS WHEN
COMPARED TO THE WASHINGTON
1 BACKBONE IN THE UPPER AND THE
LOWER RESPIRATORY TRACT
SECRETIONS AS SHOWN IN THIS
GRAPH.
SIMILARLY WHEN WE LOOKED AT
TISSUES, BOTH DELTA ORF3 VIRUS
PRESENTED MUCH LOWER VIRAL LOADS
IN THE UPPER AND LOWER
RESPIRATORY TRACT WHEN COMPARED
TO THE D614G VIRUS THIS WAS
REFLECTED IN THE PATHOLOGY AS
WELL, DELTA R3 INOCULATED
ANIMALS PRESENTED LUNG HISTOLOGY
VERY SIMILAR TO OUR CONTROL
ANIMALS, WHEN THE WASHINGTON ONE
INOCULATED ANIMALS PRESENTED
MORE SEVERE LESIONS IN THE LUNG.
WE ALSO LOOKED AT GENE
EXPRESSION LEVELS BY RNA SEQ,
AND WHAT I WOULD LIKE TO POINT
OUT HERE IS THAT WHEN WE LOOKED
AT TRANSCRIPTION FACTORS THAT
COULD BE REGULATING THESE GENES
WITHOUT THEIR GENE EXPRESSION,
WHAT WE FOUND OUT IS THAT
SEVERAL OF THE TRANSCRIPTION
FACTORS THAT WERE IDENTIFIED,
THEY BELONGED TO THIS ZNF FAMILY
OF TRANSCRIPTION FACTORS O, ANDE
SEARCHED THE LITERATURE TO SEE
WHETHER THIS HAS BEEN DESCRIBED
BEFORE AND WE FOUND THIS PAPER
WHERE THEY CONDUCTED A PRETTY
LARGE PROTEOMICS SCREEN AND THEY
ACTUALLY HAD FOUND THROUGH THIS
SCREEN THAT ORF3 INTERACTS WITH
ZNF579 ON THAT LARGE SCREEN.
THEY HAVE NOT FOLLOWED UP ON THE
CHARACTERIZATION OF THIS, SO WE
CONFIRMED THAT INTERACTION OF
ORF3 WITH THE CATH ZNF ALPHA,
AND THIS IS SOMETHING WE'RE
CURRENTLY WORKING ON TO FURTHER
CHARACTERIZE THIS.
WE'RE LOOKING AT THIS
INTERACTION IN THE CONTEXT OF
VIRUS INFECTION AND ALSO LOOKING
AT CHARACTERIZING THE EFFECT OF
THIS INTERACTION IN GENE
TRANSCRIPTION USING -- STUDIES.
SO JUST A QUICK OVERVEUL OF THE
OVERVIEW OF THEMODELS WE DISCUS.
SO THE FERRET MODEL, THERE'S
RELATIVELY LOW VIRUS SHEDDING OR
VIRUS REPLICATION WHEN COMPARED
TO THE FELINE MODEL.
THE ANIMALS, FERRETS DO NOT
DEVELOP CLINICAL SIGNS FOLLOWING
INOCULATION.
AND VIRUS RE REPLICATION IS REAY
LIMITED TO THE UPPER RESPIRATORY
TRACT WITH QUESTIONABLE
REPLICATION IN THE LUNGS, WHICH
DOESN'T REALLY REPRESENT THE
PATHOLOGY THAT WE SEE IN HUMANS.
THE CATS, ON THE OTHER HAND, THE
VIRUS REPLICATES A LOT BETTER IN
BOTH THE UPPER AND LOWER
RESPIRATORY TRACT AND WE ALSO
OBSERVE PATHOLOGY THAT RESEMBLES
THE PATHOLOGY THAT IS OBSERVED
IN HUMANS.
SO STRENGTHS, LIMITATIONS AND
OPPORTUNITIES, THIS IS MY LAST
SLIDE, THE FELINE MODEL, IT DOES
PRESENT ADVANTAGES OVER THE
FERRET MODEL AS THE VIRUS
REPLICATES IN THE UPPER AND
LOWER RESPIRATORY TRACT.
OTHER GROUPS HAVE SHOWN THAT
IT'S ALSO A VERY USEFUL MODEL TO
ASSESS TRANSMISSION OF
SARS-COV-2.
LIMITATIONS IS OBVIOUSLY SIMILAR
TO THE HAMSTER MODEL, AND MAYBE
A LITTLE LESS SO.
WE HAVE A FEW IMMUNE REAGENTS
THAT ARE AVAILABLE BUT
DEFINITELY MORE IMMUNE REAGENTS
ARE NEEDED TO ACTUALLY LOOK AT
HOST IMMUNE RESPONSES AN HOST
FACTORS.
THE OTHER LIMITATION IS THAT
STUDYING HOST GENETICS IN A
LARGE SMALL ANIMAL MODEL AS THE
FELINE MODEL IS MUCH MORE COSTLY
AND COMPLICATED THAN IT IS IN
THE SMALLER MODELS.
SO THAT IS A SIGNIFICANT
LIMITATION.
THE OTHER THING ABOUT FELINE, A
FELINE MODEL IS THAT THEY CAN
POTENTIALLY SERVE AS MODELS FOR
OTHER RESPIRATORY TRY VUSES.
VIRUSES.
WE HAVE ALSO BEEN INVOLVED IN
ALSO THE DETECTION OF HIGHLY
PATHOGENIC INFLUENZA.
IN A FEW OF THESE FARMS WE
DETECTED THE VIRUS IN DAIRY
CATTLE.
-- WERE THE FIRST TO DIE FROM
H -- YOU CAN SEE VERY HEAVY
REPLICATION OF THE VIRUS IN THE
BRAIN OF INFECTED ANIMALS, WE
ALSO SEE REPLICATION IN THE
LUNG, LIVER, AND MODERATE T -- K
THE FELINE MODEL COULD REALLY
SERVE AS A VERY GOOD MODEL TO
UNDERSTAND THE NEUROTROPISM OF
H5N1 IN MAMMALIAN SPECIES, EVEN
WITH THE LIMITATIONS OF THE LACK
OF REAGENTS THAT WE HAVE.
WITH THAT, I WOULD LIKE TO THANK
MY LAB, ESPECIALLY MOHAMMAD, WHO
DID A LOT OF THE WORK ON THE
FELINE MODEL, MATHIAS WAS THE
PERSON WHO BASICALLY STARTED THE
WORK IN THE LAB.
HE'S CURRENTLY AT PENN STATE
UNIVERSITY.
AND OBVIOUSLY FUNDING FROM NIH
AND USDA.
>> YES, SEEMA.
>> I REALLY LIKE THAT, AND I
LOVE THE FELINE MODEL, AND I AM
CURIOUS, A COUPLE OF QUESTIONS
THAT KIND OF COME TO MIND
BECAUSE I THINK IT'S SUPER
FASCINATING.
ONE HAS ANYONE LOOKED AT OMICRON
TRANSMISSION IN CATS GIVEN THE
LACK OF TRANSMISSION PHENOTYPES
IN HAMSTERS?
MY SECOND, CAN I DO THAT
AFTERWARDS.
>> NOT THAT I KNOW.
WE HAVE NOT HAD A CHANCE TO LOOK
AT -- UNFORTUNATELY WE DON'T
HAVE THE RIGHT GAUGING SYSTEM TO
DO TRANSMISSION STUDIES IN CATS.
THAT'S THE COMPLICATION OF THIS.
I'M WORKING WITH ADMINISTRATION
RIGHT NOW TO ACTUALLY GET CAGES
THAT WOULD ALLOW US TO DO
TRANSMISSION STUDIES.
>> AND THEN MY OTHER PART WAS,
COULD YOU SPECULATE SORT OF ON
THE -- I THOUGHT IT REALLY
FASCINATING DURING THE COVID-19
PANDEMIC AND NOW OBVIOUSLY
DURING H5, WHERE DURING COVID-19
WE WERE SEEING BIG CATS GET
INFECTED AT ZOOS.
TIGRRS AND LIONS, AND THEY
CLEARLY DO NOT HAVE A LOT OF
CLOSE RANGE CONTACT WITH OTHER
ANIMALS OR HUMANS, THERE'S A LOT
OF DISTANCE.
SO I'M CURIOUS WHAT YOU THINK
SORT OF THE MECHANISMS THAT
MIGHT BE FOR TRANSMISSION IN
CATS AND WHAT DO YOU THINK IS
SORT OF HAPPENING IN THE CATS'
SPACE AS THE PANDEMIC SORT OF
CONTINUED?
>> AND YOU'RE ASKING THE
QUESTION SPECIFICALLY ABOUT
SARS-COV-2?
OKAY.
SO -- YEAH.
IF WE LOOK AT THE NUMBER OF
ANIMALS THAT WERE NATURALLY
INFECTED WITH SARS-COV-2, IT'S
VERY SMALL, RIGHT?
BUT IF YOU LOOK AT THE SPECIES
THAT WERE INFECTED, I THINK CATS
WERE SECOND AFTER WHITETAILED
DEER.
SO THEY ARE HIGHLY SUSCEPTIBLE
TO INFECTION.
THEY DO GET INFECTED.
I THINK THAT THE ROLLING
TRANSMISSION IS PROBABLY
MINIMAL.
I DON'T THINK THAT THEY
TRANSMITTED SARS-COV-2 COV-2.
WITH HPAI, I DON'T KNOW.
I THINK THAT THESE CATS COULD
POTENTIALLY BE SHEDDING ENOUGH
VIRUS THAT WOULD LEAD TO
TRANSMISSION TO OTHER CATS OR
POTENTIALLY OTHER ANIMALS.
I DON'T KNOW HOW QUICKLY THEY
DIE AFTER INFECTION TOO, SO
THAT'S ANOTHER QUESTION THAT
NEEDS TO BE ADDRESSED.
THESE ARE -- THE OTHER ANIMALS
WERE FOUND DEAD AND THAT'S HOW
WE TESTED THEM.
BUT I THINK THAT, YOU KNOW,
EXPERIMENTAL STUDIES ARE NEEDED
TO REALLY ANSWER SOME OF THESE
QUESTIONS.
>> I'M GOING TO TAKE LIBERTY AND
ASK A QUESTION AND THEN I THINK
WE'LL MOVE TO THE NEXT SPEAKER.
FOR TRANSMISSION, COULD YOU TAKE
THE VIRUSES THAT ARE BEING SHED
FROM THE CATS AND THEN AT LEAST
MAYBE IN A PRIMARY CELL CULTURE
SYSTEM LIKE HUMAN EPITHELIAL
CELLS LOOK AND SEE WHAT IT DOES
TO HUMAN CELLS?
DO VIRUSES THAT ARE SHED FROM
CATS HAVE THE POTENTIAL FOR
INFECTIVITY, CELL KILLING, YOU
KNOW?
BECAUSE I REALIZE YOU'RE NOT
GOING TO BE ABLE TO DO
TRANSMISSION TO HUMANS, BUT
COULD YOU MODEL TRANSMISSION?
>> I THINK MODELING TRANSMISSION
IN VITRO IS NEARLY IMPOSSIBLE,
RIGHT?
WHAT WE KNOW IS THAT WE ARE
DETECTING SIX LOGS OF VIRUS IN
NASAL SECRETIONS IN SARS-COV-2
FROM THESE ANIMALS.
THAT KEPT THE NATURALLY INFECTED
ANIMAL THAT I SHOWED, THAT GROSS
PATHOLOGY, WE DETECTED
POTENTIALLY EIGHT LOGS OF VIRUS
IN NASAL SECRETIONS AND IN
SEVERAL OF THE --
>> BECAUSE I'M THINKING ABOUT
ONE HEALTH, YOU THINK ABOUT
THESE COMPANION ANIMALS, AND YOU
KNOW, MAYBE MORE SO ABOUT
TRANSMISSION TO HUMANS THAN TO
OTHER FELINE.
>> THERE ARE OTHER PLACES, I
MEAN, KANSAS STATE AND I THINK
THE -- GROUP IN WISCONSIN, THEY
DID TRANSMISSION STUDIES IN CATS
AND THEY TRANSMIT VERY
EFFICIENTLY.
I THINK -- SARS-COV-2.
>> THANK YOU.
>> ALL RIGHT.
I WOULD LIKE TO INVITE SEEMA
LAKDAWALA FROM EMORY UNIVERSITY
TO GIVE HER TALK ENTITLED
RE-THINKING THE FERRET MODEL TO
STUDY INFLUENZA VIRUS
TRANSMISSION.
>> THANK YOU, SABRA.
AND THANKS TO THE ORGANIZERS FOR
THIS REALLY GREAT MEETING.
I'M GLAD THAT I COULD BE HERE
AND THANKS FOR THE OPPORTUNITY
TO SPEAK.
I'LL WAIT FOR MY SLIDES.
I THINK WHAT I WANT TO TALK
ABOUT TODAY IS SORT OF LIKE
SOMETHING THAT WAS TOUCHED ON
YESTERDAY IN STACEY'S OVERVIEW
AND KIND OF I THINK THE REASON
WE'RE HERE IS TO UNDERSTAND AND
THINK ABOUT WHAT ARE THE
QUESTIONS WE'RE ASKING, AND HOW
CAN WE DESIGN THE EXPERIMENTS TO
REALLY TEST THE QUESTIONS THAT
WE ARE MOST INTERESTED IN
ASKING.
SO WE'RE GOING TO TALK ABOUT HOW
WE CAN RE-THINK TRANSMISSION.
THIS IS FINE.
I CAN DO THIS.
WE'RE GOOD.
SO I'VE BEEN STUDYING
TRANSMISSION SINCE THE 2009
PANDEMIC AND I WAS A POSTDOC.
SO I WROTE THIS REVIEW BACK IN
2012.
AND THIS IS HOW I USED TO THINK
ABOUT THIS PROBLEM, RIGHT, IS
THAT TRANSMISSION KIND OF WORKED
WITH THIS AND THERE WERE THREE
PARTS OF THE EQUATION.
I'M GOING TO KIND OF COME BACK
TO THIS WHICH IS WHERE MY MIND
HAS COME OVER A DECADE OF
STUDYING TRANSMISSION, IS WHAT
IS THE EQUATION OF TRANSMISSION,
RIGHT?
YOU GOT A DONOR, THE VIRUS HAS
TO REPLICATE IN A DONOR.
THAT VIRUS IS THEN EXPELLED INTO
A VARIETY OF AEROSOLS.
THERE'S LARGE DROPLET SPRAY, AIR
STOLES, FINE AIR SO SO AEROSOLS.
ONE OF THE THINGS VIROLOGISTS
DON'T DO ENOUGH IS ACTUALLY
THINK ABOUT THE VIRUS IN THE
VIERPT, HOW THE ENVIRONMENTAL
PARAMETERS ACTUALLY INFLUENCE
THE FATE OF THESE VIRUSES AND
HOW THAT THEN LEADS TO
SUCCESSFUL TRANSMISSION OR LACK
OF TRANSMISSION.
THEN THE THIRD PART OF THE
EQUATION IS SUSCEPTIBILITY OF
THE RECIPIENT.
WE TALKED ABOUT IMMUNITY
YESTERDAY AND VAX NAISE VACCIN.
WE ARE NOT IMMUNOLOGICALLY NAIVE
HOSTS AS WE WALK AROUND IN
SOCIETY AND OUR DAILY LIVES, SO
THINKING ABOUT WHAT THAT MEANS
IS IMPORTANT.
SO IN IS HOW I USED TO THINK
ABOUT THIS PROBLEM.
OH, IT'S VERY BINARY, RIGHT?
IT'S LIKE THERE'S A DONOR,
THERE'S THE ENVIRONMENT AND
THERE'S THE RECIPIENT.
BUT IN REALITY, AND I USED TO
ALWAYS SAY, THIS IS AN OLD
SLIDE, ANY OF THESE PARAMETERS
CAN CHANGE THE TRANSMISSION
DYNAMICS.
BUT AGAIN, THESE ARE THREE
EQUATIONS, THREE VARIABLES, YOU
MANIPULATE ANY OF THEM AND YOU
CAN SHIFT THE BALANCE OF
TRANSMISSION.
BUT ACTUALLY TRANSMISSION IS WAY
MORE COMPLICATED, RIGHT?
WE ARE NOT IN THESE ISOLATED
SCENARIOS OF A DONOR RECIPIENT
SINGLE INTERACTION.
WE LIVE IN COMPLEX SITUATIONS.
WE GO TO COMPLEK PL COMPLEX PLAE
DIFFERENT.
WE HAVE A VARIETY OF DIFFERENT
TYPES OF INTERACTIONS.
WE HAVE CLOSE RANGE CONTACTS,
THERE ARE LONG-RANGE CONTACTS,
WE SIT NEXT TO PEOPLE AT
SEMINARS, WE TALK TO PEOPLE
ACROSS TABLES AT BARS, RIGHT?
OR WE'RE SHARING A SUBWAY CAR
DISTANCES APART.
ALL OF THESE SPACES SORT OF
CHANGE TRANSMISSION DYNAMICS.
ONE OF THE THINGS I'M REALLY
GOING TO TALK ABOUT HERE IS THAT
AT CLOSE RANGE, ALL MODES OF
TRANSMISSION ARE POSSIBLE.
WE TALK ABOUT CONTACT TRAPS
MISTRANSMISSION AS THOUGH THIS S
LAKE A WAY OF ONLY LOOKING AT
TOUCHING PHYSICAL THINGS, BUT
ACTUALLY IN REALITY, CLOSE RANGE
TRANSMISSION MEANS THAT YOU ARE
BEING EXPELLED AND EXPOSED TO
ALL OF THE AIR THAT SOMEONE PUTS
OUT.
AND ALL OF THOSE SIZE RANGES
HAVE A MUCH MORE HIGHER
EFFICIENCY OF GETTING INTO YOUR
RESPIRATORY TRACT BECAUSE YOU'RE
BREATHING IN THAT PLUME
IMMEDIATELY FROM THAT SOURCE.
THEN OF COURSE LONG RANGE
TRANSMISSION K IS CAN VARY IN
TERMS OF WHAT TYPES OF --
EVERYBODY IS STARTING TO GO BACK
TO AND REALLY PUSH FORWARD THE
MODES OF TRANSMISSION, I AM
MOVING AWAY FROM THAT BECAUSE I
THINK WE HAVE DONE OURS SEFS A
DISSERVICE OF THINKING ABOUT
INDIVIDUAL MODES OF
TRANSMISSION, ISOLATING THEM AND
THEN TRYING TO DEFINE THEIR
IMPORTANCE WHEN IN REALITY WE
LIVE IN A SPACE WHERE ALL MODES
ARE FEASIBLE.
SO THAT'S JUST MY SOAP BOX.
THIS GETS BACK TO WE ARE
COMPLEX, ALL AGE RANGES OF
INDIVIDUALS HAVE DIFFERENT TYPES
OF BEHAVIORS, DIFFERENT TYPES OF
PRE-EXISTING IMMUNITY, DIFFERENT
TYPES OF MICROBIAL COMMUNITIES,
DIFFERENT TYPES OF ENVIRONMENTAL
PARAMETERS THAT WE LIVE IN.
EVERYONE HAS MINGS MENGSED, I'M
GOING TO TALK ABOUT DAYCARES, I
PROMISE BUT START OFF BY SAYING
WE HAVE TO RE-THINK THE WHOLE
SYSTEM.
SO THIS IS WHAT I'VE ACTUALLY
BEEN DOING FOR THE PAST 10
YEARS, IS THINKING ABOUT WHEN I
STARTED IN 2009, THERE WERE A
HANDFUL OF OTHER GROUPS THAT DID
TRANSMISSION AND WE ALL WERE
DOING THEM THE SAME WAY.
AND WHEN I REALIZED IN 2015, I
WAS LIKE, WE CAN'T KEEP DOING IT
THE SAME WAY.
WE'VE GOT TO REALLY RETHINK HOW
WE AS HUMANS SPREAD VIRUSES TO
OTHER HUMANS, IF THAT'S OUR
QUESTION, IF THAT'S WHAT WE'RE
TRYING TO GET AT, THEN WITH WE E
TO DESIGN OUR MODEL SYSTEMS TO
RECAPITULATE THAT.
SOME OF THE THINGS MY LAB HAS
THOUGHT ABOUT ARE HERE.
PRE-EXISTING IMMUNITY, WE'RE NOT
IMMUNOLOGICALLY NIGH EE. WE'VE
WORKED ON MICROBIAL COMMUNITIES
WITH STACEY, WE WORK ON A
VARIETY OF ENVIRONMENTAL
PARAMETERS TRYING TO UNDERSTAND
HOW THAT INFLUENCES TRANSMISSION
DYNAMICS AND THEN BEHAVIORS OF
INDIVIDUALS AND PEOPLE AND HOW
THAT CAN BE INFLUENCED.
SO WE USED THE FERRET MODEL
SYSTEM, I'M TALK ABOUT WHY,
DIEGO INTRODUCED IT, IT'S BEEN
INTRODUCED BY OTHERS, I'LL TELL
YOU WHY I LIKE IT.
BUT I WILL SAY, THIS IS
SOMETHING STACEY BROUGHT UP IN
THE OPENING AND I TOTALLY AGREE,
IS THAT SOMETIMES THE PRIMARY
MODEL IS ACTUALLY HUMANS.
AND SO I WILL SAY IN THE LAST
FEW YEARS, THE THING THAT MY LAB
IS ALSO PUSHING FORWARD IS
ACTUALLY A HUMAN TRANSMISSION
MODEL, USING CONTROLLED HUMAN
INFECTIONS TO DETERMINE HOW
VIRUSES ACTUALLY SPREAD IN
HUMANS.
SO THAT WE CAN UNDERSTAND
WHETHER OUR MODEL SYSTEMS ARE
ACTUALLY ACCURATE.
WE SPEND SO MUCH MONEY, WE SPEND
SO MUCH TIME DOING TRANSMISSION
IN ANIMAL MODEL, AND IF THAT
DOES NOT ACTUALLY RECAPITULATE
WHAT IS HAPPENING IN HUMANS,
HAVE WE REALLY PUSHED FORWARD?
THAT'S REALLY IMPORTANT TO ME.
SO TRANSMISSION IN HUMANS, WHY
DO I LIKE THEM?
THEY'RE SUSCEPTIBLE TO HUMAN
INFLUENZA VIRUSES, I CAN TAKE A
CLINICAL ISOLATE, GIVE TO
FERRETS AND LOOK AT TRANSMISSION
PHENOTYPES.
IT HAS A SIMILAR RESPIRATORY
TRACT TO HUMANS, FIVE LUNG
CLOA --SIMILAR SIGNS, THEY SNEEY
LOSE WEIGHT.
WE HAVE A REALLY NICE REVISED
SCORING SYSTEM, WE HAVE A NICE
PAPER WE PUT TOGETHER WITH
STACEY ACTUALLY THAT REALLY
HIGHLIGHTS THIS, SO YOU CAN
ACTUALLY BREAK DOWN BASED ON THE
SEVERITY OF DIFFERENT SYMPTOMS
AND HAVE A REALLY NICE BAR GRAPH
TO LOOK AT THE TYPES OF
DIFFERENT DISEASES THAT ARE
ASSOCIATED WITH VIRAL INFECTIONS
IN FERRETS.
THEN THEY EXHALE INFECTIOUS
VIRUS INTO THE AIR.
SO WHAT WE CAN DO IN MY LAB,
BECAUSE I DO CARE A LOT ABOUT
THE VIRUS IN THE ENVIRONMENT, IS
THAT WE'VE SPENT A LOT OF TIME
FIGURING OUT WAYS TO ASSESS THE
VIRUS IN THE ENVIRONMENT.
SO HERE IS OUR FERRET, IT'S IN
ONE OF OUR CHAMBER BOXES.
THIS IS A TYPE OF AEROSOL
SAMPLER CALLED A BIOSPOT.
IT IS A LIQUID CONDENSED AEROSOL
SAMPLER, SO WHAT THAT MEANS IS
THIS IS PULLING IN -- THIS IS
NOT THE VIVUS BUT IT'S AN OLDER
VERSION PULLING IN AT 2 L 2-LITS
PER MINUTE, IT GOES INTO THIS
PORT HERE, IT'S CONNECTED TO
HERE, AND THEN THE LIQUID GETS
ADDED TO ALL THE DIFFERENT
AEROSOLS AND IT GETS COLLECTED
INTO LIQUID SO WE COLLECT FOR
JUST 10 MINUTES.
WE DO IT EVERY DAY BEFORE WE
NASAL-WASH THE FERRETS.
HERE I'M WATCHING THE INFECTIOUS
VIRUS TITER JUST FROM THE AIR OF
THESE ANIMALS.
THIS IS INFECTED WITH THE
H1N1 PANDEMIC MY VUS, THIS IS --
PER TOTAL SAMPLE, WHICH IS ABOUT
750 MICRO LITERS.
WE HAVE A VARIETY OF OTHER
SYSTEMS THAT WE USE AND I'M
HAPPY TO TALK ABOUT THAT PPED A
I'LL TALK ABOUT THAT IN THE
DAYCARE SETTING TOO, BUT WE'RE
REALLY INTERESTED IN THIS
PROBLEM AND HAPPY TO HELP OTHERS
SORT OF FIGURE OUT WAYS TO
SAMPLE AIR IN THE ENVIRONMENT AS
WELL.
OKAY.
SO LET'S TALK ABOUT OUR
CLASSICAL TRANSMISSION MODELS
FOR FERRETS IS THIS SORT OF
DEVICE HERE WHERE THERE'S A
SINGLE DONOR, THERE'S A
RECIPIENT, YOU'LL SEE THIS IN MY
TALKS AS WELL, AND THERE ARE
PERFORATIONS THAT SORT OF
SEPARATE THESE ANIMALS SO THEY
DON'T HAVE ANY DIRECT COULD BE
TACT AND THERE IS DIRECTIONAL
AIR FLOW.
AT LEAST IN MY SYSTEM FROM THE
DONOR TO THE RECIPIENT.
TRADITIONALLY THESE HAVE BEEN
DONE FOR 14 CONSECUTIVE DAYS.
THIS IS A 2 BY 2 BOX.
WHEN I GIVE THESE TALKS, IMAGINE
A 2 BY 2 BOX AND IMAGINE YOU'RE
SITTING IN HALF OF THAT 2 BY
2 BOX FOR 14 DAYS AND 13 NIGHTS.
COULD YOU IMAGINE, WHO LIVES --
ANYBODY WHO HAS ACTUALLY DONE
THAT AS LIKE A THOUGHT
EXPERIMENT OR HAS LIVED THAT
LIFE, RAISE YOUR HAND.
NO ONE, RIGHT?
WE DON'T -- THAT'S NOT HOW WE
LIVE.
SO THIS IS A REALLY ARTIFICIAL
SYSTEM.
WE ALSO USE IMMUNOLOGICALLY
NAIVE ANIMALS.
THIS IS ALL THE PANDEMIC
PREPAREDNESS -- USE IMEU
KNOWLEDGEICALLY IMMUNE ANIMALS.
THEN DIRECTIONAL AIR FLOW AT
HIGH AIR CHANGE RATES.
THEY ACTUALLY DID A CROSS
CENTER, CROSS LAB EXPERIMENT
USING I THINK IT WAS 11
DIFFERENT LABS THAT DO AEROSOL
TRANSMISSION EXPERIMENTS WITH
FERRETS THAT HAVE DIFFERENT
SETUPS TO SORT OF COMPARE ACROSS
THEM, AND THE AIR EXCHANGE RATES
RANGE FROM 11 TO 35.
SO THAT IS HUGE.
I WILL SAY IN A SPACE LIKE THIS,
GIVEN THE VAULTED CEILINGS,
WE'RE PROBABLY LOOKING AT AN AIR
EXCHANGE RATE OF ABOUT -- IF WE
WERE TO TRY TO MEASURE, ABOUT 5,
RIGHT?
SO THAT'S FIVE AIR CHANGES PER
HOUR.
THAT'S ABOUT REASONABLE, IT'S
ACTUALLY PRETTY HIGH FOR AN
OFFICE BUILDING.
THAT'S WHAT IT FEELS LIKE AT
LEAST WITH THE VOLUME OF AIR IN
THIS SPACE.
MOST DAYCARE CENTERS OR OFFICE
BUILD THATION AREBUILDINGS THATE
LESS THAN 1 AIR EXCHANGE PER
HOUR AND HOSPITALS RUN AT ABOUT
6 TO 11, RIGHT?
OSHA REQUIREMENTS.
JUST TO GIVE EVERYBODY A SETUP
OF WHERE WE ARE.
SO THE WAY WE'VE DONE THIS IS
THAT WE INFECT THE FERRET, THIS
IS SORT OF TRADITIONAL HERE,
RIGHT, YOU INFECT A FERRET ON
DAY ZERO.
24 DAYS LATER, YOU PUT THE NAIVE
RECIPIENT IN THE OTHER PART OF
THE CAGE.
YOU DO THE CLASSICAL EXPOSURE
FOR 14 DAYS.
EVERY OTHER DAY YOU PULL THEM
OUT, NASAL WASH THEM, WEIGH
THEM, AND DAY 14 YOU COLLECT
SERUM.
THIS WAS PUBLISHED IN A PAPER IN
2012 -- SORRY, 2021, OH GOSH,
THAT WE LOOKED AT THE DIFFERENT
TIMINGS OF EXPOSURE.
SO WE DID A 7-DAY -- WE THOUGHT
14 WAS RIDICULOUS, THEN WE DID A
TWO-DAY SHORT, WHICH I CONSIDER
BEING A WEEKEND.
SAY YOU'RE STUCK IN A 2-BEDROOM
CONDO WITH SOMEBODY FOR THE
WEEKEND.
I CALLED IT A SLEEPOVER, BUT
BASICALLY WE DID A SLEEPOVER,
BUT WE DID IT FOR FIVE NIGHTS.
WHAT WE WERE TRYING TO MIMIC
HERE WAS LIKE I GO TO WORK AND I
SHARE A SMALL OFFICE WITH
SOMEONE EVERY DAY FOR FIVE DAYS,
FROM MONDAY TO FRIDAY.
BUT SINCE FERRETS ARE NOCTURNAL,
WE DID IT AT NIGHT, SO THAT'S
WHY IT'S LIKE A SLEEPOVER.
WE DID THIS FOR THE SEASONAL
2009H1N1.
AND SEASONAL -- I'M JUST GOING
TO SUMMARIZE THIS HERE.
WHAT WE FOUND IS THAT IN THE
2009 PANDEMIC, ALL OF -- IN
EVERY SYSTEM WE TESTED THAT
VIRUS LOVES TO TRANSMIT, THIS IS
WHY FOR A VIRUS THIS IS
FANTASTIC IN FERRETS.
IT'S REALLY HARD TO GET THAT
VIRUS NOT TO TRANSMIT.
THEN THE SEASONAL -- ALSO HAD
REALLY HIGH EFFICIENT
TRANSMISSION.
I WILL SAY WE'VE MOVED TO TWO
DAYS SHORT BECAUSE I THINK IT
STREAMLINES EVERYTHING IN TERMS
OF HUSBANDRY, IN TERMS OF
PERSONNEL, SO WE'VE NOW MOVED TO
DOING TWO-DAY EXPOSURES FOR OUR
STUDIES, SO I'LL TALK ABOUT
THAT.
WE THINK THAT'S MAYBE NOT AS
GREAT AS WHERE WE ARE WHAT
HUMANS LIVE IN, BUT IT'S BETTER
THAN 14.
AND THEN THE OTHER THING I'LL
MOVE ON TO IS PRE-EXISTING
IMMUNITY.
THIS IS A PICTURE OF MY KIDS
AROUND THE AGE OF 5, SO WE KNOW
FROM REALLY NICE COHORT STUDIES
DONE BOTH IN EUROPE AND THE U.S.
THAT PREDOMINANTLY BY THE AGE OF
5, WE'VE ALL EXPERIENCED
INFLUENZA, EITHER FROM NATURAL
INFECTION OR FROM VACCINATION.
SO THIS WORK DONE BY KATIE
GOSTIC AND SARAH COBE AND JAMIE
LLOYD SMITH AND OTHERS REALLY
NICELY SORT OF GIVE THE
PROBABILITY BASED ON YOUR BIRTH
YEAR OF WHAT YOUR INITIAL VIRAL
INFECTION PO POTENTIALLY WAS.
SO I REALLY LIKE THIS BECAUSE IT
SORT OF HIGHLIGHTS THAT IF YOU
WERE LIKE ME AND YOU'RE BORN IN
THE 80s, YOU PREDOMINANTLY
HAVE AN H3N2 INITIAL PRIMING
IMMUNE RESPONSE VERSUS AN H1N1.
BUT IF YOU WERE BORN MAYBE IN
2009, THESE YOUNG ONES HERE,
THEY HAVE A MUCH MORE
PREDOMINANT AGE 1 PRIMARY
INFECTION VERSUS THE ELDERLY.
SO THIS IS IMPORTANT BECAUSE
TRANSMISSION OF BOTH SEASONAL
AND PANDEMIC INFLUENZA VIRUSES
OCCURS IN THE CONTEXT OF
PRE-EXISTING IMMUNITY FROM
SEASONAL VIRUSES, RIGHT?
LIKE THAT IS -- THAT SHOULD BE
OBVIOUS TO ESH.
EVERYBODY.
SO WHEN WE TALK ABOUT
TRANSMISSION FITNESSES OF BOTH
SEASONAL VIRUSES AND EMERGING
PANDEMIC THREATS, THEY SHOULD
ALWAYS BE DONE IN THE CONTEXT OF
PRE-EXISTING IMMUNITY.
SO WE STARTED WITH THIS IDEA OF
SEASONAL INFECTIONS, BECAUSE
OBVIOUSLY H3N2 STILL TRANSMITS
AND H1N1 STILL TRANSMITS EACH
YEAR IN ALL THIS PRE-EXISTING
IMMUNITY IN THE POPULATION.
SO WE DID THIS EXPERIMENT AGAIN
WHERE WE HAVE OUR TRANSMISSION
SETUP HERE, AND THIS IS WHAT OUR
CAGING SYSTEM LOOKS LIKE AND
I'LL SHOW YOU OUR STUDY DESIGN
IN A MINUTE, BUT WE HAVE THESE
NICE ALLENTOWN CAGES, I DESIGNED
THEM WHEN I WAS A POSTDOC AT THE
NIH AND WE STILL USE THEM NOW.
I MADE SOME MODIFICATIONS, BUT
THE PERFORATION BARRIERS ARE ONE
INCH APART AND THE PERFORATIONS
ARE OFFSET SO THAT THERE IS
REALLY WHAT WE'RE SHOWING HERE
IS LARGER AEROSOL DYNAMICS.
WE'VE HAD SOME AEROSOL
COLLABORATORS WORK WITH US TO
TRY TO FIGURE OUT WHAT THE
AEROSOL SIZE IS THAT REALLY GETS
ACROSS BUT IT PROBABLY ANYTHING
THAT'S SMALLER THAN 10 OR
15 MICRONS IS GETTING ACROSS.
SO WHAT WE DECIDED TO DO TO TEST
THE IMPACT OF PRE-EXISTING
IMMUNITY ON TRANSMISSION IS TO
LOOK AT -- TAKE DONOR AND
RECIPIENT ANIMALS AND GIVE THEM
A PRIMARY INFECTION.
SO AS STACEY MENTIONED, WE
PRESCREEN FERRETS TO MAKE SURE
THEY'RE SERONEGATIVE, AND THAT'S
BECAUSE I'D LOVE TO START BUYING
THE IMMUNE ONES THAT NOBODY ELSE
WANTS BUT I WANT TO KNOW THE
TIMING OF PRIMARY INFECTION TO
SECONDARY INFECTIONS CURRENTLY.
SO WE DO A PRIMARY INFECTION AND
THEN WE WAIT -- WE USED TO WAIT
60 DAYS, WE NOW WAIT UPWARDS OF
84 DAYS.
SO THIS IS THREE MONTHS.
THIS IS -- WE HAVE A FERRET
DAYCARE AS WELL AS A FERRET
STAYCATIO INSURE, A LITTLEN, A T
COMMITTEE WE'RE RUNNING.
SO WE HAVE THE ANIMAL, GIVE THEM
A PRIMARY INFORECAST, THREE INFE
MONTHS LATER, BRING THEM BACK
IN, GIVE THE DONOR A SECONDARY
INFECTION AND RECIPIENTS ARE
PLACED INTO A CAGE 24 LATER, AND
THEN WE DO A 2-DAY EXPOSURE.
WE STARTED OFF BY LOOKING --
ASKING THIS QUESTION ABOUT
HETEROLOGOUS IMMUNITY.
IF YOU HAVE A PRIMARY
H1N1 INFECTION WHAT IS YOUR
SUSCEPTIBILITY TO
HDN2 TRANSMISSION AND VICE
VERSA.
SO THIS VIRUS IN 2009 EMERGED IN
THE CONTEXT OF BOTH PRE-EXISTING
H1N1 IMMUNITY FROM PRE2009 AND
H3N2 IMMUNITY.
SO I'M JUST GOING TO WALK
THROUGH SOME OF THE GRAPHS HERE.
SO THIS IS NO PRIOR IMMUNITIES
OF THE DONOR ANIMALS.
EACH BAR IS AN INDIVIDUAL
ANIMAL.
WE DID FOUR PAIRS.
AND HERE YOU'LL SEE THAT THIS
VIRUS TRANSMITS 100% -- SORRY,
THIS IS THREE PAIRS -- TO 100%
OF THE NAIVE RECIPIENTS AND EACH
LINE IN BLUE IS A RECIPIENT.
THEN THESE ANIMALS HERE ON THE
RIGHT-HAND SIDE HAVE A PRIOR
H3N2 IMMUNITY, BOTH THE DONORS
AND THE RECIPIENTS.
SO YOU'LL SEE FIRST THAT IF YOU
PUT A LOT OF VIRUS UP A FERRET
NOSE WITH PRE-EXISTING IMMUNITY,
THEY'LL SHED A LOT OF VIRUS
STILL FOR LOTS OF DAYS, BUT THEY
DO CLEAR THE VIRUS FASTER, VIRAL
CLEARANCE IS SHORTENED, THE
TIMING TO VIRAL CLEARANCE.
SO DAY 5, YOU'LL SEE THERE'S
LESS VIRUS IN PREIMMUNE FERRETS
COMPARED TO THOSE THAT DON'T
HAVE ANY PRE-EXISTING IMMUNITY.
AND THEN IN THIS EXPERIMENT THAT
WE'VE DONE, WE'VE DONE THIS MANY
TIMES AND TRANSMISSION
EFFICIENCIES HAVE ONLY GONE UP.
WE'VE GOT 2 OUT OF 3, IN
SUBSEQUENT ONES WE'VE GOT 3 OUT
OF 3.
BUT THIS IS CLASSIC REALLY
EFFICIENT TRANSMISSION.
SO H1N1 TRANSMITS IN THE CONTEXT
OF PRE-EXISTING IMMUNITY AGAIN
AFTER A TWO-DAY EXECUTIVE
EXPOSURE.
WHAT ABOUT H3N2?
SO HERE IS OUR NAIVE --
IMMUNOLOGICALLY NAIVE FERRETS,
SO ALL OF THE DONOR ANIMALS SHED
VIRUS, ALL OF THE RECIPIENTS, WE
DID IN GROUPS OF 4, ALSO SHED
VIRUS AND WERE INFECTED.
AND THEN IF YOU HAVE
PRE-EXISTING H1N1 I ME1 IMMUNITH
IN THE DONORS AND RECIPIENTS,
YOU HAVE INCREASED CLEARANCE BY
DAY 5, YOU DO SHED A LOT OF
VIRUS IN THE DONORS BUT YOU HAVE
NO TRANSMISSION TO THE
RECIPIENTS.
IT'S REALLY FASCINATING.
THIS IS AIRBORNE TRANSMISSION.
SO WE ALSO THEN FURTHER MAP THIS
TO THE RECIPIENT IMMUNITY
SPECIFICALLY.
YOU COULD HAVE DONORS THAT ARE
IMMUNOLOGICALLY NAIVE AND THE
RECIPIENTS WERE THE ONES THAT
WERE CAUSING THIS GLOBAL
PROTECTION.
BUT YOU'RE THINKING IN YOUR
HEAD,S THAT INTERESTING BUT WE
GET H2N2, IT TRANSMITS ALL THE
TIME.
2017, 2018.
HUGE H3N2 SEASON, ALMOST 80,000
DEATHS THAT YEAR.
IT WAS A BAD YEAR.
SO WHAT HAPPENED?
SO LET'S THINK AGAIN ABOUT HOW
DO WE LIVE IN SPACES?
WE LIVE WITH COMPLEX BEHAVIORS,
WE LIVE IN COMPLEX ENVIRONMENTS.
SO AGAIN AT CLOSE RANGE, ALL
TRANSMISSION MODES ARE POSSIBLE.
SO WHAT WE DID WAS WE JUST DID
THE SAME EXPERIMENT BUT NOW
REMOVE THE BARRIER SO IT'S A
CONTACT TRANSMISSION EXPERIMENT,
TWO DAYS, YOU GET EFFICIENT
TRANSMISSION.
SO WHEN YOU HAVE ALL MODES OF
TRANSMISSION, THE BARRIER TO
INFECTION FROM PRE-EXISTING
IMMUNITY CAN BE OVERCOME.
YOU'RE GOING TO ASK ME IS IT A
DOSE ISSUE, IT'S NOT.
WE TRIED TO DO FD50 STUDIES AND
WE DON'T HAVE A BIG DIFFERENCE
IN TERMS OF THE INFECTIOUS DOSE
INTRANASALLY INOCULATED INTO AN
ANIMAL, BUT AGAIN, TRANSMISSION
IS NOT A BIG BOLUS OF VIRUS UP A
FERRET'S NOSE.
IT IS LOTS OF DIFFERENT AEROSOL
PARTICLES FALLING AT DIFFERENT
SITES WITHIN THE RESPIRATORY
TRACT AND INITIATING INFECTIONS
AND BEING SUCCESSFUL.
SO THERE'S A LOT OF COMPLEXITY
HERE.
SO BEFORE I MOVE ON TO THE
FERRET DAYCARE, WHAT THIS MEANS
I THINK THAT'S REALLY IMPORTANT
IS THAT SEASONAL VIRUSES HAVE
DIFFERENT AIRBORNE TRANSMISSION
IN DIFFERENT -- FORECASTING AND
PREDICTION MODELING AND ALL OF
THAT, WE SORT OF LUMP EVERYTHING
TOGETHER FOR INFLUENZA.
ALSO WE HAVE STARTED USING THIS
IN OUR NEW RISK ASSESSMENT
PIPELINE.
WE HAVE A NEW TRIAGE MANUSCRIPT
OUT FOR SWINE VIRUSES AND WE'VE
DONE IT NOW WITH H5s, IS
LOOKING AT THE ROLE OF THESE
PRE-EXISTING IMMUNITY IN THE
PATHOGENESIS AND TRANSMISSION OF
EMERGING PANDEMIC THREATS.
SO WE SEE DEFINITELY INFECTION
HAPPENS BUT REDUCED SEVERITY OF
DISEASE.
SO COMING BACK TO THIS, HOW DO
WE CHANGE THINGS?
WE'VE GOT TO CHANGE EXPOSURE
TIMES 100%, WE'VE GOT TO START
USING ANIMALS WITH PRE-EXISTING
IMMUNITY BECAUSE WE ARE NOT
IMMUNOLOGICALLY NAIVE.
MAYBE YOU COULD ARGUE THAT WE
WERE FOR SARS-COV-2, BUT WE ALSO
HAD SOME PRE-EXISTING
CORONAVIRUS IMMUNITY THAT I
THINK IS IMPORTANT TO IMPLEMENT
WHEN WE DO THESE SORT OF
STUDIES.
AND THEN LET'S TALK ABOUT AIR
FLOW.
I'M HOPING I'M ON TIME.
I ACTUALLY HAVE NO IDEA.
AM I REALLY?
I'M SO SORRY.
OH, IS THAT THE TIMER?
OKAY.
SO WE BUILT A FERRET PLAY PEN.
WE'VE GOT A ROBOT, AIR SAMPLING
AT THE SOURCE.
WHAT WE CAN DO IS HAVE REALISTIC
BEHAVIORS.
EVERYONE LOVES THIS, THE FERRETS
LOVE TO RIDE ON THESE ROBOTS.
WE'VE GOT AIR SAMPLERS AROUND
THE PERIMETER AND THEY HAVE
REALISTIC BEHAVIORS.
WHAT WE END UP DOING IS WE DO
ANYWHERE FROM ONE TO FOUR HOURS
OF EXPOSURE AND THEN WE
INDIVIDUALLY HOUSE FERRETS.
WE ARE LOOKING AGAIN AT THIS
EQUATION OF TRANSMISSION THAT'S
REALLY IMPORTANT TO ME.
SO WHAT WE'RE DOING IS NOW
BEHAVIOR TRACKING THESE ANIMALS
AND WE'RE USING BEHAVIOR THINGS
TO DEFINE WHAT ARE ALL THE
DIFFERENT INTERACTIONS THAT
FERRETS MAKE AND YOU WOULD
EXPECT THAT OF COURSE FERRETS
WOULD HAVE THE TOTAL DURATION OF
DONOR TOUCHING WOULD BE WHAT
WOULD LEAD TO SUCCESSFUL
TRANSMISSION EVENT, AND IN
REALITY IT'S NOT, SO THESE ARE
TWO RECIPIENT ANIMALS THAT GOT
INFECTED AND THEY HAVE THE
LOWEST NUMBER OF TOTAL DURATION
IN ONE OF OUR STUDIES.
WE ALSO CAN LOOK AT INFECTION IN
THE ENVIRONMENT, SO WE SWAB A
VARIETY OF SURFACES.
THIS IS ERNIE, OUR ROBOT
SAMPLER.
SO THESE ARE THINGS WE FOUND
INFECTIOUS VIRUS ON AND WE FOUND
MORE WITH RNA.
MY BIG THING IS THAT WE SHOULD
BE RE-THINKING ANIMAL MODELS,
AND WE'LL TALK ABOUT THAT MORE.
SO THIS IS LIKE WHERE WE SHOULD
BE FOCUSING ON THINGS THAT ARE
MORE REALISTIC IN TERMS OF
EXPOSURE TIMES, VENTILATION
RATES AND NETWORKS OF HOW PEOPLE
INTERACT.
AND THIS IS MY TEAM.
WE'RE REALLY FORTUNATE ALL OF
THE PLAY PEN STUFF WAS FUNDED BY
FLU LAB BECAUSE IT'S HARD TO GET
NIH FUNDING FOR THAT SORT OF
STUFF.
YEAH, THIS YEAR, OTHER FOLKS AND
MY TEAM.
THANKS.
I'M SORRY.
>> HI, SEEMA.
REALLY COOL TALK.
I REALLY LIKE WHAT YOU'RE DOING
WITH THE PRE-EXISTING MODEL.
I THINK IT'S SUPER IMPORTANT.
I HAVE TWO QUESTIONS RELATED TO
THAT.
DO YOU HAVE ANY UNDERSTANDING OF
THE CORRELATES FOR PROTECTION?
I KNOW YOU SAID IT'S SENTINEL
BUT DO YOU KNOW WHAT IS GOING ON
IN THIS ANIMAL?
SECONDLY, DO YOU THINK THAT YOUR
TIMING IS GOOD ENOUGH, LIKE, TO
84 DAYS, WOULD YOU -- FOR
EXAMPLE, IF YOU WAIT A YEAR, IS
THAT MORE LIKE WHAT WE SEE IN A
HUMAN POPULATION?
>> THOSE ARE BOTH GOOD
QUESTIONS.
THE FIRST ONE IS DO WE HAVE ANY
IDEA WHAT THE CORRELATIVE
PROTECTION IS?
WE KNOW IT IS NOT NEUTRALIZING
ANTIBODIES, WE KNOW IT'S NOT
NEUROMINIDACE AND BODIES, WE
DON'T THINK IT'S ANY SORT OF
ANTIBODIES AGAINST THE HUMAN
GLUTE NN, WE'RE DOING DIFFERENT
ROUTES OF INOCULATION FOR THE
VACCINES TO TEASE APART
IMMUNE -- THEORETICALLY TICKLING
OF IMMUNE RESPONSES, T CELLS
VERSUS B CELLS, GERNTION WE
DON'T HAVE THE REAGENTS FOR
FERRET TO REALLY TEASE THAT
APART AND I'M NOT AN
IMMUNOLOGIST SO EVEN IF WE DID,
I'D BE LOST.
SO WE HAVE FOUND THAT
INACTIVATED VACCINES
ADMINISTERED INTRANASALLY IS NOT
PROTECTIVE.
IF I CAN GIVE AN H1N1 INTRANASAL
MONOVALENT INACTIVATED VACCINE
WITH ADJUNCT IT DOES NOT PROTECT
AGAINST H2N2 NATURAL INFECTION.
SO TRANSMISSION NOT UP THEIR
NOSE BUT AS A RECIPIENT.
HOWEVER, IF WE GIVE LA IV, WE
ARE STARTING TO SEE PROTECTION.
SO WE DO THINK THAT'S SOMETHING
TO DO ABOUT THE REPLICATION AND
THAT INDUCING POTENTIALLY MORE
MUCOSAL T CELLS RATHER THAN
ANTIBODIES, BUT WE'RE TRYING TO
FIGURE ALL THAT OUT.
SO WE'RE WORKING ON THAT.
BECAUSE OF THE LACK REAGENTS,
WHAT WE'RE DOING WITH SCOTT
BOYD'S GROUP IS DOING SINGLE
CELL RNA SEQ WITH ALL OF THESE
STUDIES WITH PRE-EXISTING
IMMUNITY AFTER THE PRIMARY IB
INFECTION, AFTER THE SECONDARY
INFECTION TO SEE WHAT TYPE OF
RESPONSES WE GET.
THEN YOUR SECOND QUESTION ABOUT
TIMING, WE HAVE NOT WAITED A
YEAR BUT WE HAVE WAITED OUT TO
160 DAYS, I THINK IS THE LONGEST
WE'VE GONE, MOSTLY BECAUSE THESE
FERRETS, WE HANG THEM OUT,
REALLY IT'S A RETIREMENT
COMMUNITY THAT WE'VE BEEN
RUNNING UP AT PITTSBURGH AND
VALERIE IN MY GROUP AND -- ROCKY
DID A LOT OF THIS WORK THAT I
PRESENTED HAS REALLY BEEN
MAINTAINING THAT IN PITTSBURGH
AND SO YEAH, WE HAVEN'T GONE OUT
TO A YEAR, IT'S INTERESTING TO
THINK ABOUT.
WHAT WE HAVE DONE ARE THESE
TERTIARY INFECTIONS WHERE WE DO
A PRIMARY WITH ONE H1, SECONDARY
WITH AN H3N2 THROUGH NATURAL
INFECTION, THESE RECIPIENTS, AND
BRING THEM BACK IN FOR TERTIARY
TO SEE IF WE CAN GET ANOTHER
THIRD REPLICATION OR THIRD
INFECTION AND THROUGH NATURAL
ROUTES AS RECIPIENTS.
>> WE'LL TAKE ONE MORE.
>> I LOVE THE FERRET DAYCARE
PLAY PEN MODEL FOR TRANSMISSION.
IT'S SO MUCH MORE RELEVANT TO
HUMAN BEHAVIOR.
HAVE YOU EVER THOUGHT ABOUT HOW
THIS COULD BE POSSIBLY ADAPTED
OR SPOKEN WITH EHS OR OTHER
PEOPLE ABOUT LIKE IF YOU'RE
DEALING WITH A HIGHER
CONTAINMENT PATHOGEN, IS THERE
ANY WAY TO REPLICATE THAT?
>> THEY'RE OWE C OKAY WITH US D-
WE ALREADY HAVE PLANS IN PLACE
TO DO THIS.
WHAT WE HAVE DISCUSSED WITH
THEM, SO WE HAVE THE SYSTEM IS
SORT OF CONTAINED BUT ALSO WE
WEAR PPE AS THE INDIVIDUALS IN
THE ROOM TO PROTECT US FROM THE
EXPULSION OF THE AIR OF THE
ANIMALS WHEN THEY ARE OUT
PLAYING IN THAT SPACE.
AND SO IT'S THE SAME THING AS
LIKE MOVING AN ANIMAL OUTSIDE OF
THEIR CAGING BETWEEN, YOU KNOW,
I TAKING THEM OUT OF THE CAGE,
PUTTING THEM INTO A BOX IN THE
BIOSAFETY CABINET.
AND SO THEY SEEM TO BE OKAY WITH
US DOING THIS, AND AS LONG AS
LIKE NOBODY ELSE IS COMING INTO
THE ROOM.
THE ROOM IS NEGATIVE PRESSURED,
AS LONG AS EVERYTHING IS
CONTAINED IN THE ROOM,
PROTECTED, AND THERE'S OBVIOUSLY
A DWELL TIME BEFORE WE OPEN THE
DOOR SO THAT EVERYTHING CAN BE
EXCHANGED.
THEY SEEM TO BE ALL RIGHT WITH
THAT.
>> THAT'S AMAZING.
>> I'M GOING TO TURN TO MICHELLE
REALLY QUICK.
DO YOU WANT TO KEEP GOING WITH
QUESTIONS OR --
>> WE HAVE TIME IN THE PANEL
DISCUSSION.
SO PLEASE SAVE YOUR QUESTIONS
AND BRING THEM UP THEN.
YEAH, LET'S JUST TAKE -- THANK
YOU, SEEMA.
>> THANK YOU.
[APPLAUSE]
YOU ANDAND WE HAVE A BREAK UNTI,
SO WE'LL RECONVENE AT 10:20.
>> I'M GOING TO ENCOURAGE
EVERYBODY TO GET SETTLED AS I
WELCOME PETER HALFMANN FROM THE
UNIVERSITY OF WISCONSIN TO THE
PODIUM TO GIVE A TALK ENTITLED
INFLUENZA VIRUS AND SARS-COV-2
TRANSMISSION IN SMALL ANIMAL
MODELS.
AN OVERVIEW.
>> THANKS FOR THE INTRODUCTION.
SO THE PURPOSE OF THE
PRESENTATION TODAY WAS TO TRY TO
GIVE A TRANSMISSION 101 TALK ON
INFLUENZA VIRUSES, WHAT SUBTYPES
TRANSMIT, WHAT SUBTYPES DON'T,
AND THEN ALSO ON SARS-COV-2
VARIANTS, OF WHICH VARIANTS DO
TRANSMIT AND DON'T.
AND THEN LOOKING AT DIFFERENT
ANIMAL MODELS, PARTICULARLY JUST
FOCUSING ON THE RODENT MODEL OF
TRANSMISSION, WHAT RODENT MODELS
SUPPORT TRANSMISSION AND ONES
THAT DON'T.
AND YES, WE'RE GOING TO TALK
ABOUT ROUTES OF TRANSMISSION,
AIRBORNE TRANSMISSION, DIRECT
CONTACT, AND ALSO LOOKING AT
VERTICAL TRANSMISSION.
HERE WE GO.
SO FOR CONSIDERATIONS WITH
TRANSMISSION, THERE'S A LOT TO
THINK ABOUT.
I'M ONLY GOING TO SCRATCH THE
SURFACE, BUT THE VIRUS TITERS,
THE AMOUNT OF SHEDDING YOU HAVE,
THE TIME LAPSE WHEN YOU ADD THE
CONTACT OF THE NAIVE WITH THE
INFECTED ANIMAL.
WHAT ARE YOUR READOUTS FOR
EXPOSURE?
ARE YOU LOOKING AT FOR AN
INFECTIOUS VIRUS, ARE YOU
LOOKING FOR VIRAL RNA, DO YOU
WAIT THE 21 DAYS, 28 DAYS AND
LOOK FOR SEROCONVERSION.
DURING THE BEGINNING OF THE
PANDEMIC, WE SPENT A LOT OF TIME
WORKING WITH OUR MECHANICAL SHOP
MAKING DIFFERENT CAGES FOR
TRANSMISSION STUDY.
EVERYONE HAS THEIR OWN CAGING
SYSTEM THAT'S NOT REALLY
UNIVERSAL.
AIR FLOW IS IMPORTANT FOR THE
ENVIRONMENT.
THERE'S WORK DONE THAT IF YOU
LOOK AT TRANSMISSION AT
5 DEGREES VERSUS 20 DEGREES, YOU
SEE DIFFERENCES, AND WHAT KIND
OF BEDDING ARE YOU USING IN THE
CAGES?
DO YOU HAVE AIR FLOW THAT'S
BLOWING YOUR CORN COULD BE
BEDDING ACROSS TO THE NEXT CAGE
AND YOU'RE LOOKING AT AIRBORNE
TRANSMISSION, YOU ARE REALLY
LOOKING AT AIRBORNE
TRANSMISSION?
THAT KIND OF GOES INTO THEN WHAT
IS BEING TRANSMITTED, YOU COULD
SPEND THE WHOLE AFTERNOON
TALKING ABOUT MECHANICS OF
TRANSMISSION SIZES, ARE YOU
LOOKING AT RESPIRATORY DROP
LEATHER DROPLEATHERS BEING TRAND
OR AEROSOL FOMITES THAT ARE
BEING TRANSMITTED.
SO TRANSMISSION AND THE USE OF
ANIMAL MODELS FOR INFLUENZA
STARTED BACK IN THE 1930s.
THEY LOOKED IN THE 1940s AT
TRANSMISSION OF FLU VIRUSES WITH
INFLUENZA VIRUS.
AND EVEN BACK THEN, THEY COULD
SEE AT A DISTANCE OF 3 FEET THAT
YOU COULD SEE TRANSMISSION
STRAIGHT DOWN THE CORRIDOR.
IF YOU HAVE LIKE A BEND IN YOUR
TRANSMISSION UNIT, YOU COULD
STILL SEE TRANSMISSION
HAPPENING.
THE SAD PART HERE IS THAT THIS
IS 80 YEARS AGO AND WE HAVEN'T
REALLY CHANGED MUCH LOOKING AT
TRANSMISSION UNFORTUNATELY.
THE TALK RIGHT BEFORE THE BREAK
IS EXCELLENT WAYS TO START
THINKING ABOUT HOW LOOKING AT
TRANSMISSION AGAIN.
HOWHAT COSTSAND HOW MANY LABS CY
DO THAT KIND OF TRANSMISSION
STUDY IS ANOTHER CONSIDERATION.
NOT EVERYONE CAN USE FERRETS FOR
TRANSMISSION, SO OF COURSE THERE
WAS EFFORTS IN THE 60s TO LOOK
AT CAN WE LOOK AT MACE FOR MICET
WAS ALLUDED YESTERDAY THAT MICE
ARE A POOR MODEL FOR
TRANSMISSION.
A GROUP DID LOOK AT
H2N2 VIRUSES, BOTH MOUSE-ADAPTED
VIRUSES AND NON-MOUSE ADAPTED
H2N2.
WITH MICE, YOU COULD SEE 50%
TRANSMISSION, THEY WERE USING
THE SWISS WEBSTER MICE.
THESE WERE MALES, THEY DID
24-HOUR CONTACT.
OF COURSE WE DON'T HAVE
H2N2 CIRCULATING IN HUMAN
POPULATION ANYMORE, AND
CURRENTLY NO OTHER SUBTYPES ARE
IDENTIFIED FOR TRANSMISSION WITH
MICE, ADULT MICE, I SHOULD SAY.
DIRECT CONTACT TRANSMISSION HAS
ITS MERITS.
OF COURSE THIS IS AGAIN
SUBTYPE-SPECIFIC.
A GROUP LOOKING AT H1 VERSUS
H3N2 VIRUSES SHOWED THAT WHEN
YOU LOOKED AT THE H1N1s
INCLUDING PRA, YOU DIDN'T SEE
ANY DIRECT CONTACT TRANSMISSION
INVOLVE -- C FEMALES.
BUT YOU DID SEE ISOLATE SPECIFIC
SUBTYPE SPECIFIC WITH
H2N2 VIRUSES.
THEY WENT ON TO SHOW THAT A
FACTOR THAT I BROUGHT UP OF THE
TIMING IN THE VIRAL LOAD IS
IMPORTANT WHEN YOU LOOK AT
TRANSMISSION FOR DIRECT CONTACT.
THIS LAG BETWEEN WHEN YOU PUT A
CONTACT MOUSE IN WITH THE
INFECTED MOUSE WILL SKEW OR WILL
CHANGE YOUR TRANSMISSION
PERCENTAGES.
AND THEY SHOWED THAT IF YOU LOOK
AS LATE AT 56 HOURS, YOU NEVER
SAW TRANSMISSION WHEN YOU PUT
THE NAIVE IN, AND THAT PEAKED AT
30 HOURS AFTER CONTACT TIME.
AND YOU COULD SEE SOME
TRANSMISSION WITHIN SIX HOURS,
PUTTING THE NAIVE CONTACT IN.
AND THEN THEY ACTUALLY WENT
FURTHER TO SHOW THAT THE HIGHER
THE VIRUS TITER IN THE I SALIVA,
THESE ARE THE H3N2s IN GREEN,
YOU HAD HIGHER PERCENTAGE OF
INFECTION IN THE CONTACTS.
THE VIRUSES THAT ARE IN THE
GREY, THE WSN, THE H1N1s AND
MEMPHIS HAD LOWER TITERS IN THE
SALIVA AND THESE WERE
NON-TRANSMISSIBLE.
WITH THE DISCOVERY OF H5N1 IN
COWS AND INFECTED COW MILK, WE
WERE INTERESTED IN KNOWING COULD
THE MOUSE MODEL BE USED TO LOOK
AT VERTICAL TRANSMISSION.
TRYING TO ANSWER BASIC QUESTIONS
OF DO YOU ACTUALLY FIND
INFECTIOUS VIRUS IN THE MILK OF
MICE LIKE YOU DO WITH COWS, AND
THEN CAN THAT BE TRANSMITTED TO
PUPS.
SO IN OUR STUDY, WE USED BALB
Cs, LACTATING FEMALES.
THEY GAVE BIRTH 5 TO 7 DAYS AND
THEN WE INFECTED THEM WITH 100
PFU OF BOVINE H5N1 VIRUS AND
THEN WE WAITED 7 OR 9 DAYS TO
LOOK AT EITHER FOR THE PUPS, DO
WE FIND INFECTIOUS VIRUS IN THE
TISSUES, AND WHEN WE ALLOWED THE
MOTHER TO TAKE CARE OF THE PUPS
FOR NINE DAYS, WE SAW AROUND 37%
TRANSMISSION TO THE PUPS,
LOOKING AS SOON AS FOUR DAYS
AFTER THE INFECTION OF THE
MOTHER TO THE PUPS, WE ACTUALLY
SAW NO TRANSMISSION.
AND THIS CORRELATED TO HAVING
VIRUS POSITIVE SAMPLES OF THE
MOUSE MILK, AND ALSO POSITIVE
MAMMARY GLANDS IN THESE MOTHERS.
WHEN WE DID THIS SAME EXACT
EXPERIMENT BUT INSTEAD OF
PUTTING -- HAVING THE PUPS STAY
IN THE CAGE, WE INTRODUCED ADULT
FEMALES, AND WAITED FOR THE
EXACT SAME DAYS.
WE SAW NO DIRECT CONTACT
TRANSMISSION IN THOSE ADULT GUY
FEMALES, SUGGESTING THAT IT'S
THE INFECTED MILK THAT WE SEE
THE TRANSMISSION AND NOT SOME
KIND OF FOMITES IN THE CAGE.
SO WITH A MOUSE MODEL, THAT'S
VERY SUBTYPE-SPECIFIC.
TIMING IN VIRAL LOADS CAN PLAY A
BIG FACTOR TO DIRECT
TRANSMISSION, ALONG WITH
ASSOCIATION WITH THE HA PROTEIN
FOR TRANSMISSION.
AND WITH H5, WE SUSPECT THAT
ANOTHER HIGH PATH AVIAN
INFLUENZA LIKE H7N9 COULD ALSO
BE TRANSMITTED BY VERTICAL
TRANSMISSION.
THE GREAT MODEL, INSTEAD OF
USING MICE, GUINEA PIGS HAVE
BEEN A GREAT MODEL OF
TRANSMISSION.
A GROUP HAS SHOWN THAT
TEMPERATURE IS A FACTOR WHEN YOU
LOOK AT TRANSMISSION.
WHEN THEY USED A B VIRUS TO LOOK
AT DIRECT TRANSMISSION BETWEEN
TWO GUINEA PIGS, THEY SAW 100%
TRANSMISSION FOUR PAIRS, SPI
SPIKE -- HOWEVER WHEN THEY
LOOKED AT AIRBORNE TRANSMISSION,
ONLY 50% OF THE PAIRS HAD
TRANSMISSION WHEN IT WAS
20 DEGREES AND 100% OF THE
ANIMALS TRANSMITTED AT
5 DEGREES.
YOU CAN ASSUME THIS CORRELATES
TO THE TRANSMISSION OF FLU
VIRUSES IN THE WINTERTIME.
THIS WAS AN N OF 4 BUT THEY ALSO
REPEATED IT WITH ANOTHER LINEAGE
OF B VIRUS AND ALSO SAW THE SAME
KIND OF TRANSMISSION KINETICS.
WITH GUINEA PIG, YOU CAN ALSO
LOOK AT MUGS PEL DIFF MULTIPLE F
INFLUENZA A VIRUS AND YOU CAN
ALSO START PULLING APART VIRAL
DETERMINANTS AND PROBABLY
MORPHISMS THAT CORRELATE TO
TRANSMISSION.
IN FEMALE GUINEA PIGS, FOR
DIRECT TRANSMISSION WITH H1N1,
WE SEE 100% TRANSMISSION.
SIMILARLY YOU SEE 100%
TRANSMISSION WITH THE AIRBORNE
TRANSMISSION MODEL.
THIS WAS DONE WITH THE
CALIFORNIA ZERO NINE BUT IT WAS
ALSO SIMILAR RESULTS WITH
ANOTHNETHERLANDS PANDEMIC VIRUS.
ALSO H3N2 VIRUSES CAN BE
TRANSMITTED IN GUINEA PIGS.
100% TRANSMISSION WITH THE
PANAMA VIRUS.
THE GROUP THEN LOOKED AT WOULD
THE ABILITY FOR THE VIRUS TO
REPLICATE AT HIGH EFFICIENCY DUE
TO THE PB2 AMINO ACIDS KIND OF
CHANGE THIS, SO IF YOU MUTATED
THE PB2 AMINO ACIDS AT 627 AND
701, PARTICULARLY AT 627 -- BACK
AGAIN CLOSE TO 100% WHEN YOU GO
IN WITH A MUTATION AT THE
709 POSITION.
THEY WERE ABLE TO SHOW THAT WITH
VIETNAM 1203, WHICH DOES NOT
HAVE AIRBORNE TRANSMISSION IN
GUINEA PIGS BUT THERE'S DIRECT
CONTACT.
THE SAME KIND OF AMINO ACID
POLYMORPHISMS WILL CHANGE THE
REPLICATION POTENTIAL.
LIKE JACCO MENTIONED EARLIER,
AIRBORNE TRANSMISSION IS ALSO IN
HAMSTERS.
THIS WAS DONE BY THE KIYOKO
GROUP IN JAPAN.
THEY LOOKED AT SYRIAN HAMSTERS
IN FEMALES, AND THEY SAW 100%
TRANSMISSION OR CLOSE TO IT WITH
THE H1N1 SUBTYPES.
THIS FALTERED WITH THE
H3N2 SUBTYPES AND THEY SAW NO
TRANSMISSION WITH THE B VIRUSES.
SO FOR THE RODENT MODELS FOR
AIRBORNE TRANSMISSION WITH FLU,
I WOULD SUGGEST THAT WE PUT OUR
EFFORTS INTO MAKING REAGENTS FOR
GUINEA PIGS SINCE THE H1s, THE
H3s TRANSMIT ALONG WITH THE B
VIRUSES.
IT'S STILL UNKNOWN ABOUT
MAMMALIAN TRANSMISSIBLE
H5 VIRUSES OR POTENTIAL H5N1s
FROM COW VIRUSES ARE BIGGER THAN
A TRANSMIT IN GUINEA PIGS.
SO THE REST OF THE TALK WILL BE
ABOUT TRANSMISSION OF
SARS-COV-2.
WHEN THE ORIGINAL SARS
COV1 OUTBREAK OCCURRED HAMSTERS
WERE KNOWN TO BE GREAT
TRANSMISSION MODEL FOR THAT
VIRUS.
WE PUT A LOT OF WORK INTO
SHOWING TRANSMISSION OF COV2 IN
HAMSTERS.
WE HAD TWO DIFFERENT SETUPS FOR
TRANSMISSION IN THE BEGINNING.
WE HAD ANIMALS THAT WERE IN THE
SAME CAGE TOGETHER WITH A WIRE
MESH SEB RATING SEPARATING THER
FLOW GOING FROM THE INFECTED TO
THE CONTACT.
WE ALSO HAD NON-DIRECTIONAL AIR
FLOW SETUPS TOO WHERE THE
ANIMALS WERE SIDE BY SIDE AND
AIR FLOW WAS GOING IN A
NON-DIRECTIONAL MANNER.
THESE ARE OUR SETUPS AT MADISON
FOR TRANSMISSION.
WE CAN EITHER PUT ONE CAGE IN
FOR THE DIRECTIONAL AIR FLOW AND
THE SIDE BY SIDE.
WE CAN PUT SIX PAIRS OF HAMSTERS
TO DO OUR STUDIES.
WE SAW RIGHT AWAY WHAT THE DELTA
VARIANT WAS, IT WAS DIFFICULT
NOT TO HAVE TRANSMISSION GOING
ON.
SO YOU COULD PAIR THESE ANIMALS
TOGETHER FOR 24 HOURS OR AS
SHORT AS 6 HOURS AND WE SAW 100%
TRANSMISSION IN OUR DIRECTIONAL
AIR FLOW SETUP.
SO TRIED TO MAKE THINGS A LITTLE
MORE STRINGENT, WE WENT AHEAD
AND TRIED NON-DIRECTIONAL AIR
FLOW.
AGAIN AT 24 HOURS, WE SAW 100%
TRANSMISSION, AND THAT STARTED
CREEPING DOWN TO NO TRANSMISSION
IF YOU DID JUST AN HOUR OF
CONTACT TIME.
AGAIN JACCO MENTIONED THIS
MORNING THIS ALL KIND OF CHANGED
WITH THE BA1 INTRODUCTION.
NON-DIRECTION OR DIRECTION AIR
FLOW DIDN'T MATTER, WE COULD
NEVER GET AIRBORNE TRANSMISSION
WITH BA1.
WE TRIED A COUPLE DIFFERENT
PARAMETERS CHANGING.
WE USED 72 HOURS FOR CONTACT
TIME BUT STILL NO TRANSMISSION.
WE ALSO INCREASED THE DOSE THAT
WE GAVE FROM 10 TO THE THIRD TO
10 TO THE FIFTH AND STILL SAW NO
TRANSMISSION IN THESE ANIMALS.
WE WORKED WITH ZONDA AND USED
THIS TRANSGENIC ACE 2 HAMSTERS
AND REALLY INCREASED THE CONTACT
TIME TO 96 HOURS.
BUT STILL SAW NO TRANSMISSION.
IT WAS ONLY WHEN WE DID DIRECT
CONTACT STUDIES WITH BA1 THAT WE
SAW THE TRANSMISSION.
OOSK TOF COURSE THE VIRUS HAS BN
MUTATING THROUGHOUT THESE LAST
FOUR YEARS.
WUHAN, DELTA, MU, WE ALWAYS SAW
100% TRANSMISSION.
WITH BA1 ON TO BA5, THERE WAS
NEVER ABLE TO SEE AIRBORNE
TRANSMISSION WITH THESE
VARIANTS.
IT WAS INTERESTING THAT WHEN A
NATURALLY OCCURRING RECOMBINANT
OF DELTA THAT POSSESSED THE
BA1 SPIKE EMERGED, WHICH WAS
CALLED XD, AGAIN WE SAW 100%
TRANSMISSION.
SO A SPIKE FROM BA1 THAT IS NOT
TRANSMISSIBLE PUT INTO THE DELTA
BACKGROUND DID RESULT IN A
TRANSMISSIBLE VIRUS THEN.
AS THE VIRUS KEPT EVOLVING, WE
GOT MORE MUTATIONS IN THE SPIKE
AND OTHER OPEN READING FRAMES,
WE DID SEE TRANSMISSION CREEP UP
FROM XBB.1.16 THAT WE SAW 50%
TRANSMISSION, AND THEN 50 AND
60% TRANSMISSION WITH
XBB.1.5 AND EG.5.1.
WE STARTED DOING VACCINATION
STUDIES TO SEE IF VACCINES WOULD
CHANGE TRANSMISSION KINETICS,
AND ALSO A NEW VARIANT CAME OUT,
JN.1, WHICH HAD ANOTHER 30 TO 40
AMINO ACID DIFFERENCES IN ITS
GENOME.
AND THAT VIRUS NOW WAS AGAIN
DEAD IN OUR AIRBORNE
TRANSMISSION STUDIES.
NOW WE WORKED TOGETHER TO LOOK
AT SOME CORONAVIRUS OF ENDEMIC
POTENTIAL.
THIS INCLUDED THE PENG LIN
CORONAVIRUS THAT WE SAW 17%
TRANSMISSION AND WIVSH014 AND
BANAL WHICH DON'T SHOW ANY
TRANSMISSION IN THE HAMSTER
MODEL.
WHICH OF COURSE IS GOOD SINCE
THESE DON'T TRANSMIT IN HUMANS
AND THERE NEEDS TO BE SOME KIND
OF BARRIER THAT NEEDS TO
OVERCOME FOR THESE VIRUSES TO
TRANSMIT.
SO WHAT ARE THE ALTERNATIVES
THAT WE HAVE?
SINCE IT'S PROBABLY NOT GREAT TO
PASSAGE ANY OF THESE VIRUSES TO
GET THEM TO TRANSMIT, LET MOTHER
NATURE DO THAT, WE'RE LOOKING AT
ALTERNATIVE ANIMAL MODELS.
WE THOUGHT FERRETS WOULD BE
GOOD.
SO WE INFECTED FEMALES 10 TO THE
6 PFU DELTA OR EG.5.1.
THE DELTA VARIANT GREW HIGH
TITERS IN THE NASAL TURBINATE
AND IN THE SOFT PA PAL L PALAT
LUNGS.
IF WE WANT TO STUDY GN1 OR OTHER
VARIANTS THAT DON'T TRANSMIT IN
HAMSTERS, THE FERRET MODEL DOES
NOT SEEM TO BE USEFUL HERE.
WE HAVE STARTED LOOKING AT DWARF
HAMSTERS.
THIS CAME AFTER A GERMAN
PUBLICATION THAT THESE DWARF
HAMSTERS ARE EXTREMELY
SUSCEPTIBLE TO COV-2.
THEY SHAKE, THEY LOSE WEIGHT,
THEY WILL DIE.
SO WE DECIDED TO INFECT THESE
DWARF HAMSTERS WITH A BUNCH OF
DIFFERENT OMICRON VARIANTS, AND
YOU CAN SEE ON THE LEFT-HAND
SIDE THE NASAL TURBINATE TITERS,
THIS VIRUS REPLICATES GREAT.
THIS IS DIFFERENT FROM WHAT WE
SAW WITH BA1 AND SYRIAN HAMENTS
TERSE, YOU SO YOU NO OR VERY
POOR NON-CONSISTENT REPLICATION
OF BSM A1 AND THE SYRIAN
HAMSTERS WITH LOW TITERS AND THE
NASAL TURBINATE TO ALMOST NO
DETECTABLE VIRUS TITERS IN THE
LUNG.
THIS EXTENDS FROM BA1 TO BA5 TO
XBB.1.16.
SO WE SET UP A PILOT STUDY JUST
LOOKING AT DIRECT TRANSMISSION
OF DELTA, DELTA WILL TRANSMIT
ALL THE TIME.
SO WE INFECTED THESE DWARFS WITH
10 TO THE THIRD PFU, WE DIDN'T
WANT TO KILL THEM.
WE WAITED 48 HOURS FOR
TRANSMISSION TO HOPEFULLY
HAPPEN, AND YOU CAN SEE FROM THE
INFECTED ANIMALS WE HAD GREAT
VIRUS REPLICATION IN THE NASAL
TURBINATES AND THE LUNGS, BUT WE
WERE NEVER ABLE TO DETECT DIRECT
CONTACT TRANSMISSION IN THE
DWARFS.
WE HAVE NOT GONE OUT TO 21, 28
DAYS TO LOOK AT SERO CONVERSION,
JUST BASING THIS ON NO
INFECTIOUS VIRUS ON DAY 3.
ANOTHER ALTERNATIVE IS WHAT WE
CALL THE BUBBLE BOY HAMSTERS.
THESE HAMSTERS LACK IL2 RECEPTOR
GAMMA.
IL2 IS IMPORTANT FOR YOU TO
DEVELOP T CELLS, B CELLS AND --
CELLS.
AGAIN THESE ARE FROM ZONDA'S
LAB.
WE CAN TAKE THESE ANIMALS OUT TO
DAY 100 WHEN WE INFECT THEM WITH
10 TO THE FOURTH PFU SO IT'S A
PERSISTENT MODEL OF INFECTION.
WE ONLY SAW TETHER IN ONE ANIMAL
AROUND DAY 97.
DESPITE THE INCREASE IN WEIGHT
GAIN, YOU CAN SEE THAT FROM ORAL
SWABS, WE HAVE DECENT VIRUS
TITERS FOR THAT STUDY.
WE HAVE LOOKED AT VIRUS TITERS
ON DAY 100 AND YOU CAN SEE THAT
WE HAVE VERY NICE REPLICATION IN
THE LUPG AND NASAL TURBINATE,
AND NOW WHAT WE'RE WORKING ON
WITH ZONDA RIGHT NOW IS LOOKING
WOULD WE BE ABLE TO USE THESE LR
TRANSMISSION STUDIES.
MAYBE NOT TO LOOK AT EVERY
VARIANT THAT'S OUT THERE, BUT
LOOK AT TRANSMISSION DYNAMICS
AND MAYBE MORE IMPORTANTLY LOOK
AT VIRUS POPULATIONS.
WHAT'S THE VIRUS POPULATION IN
THE INFECTED ANIMAL VERSUS
WHAT'S THE VIRUS POPULATION IN
THE TRANSMITTED ANIMAL, IF THIS
VIRUS WILL TRANSMIT.
ALSO I DON'T THINK WE SHOULD
TURN OUR BACKS ON MERS.
THERE'S A PRE-PRINT OUT ON USING
THE TRANSGENIC HUMAN
DPP4 TRANSGENIC MICE.
THE MICE ARE POOR OR VERY
INEFFICIENT FOR TRANSMISSION.
THAT'S BOTH DIRECT AND AIRBORNE
TRANSMISSION.
WE WORKED WITH ZONDA TO DO SOME
PRELIMINARY STUDIES WITH
TRANSGENIC HAMSTERS NOW
INFECTING THEM WITH MERS AND
LOOKING AT DAY THREE, WE CAN SEE
VIRUS REPLICATION LOWER IN THE
LUPG LURNG AND HIGHER IN THE NAL
TURBINATES AND IN THE NEXT
COUPLE MONTHS WE'LL LOOB AT
TRANSMISSION WITH THESE ANIMALS.
SO IN SUMMARY, THE GUINEA PIG
WOULD BE MY VOTE FOR THE
TRANSMISSION MODEL FOR INFLUENZA
VIRUSES, IF WE'RE TALKING ABOUT
A RODENT MODEL, THERE'S BEEN
GREAT STUDIES TO LOOK AT VIRUS
DETERMINANTS OF INFECTION,
LOOKING AT AMINO ACIDS AND
PROTEINS THAT ARE ESSENTIAL FOR
TRANSMISSION.
WITH SARS-COV-2, HAMSTERS ARE
SUITABLE BUT WE'RE LACKING THE
AIRBORNE TRANSMISSION ROUTES IN
SOME OF THESE.
AND WE SHOULD START LEARNING
WHAT THE INFLUENZA GROUP HAS
DONE FOR TRANSMISSION STUDIES AS
WE START BRINGING THIS INTO
SARS, SARS-COV-2 TRANSMISSION
STUDIES, MAINLY LOOKING AT WHAT
ARE THE VIRAL DETERMINANTS THAT
WE'RE SEEING WITH THIS
DIFFERENCE BETWEEN PREOMICRON
AND POST OMICRON VARIANT AND OF
COURSE DOING LOSS OF FUNCTION
STUDIES.
FOR THAT, THNGS FOR YOUR TIME.
THANKS FOR YOUR TIME.
>> TIME FOR QUESTIONS.
>> THIS IS TONY WONG FROM FDA.
VERY BEAUTIFUL WORK.
I'M JUST WONDERING FOR THOSE
SOME OF THE OMICRONS THAT DO NOT
TRANSMIT VERY WELL IN HAMSTER
MODEL, DID YOU TRY TO CORRELATE
THE VIRAL LOAD IN THE NASAL
CAVITY VERSUS TRANSMISSIBILITY?
>> THIS IS A STUDY WE HAVE
PLANNED HERE, NOT ONLY
CORRELATION BETWEEN TITERS BUT
WE ALSO WANT TO SEE WHAT'S BEING
EXPELLED BY CATCHING VIRUSES ON
CELL CULTURE PLATES AND THEN
ALSO I THINK IT'S WORTH DOING
JUST DIRECT TRANSMISSION WITH
ALL OF THESE VARIANTS, AS MANY
AS WE CAN, TO SEE IF THERE'S ANY
DIFFERENCES THERE.
BUT YEAH, THAT'S A STUDY THAT
WE'RE LOOKING AT
>> DO YOU THINK THE REASON THAT
SOME OF THE OMICRON SUBVARIANTS
DO NOT TRANSMIT VERY WELL IN
HAMSTER MODEL IS BECAUSE THEY
JUST SIMPLY DON'T REPLICATE VERY
WELL IN NASAL CAVITY?
>> I WOULD SAY THE OMICRONS
REPLICATE BETTER IN THE NASAL
TURBINATES --
>> EVEN IN HAMSTERS?
>> YEAH.
SO IT'S KIND OF ACTUALLY
DIFFERENT, SO WITH THE DELTA YOU
SEE HIGHER IN THE LUNG AND LOWER
IN THE NASAL TURBINATES WITH
OMICRONS AND AFTER, IT'S THE
OTHER WAY AROUND.
>> WE KIND OF -- I MEAN, THE
LITERATURE, MY RECOLLECTION, IS
SOME LABS SHOW THAT OMICRON
REPLICATES WELL IN NASAL
TURBINATES, BUT SOME LABS DON'T.
LIKE IN OUR OWN GROUP, OMICRON
SUBVARIANTS ALWAYS REPLICATE
WORSE THAN THE PREOMICRON EVEN
IN SYRIAN HAMSTERS.
>> MALES OR FEMALES?
>> ACTUALLY BOTH.
>> INTERESTING.
COULD BE VENDOR-SPECIFIC TOO.
>> SURE.
THANKS.
>> STANLEY.
>> SO ALONG THE SAME LINES, I
THOUGHT THE EXPERIMENT YOU
BRIEFLY MENTIONED BETWEEN THE XD
AND BA1 AND DELTA VARIANTS WAS
VERY INTERESTING BECAUSE YOU SAW
THAT TRANSMISSION BASICALLY
WASN'T DEPENDENT ON THE SPIKE
PROTEIN.
SO ARE YOU GOING TO BE
INVESTIGATING THOSE MORE?
>> WE'RE KIND OF FOLLOWING MATT
FRIEDMAN'S LEAD ON LOOKING AT
SOCK KNOCKOUTS, # OR 6.
MAYBE THINKING ABOUT AMINO ACID
CHANGES IN THE ORFs --
>> WONDERING IF IT'S THE ORFs
OR MSPs OR --
>> YEAH.
>> SO IN YOUR MODEL TO PUP
TRANSMISSION MODEL OF HN1 YOU
SAID SOME OF THEM WERE
SUCCESSFUL.
WHEN YOU LOOK AT THE LITTERS, IS
IT REALLY LITTER-SPECIFIC, LIKE
MOMS THAT TRANSMIT BETTER THAN
OTHER MOMS?
>> NO, THERE WAS NO CORRELATION.
SO WITH THE NUMBERS THAT WE DID,
YOU WOULD HAVE A LITTER OF FIVE
PUPS, ALL OF THEM WOULD BE
INFECTED.
SOMETIMES YOU'D HAVE ONE OR TWO.
IF YOU TRY TO CONSIDER LATE THAT
WITH WAS THE MAMMARY GLAND
POSITIVE FOR VIRUS, WE HAD SOME
MOMS THAT DID HAVE POSITIVE
MAMMARY GRANDZ BU GLANDS BUT NOT
WERE AFFECTED.
SO IT SEEMS TO BE A TIMING
THING, HOW WELL THE VIRUS CAN GO
FROM THE LUNGS TO THE REST OF
THE BODY.
NINE DATES IS AS FAR AS WE CAN
TAKE IT OUT BECAUSE OF THE
LETHALITY AT 100 PFI OF THAT
VIRUS.
>> THANK YOU.
I'D LIKE TO INVITE MILA ORTIGOZA
FROM NYU LANGONE HEALTH, SHE
WILL BE TALKING ABOUT THE INFANT
MOUSE MODEL TO STUDY INFLUENZA
VIRUS AND SARS-COV-2
TRANSMISSION.
>> I WANT TO THANK THE
ORGANIZERS FOR INVITING ME.
I'M REALLY EXCITED TO TALK ABOUT
THE MOUSE MODEL FOR
TRANSMISSION.
I THINK IN THE PAST 36 HOURS,
EVERY SLIDE I'VE SEEN, THERE IS
A LOT OF PEOPLE SAYING MICE
DON'T TRANSMIT, SO I HOPE THAT I
CAN CONVINCE YOU OTHERWISE.
AND WE'LL HAVE A DIALOGUE THAT
WE NEED DIVERSITY OF MODELS,
DEPENDING ON THE QUESTIONS WE
ASK, WE CAN LEARN A LOT ABOUT
TRANSMISSION.
SO IN OUR LAB, WE THINK OF
TRANSMISSION IN THREE DIFFERENT
STAGES.
STAGE I IS THE UPPER RESPIRATORY
TRACT REPLICATION OF THE VIRUS
AND THAT HOST IMMUNE RESPONSE TO
INFECTION.
STAGE II IS SHEDDING OF THAT
VIRUS INTO THE ENVIRONMENT.
AND STAGE III IS VIRUS
ACQUISITION OF A NAIVE HOST.
I DO ALSO WANT TO BRING UP STAGE
IV THAT'S OFTEN STUDIED IN
INFLUENZA VIRUSES, IT'S THE
STAGE OF PATHOGENESIS WHERE
VIRUS IN THE UPPER RESPIRATORY
TRACT TRANSITIONS TO THE LOWER
RESPIRATORY TRACT, CAUSING
INVASIVE DISEASE, IN THIS CASE,
PNEUMONIA.
THIS IS NOT THE FOCUS OF OUR
STUDY.
ALTHOUGH VERY IMPORTANT FOR
VACCINE WORK, WE KNOW THAT
VACCINES PROTECT HOSTS FROM
DISEASE, STAGE IV, BUT WE ALSO
KNOW THAT WE CAN DO BETTER
BECAUSE CURRENT VACCINES DO NOT
PROTECT AGAINST TRANSMISSION
BECAUSE THEY DON'T INDUCE A
POTENT MUCOSAL IMMUNITY IN THE
UPPER RESPIRATORY TRACT.
SO WE DO FOCUS ON STAGE I
THROUGH STAGE III, FOCUSED ON
THE ANATOMY OF THE UPPER
RESPIRATORY TRACT IN OUR STUD
STUDIES.
THE TWO FUNDAMENTAL QUESTIONS
THAT WE REALLY WANT TO ANSWER IS
WHAT DRIVES HOST INFECTIOUSNESS,
AND WHAT DRIVES HOST
SUSCEPTIBILITY TO VIRUS
INFECTION AFTER AN EXPOSURE, AND
THIS REQUIRES REALLY AN IN DEPTH
ANALYSIS OF STAGE I, II AND II
BOTH FROM THE VIRUS PERSPECTIVE
AS WELL AS FROM THE HOST
PERSPECTIVE.
WITH THAT, WE NEED TO CHOOSE AN
ADEQUATE ANIMAL MODEL THAT NOT
ONLY ALLOWS US TO STUDY VIRAL
BIOLOGY BUT ALSO HOST BIOLOGY.
I'M A PHYSICIAN.
THERE ISN'T ANYTHING THAT I WANT
TO DO WITH HUMANS.
HUMANS ARE THE PERFECT MODELS
FOR STUDYING TRANSMISSION, BUT
THEY'RE ALSO VERY CHALLENGING TO
STUDY THE HOST REQUIREMENTS FOR
TRANSMISSION.
SO ARE FERRETS AND GUINEA PIGS.
THEY'RE FANTASTIC MODELS.
YOU CAN USE CLINICAL ISOLATES
AND ASK QUESTIONS ABOUT VIRAL
TRANSMISSIBILITY VERY EASILY,
BUT THEY DO HAVE DRAW BACKS.
THEY'RE COMPLEX, THEY'RE
EXPENSIVE, THEY REQUIRE SPECIAL
HOUSING.
OFTEN TRANSMISSION STUDIES ARE
LARGELY UNDERPOWERED, AND MOST
IMPORTANTLY, RIGHT NOW I THINK
THE TOPIC OF THIS SESSION,
LIMITED REAGENTS, WE CAN DO MUCH
BETTER ABOUT REAGENTS IN THESE
MODELS.
ALL OF THESE DRAWBACKS IN THESE
COMPLEX MODELS ARE ACTUALLY
ADVANTAGES IN THE MOUSE MODEL,
BUT AS WE KNOW HISTORICALLY FOR
THE PAST 70 YEARS, ADULT MICE
HAVE DEMONSTRATED INCONSISTENT
TRANSMISSION AND WE WERE VERY
INTERESTED IN UNDERSTANDING WHY
ADULT MICE DON'T TRANSMIT
BECAUSE WE REALLY WANTED TO
STUDY HOST BIOLOGY AND HOST
DETERMINANTS OF TRANSMISSION.
SO FOR THE PAST SIX YEARS, WE'VE
BEEN OPTIMIZING AND VALIDATING
THE MOUSE MODEL, WE IDENTIFIED
INFANT MICE TO BE VERY RELIABLE,
VERY EFFICIENT, VERY ROBUST
MODEL TO STUDY FLU TRANSMISSION.
BUT NO MODEL IS PERFECT, AND THE
INFANT MOUSE MODEL IS NOT
UNIQUE.
ONE OF THE MAJOR DRAWBACKS IS
THAT THEY COME AS A UNIT, SO THE
MOTHERS IN THE CAGE, YOU CANNOT
SEPARATE THE PUPS FROM EACH
OTHER, YOU CANNOT SEPARATE THE
MOM FROM THE PUPS SO THAT LIMITS
THE KIND OF QUESTIONS WE CAN ASK
WITH THIS MODEL SUCH AS THE
MODES OF TRANSMISSION, AND LIKE
SEEMA MENTIONED PREVIOUSLY, WHEN
IT COMES TO LIKE A HOUSEHOLD
KIND OF MODEL TRANSMISSION, ALL
TRANSMISSION MODES ARE VALID.
SO THIS IS BASICALLY WE VIEW IT
AS A HOUSEHOLD KIND OF
TRANSMISSION, YOU KNOW, ASSAY.
WE CANNOT REALLY ASK QUESTIONS
ABOUT ROLE OF IMMUNITY BECAUSE
OBVIOUSLY WE CANNOT SEPARATE THE
PUPS FROM THE MOTHER.
THEY'RE CONSTANTLY FEEDING SO WE
CANNOT ISOLATE THE EFFECTS OF
THE IMMUNITY FROM THE MOTHER
BEING TRANSMITTED THROUGH THE
MILK.
AND MOST IMPORTANTLY, MICE ARE
STRAIN-DEPENDENT, SO THERE ARE A
LOT OF -- ISOLATES THAT JUST
DON'T AFFECT MICE.
OTHER THINGS ABOUT THIS MODEL I
WANT TO MENTION AND I GET THESE
QUESTIONS A LOT SO I WANT TO
JUST BRING IT FORTH, DOES THE
MOTHER GET INFECTED WHEN WE
INFECT THE PUPS, AND THE ANSWER
IS YES.
BUT THE PUPS TRANSMIT TO THE
MOTHER VERY INEFFICIENTLY, LESS
THAN 10% OF EXPOSURES.
OTHER QUESTIONS WE GET ASKED IS,
WELL, IS THIS A FOMITE KIND OF
MODE OF TRANSMISSION IN THIS
MODEL.
OBVIOUSLY THE INFANTS ARE
SUCKLING SAME NIPPLES AND THE
BEDDING IS CONSTANTLY THERE, BUT
WHEN WE DO CROSS DB EXPERIMENTS
OR CHANGE THE CAGES,
TRANSMISSION DOES OCCUR, BUT
AGAIN, VERY INEFFICIENT.
SO IN THIS MODEL, TRANSMISSION
IS DRIVEN FROM PUP TO PUP VIA
UPPER RESPIRATORY TRACT
SECRETIONS.
AND WE DEMONSTRATE THIS HERE IN
THIS GRAPH OF INFANT SHEDDING,
WHERE WE HAVE INFECTIOUS TITERS
OVER TIME, AND EACH DOT IS A
SHEDDING EVENT FROM A SINGLE
PUP, AND IN BLACK WE SEE TH THE
INDEX SHEDDING CORRESPONDS WITH
THE ACQUISITION OF SHEDDING
CONTACTS IN RED, AND YOU SEE THE
TIMING OF INDEX SHEDDING
CORRESPONDS WITH TRANSMISSION BY
THE CONTACTS.
HOW WE COLLECT SHEDDING IN THIS
ASSAY IS SIMPLY BY DIPPING THE
NOSTRILS AND ONLY THE NOSTRILS
OF THE PUPS IN VIRAL MEDIA, AND
WE CAN EASILY TRACK THE KINETICS
OF TRANSMISSION OVER TIME
WITHOUT THE NEED OF SACRIFICING
THE ANIMAL OR EUTHANIZING OR
USING ANESTHESIA.
BUT ALSO WE CAN SELECT DIFFERENT
TIME POINTS AND ASSESS FOR
REPLICATION IN THE UPPER
RESPIRATORY TRACT, AND ASSESS
FOR TRANSMISSION, AND THIS IS
HOW WE DEFINE TRANSMISSION.
SO FOR EXAMPLE, AT DAY # 4, WE
KNOW WE ACHIEVE 100%
TRANSMISSION ALREADY IN THIS
MODEL.
SO I DON'T WANT TO SPEND TOO
MUCH TIME TALKING ABOUT THE
MODEL ITSELF, BUT MORE ABOUT
WHAT WE'VE LEARNED FROM DOING
THESE ASSAYS, WHAT ARE THE
FACTORS THAT ARE IMPORTANT IN
INFLUENZA TRANSMISSION IN MICE.
SO WE'RE GOING TO TALK ABOUT
HOST AGE AND SEX, EXPOSURE
PERIOD, THE UPPER RESPIRATORY
TRACT ENVIRONMENT, WHICH
INCLUDES NOT ONLY SIALIC ACIDS
BUT THE MICROBIOTA, THE
INFLAMMATORY STATE AND OBVIOUSLY
WE'LL FINISH UP WITH OUR
FAVORITE TOPIC, WHICH IS VIRUS
STRAIN.
SO ONE OF THE FIRST THINGS WE
LEARNED WAS THAT TRANSMISSION
EFFICIENCY IS DEPENDENT ON HOST
AGE.
HERE AGAIN WILL SHOW VIRUS --
EACH PANEL REPRESENTS EACH STAGE
OF TRANSMISSION, SO STAGE ONE
THE UPPER RESPIRATORY TRACT
REPLICATION, STAGE II, SHEDDING,
AND STAGE III, CONTACT
ACQUISITION.
AS WE CAN SEE COMPARED IN
INFANTS AND ADULTS, DESPITE
EQUAL REPLICATION IN THE UPPER
RESPIRATORY TRACT, WE SEE VERY
DIFFERENT KINETICS OF SHEDDING,
WHEREBY INFANTS SHED HIGHER AND
MORE EFFICIENTLY THAN ADULTS AND
CORE SPORINGLY, THEY TRANSMIT
BETTER COMPARED TO ADULTS.
AND ALSO INFANTS ARE MORE
SUSCEPTIBLE TO ADULTS IN
TRANSMISSION, SO THERE IS AN
ADVANTAGE TO STAGE TWO, WHERE
THEY'RE MORE INFECTIOUS, AND
THERE'S ALSO -- IN STAGE III
WHERE THEY'RE MORE SUSCEPTIBLE.
WHEN WI WE LOOK AT CORRELATION F
AGE COMPARED TO TRANSMISSION
EFFICIENCY, WE SEE THAT THE
YOUNGEST TRANSMIT MORE
EFFICIENTLY AND THE OLDEST
TRANSMIT VERY INEFFICIENT, SO
THERE'S A VERY BEAUTIFUL
CORRELATION WITH AGE.
OTHER THINGS THAT WE'RE STARTING
TO LOOK AT, YES, THE FANT MOUSE
MODEL IS BEAUTIFUL, BUT THERE IS
AN ADVANTAGE AT LOOKING AT ADULT
MICE WHERE THEY'RE NOT
SUSCEPTIBLE TO INFLUENZA VIRUS
TRANSMISSION, BUT WE DID LEARN
THAT SEX IS AN IMPORTANT
BIOLOGICAL VARIABLE THAT WE NEED
TO TAKE INTO ACCOUNT.
AND WHEN WE DID THAT IN HERE,
WE'RE USING ADULT ANIMALS THAT
MALES ARE MORE SUSCEPTIBLE TO
ACQUISITION OF INFLUENZA VIRUS
COMPARED TO FEMALES, SO NOW WE
NOT ONLY HAVE AN INFANT MOUSE
MODEL OF TRANSMISSION, WE ALSO
HAVE AN ADULT MODEL OF
TRANSMISSION IF WE USE THE RIGHT
HOST.
BUT ALSO NOT ONLY USING THE
RIGHT HOST, WE CAN USE RIGHT
CONDITIONS.
SO THINGS THAT WE'VE LEARNED
FROM HUMANS, WE'RE NOW TRYING TO
APPLY THIS IN OUR MODEL.
AGAIN, ADULT HERE SHEDDING, SO
INDEX SHEDDING IN BLACK CORE
SPONTS TCORRESPONDS TO ACQUISITF
THE VIRUS IN THE CONTACTS IN
RED.
AS YOU CAN SEE IF YOU REMEMBER
FROM THE INFANT GRAPH, THE ADULT
ACQUISITION IS A BIT DELAYED AND
IF WE SELECT DIFFERENT TIME
POINTS AND ASSESS FOR VARIABLE
REPLICATION IN THE UPPER
RESPIRATORY TRACT, WE SEE THAT
BY DAY FOUR, INEFFICIENT
TRANSMISSION BUT LET THEM HANG
OUT A LITTLE BIT TO FORM THAT
MEANINGFUL EXPOSURE.
WE SEE THAT TRANSMISSION IS
NEARLY 100% IN THE ADULT ANIMALS
BECAUSE WE ARE EXPOSING THEM FOR
A LONGER PERIOD OF TIME.
SO KIND OF LIKE CORRELATES WHEN
YOU'RE WALKING BY SOMEBODY IN
THE HALLWAY, THAT EXPOSURE IS
NOT VERY MEANINGFUL COMPARED TO
WHERE YOU LIVE WITH THAT PERSON
OR YOU'RE SITTING IN A CLOSED
OFFICE.
SO THE FACT THAT MICE DON'T
TRANSMIT, I THINK THAT NARRATIVE
WE NEED TO CHANGE THAT.
IT'S MORE ABOUT THE RIGHT HOST,
THE RIGHT VIRUS AND THE RIGHT
CONDITIONS.
OTHER THINGS I WANT TO MENTION
IS TRANSMISSION EFFICIENCY
DEPENDENT ON THE UPPER
RESPIRATORY TRACT ENVIRONMENT.
TWO POINTS I WANT TO MAKE HERE.
SO FIRST THAT THE INFLUENZA
RECEPTORS IS INDEED PRESENT IN
THE UPPER RESPIRATORY TRACT FOR
MICE AND WE LOOKED USING
IMMUNOHISTOCHEMISTRY OF THE
UPPER RESPIRATORY TRACT OF
INFANTS AND ADULTS, AND WE SEE
HERE BEAUTIFULLY TERMINAL ALPHA
26 AND ALPHA 2-3 LING -- IN THE
OLFACTORY AND THE RESPIRATORY
EPITHELIUM IN -- IN THE LUNGS AS
SHOWN BY PETER, HOWEVER THE ONE
DIFFERENCE BETWEEN MICE IS THEY
DON'T EXPRESS AL VA 2-6 IN THE
TRACHEA BUT THEY DO EXPRESS
ALPHA 2-3.
SO IF YOU'RE THINKING ABOUT
STUDIES WHERE YOU'RE LOOKING AT
TRANSITION OF VIRUS FROM THE
UPPER TO THE LOWER, YOU HAVE TO
TAKE INTO ACCOUNT VIRUSES THAT
REQUIRE OR PREFERENTIALLY BIND
TO 2-6 LENGTH SIALIC ACIDS
BECAUSE THEY -- THEY'RE NOT
EXPRESSED IN THE TRACHEA.
BUT LOOKING AT THE UPPER
RESPIRATORY TRACT THAT SEEMS TO
BE LESS OF A CONCERN.
THE SECOND THING I WANT TO
MENTION IS THESE RECEPTORS ARE
NOT ONLY EXPRESSED, THEY'RE NOT
FUNCTIONAL.
DOING A SIMPLE TRANSMISSION
ASSAY AND INOCULATED OR
INTRANASALLY ADMINISTERING
BROADLY ACTIVE NEURO -- WE USE
DAS181 WHICH EXPRESSES -- JUST
TO PROVE A POINT THAT IF WE
TREAT THESE ANIMALS WITH DISIGH
AL LATE IN VIVO WE SEE A DEFECT
IN SHEDDING AND ALSO A DEFECT IN
TRANSMISSION.
WHICH AGAIN PROVES A POINT THAT
THESE VIRUSES IN THE SYSTEM,
SIALIC ACIDS ARE REQUIRED TO
ESTABLISH A TRANSMISSION EVENT.
AND A SIDE NOTE, IF YOU WANT TO
TEST DIFFERENT THERAPEUTICS FOR
THEIR EFFECT ON TRANSMISSION,
THIS MODEL IS ALSO VERY
EFFICIENT FOR THAT.
WE CAN DO THE SAME THING USING A
COMPLETELY DIFFERENT ASSAY
ACHIEVING THE SAME RESULTS.
IT'S IMPORTANT TO ACKNOWLEDGE AS
MENTIONED PREVIOUSLY THAT
INFLUENZA VIRUS DOES NOT INFECT
A STERILE ENVIRONMENT.
SO WE WANTED TO ADD A LITTLE BIT
OF COMPLEXITY TO THIS MODEL BY
DOING COINFECTIONS.
AND WE KNOW FROM INFANTS THAT
ONE OF THE MAJOR COLONIZERS OF
THE UPPER RESPIRATORY TRACT IS
STREPTOCOCCUS PNEUMONIA, AND SO
WE ESTABLISH A COINFECTION MODEL
WHERE ALL OF THE LITTERS ARE
COLONIZED FIRST WITH STREPT
COCOST PNESTREPTOCOCCUS PNEUMONN
ASK HOW DOES COLONIZATION OF THE
BACTERIA AFFECT TRANSMISSIBILITY
OF INFLUENZA.
HERE WE'RE SPECIFICALLY
INTERESTED IN BACTERIAL
NORMALIZATION OF THOSE EFFECTS
AND WE COLONIZE ONE COHORT WITH
WILD TYPE BACTERIA WHICH
EXPRESSES -- AND A SECOND
BACTERIA THAT DOES NOT EXPRESS
UNDER THE SURFACE.
AS WE CAN SEE FROM THE RESULTS,
BOTH UPPER RESPIRATORY TRACT
REPLICATION AND SHEDDING ARE
CO-INFECT BID COLONIZATION OF
THE TWO STRAINS BUT THE DEFECT
IS HERE AT THE PART OF
ACQUISITION WHERE THE BACTERIA
THAT HAS -- EXPRESSED LIMITS THE
CONTACT ACQUISITION OF INFLUENZA
VIRUSES WHERE THAT IS NOT
PRESENT WHEN THEY'RE COLONIZED
WITH THE MUTANT BACTERIA.
AGAIN, PROVING THE POINT THAT
DESIALYLATING IN VIVO BLOCKS
TRANSMISSION BUT ALSO A BIGGER
PICTURE IS THAT YOUR
SUSCEPTIBILITY CAN CHANGE
DEPENDING ON WHAT YOU'RE
COLONIZED WITH.
SO WE'RE TRYING TO DO SIMILAR
EXPERIMENTS USING DIFFERENT
BACTERIA, PARTICULARLY STAPH
AUREUS AND SEEING IF WE SEE A
SIMILAR EFFECT.
LAST POINT I WANT TO MAKE IS
ABOUT VIRUS STRAIN.
SO I WAS ONLY ALLOTTED 15
MINUTES SO I'LL BE A COUPLE
MINUTES OVER.
VIRUS TRANSMISSION IN MICE IS
DEPENDENT ON VIRUS STRAIN.
THAT IS NOT NEW TO US, AND I
ONLY HAVE A FEW THAT I'VE TESTED
SO IF YOU HAVE YOUR FAVORITE
STRAIN AND YOU WANT TO TEST IT
IN THE SYSTEM, I'M HAPPY TO TAKE
IT AND TEST IT, BUT IN GENERAL,
H1N1 VIRUSES SHED LOWER THAN
H3N2 VIRUS AND CORRESPONDINGLY
THEY TRANSMIT TO A LOWER
EFFICIENCY COMPARED TO
H3N2 VIRUS WHICH IS INTERESTING
BUT IT ALSO GIVES US GOOD TOOLS
TO WORK WITH.
SO OUR FERRET VI FAVORITE VIRUSS
AN OPPORTUNITY TO STUDY BIOLOGY
OF THE HOST THROUGH VIROLOGY.
AND OBVIOUSLY THEY'RE ICEGENICS
OISOGENICSOF ONE ANOTHER SO WE E
DIFFERENT COMBINATIONS AND ASK
WHAT IS THE TRANSMISSION
EFFICIENCIES OF THESE DIFFERENT
CLONES.
SO WHEN WE TEST ALL OF THESE
VIRUSES, WE SEE THE UPPER
RESPIRATORY TRACT REPLICATION IS
SIMILAR ACROSS ALL OF THESE
STRAINGS.
STRAINS.
SHEDDING IS VERY DIFFERENT, SO
IF YOU HAVE VIRUS THAT CONTA
CONTAINS -- THAN H -- VIRUSES
CONTAINING H1s AND
CORRESPONDINGLY THEY TRANSMIT
MUCH EFFICIENTLY COMPARED TO
VIRUSES CONTAINING H1s.
CLEARLY THIS TYPE OF VIROLOGY,
YOU CAN DO IN GUINEA PIGS,
FERRETS, YOU CAN EVEN DO IN
HUMANS, BUT THIS WAS INTERESTING
TO US BECAUSE NOW WE HAVE TOOLS.
WE HAVE HIGHLY TRANSMISSIBLE
VIRUSES, WE HAVE LOW
TRANSMISSIBLE VIRUSES AND WE CAN
NOW APPLY THESE IN OUR SYSTEM
AND USE THE POWER OF THE MICE
UNDERSTAND HOST REQUIREMENTS FOR
TRANSMISSION.
SO THE ONE -- ONE OF THE
FUNDAMENTAL QUESTIONS WE WANTED
TO ASK IS WHAT MAKES A HOST
INFECTIOUS?
SO WHAT WE DID IS WE DID A
COMPREHENSIVE UPPER RESPIRATORY
PROFILING OF INDEX MICE AND FROM
THEIR SCREATIONS, WE DID SEVERAL
ANALYSES.
ONE OF THE THINGS WE LEARNED IS
THAT HIGHLY TRANSMISSIBLE
VIRUSES INDUCE A HIGHER
INFLAMMATORY CYTOKINE RESPONSE,
THEY INDUCE HIGHER JEAN
EXPRESSION, INFLAMMATORY GENE
EXPRESSION.
THEY RECRUIT ABUNDANCE OF
MACROPHAGE AND NEUTROPHILLING TO
THE UPPER RESPIRATORY TRACT AND
THEY PRODUCE MORE MUCUS.
SO IN FOUR DIFFERENT ASSAYS, WE
SEE A SIMILAR TREND.
THE SIMILAR TREND IS THAT
INFLUENZA TRANSMISSION IS MORE
EFFICIENT WHEN THE INDEX HOST
HAS A STRONG INFLAMMATORY
RESPONSE.
SO IF YOU ARE PUZZLED ABOUT WHAT
INFLAMMATION MEANS, SO IN
HUMANS, WHEN YOU'RE INFECTED
WITH A VIRUS, YOU CREATE A VERY
ROW WROBUST INFLAMMATORY RESPONE
AGAINST THAT.
YOU BECOME SYMPTOMATIC AND WE
KNOW FOR A FACT THAT WE ARE
HIGHLY INFECTIOUS AND MORE
TRANSMISSIBLE WHEN WE ARE
SYMPTOMATIC.
SO THIS IS HOW WE MEASURE
SYMPTOMS IN THE MICE, IT'S HARD
TO ASK THEM HOW THEY FEEL.
AN NOW WEAR TRYING T WE'RE TRYIA
LITTLE DEEPER WITH KNOCKOUT
MODELS AND THERAPEUTICS TO SEE
WHICH HOST PATHWAYS ARE
IMPORTANT FOR THE
TRANSMISSIBILITY OF FLIE.
FLU.
SO WITH THAT, I JUST WANT TO
SUMMARIZE THAT I HOPE I
CONVINCED YOU THAT MICE ARE AN
ALTERNATIVE MODEL TO STUDY HOST
BIOLOGY AND TRANSMISSION,
PARTICULARLY FACTORS THAT ARE
IMPORTANT FOR INFECTIOUSNESS AND
SUSCEPTIBILITY.
THERE ARE BIOLOGICAL FACTORS WE
NEED TO TAKE INTO ACCOUNT,
OBVIOUSLY AGE, SEX AND THE UPPER
RESPIRATORY TRACK ENVIRONMENT
AND VIRUS STRAINS MATTER.
AND IT AFFECTS HOW WE RESPOND TO
THE INFECTION AND HOW WE
TRANSMIT THE VIRUS.
SO THANK YOU FOR YOUR ATTENTION.
I WOULD LIKE TO THANK MY LAB.
WE'RE SMALL BUT WE'RE FEISTY,
AND OUR COLLABORATORS AND OUR
FUNDING.
>> OKAY.
WE'VE GONE A BIT OVER.
DOES ANYONE HAVE A BURNING
QUESTION?
BECAUSE I CANNOT LET A SPEAKER
GO WITHOUT --
>> SO WHAT IS REALLY THE
DIFFERENCE BETWEEN IN YOUR MOUSE
MODEL APR8 AND X31, SINCE
X31 HAS THE APR8 INTERNAL GENES?
>> WELL, X31 IS PR8 WITH
EXCEPTION OF TWO SEGMENTS, THE
HEMAGGLUTININ AND NEUROMEN DAYS,
WHICH EYE RIFED FROM THE HONG
KONG 1968 PANDEMIC VIRUS.
SO THEY'RE ICEGENIC OF ONE
ANOTHER WITH THE EXCEPTION OF
TWO SEGMENTS.
>> SO WHERE IS THE TRANSMISSION
BLOCK REPLICATION FROM INTERNAL
GENES OR INNATE IMMUNE RESPONSE?
>> WE HAVE IT DOWN TO BEING THE
H3 COMPONENT.
WE HAD OUR MONEY ON THE NEUROMIN
DAYS COMPONENT, IT WAS
SURPRISING THAT H3 WAS A DRIVER
OF TRANSMISSION, SO WE'RE DOING
FOLLOW-UP STUDIES WITH WHAT
SECTION OF H3 DRIVES THAT, BUT
OBVIOUSLY THOSE ARE EXPERIMENTS
THAT YOU CAN DO BETTER WITH
OTHER MODEL SYSTEMS, BUT
OBVIOUSLY WE'RE DOING IT IN OUR
SYSTEM AS WELL.
BUT WE'RE USING THESE VIRUSES AS
TOOLS TO LEARN MORE ABOUT WHAT
OTHER SYSTEMS CAN ANSWER, WHICH
IS HOST BIOLOGY AND
INFLAMMATION.
>> ALL RIGHT.
WELL, I WOULD LOVE TO INVITE ALL
OF THE SPEAKERS, AND I ASSUME
WE'LL BE ABLE TO BRING JACCO UP
ON THE SCREEN.
I'M HOPING WE CAN --
>> THIS IS AV.
IF YOU CAN UNMUTE YOUR
MICROPHONE AND TURN ON YOUR
CAMERA.
>> I DID.
CAN PEOPLE HEAR ME?
>> WE CAN SEE AND HEAR YOU.
THANK YOU.
>> ALL RIGHT.
WE ARE READY TO GO.
SO LET'S BEGIN OUR DISCUSSION
ABOUT THE UTILITY OF THESE
ANIMAL MODELS.
IS THERE ANYBODY WHO WOULD LIKE
TO START OFF WITH A BROAD
QUESTION?
>> SO WHEN YOU LOOK AT THESE
TRANSMISSION MODELS, HOW MUCH OF
THIS DO YOU BENCHMARK AGAINST
THESE HOUSEHOLD TRANSMISSION
STUDIES THAT PEOPLE HAVE DONE
AND TRIED TO CORRELATE OR
ASSOCIATE SOME OF THOSE FINDINGS
WITH SYMPTOMATIC VERSUS
ASYMPTOMATIC TRANSMISSION?
>> I CAN START WITH THAT.
I THINK FACING EVERYTHING IN
WHAT WE UNDERSTAND FROM HUMANS
IS REALLY IMPORTANT.
I THINK HOUSEHOLD TRANSMISSION
STUDYS ARE FASCINATING, BUT THEY
SORT OF LACK A LOT OF CLARITY,
WHICH YOU SORT OF NEED IN ORDER
TO BENCHMARK, SPECIFICALLY WHAT
WAS THE INITIAL INFECTION
INITIATION, WHAT WAS THE
EXPOSURE DURATIONS.
THERE'S A LOT OF COMPLEXITY IN
THE SYSTEM.
I THINK THAT -- SO I THINK WHAT
I HAVE REALIZED ABOUT
ASYMPTOMATIC INFECTIONS AND
POTENTIAL TRANSMISSION
EFFICIENCIES, AND SORT OF WHAT
THAT MEANS, HAS REALLY COME FROM
LOOKING AT TRANSMISSION FITNESS
IN ANIMALS THAT ARE
ASYMPTOMATIC.
AND YOU CAN CORRELATE VIRAL
LOADS TO EXPULSION TO WHAT'S
BEING RELEASED INTO THE AIR TO
THEN A FORWARD TRANSMISSION
EVENT, SO I THINK THAT'S WHAT'S
IMPORTANT.
BUT I THINK THAT IT'S A GREAT
QUESTION TO SORT OF CONTINUE TO
THINK ABOUT WHAT HAPPENS IN
HUMANS AND HOW YOU RELATE THAT
BACK.
THAT'S JUST MY TAKE.
>> I JUST WANTED TO ADD TO THAT.
WITH RETROSPECTIVE STUDIES IN
HUMANS HAVE DEMONSTRATED THAT
ASYMPTOMATIC OR PRESYMPTOMATIC
IS THOSE WHO DON'T YET
EXPERIENCE SYMPTOMS BUT GO ON TO
DEVELOP SYMPTOMS LATER.
THEY DO SHED VIRUS.
BUT THEY DON'T SHED VIRUS TO
HIGHER AMOUNTS COMPARED TO
SYMPTOMATIC.
SO THE POTENTIAL FOR
TRANSMISSION IS THERE, BUT AT
THE TIME, LA LIKE SEEMA SAID, IS
PRETTY UNCONTROLLED, YOU DON'T
HAVE SYMPTOMS, YOU DON'T HAVE
THOSE METRICS.
BUT WHAT WE KNOW FROM HUMAN
STUDIES IS THAT ONCE SYMPTOMS
ARE PRESENT, WHEN THE PATIENT
EXPERIENCES SYMPTOMS, THEY'RE
VERY INFECTIOUS.
>> THANK YOU.
ANYBODY --
>> OVER HERE.
SO MY LAB, WE DO ANIMAL STUDIES
BUT WE WE ALSO DO STRUCTURAL
BIOLOGY AND CHEMISTRY.
SO AS FAR AS TRANSMISSION
STUDIES, IS THERE A ROLE FOR
NON-TRADITIONAL ANIMAL OR FLU
PEOPLE EITHER FROM STRUCTURAL
BIOLOGY, CHEMISTRY, ARTIFICIAL
INTELLIGENT MODELING?
WHERE ARE SOME OF THE GAPS THAT
PEOPLE WHO HAVE EXPERTISE IN
OTHER FIELDS CAN COME IN AND
HELP?
JUST A GENERAL QUESTION.
WHAT ARE SOME OF THE GAPS THAT
YOU GUYS NEED FROM OTHER PEOPLE
IN OTHER FIELDS THAT COULD HELP?
>> I THINK A.I. WILL BE HELPFUL.
HOW MANY MUTANT CORONAVIRUS DO
WE WANT TO MAKE TO START TEASING
APART WHY THE TRANSMISSION IS
NOT WORKING VERSUS WHY IT'S
WORKING?
AT LEAST IN THE HAMSTERS.
SO I DON'T KNOW IF WE HAVE
ENOUGH SEQUENCE INFORMATION WITH
THE DIFFERENT VARIANTS THAT WE
HAVE FOR COV-2, BUT ARTIFICIAL
INTELLIGENCE WOULD BE GREAT,
LARGE LANGUAGE LEARNING, TO
START DIVING DEEP INTO THESE.
IT DOESN'T HAVE TO BE
TRANSMISSION.
IT COULD BE ANY KIND OF
PHENOTIME THAT PHENOTYPE THAT WE
VIRUS IN A CERTAIN ANIMAL.
>> WOULD YOU LIKE TO ADD
ANYTHING ABOUT THE STRUCTURAL
BIOLOGY AND HOW YOU THINK THAT
MIGHT -- LIKE WHAT YOU WERE
THINKING.
>> WELL, PEOPLE WHO STUDY VIRAL
GLYCOPRETEENS OR STRUCTURAL
PROTEINS, PARTICULARLY WHEN
SEEMA WAS GIVING HER TALK RS I
WAS THINKING, OH, YOU SHOULD
SEND ME YOUR FLU VIRUSES SO WE
CAN LOOK AT THE MORPHOLOGY TO
SEE, YOU KNOW, WHAT YOU GAVE THE
FERRETS AND WHAT CAME OUT.
ARE THERE MORE FILAMENTOUS, HOW
IS MORPHOLOGY STRUCTURED, AND
WITH THE LOSS OF FUNCTION OR
GAIN OF FUNCTION WITH
CORONAVIRUS, DOES THE VIRUS HAVE
MORE HOST FUSION MORE POST FUSIR
PREFUSION SPIKES?
THAT'S JUST MY WORLD WHERE WE
LIVE?
>> I ALSO THINK THERE'S A ROLE
FOR MODELERS IN ALL OF THIS.
THERE IS SO MUCH DATA AND SO
MANY DIFFERENT ANIMAL MODELS AND
THE COMPLENGSITY AS SEEMA AND
OTHERS HAVE ALLUDED TO IS VERY
HIGH IN TERMS OF DURATION, WHEN
THE EXPOSURE, WITH OR WITHOUT
SYMPTOMS, PRE-EXISTING IMMUNITY.
I THINK HAVING MODELERS TAKE ALL
THE DATA, AI TAKE ALL THE DATA
THAT IS OUT THERE AND GENERATE
SOME TESTABLE HYPOTHESES, I
THINK WILL BE EXTREMELY HELPFUL
GOING FORWARD.
>> BRIAN, I SEE YOUR HAND UP.
>> YEAH, THAT'S GREAT.
THANK YOU, SABRA.
HI, EVERYBODY.
IT'S BEEN -- CAN EVERYONE HEAR
ME ALL RIGHT?
>> YES, VERY WELL.
>> OKAY.
IT'S HARD TO TELL ON THIS END.
THANK YOU, MICHELLE, FOR A
FANTASTIC WORKSHOP.
SO TRIED TO ASK A QUESTION
YESTERDAY WHEN THINKING ABOUT
MUCOSAL IMMUNITY, AND I THINK IT
GOES INTO THIS TRANSMISSION AS
WELL.
THINKING ABOUT SEEMA'S QUESTION
ABOUT PRE-EXISTING IMMUNITY IN
SOME OF THESE ANIMALS, HOW
THAT'S INFLUENCING TRANSMISSION.
SO I GUESS WHAT I WANT TO FORM
IS LIKE A HYPOTHESIS AND I WANT
TO SEE HOW THESE ANIMALS MODELS
ARE BEST CHALLENGING THIS, THAT
THE MUCOSAL IMMUNITY IN THE
UPPER RESPIRATORY TRACT, I THINK
WE KIND OF GOT TO YESTERDAY IN
THAT IT DOESN'T REALLY ALLOW
FOR, IT'S NOT EVOLVED FOR
STERILIZATION.
ITS GOAL IS NOT MEMORY.
THAT COULD JUST BE BECAUSE OF
THE EVOLUTION OF THE VIRUSES,
ITS ANTIGENIC EVOLUTION, BUT
BECAUSE OF CARRIAGE OF
COMMENSALS AND EVERYTHING ELSE
TOO, THAT WE'RE NOT HAVING A
LARGE IMPACT ON LIKE MEMORY.
SO DO WE REALLY FEEL LIKE WE CAN
TARGET TRANSMISSION IN A
VACCINATION SCHEME AND GETTING
AT THE HYPOTHESIS, WHERE IN OUR
ANIMAL MODELS DO WE REALLY HAVE
A CLEAR UNDERSTANDING OF WHERE
THAT PREVIOUS IMMUNITY IS GOING
TO REALLY HAVE AN IMPACT ON
TRANSMISSION AS OPPOSED TO
SETTING ASIDE THE ISSUE OF
SYSTEMIC MEMORY?
>> GREAT QUESTION.
ANYBODY WANT TO START TO ADDRESS
THAT?
>> SURE, I'LL TAKE A STAB.
ABSOLUTELY, I THINK WE CAN
ACHIEVE, FIRST OF ALL,
UNDERSTAND A LITTLE BIT MORE
ABOUT THE UPPER RESPIRATORY
TRACT.
A LOT OF THESE STUDIES
HISTORICALLY FROM RESPIRATORY
VIRUSES HAVE EMPHASIZED A LOT IN
THE LUNGS, INFLUENZA AS AN
EXAMPLE.
EVEN SARS-COV-2, WE HAVE SEVERAL
MOUSE MODELS, AND MOST OF THE
FIGURES AND PAPERS ARE ABOUT THE
LUNGS AND EVEN IN MIGRANTS, WHEN
I TALK ABOUT THE UPPER
RESPIRATORY TRACT, REVIEWERS ARE
YOU BE SED WITH THE LUNGS, WHICH
IS ABSOLUTELY IMPORTANT, IS A
MODEL OF PNEUMONIA BUT NOT A
MODEL OF TRANSMISSION.
SO WE KNOW ABSOLUTELY NOTHING
ABOUT THE UPPER RESPIRATORY
TRACT, SO I THINK THAT'S STEP
ONE.
LET'S FOCUS A LITTLE BIT MORE
ABOUT THE UPPER RESPIRATORY
TRACK SO WE CAN LEARN A LITTLE
MORE ABOUT THE STRUCTURE, THE
IMMUNITY, THE CELLS, HOW THEY
RESPOND TO VIRUSES AND THEN WE
CAN APPROACH THE HE QUESTION
ABOUT HOW TO DESIGN BETTER
THERAPEUTICS, BETTER VACCINES
THAT CAN INDUCE THAT LONG TERM
MUCOSAL IMMUNITY, ABSOLUTELY.
IT'S POSSIBLE.
WE JUST AS A GROUP NEED TO KIND
OF TAKE TO THE NEXT LEVEL WITH
INTENTION.
>> WOULD ANYONE ELSE LIKE TO ADD
ANYTHING?
>> I'D LIKE TO ADD IN SOMETHING.
I THINK SEVERAL GROUPS NOW HAVE
LOOKED AT MUCOSAL VERSUS
SYSTEMIC VACCINES AND ITS ROLE
ON PROTECTION AGAINST
TRANSMISSION AND EVEN ONWARDS
TRANSMISSION.
THERE'S A CLEAR PHENOTYPE THAT
MUCOSAL VACCINES AND PRIOR
INFECTION ARE VASTLY SUPERIOR IN
REDUCING UPPER AND LOWER AIRWAY
TITERS AS WELL AS PREVENTING
ONWARDS TRANSMISSION.
WHAT I THINK IS THE FUTURE HERE
IS UNDERSTANDING WHAT ARE THE
CORRELATES OF THAT PROTECTION?
IS IT T CELLS, IS IT SECRETORY
IGA, AND HOW DOES DRIFT PLAY IN
ALL OF THIS, BECAUSE WE
SHOULDN'T FORGET A LOT OF THE
REINFECTIONS AND ESPECIALLY FOR
FLU AND SARS-COV-2, A LOT OF THE
REINFECTION POTENTIAL ACTUALLY
COMES THROUGH EVASION OF
PRE-EXISTING IMMUNITY,
SPECIFICALLY NEUTRALIZING
ANTIBODIES.
SO YES, I AGREE THAT THE UPPER
HAIR WAY MAY NOT BE THERE TO
MAINTAIN STERILIZING IMMUNITY,
BUT IT'S STILL PRETTY GOOD AT
CONTROLLING THE VIRUS, AND EVEN
PREVENTING REINFECTION WITH
HOMOLOGOUS STRAIN.
SO THOSE ARE MY THOUGHTS ON THAT
TOPIC.
>> THANK YOU.
ANYONE ELSE?
>> I DON'T HAVE ANYTHING MORE TO
ADD ON MUCOSAL IMMUNITY, BRIAN,
BECAUSE IT'S REALLY HARD, BUT I
WANTED TO JUST ADD ON TO LIKE
WHAT I THINK IS MISSING, BECAUSE
REALLY I THINK IT'S SO
IMPORTANT.
YOU KNOW, WE'RE VIROLOGISTS AND
TALKING ABOUT ANIMAL MODELS AND
THERE'S A BIG INTEREST IN
IMMUNOLOGY, BUT WHAT WE ARE
LACKING AND WHAT WE REALLY NEED
TO DO IS BRING IN MORE
ENGINEERS, ESPECIALLY AEROSOL
SCIENTISTS AND PEOPLE WHO DO
DEVELOPMENT OF SPACES, TALKING
ABOUT CIVIL ENGINEERS.
WE DON'T HAVE AN APPRECIATION
FOR A LOT OF THESE THINGS FOR
DETECTION OF VIRUSES IN THE
ENVIRONMENT, FOR EXPULSION OF
VIRUSES, FROM ANIMAL MODELS AND
HUMANS, FROM THE ORAL CAVITY,
WE'RE TALKING ABOUT THE UPPER
RESPIRATORY TRACT, THE ORAL
CAVITY IS A REALLY BIG SITE HERE
TOO FOR EXPULSION IN ALL OF
THESE MODEL SYSTEMS.
I THINK THAT TO ME HAS ALWAYS
LACKED A LOT, AND I WOULD LIKE
FOR US TO BE MORE THOUGHTFUL
MOVING FORWARD.
>> THANK YOU FOR ADDING THAT.
SO I THINK WE'VE HEARD ABOUT
LOTS WONDERFUL MODELS FOR
TRANSMISSION, AND I WELCOME THE
PANEL DISCUSSING WITH US, DO WE
NEED ALL THESE MODELS, IS THAT
CONVERGING EVIDENCE, DOES THAT
GIVE STRENGTH?
ARE THERE AT TIMES SOME OF THESE
MODELS ARE BETTER THAN OTHERS,
AND YOU EACH KIND OF PRESENTED
ON YOUR UNIQUE MODELS AND GAVE
YOUR INDIVIDUAL OPINIONS, BUT
THAT DIDN'T RESULT NECESSARILY
IN SOME DEBATE AND DISCUSSION.
I LEFT EACH OF YOUR TALKS
THINKING IT'S HAMSTERS, NO, IT'S
GUINEA PIGS, NO, IT'S DEFINITELY
FERRETS, IT'S INFANT MICE.
SO I THINK IT COULD BE OF VALUE
AS A COMMUNITY TO HAVE A LITTLE
BIT OF A DEBATE AND DISCUSSION
ABOUT WHERE WE REALLY SEE SOME
OF THE STRENGTHS, WHERE MAYBE
EACH OF YOU AS EXPERTS IN THIS
AREA SAW SOME CONVERGING
EVIDENCE ACROSS YOUR MODELS OR
PLACES WHERE YOU'RE NOT SEEING
THESE THINGS SO WE CAN
APPRECIATE MAYBE SOME OF THE
LIMITATIONS.
AND I KNOW THAT'S A BIG ASK, BUT
I THINK THAT'S WHERE THIS PANEL
NEEDS TO GO.
>> ALL RIGHT.
SO EXCELLENT POINT, AND I HOPE
OTHERS WOULD JUMP IN AS WELL,
BUT MY TAKE ON THIS IS THE
QUESTIONS, RIGHT?
SO THE QUESTIONS YOU ASK,
WHATEVER YOU'RE INTERESTED IN,
WHATEVER YOU'RE PASSIONATE ABOUT
WILL DETERMINE WHAT MODEL YOU
USE.
IF YOU WANT TO STUDY THE
DYNAMICS OF TRANSMISSION AND
AEROSOL BIOLOGY, OBVIOUSLY YOU
HAVE TO CHOOSE A MODEL THAT
ALLOWS FOR THAT HUMANS, FERRETS,
FANTASTIC FOR THAT, EVEN GUINEA
PIG, I GUESS.
IF YOU WANT TO STUDY HOST
BIOLOGY, AND WE DON'T HAVE
ENOUGH REAGENTS IN THE FERRET
MODEL OR GUINEA PIGS THAT YOU
WANT TO USE, MICE ARE ADEAL FOR
THAT.
IF YOU WANT TO TEST PRE-EXISTING
IMMUNITY, WE CAN MODEL THAT IN
ANY OF THESE MODELS, BUT
ULTIMATELY IT WOULD BE THE HUMAN
MODEL THAT WOULD BE THE MOST
USEFUL FOR THAT.
SO THAT'S MY TAKE ON IT.
>> I WILL JUST ADD THAT I THINK
I HAVE NOT EVER BEEN AS HUMBLED
ABOUT TRANSMISSION AS I HAVE
BEEN WHEN I STARTED USING
HUMANS.
I'VE LEARNED MORE IN THE LAST
TWO YEARS ABOUT TRANSMISSION
THAN I HAVE IN OVER A DECADE OF
STUDYING IT AND READING
MANUSCRIPTS ON IT.
SO I THINK WHILE I DON'T HAVE AN
ANSWER, I MEAN, EVERY ANIMAL
MODEL IS FLAWED.
YOU KNOW?
FERRETS ARE FLAWED, MICE ARE
FLAWED, HAMSTERS ARE FLAWED,
GUINEA PIGS ARE FLAWED, CATS ARE
FLAWED, ALTHOUGH I REALLY LIKE
THE CATS.
I JUST THINK THAT, YOU KNOW, WE
ARE -- ANIMAL MODELS ARE FLAWED
AND WE REALLY DO, LIKE SAID AT
THE BEGINNING, WE SHOULD BE
CONSTANTLY THINKING ABOUT OUR
QUESTION, WHAT THAT IS,
ESPECIALLY FOR TRANSMISSION IN
TERMS OF HUMAN ASPECTS.
>> SEEMA, CAN I REALLY QUICKLY
EXPAND ON THAT?
ARE THERE FLAWS TO THE HUMAN --
CONTROL HUMAN INFECTION STUDIES?
>> ABSOLUTELY THERE ARE FLAWS IN
CONTROLLED HUMAN INFECTION
MODELS, AS YOU KNOW, SOME OF THE
PUBLISHED DATA, AND HOURS --
THERE IS LIMITED TRANSMISSION.
I JOKE AROUND A LOT, DOES FLU
ACTUALLY TRANSMIT?
LIKE I AM CONSTANTLY AMAZED WHEN
I SIT PEOPLE IN A TENT AND I
STARE AT THEM AND THEY DO CRAZY,
KREAS THINGS TO TRY TO GET
TRANSMISSION AND IT DOESN'T
WORK, IT IS ASTONISHING TO ME.
BUT I THINK WHAT I HAVE LEARNED
IS THAT THERE MAY BE A LOT OF
STOCHASTICITY IN HUMANS THAT WE
DO NOT RECAPITULATE IN HUMANS AT
ALL AND THAT WE REALLY NEED TO
UNDERSTAND A BIT MORE TO THEN
BUILD ANIMAL MODELS THAT ARE
MORE EFFICIENT AT HELPING US
STUDY IMPORTANT ASPECTS OF HUMAN
TRANSMISSION.
DOES THAT MAKE SENSE?
YEAH.
>> DIEGO, I KNOW YOU --
>> I WAS JUST GOING TO ADD THAT
I'M THE ODD BALL HERE BECAUSE
I'VE DONE WORK ON TRANSMISSION
BUT IF WE THINK ABOUT
PATHOGENESIS, I DON'T THINK
THERE'S ONE ANIMAL MODEL THAT
FITS ALL AS WELL.
AND I DO THINK THAT WE DO NEED
TO COMPARE AND CONTRAST TO
REALLY DRAW RELEVANT CONCLUSIONS
ABOUT INFECTION BIOLOGY AND
VIRAL BIOLOGY IN GENERAL.
>> PETER.
>> I WAS JUST GOING TO SAY THAT
WE CAN TRY TO HUMANIZE THE
ANIMAL MODELS TO TRY TO MAKE
MORE TRANSGENICS, BUT IT MIGHT
COME DOWN TO THE FACT THAT THIS
IS WHAT THE HUMAN FACE, THE
HUMAN RESPIRATORY -- LOOKS LIKE
AND IT'S GOING TO DIFFER FROM A
GUINEA PIG, IT'S GOING TO DIFFER
FROM A HAMSTER, SO IT MIGHT HAVE
TO LEARN TO LIVE WITH SOME OF
THE ISSUES THAT WE'RE DEALING
WITH.
>> I AGREE.
>> ANOTHER QUESTION BUT THIS IS
VERY GRANULAR.
YESTERDAY WE SORT OF TALKED
ABOUT DIRTY MICE, THE SORT OF
RAISED A THOUGHT IN MY HEAD
ABOUT THE ROLE OF COMMENSAL
BACTERIA.
ONE OF THE THINGS THAT AT LEAST
WE'VE LEARNED FROM SARS-COV-2,
I'M SURE THIS HAS BEEN THERE FOR
FLU I'M JUST NOT AS FAMILIAR
WITH THE FIELD, IS THAT THERE'S
CLEARLY CHANGES IN THE NASAL
MICROBIOTA DURING SARS-COV-2
INFECTION.
WHEN YOU LOOK AT THESE AIRBORNE
DROPLETS OF TRANSMISSION, DO
THEY CONTAIN BACK T BACTERIA?
AND WHEN YOU THINK ABOUT YOUR
TRANSMISSION MODEL, HOW MUCH
DOES THAT POTENTIALLY INFLUENCE
THE SEEDING OF VIRUS IN ANOTHER
NASAL CAVITY AND POTENTIAL
INNATE IMMUNITY, INDUCTION AND
THOSE TYPES OF THINGS?
>> I WILL TOUCH ON THIS BECAUSE
I THINK IT'S REALLY IMPORTANT.
WE ARE NOW DEEP SEQUENCING AS
MUCH AS WE CAN THINGS THAT ARE
BEING EXPELLED FROM FERRETS AND
HUMANS, TO SAY THAT.
BUT WHEN WE HAVE COINFECTION
STUDIES AND WE'VE LOOKED
SPECIFICALLY FOR BACTERIA KIND
OF SIMILAR TO WHAT -- WE
DEFINITELY SEE IT IN THE AIR,
STREP PNEUMO AND FLU, THEY'RE
CONTAINED IF DIFFERENT SIZE
AEROSOL CONTAINMENTS.
SO YOU CAN FRACTIONATE THE AIR
USING SOME OF THESE NIOSH
SAMPLERS THAT CAN DO A 1 MICRON
4-MICRON SPLIT YOU COULD SEE
THAT BACTERIA AS YOU WOULD
EXPECT ARE IN THESE LARGER
AEROSOL FRAGMENTS THAN THOSE
SMALLER ONES.
SO WE'RE WORKING ON SOMETHING WE
CALL A PAYLOAD EXPERIMENT WHERE
DIFFERENT SIZE AEROSOLS TRY TO
UNDERSTAND HOW MANY VIRAL
PARTICLES AND POTENTIALLY OTHER
PATHOGENS ARE IN EACH AEROSOL OF
DIFFERENT SIZE RANGES, BECAUSE
WE CAN -- THIS IS AMAZING THINGS
YOU CAN DO WHEN YOU WORK WITH
ENGINEERS, ACTUALLY FIGURE OUT
WAYS TO TUNE AEROSOL DELIVERY
DEVICES AS SPECIFIC SIZE
FRACTIONS CAN BE MADE.
BUT THESE ARE IMPORTANT
QUESTIONS.
YOU DO FIND THEM.
>> SO THE SWAFER HAMSTERS AND
THE SYRIAN HAMSTERS, THE DWARFS
ARE DIRTY.
THEY'RE BASICALLY COMING FROM
WILD AND BEING COLLECTED.
SO YOU SEE WHAT THE DWARFS, YOU
SEE THE LETHALITY, YOU SEE THE
WEIGHT LOSS YOU DON'T SEE IN THE
SYRIANS.
YOU ALSO DON'T SEE TRANSMISSION.
SO IF WE COULD START TRYING TO
CLEAN UP THE DWARFS, HAVE
SEPARATE SETS THAT ARE DIRTY,
SEMI CLEAN OR CLEAN AND SEE IF
WE SEE CHANGES, IT WOULD BE
INTERESTING TO LOOK AT.
IF WE CAN CLEAN THEM UP.
>> MICHAEL.
>> TO PICK UP ON WHAT SEEMA
SAID, DEPENDING ON WHERE IN THE
RESPIRATORY TRACTS THAT THE
AEROSOL IS MADE, YOU WILL HAVE
DIFFERENT PARTICLE SIZE, RIGHT,
AND THIS WILL AFFECT AEROSOL
TRANSMISSION, SO IS THERE A
DIFFERENCE BETWEEN THE ANIMAL
MODELS, AND WHICH ONE IS
REFLECTING BEST WHAT HAPPENS IN
HUMANS, FOR EXAMPLE, AFTER
SNEEZE.
>> I WILL SAY ACTUALLY I THINK
THAT THIS IS WHERE WE THINK A
LITTLE BIT BINARY ABOUT
AEROSOLS, SO YOU'RE ABSOLUTELY
RIGHT THAT AEROSOLS CAN BE
GENERATED BOTH FROM THE ORAL AND
NASAL CAVITY DURING A VARIETY OF
EXPULSIONS.
HOWEVER, THERE IS REALLY POOR
DATA, EMPIRICAL DATA ON THE
DIFFERENT AEROSOL FRACTIONS THAT
EMERGE FROM THE ORAL VERSUS
NASAL CAVITY.
MOST OF IT IS DONE THROW
MODELING AND TRYING TO
UNDERSTAND HOW THESE THINGS ARE
POTENTIALLY BEING FORMED, WHICH
IS WHY PEOPLE SAY THAT YOU
GENERATE LARGER AEROSOLS FROM
THE ORAL CAVITY VERSUS THE NASAL
CAVITY.
I DON'T THINK THERE'S GOOD DATA
ON THAT IS WHAT I WILL SAY, LIKE
ACTUAL SHARE MENTAL DATA.
BUT THE OTHER THINK I THINK IS
REALLY IMPORTANT WE IG NOOR IS
THAT AEROSOL THAT GET EXPELLED
ARE NOT -- THEY ARE NOT --
THEY'RE VERY DYNAMIC IN THE
ENVIRONMENT AS WELL, SOLID YA'S
GROUP AT MIT HAS DONE SOME
BEAUTIFUL WORK USING FLUID
DYNAMIC MODELS TO SHOW HOW
AEROSOLS ACTUALLY CHANGE SIZE
AND SHAPE IN THE AIR SO
DEPENDING UPON HUMIDITY AND
TEMPERATURE THEY CAN DESICCATE
AT FASTER RATES.
ALSO LARGER, SMALLER AEROSOLS
WILL COME OFF, THEY'RE NOT JUST
GOING TO STAY AT THAT ONE SIZE
RANGE WHEN IT GETS EXPELLED.
SO IT'S REALLY IMPORTANT TO
THINK ABOUT WHEN WE THINK ABOUT
AEROSOLS, IS THAT THEY ARE SUPER
DYNAMIC, AND OF COURSE THE
LARGER ISSUE IS WHEN A RECIPIENT
BREATHES THEM IN, DEPENDING UPON
THE SIZE AND WHAT ELSE IS IN
THEM, DEPOSIT AND BE SUCCESSFUL
SOMEWHERE ELSE IN THE
RESPIRATORY TRACT.
>> IS IT NORMAL -- A FERRET THAT
SNEEZES VERSUS A GUINEA PIG?
>> SO FERRETS ARE SIMILAR TO
HUMAN, IF YOU LOOK AT TOTAL
PARTICLE EXOWNTS, WHAT I CALL
THE DARK MATTER OF AEROSOL
BIOLOGY, IS THAT ALL OF OUR
DETECTION SYSTEMS THAT STUDY
AEROSOL SIZES IS REALLY GOOD AT
25 MICRONS AND SMALLER.
UNDERSTANDING AND QUANTIFYING
LARGER AEROSOLS GREATER THAN
25 MICRONS IS REALLY HARD.
IF YOU READ OLD AEROSOL PAPERS
TO UNDERSTAND HOW THEY DO THAT,
IT'S LIKE MAKE SOMEBODY BUT DYE
IN THEIR MOUTH AND SPLATTER IT
ON TO A PIECE OF PAPER AND LOOK
AT SIZE FRACTIONS FROM DIFFERENT
EXPULSIONS.
THAT IS LIKE SO CRUDE, IT IS
REALLY A CHALLENGE HERE TO STUDY
THINGS.
SO IF YOU LOOK AT -- THAT IS TO
SAY IF YOU LOOK AT THESE
PARTICLE COUNTERS THAT GO
25 MICRONS AND SMALLER, FERRETS
LIKE HUMANS, WE MAKE
PREDOMINANTLY REALLY SMALL
AEROSOLS, BUT A LOT OF THAT IS
CONTAMINATED STUFF FROM THE AIR.
YOU HAVE TO GET DOWN TO CLEAN
ROOM SIZES TO GET TO THAT, BUT
YOU STILL SEE A LOT OF SMALL
PARTICLES BEING GENERATED.
DOES THAT MAKE SENSE?
I DON'T KNOW IF ANYBODY ELSE
WANTS TO ADD ANYTHING TO THAT.
>> THANK YOU.
I WANT TO LET ZONDA HAVE A
MOMENT TO ASK A QUESTION.
>> YES, ON TRANSMISSION MODELS,
WE HAVE DISCUSSED A LOT ABOUT --
INFECTION HISTORY, CLEAN AND
DIRTY MICE LIKE THAT, AND WE
ALSO TOUCH ON AGING,
AGE-DEPENDENT, ABOUT HOW ABOUT
OTHER CONDITIONS LIKE METABOLIC
CONDITIONS?
SHOULD WE ADDRESS THAT?
>> TBD, WE'RE DOING THOSE
EXPERIMENTS.
AT LEAST IN THE MOUSE MODEL.
ABSOLUTELY.
METABOLIC CONDITIONS, AT LEAST
IN HUMANS, PATIENTS WITH
DIABETES, AS WELL AS THOSE WITH
HEART DISEASE TEND TO BE MORE
SUSCEPTIBILITY TO FLU AND THEY
ALSO SHED FLU FOR PROLONGED
PERIOD OF TIME,
IMMUNOCOMPROMISED PATIENTS, SO
WE'RE TRYING TO MODEL ALL OF
THOSE IN MICE.
>> I KNOW YOU LIKE MICE BUT --
>> FERRETS OVER HERE.
>> METABOLIC SYNDROMES, I REALLY
RECOMMEND YOU USE HAMSTERS.
TODAY I WOULD TALK A LITTLE BIT
ABOUT THE ADVANTAGE OF HAMSTER
IN TERMS OF MODELING METABOLIC
DISEASE.
>> SO IT WOULD BE GREAT TO SEE
YOUR HAMSTERS AND SEND THEM TO
WISCONSIN MAYBE.
>> I LOVE IT.
ARE THERE ANY QUESTIONS ONLINE
SINCE I KNOW I'VE BEEN REMISS
ASKING THAT?
ANY LAST BURNING QUESTION?
YES, STANLEY.
>> SO APOLOGIZE, THIS IS
PROBABLY NOT A BURNING QUESTION.
BUT WHEN YOU THINK ABOUT
TRANSMISSION, SO IF WE THINK
ABOUT THE PERSON WHO'S THE DONOR
AS IT WERE, DO WE THINK THAT
THAT PERSON HAS UNIFORM VIRAL
INFECTION ACROSS HIS OWN NASAL
CAVITY OR ARE WE THINKING
DIFFERENT PEOPLE, VIRUS MIGHT BE
IN ONE PLACE IN ONE PERSON,
DIFFERENT PLACE IN ANOTHER
PERSON AND THIS MIGHT CONFOUND
TRANSMISSION STUDIES BECAUSE
MAYBE ONE PART DOESN'T TRANSMIT
AS WELL?
>> I'M HAPPY TO TAKE THAT.
SO IF I'M USING CHIM STUDIES TO
JUST FLU H3 IN OUR CONTROLLED
HUMAN INFECTION MODELS WE SEE A
HUGE AMOUNT OF HETEROGENEITY IN
TERMS OF VIRAL LOAD IN THE NASAL
CAVITY VERSUS THE ORAL CAVITY.
SO WE SAMPLE THE NASOPHARYNGEAL
SITE, WE DO ORAL SWABS, SALIVA
COLLECTION THROUGH SPIT, WE DO
THROAT SWABS AND WE DO LIKE SORT
OF THE GCF TO SORT OF LIKE
BRUSHING YOUR TEETH, AND WE FIND
VARYING VIRAL LOADS IN ALL OF
THOSE SPACES.
IN FERRETS, WE SEE HIGH
CORRELATION BETWEEN NASAL TITERS
AND ORAL TITERS.
WE ALSO COLLECT SALIVA IN THE
FERRETS, I FORGOT TO MENTION
THAT, BUT I THINK IN HUMANS,
IT'S MUCH MORE HETEROGENEOUS.
>> WHAT ABOUT WITHIN EACH OF
THOSE AREAS, WITHIN THE NASAL OR
ORAL CAVITY, DO YOU THINK THAT'S
UNIFORM?
>> WE'RE ACTUALLY NOW DOING
SAMPLING TWO TIMES A DAY TO SEE
WHAT IS THE VAIR YITION OVER A
DAY WITHIN A SINGLE INDIVIDUAL.
I WILL TELL YOU WHAT I THINK IS
REALLY FASCINATING, THIS IS
WHERE I'M ALSO -- BEN CALLEN
TOLD ME THIS REALLY NIGHT TIDBIT
ONCE, IS THAT A SUPER SPREADER
IS ONLY A SUPER SPREADER ONCE,
RIGHT?
SO WHEN THEY'VE LOOKED AT HONG
KONG AND THEY'VE LOOKED AT THESE
SUPER SPREADERS AND I FIND
SOMEBODY SEEDED 200 INFECTIONS
AND THEN WENT TO ANOTHER SPACE
LATER THAT DAY, THEY DID NOT
SEED ANY FURTHER INFECTIONS SO.
STOCHASTICKITY IS SUPER
INTERESTING.
>> SO WE'VE ONLY GONE 2 MINUTES
OVER SO I THINK WE'VE HAD A
RATHER SUCCESSFUL SESSION.
PLEASE JOIN ME IN THANKING OUR
PANELISTS.
[APPLAUSE]
AND ACCORDING TO THE SCHEDULE,
WE NOW HAVE LUNCH UNTIL 12:45.
SO THAT'S TRUE FOR ALL OF OUR
ONLINE PARTICIPANTS.
THANK YOU.
>> I JUST WANT TO MAKE NOTE, I
THINK BECAUSE LUNCH IS A LITTLE
EARLIER TODAY, THEY'RE ON THEIR
WAY BUT IT'S NOT HERE IN THE
CAFETERIA THIS EXACT MOMENT, BUT
WE EXPECT THEM VERY SOON. THANKS.
IN THE INSTITUTE FOR ANTIVIRAL
RESEARCH, WE HAVE A GROUP OF
ABOUT 50 PEOPLE, AND REALLY
WE'VE HAD -- WE'VE BEEN SO
BLESSED WITH WORKING WITH THE
NIH, WE HAVE BEEN PART OF THE
COLLABORATIVE ANTIVIRAL TESTING
GROUP, REALLY FOR MORE THAN 25
YEARS.
WE APPRECIATE THE FUNDING ALL
THAT TIME FROM THE RESPIRATORY
DISEASES BRANCH, THE VIROLOGY
BRANCH.
WE DO BOTH IN VITRO EVALUATION,
SO IN VITRO SCREENING.
THIS IS A VALID ASSAY, WE JUST
CANNOT EVALUATE ALL THE
COMPOUNDS IN ANIMALS.
IT WOULD JUST BE TOO EXPENSIVE.
SO IN VITRO SCREENING STILL
PLAYS AN IMPORTANT PART.
15 YEARS AGO, WE WERE RUNNING
5,000 COMPOUNDS AGAINST
INFLUENZA EVERY YEAR.
THAT'S GREATLY DECREASED BUT WE
STILL ARE RUNNING THOUSANDS OF
COMPOUNDS EVERY YEAR.
THERE ARE ABOUT 40 DIFFERENT IN
VITRO ASSAYS THAT WE RUN.
WE ACTUALLY HAVE ANIMAL MODELS
FOR PROBABLY HALF OF THESE
VIRUSES.
DISTRIBUTED AMONG DIFFERENT
PIs, WE KIND OF DO SIMILAR
THINGS BUT WITH DIFFERENT
VIRUSES.
WE OFTEN HAVE SPONSORS, THESE
PEOPLE WHO HAVE HAD THEIR
COMPOUND APPROVED FOR TESTING TO
THE NIH.
IT'S THN GIV THEN GIVEN US TO.
THEY RECEIVE THE RESULTS, THEY
MAINTAIN ALL THE PROPRIETARY
RIGHTS AND THE NIH PAYS US.
A DEAL FOR ALL AROUND, ISN'T IT?
I'M OFTEN ASKED BY SPONSORS,
THEY HAVE A COMPOUND THAT'S VERY
EFFECTIVE IN VITRO AND IT FAILS
IN THE ANIMAL.
THEY'LL SAY, WHAT HAPPENED?
WHY DIDN'T THIS WORK?
AND I READ THIS LITTLE
STATEMENT, THE ONLY LATIN I
KNOW, OKAY, INSILICO -- IN VIVO
VERITAS.
I'VE ADDED IN VITRO IN THERE.
SO IN COMPUTERS AND CELL
CULTURE, WE MAKE PREDICTIONS.
IT'S IN THE ANIMAL WE LEARN THE
TRUTH.
THAT'S THE GOAL AND I HAVE TO
SAY THE TRUTH OFTEN FALSE FAR FR
SHORT OF THE GOAL.
I LOVE THIS QUOTE BY GEORGE BOX,
STATISTICIAN WHO SAYS ALL MODELS
ARE WRONG, SOME ARE USEFUL.
I THINK IS POINT IS WE SHOULD
FOCUS ON SOMETHING THAT CAN BE
APPLIED IN A USEFUL MANNER
RATHER THAN DEBATING IF IT'S
CORRECT IN ALL CASES.
AND REA PRODUCIBILITY, I THINK
IS ABSOLUTELY KEY.
I THINK ABOUT WHAT I CAN TALK
ABOUT IN THIS LITTLE SHORT
PRESENTATION TODAY, AND I'M
GOING TO TALK ABOUT LESSONS
LEARNED, OFTEN LESSONS LEARNED
THE HARD WAY IN MY LIFE, OKAY,
THAT REPRODUCIBILITY, ABSOLUTELY
KEY TO ALL THE MODELS THAT WE'RE
PRODUCING.
A LITTLE BIT OF A DIFFERENCE
BETWEEN A QUALIFICATION OR A
QUALIFIED ASSAY AND A VALIDATED
ASSAY.
WHEN I WORKED IN THE VACCINE
INDUSTRY, WE WOULD OFTEN DEVELOP
OR VALIDATE ASSAYS AS PART OF
THE LICENSING PROCESS.
VALIDATION IS A VERY HIGH
STANDARD.
QUALIFICATION MEANS IT PROVIDES
A HIGH DEGREE OF CONFIDENCE THAT
A PROCESS OR A MODEL IS
REPRODUCIBLE UNDER ANTICIPATED
VARIABLE RANGES.
ANTICIPATED BEING THE KEYWORD,
ANTICIPATED OR EXPECTED.
VALIDATION DESCRIBES PERFORMANCE
UNDER VERY HIGHLY CONTROLLED
CONDITIONS.
USUALLY BEYOND WHAT WE DO WITH
ANIMAL STUDIES.
IN PRE-CLINICAL TESTING FOR
ANTIVIRALS FOR VACCINES, WE
NORMALLY USE MODELS THAT ARE NOT
FULLY VALIDATED BUT QUALIFIED.
IT MEANS WE'VE DETERMINED WHERE
THE VARIABILITY IS FOUND.
SO LESSONS LEARNED, 25 YEARS A
AGO, HARD TO BELIEVE THAT, 1999,
HORSES IN NEW YORK WERE
DIAGNOSED WITH WEST NILE VIRUS,
A VIRUS THAT HAD NEVER BEFORE
BEEN IDENTIFIED IN THE WESTERN
HEMISPHERE.
PEOPLE DIDN'T BELIEVE IT AT THE
TIME, IT WAS TRUE.
SO LESSON LEARNED, WHAT IS
CONSIDERED A FOREIGN DISEASE
AGENT TODAY MAY BECOME AN
ENDEMIC AGENT TOMORROW.
YOU SEE, DUH, OBVIOUS.
AT THE TIME IT WAS NOT.
THIS WAS ASTONISHING.
IT TOOK FIVE YEARS FOR WEST NILE
TO GO FROM THE WEST V WEST COASE
EAST COAST.
MOSQUITO-BORNE.
THIS WAS ASTONISHING TO
BIOLOGISTS, EPIDEMIOLOGISTS, TO
PUBLIC HEALTH PEOPLE.
WE'D NEVER SEEN THIS BEFORE.
WITH REGARDS TO VACCINE
DEVELOPMENT, WE ALWAYS THINK
PROS AND CONS.
WE WANT TO BE FIRST.
FOR A NOVEL PRODUCT AT THAT
TIME, WE WERE DEVELOPING ONE OF
THE VERY FIRST RECOMBINANT
VACCINES FOR USE IN VETERINARY
MEDICINE.
SO THE PRO, WE CAN SET THE
STANDARD TO WHERE WE WANT TO BE.
CON, THE REGULATORY GUIDELINES
MAY NOT BE IN PLACE FOR
SOMETHING THAT NOVEL.
WE STITTED SUBMITTED AN ADDITIOL
DOSSIER FOR LICENSING THE
VACCINE.
IT TOOK THE USDA18 MONTHS TO
COME BACK TO US TO SAY THIS IS A
CHIMERIC VIRUS.
WE KNOW.
THEY DIDN'T HAVE THE EXPERTISE
AT THAT TIME.
SO I STARTED WORKING ON EQUINE
VACCINES, ACTUALLY DEVELOPED TWO
EQUINE VACCINES IN HORSES, A FEW
VACCINES, AND AT THAT TIME IN
2004, SO ABOUT 20 YEARS AGO,
EQUINE INFLUENZA JUMPED FROM
RACING HORSES TO RACING DOGS IN
FLORIDA.
THAT WAS KIND OF ASTONISHING,
LESSON LEARNED.
SO A DISEASE AGENT MAY JUMP FROM
ONE SPECIES TO ANOTHER.
AGAIN, DUH, OF COURSE.
SURPRISING AT THE TIME, NOT
EXPECTED 20 YEARS AGO.
AGAIN, WE DEVELOPED A NOVEL
PRODUCT, TRYING TO BE FIRST ON
THE MARKET, BEING FIRST, YOU CAN
SET THE STANDARD.
BUT IN THIS CASE THE CON, THE
REGULATORY AUTHORITIES, MAY NOT
HAVE THE SPEAR TE EXPERTISE, SON
TO OUTSIDE EXPERTS THAT MAY HAVE
INCORRECT BIASES.
WE SUBMITTED AN EMERGENCY USE
AUTHORIZATION DOSSIER.
THEY GOT IT, THEY SENT IT OUT TO
AN EXPERT.
THEY DIDN'T HAVE EXPERTISE IN
CANINE INFLUENZA.
THE EXPERT WAS AN EXPERT IN
BACTERIAL RESPIRATORY DISEASES
OF DOGS AND HE SAYS THIS IS
KENNEL COUGH.
WE SAID WE KNOW THIS IS NOT
KENNEL COUGH.
WE GOT HIM BACK, GOT ALL THE
TESTS, AGAIN SUBMITTED THE
DOSSIER AND YES, IT WAS LICEN
LICENSED.
SO LET ME GO TO THE MOUSE.
USU IN 2008.
IN 2009, ENDEMIC H1N1 HIT.
THAT WAS A PANDEMIC.
WE DODGED THE BULLET AT THAT
TIME, WE DID NOT LEARN THE
LESSON THAT WE NEEDED TO LEARN
FOR 2020, SARS-COV-2.
DON SNEAD AND MYSELF WERE
WORKING ON THE PANDEMIC FLU AT
THAT TIME AND HELPING RESPOND TO
THIS THREAT, THIS PANDEMIC.
WE REALLY DIDN'T HAVE TIME TO GO
THROUGH A LICENSURE PROCESS.
SO WHAT WE WERE FOCUSING ON WAS
COMBINATION DRUG STUDIES, TRYING
TO TAKE TWO LICENSED DRUGS,
SEEING IF WE COULD INCREASE THE
EFFICACY OF THE EXISTING
ANTIVIRAL DRUGS.
JUST KIND OF LOOKING HERE
QUICKLY AT THE MOUSE, WE HAVE
WEIGHT LOSS, KIND OF METRICS, I
LIKE TO CALL THEM.
THEY'RE PARAMETERS WE CAN
EVALUATE TO EVALUATE ANTIVIRAL
EFFICACY, VACCINE EFFICACY.
WEIGHT LOSS, MORTALITY ARE GOOD
METRICS.
PNEUMONIA, WE CAN LOOK AT LUNG
SCORE.
THIS IS SOMETHING VERY SIMPLE,
WORKING AT SAY FOUR QUADRANTS IN
THE LUNG, HOW MUCH OF THE LUNG
IS SHOWING A PURPLE COLOR
CONSOLIDATION?
WHITE ARE A GOOD NUMBER.
VIRUS TITERS, LUNG HI, STO.
CYTOKINE MEASUREMENTS, NOT
REALLY SO MUCH FOR ANTIVIRALS
BUT DEFINITELY FOR VACCINES.
SO A FEW LESSONS LEARNED, VIRUS
STRAIN DIFFERENCES.
SEASONAL INFLUENZA HAS BEEN
TALKED ABOUT.
IN FACT, THERE'S BEEN SO MANY
THINGS THAT HAVE BEEN DISCUSSED,
I CAN JUST STAND UP HERE AND SAY
DITTO AND GO BACK AND SIT DOWN.
SEASONAL INFLUENZA A REQUIRES
VIRUS ADAPTATION TO CAUSE
LETHALITY IN MICE.
THANK GOODNESS HIGHLY PATHOGENIC
AVIAN FLU DOES NOT REQUIRE
ADAPTATION.
SO DON'T HAVE TO DEAL WITH ADAP
TITION TO GAIN OF FUNCTION-TYPE
THINGS.
TALK ABOUT H3N2 VIRUSES, VERY
DIFFICULT TO ADAPT.
THEY ARE HEAVILY GLYCOSYLATED.
WHEREAS WE FIND WITH MOUSE
ADAPTATION OF FLU VIRUSES, IF
THEY ARE GLYCOSYLATED, THEY WILL
LOSE GL GLYCOSYLATION.
IT'S WHAT MAKING IT SO DIFFICULT
TO ADAPT.
2009, PANDEMIC H1 WAS JUST
DIFFERENT.
IT WAS JUST DIFFERENT.
THESE LITTLE CHARTS THAT COME
OUT OF THE WOR WORLD HEALTH
ORGANIZATION, THIS WAS FOR THE
END OF JUNE.
IF YOU'VE NOT LOOKED AT THESE,
THE LITTLE PIE CHART SHOWS
GEOGRAPHICAL AREAS AROUND THE
WORLD WHERE THEY'RE GETTING
SAMPLES OF DIFFERENT COLORS IN
THE LITTLE PIE CHART SHOWING
WHICH FLU SUBTYPES.
SO THE LIGHT GREEN IS H1.
MEDIUM COLORED GREEN IS JUST
INFLUENZA A, THEY'RE NOT
DISTINGUISHING BETWEEN H1 OR H3.
THE DARK GREEN IS NOW THE
COMBINATION, THE MIDDLE GREEN IS
H3.
THE RUST COLOR ORANGE, WHATEVER
COLOR YOU WANT TO CALL IT,
THAT'S INFLUENZA B.
WHAT WAS REMARKABLE IS LOOKING
AT THIS LITTLE CHART, AND IT
CHANGES, IT'S ALWAYS BEEN
INTERESTING TO ME HOW DIFFERENT
REGIONS OF THE WORLD, DIFFERENT
FLU VIRUS SUBTYPES ARE
CIRCULATING.
WHEN THE PANDEMIC CAME IN,
PANDEMIC H1 BACK IN 2009, IT
PUSHED EVERYTHING ELSE OUT.
ALL THESE PIE CHARTS WERE ONE
COLOR.
THE PANDEMIC, 2009, H1N1, WAS
DIFFERENT.
IT NEVER EXISTED IN THE WORLD
BEFORE.
SO JUST KIND OF A REMARKABLE
THING.
I DON'T WANT TO BO BEAT THIS TO
DEATH, IF I WORK FOR THE FDA AND
SOMEBODY CAME UP TO ME TO BRING
A DOSSIER SHOWING EFFICACY FOR
PR8, I WOULD SAY THIS IS VERY
NICE, NOW PLEASE DO IT IN A
CLINICALLY RELEVANT MODEL.
I WENT TO A KEYSTONE MEETING
HERE A COUPLE YEARS AGO ON
INJURY AND INFLAMMATION IN THE
LUNG.
SO THESE WERE NON-VIROLOGISTS.
THEY WERE USING PR8 AND I
THOUGHT, I LIKE IT.
THEY WERE USING PR8 AS THE
INSULT.
THAT WAS THE INJURY TO THE LUNG
TO LOOK AT INJURY AND
INFLAMMATION.
THEY WERE NOT VIROLOGISTS, THEY
DIDN'T CARE WHAT -- THEY COULD
BE ASSUMING ASBESTOS.
BUT I THOUGHT IN THAT SITUATION,
I THOUGHT THAT WAS A GREAT
MODEL.
SO I RECOGNIZE THERE ARE
DIFFERENT LOCATIONS WHERE THAT
COULD BE A GOOD M MODEL.
INFLUENZA B, THIS LITTLE CHART
HERE, IMAGINE IF IT WAS SARS,
BECAUSE WE WERE WEARING MASKS
AND OF COURSE NO ONE BELIEVES
MASKS WERE EFFECTIVEMENT WE KNOW
TEFFECTIVE.WE KNOW THAT THEY WE.
SO EFFECTIVE THAT IT'S PROBABLY
DRIVEN THE MAMAGATA VICTORIA
LINEAGES OF INFLUENZA B,
SPORADICALLY IDENTIFIED SINCE
SARS 22, IT'S BEEN PUSHED TO
EXTINCTION.
I BELIEVE, SOMEONE ELSE CAN
CORRECT ME, THAT THIS YEAR IT'S
NOT A QUADRA VALENT VACCINE.
PRIOR TO THIS WE'VE HAD H1H3,
INFLUENZA A, AND BOTH VICTORIA
AND A MAMAGATA FOR THE INFLUENZA
Bs, THIS YEAR THEY WENT BACK
TO H1, H3 AND VICTORIA.
THE YAMAGATA IS BEING PUSHED
OUT.
WHICH IS KIND OF INTERESTING.
OKAY.
VIRUS CHALLENGE DOSE.
WE WILL DO A 3XLD50 AS PART OF
OUR QUALIFIED ANTIVIRAL TESTING
MODELS.
IF YOU THINK A LOG SCALE PAPER,
HALF A LOG IS NOT HALFWAY
BETWEEN THE SCALES.
IT'S A THIRD OF THE WAY.
SO 3 IS ABOUT A HALF LOG.
SO L3XLD50 IS ABOUT A HALF LOG
HIGHER THAN LD50.
IT'S THEORETICALLY A LITTLE
HIGHER THAN LD90.
BUT WHAT WE'RE WANTING TO DO,
WE'RE WANTING TO CAUSE LETHALITY
IN ALMOST 100% OF THE MICE.
BUT WE DON'T WANT TO
OVERCHALLENGE BECAUSE WE WANT TO
GIVE THE ANTIVIRAL -- OF THE
VACCINE THE OPTIMAL CHANCE OF
SHOWING EFFICACY.
SO WE DON'T WANT TO
OVERCHALLENGE IT.
THE DOSE IS PART OF THE
QUALIFIED MODEL.
WHY I'M STRESSING THAT, I WILL
HAVE SPONSORED FROM THE NIH HAS
APPROVED, YES, WE'D LIKE TO MORE
OR LESS HOLD THE VACCINE DOSE
CONSTANT AND HAVE US CHANGE THE
VIRUS CHALLENGE DOSE.
AND I HAVE TO BE VERY TIP LOMA
TICK ADIPLOMATICAND TELL THEM T.
THE CHALLENGE DOSE IS PART OF
THE MODEL.
IF THE VACCINE DOESN'T WORK, I'M
SORRY, IT'S NOT EFFICACIOUS, BUT
THE MODEL STANDS.
SOMETHING ELSE I'VE LEARNED FROM
REALLY JUST INS TEUGAL MEMORY
WHEN I WORKED IN THE VACCINE
INDUSTRY, IT WAS A VERY GOOD
COMPANY, WE HAD A LOT OF
EXPERTISE.
I WILL HAVE SPONSORS COME TO US,
TWO, THREE, FOUR VACCINATIONS
FOR EFFICACY, AND WHAT I WAS
TAUGHT VERY EARLY ON, WE WANT TO
DO A SINGLE DOSE IN A
PRELIMINARY STUDY, THAT IF I'M
USING MORE THAN ONE DOSE OF
VACCINE, IT CAN MASK A VERY WEAK
IMMUNOGEN.
AND THE COMPANY I WAS WORKING
FOR, WE WANT TO KNOW IF IT'S A
PORE IMMUNOGEN BECAUSE WE WANT
TO CUT OUR LOSSES BEFORE WE GO
ON.
WHY KEEP CHASING, WE'RE KICKING
A DEAD COURSE AS THEY MIGHT SAY.
YOU WANT TO KEEP WORKING ON TH
THIS -- YOU DON'T WANT TO KEEP
WORKING ON THIS IF IT'S NOT
GOING TO BE EFFECTIVE.
FROM THERE, WE CAN GO ON TO DO
LOTS OF OTHER THINGS.
REGARDING ADJUVANT STUDY,
THERE'S A REASON I'M STARTING
WITH THIS.
I OFTEN WILL CALL THIS KIND OF
THREADING THE NEEDLE.
SO I WANT TO GET A SUBOPTIMAL
DOSE OF A VACCINE THAT DOES NOT
PRODUCE STATISTICALLY
SIGNIFICANT EFFICACY BY ITSELF,
BUT NOW THAT I HAVE AN ADJUVANT,
THEN I CAN SEE STATISTICAL
SIGNIFICANCE, AND IT'S SHOWING
ME THAT THE ADJUVANT IS WORKING.
KIND OF A CHALLENGE THERE.
VERY SIMILAR TO AN ANTIVIRAL
COMBINATION DRUG STUDY.
BUT YOU TAKE TWO EFFICACIOUS
DRUGS, PUT THEM TOGETHER, YOU
CAN'T SEE IF THERE'S SYNERGY OR
NOT.
SO WHAT WE'LL DO, WE DO A
SUBOPTIMAL DOSE OF BOTH, PUT
THEM TOGETHER AND THEN LOOK FOR
SIGNIFICANCE AND IT WILL TELL
YOU IF THERE'S SYNERGY OR NOT.
BREATHING ANTAGONISM.
SO MODEL TO EVALUATE AND DEVELOP
THERAPEUTICS LETHAL VERSUS
NON- LETHAL OBVIOUS QUESTION.
MANY TIMES I'VE BEEN CRITICIZED
FOR LETHAL MODELS, THEY SAY WHAT
POPULATION ARE YOU REALLY
MODELING?
WELL, PEOPLE THAT ARE SEVERELY
INFECTED IN HOSPITALS, THAT'S
PROBABLY NOT TOO BAD A
POPULATION.
IF WE CAN DEVELOP AN ANTIVIRAL
THAT WILL PROTECT THEM, IT WILL
DEFINITELY PROTECT SOMEONE AT
HOME.
BUT THIS IDEA OF THE NON-LETHAL
MODEL IS IMPORTANT AND I'M GOING
TO COME BACK TO THAT IN JUST A
MINUTE.
MORE GENERALLY, WHEN I THINK
ABOUT DEVELOPING A MODEL, I'M
DEVELOPING SOME NEW ONES RIGHT
NOW FOR HE COVIRUSES IN
CHILDREN, WHAT AM I WANTING TO
MODEL?
THE CLINICAL SIGNS IN THAT
POPULATION, YOUNG CHILDREN, AND
OFTENTIMES WE CAN'T MODEL IT,
BUT I SEE WHAT CAN WE PRODUCE,
WHAT CLINICAL SIGNS CAN WE
OBSERVE IN OUR MODEL IN THE
ANIMAL AS BEING OBSERVED IN THE
CLINIC, WITH THE IDEA OF COMING
UP WITH SOME KIND OF A METRIC,
BECAUSE I LIKE TO CALL IT A
PARAMETER THAT WE CAN EVALUATE
TO SEE IF THAT ANTIVIRAL DRUG IS
HELPING OR ALLEVIATING THAT
METRIC.
I FOUND GOOD POSITIVE CONTROL IS
SO IMPORTANT.
A FEW YEARS AGO WE DEVELOPED
ENTEROVIRUS MODELS FOR
ENTEROVIRUS 68 #, ENTEROVIRUS
71.
THE D68 WENT WELL, WE HAD A
GREAT POSITIVE CONTROL, WE WERE
HAPPY MOVING ON, JUST GO AHEAD,
WORK ON THE 71 MODEL.
GOT IT WAY DOWN THE LINE, NOW
STARTED WORKING LOOKING FOR
POSITIVE CONTROL, GOODNESS,
NOTHING WORKED.
SO I HAVE TO START LOOKING FOR
POSITIVE CONTROL VERY EARLY ON
BECAUSE IDENTIFYING THOSE
METRICS FOR EVALUATION, I CAN
SEE HOW THE POSITIVE CONTROL IS
WORKING IN THOSE CLINICAL
PARAMETERS.
IS IT REDUCING PARAMETERS IF
IT'S HIGH, IS IT INCREASING AND
IF IT'S LOW, IS IT A BENEFIT.
THANK YOU FOR THE INVITATION TO
COME AND TALK.
THIS IS SOMETHING I'VE BEEN
THINKING ABOUT, THIS MIDDLE
DIAMID LITLITTLEDIAGRAM, WHY H1,
SARS WAS DIFFERENT.
SO IT WAS KIND OF FUN TO PUT
THIS TOGETHER.
I'VE DRAWN THIS ON BOARDS, I'VE
NEVER ACTUALLY HAD IT IN A
POWERPOINT.
SO HERE WE HAVE AN INFECTION,
THIS IS FLU, DAY 3, DAY 6, IT'S
NOT EXACTLY PERFECT BUT IT'S
PRETTY CLOSE.
SO IF I DRAW A SECOND LINE, THIS
WOULD BE THE HISTOPATHOLOGY.
SO FIRST WE HAVE VIRUS TITERS
COMING UP.
THE VIRUS IS DEPI -- CYTOPATHIC
SPREAD WE'RE SEEING LESIONS IN
THE TISSUE, AS THE LESIONS IN
THE TISSUE COME UP, THEN WE
BEGIN TO SEE CLINICAL SIGNS.
AND JUST PARD3 MY MAKING A
COMPLEX VERY SIMPLE BUT IN MY
MIND I'M KIND OF SIMPLE, I'M
SEEING THESE THINGS COME UP AND
THEN THEY'RE GOING DOWN.
FIRST TITER GOES DOWN, HEALING
BEGINS, THE TISSUES ARE STARTING
TO HEAL.
CLINICAL SIGNS ARE THEN
FOLLOWING.
AND SO WE SEE THIS NICE PRETTY
PARALLEL LINES.
TIME TO MORBIDITY IS IMPORTANT
FOR A TREATMENT WINDOW, LOOKING
AT ANTIVIRAL DRUGS.
AGAIN, WE WILL HAVE A SPONSOR
COME AND THIS IS QUITE ROUTINE,
I WANT TO DOSE MICE TWICE A DAY
FOR 10 DAYS.
OKAY, LOOK AT MY TIMELINE HERE.
IF I'M DOSING CLEAR OUT HERE AT
10, I'LL PROBABLY START SEEING,
AGAIN, BASED UPON H1, H3, B, H5,
THAT TIME OF MORTALITY CAN SHIFT
A COUPLE DAYS, THEY'RE STILL
PRETTY CLOSE.
THEY'RE WANTING TO DOSE TO DAY
10 SOME OF THESE ANIMALS ARE
DYING.
BECAUSE THEY'RE ALREADY IN THE
PERIOD WHERE THERE'S SOME PRETTY
SEVERE CLINICAL SIGNS.
THIS TIME PERIOD BEFORE 5, I
WANT TO MAKE SURE TREATMENT IS
EXREETED BEFORE DAY 6.
I DON'T WANT TO HAVE A COFACTOR
OF THE SICKNESS OF THE ANIMAL
BEING INTERFERED WITH BY THE
DOSING OF THE DRUG.
SO I'LL COME BACK TO THAT IN
JUST A FEW MINUTES HERE.
THAT'S KIND OF INTERESTING.
THIS IS A ROUTE OF THERAPEUTIC
ADMINISTRATION.
THIS IS A VERY COMPLEX SLIDE BUT
I'M GOING TO MAKE IT VERY
SIMPLE.
OF MICE AND MEN, MICE ARE NOT
MEN.
THIS IS A LICENSED DRUG USED IN
THE CLINIC.
IT'S ADMINISTERED BY NEBULIZER
TO PEOPLE BECAUSE THE
PHARMACODYNAMICS WERE NOT GOOD
ENOUGH BY ORAL ROUTE OR ANY
OTHER MEASUREMENT SO IT WOULD
HAVE TO BE ADMINISTERED BY
NEBULIZER TO PEOPLE.
THIS IS SHOWING YOU MICE ARE NOT
MEN.
SO THIS NOW LOOKING SANAMOVIR,
10 MILLIGRAMS PER KILOGRAM PER
DAY FOR FIVE DAYS DOWN TO THREE,
1.3, THESE ARE THE DOSE RANGES,
INTRAPERITONEAL, INTRAMUSCULAR,
BY ORAL, AND WHICH MIMICS
NEBULIZER.
YOU CAN SEE THESE ARE ALL PRETTY
POTENT REGARDLESS OF THEIR ROUTE
OF ADMINISTRATION.
THIS IS NOT OBSERVED IN HUMANS.
SO I'LL HAVE SOMEONE COME TO ME
AND SAY WE WANT TO GO FOR NEB
LASER ROUTE, CAN YOU DO THAT?
I'LL SAY WE CAN, BUT IN A MOUSE,
WE CAN LOOK AT A DIFFERENT ROUTE
AND SEE IF IT'S GOING TO WORK.
AND I'LL COME BACK TO THAT IN A
MINUTE.
ANOTHER POINT HERE KIND OF
INTERESTING, SO IF YOU LOOK UP
HERE AT THIS HIGH DOSE, 10 MG
DOSE, THEY'RE 11 MOUSE OF EACH
OTHER ON A DIFFERENT ROUTE.
WHAT I FOUND, WHAT ALL OF US
HAVE FOUND OVER AND OVER, I
CONTEND TWO GROUPS OF MICE,
ORDERED THEM AT THE SAME TIME,
ARRIVED THE SAME TIME, I STARTED
ON TWO DIFFERENT DAYS, SAME
TECHNICIANS, SAME TECHNIQUE,
SAME DRUG, SAME VIRUS, AND THERE
CAN BE ONE MOUSE DIFFERENCE IN
THE GROUP OF 10 BETWEEN THOSE
TWO STUDIES.
SO THIS IS PRETTY GOOD HERE.
NOW IF WE GO DOWN TO THREE,
HOPEFULLY YOU CAN SEE THIS, THE
BLUE HIGHLIGHT, EFFICACY REALLY
DROPPED, AND NOT ONLY DOES THE
EFFICACY DROP, VARIABILITY IS
INCREASING.
IT'S FROM HAVING A SINGLE MOUSE,
NOW THERE'S TWO MICE.
IF I DROP AGAIN, NOW I'M DOWN TO
30% VARIABILITY.
I'VE NOT PUBLISHED THIS BUT THIS
IS SOMETHING I'VE FOUND FROM
MANY, MANY ANTIVIRAL STUDIES, I
SEE AIN CRE AN INCREASE IN VARIY
AT THE LOWER LEVEL OF ANTIVIRAL
EFFICACY.
IT DOESN'T MEAN THE DOSE.
SPONSOR COMES TO US, THEY HAVE
THE DRUG, VERY FIRST THING WE
DO, WE DO WHAT WE CALL A MAXIMUM
TOLERATED DOSE STUDY.
WE EVALUATE THE DOSE RANGE OF
THAT DRUG, NO VIRUS CHALLENGE,
WE LOOK FOR WEIGHT LOSS,
SEIZURE, TOXICITY.
I CAN GO WITH THE VERY HIGHEST
DOSE OF DRUG AND SEE VERY HIGH
VARIABILITY, AND THAT SUGGESTS
TO ME THAT THAT DRUG IS PROBABLY
AT ITS LOWEST LIMIT.
SO AGAIN THIS WAS KIND OF
INTERESTING.
THIS WAS A 5-DAY TREATMENT.
WE FOUND THAT WE WERE USING -- I
MUST HAVE STARTED LATE.
WE FOUND THAT THE INTRANASAL
SALINE TREATMENT OR INTRANASAL
ADMINISTRATION OF THE DRUG
EXACERBATED THE INFECTION.
SO WE WENT BACK AND EVALUATED
THIS.
HERE'S CALIFORNIA, HERE'S THE
PANDEMIC H1, HERE'S ACTUALLY A
RESISTANT H1, HERE'S A
HISTORICAL H1, KIND OF LIKE THE
PR8.
HERE'S VICTORIA H3N2, H5N1 LOW
PATH, AND THESE ARE NOW LOOKING
AT THE INTRANASALLY TREATED, AND
THE UNTREATED MICE.
I WANT TO PICK OUT THIS ONE IS
THE HIGHEST HERE.
THE MLD50 FOR THE TREATED MICE,
TREAT DADELY FOR FIVE DAYS, 1.5.
4.5 FOR UNTREATED.
AND AGAIN, THIS IS NOT A
THREE-FOLD INCREASE IN VIRUS.
THIS IS A THOUSAND FOLD INCREASE
IN VIRUS.
UP HERE YOU CAN SEE THAT LOW
DOSE VIRUS WITHOUT INTRANASAL
TREATMENT OF LIQUID, THEN WE ADD
THE SALINE TO IT, WE CAN SEE
THAT THAT LOW DOSE CHALLENGE IS
BEHAVING JUST LIKE A HIGHER
CONCENTRATION OF VIRUS.
I'M WAY OUT OF TIME, I'M SORRY.
THIS IS A COMBINATION DRUG STU
STUDY.
WE HAD STANDING ORDERS FOR 350
MICE PER WEEK AND WE WERE STILL
ORDERING MORE MICE.
THIS IS 28 GROUPS OF 10 MICE
EACH LOOKING AT A COMBINATION
DRUG STUDY.
WE WERE SEEING IF THERE WAS
SYNERGY BETWEEN TWO DRUGS IF
THEY COULD BE ADDED TOGETHER.
SO WE'RE ADDING SUBOPTIMAL
DOSES, THIS IS USING A SYNERGY
SOFTWARE DEVELOPED BY PRITCHARD
AND CHIPMAN AND THE DRUG-DRUG
INTERACTION THAT HAD BEEN REALLY
MODELED IN A GROUP AT SAINT
JUDE'S WHERE LESS THAN 25% IS
INSIGNIFICANT SYNERGY
ANTAGONISM, 25 TO 50 IS MINOR
SYNERGY, 50 TO 100 MODERATE,
STRONG SYNERGY.
THIS ACTUALLY IS MEASURING AREA
UNDER A CURVE DRUG DOSING
SHOWING DRUG-DRUG INTERACTIONS.
WHAT I'M GETTING TO, THIS IS A
CONCERN I HAVE IN THE FIELD OF
INFLUENZA.
THAT WE WILL SEE IN A PUBLISHED
WORK SOMEONE IS USING C57 BLACK
AND THEY'RE USING PR8 AND
THEY'RE TELLING YOU THEIR
MORTALITY BUT THEY'VE NOT TOLD
ME THEIR WEIGHT LOSS CUTOFF.
SOMEONE ELSE IS LOOKING AT
H3N2 BALB C AND USING A
DIFFERENT WEIGHT LOSS CUTOFF OR
DIFFERENT MORTALITY, HOW DO WE
COMPARE THESE THINGS?
IT'S SHOWING THE ROBUSTNESS IN
THE FIELD THAT THEY'RE WORKING,
BUT HOW DO WE COMPARE THEM?
THIS STUDY WAS DONE, THIS WAS
TRUE MORTALITY, AND WE FIND --
THEY HAVE THE SAME MECHANISM OF
ACTION SO I WOULD KIND OF EXPECT
MINOR ANTAGONISM OR MINOR
SYNERGY AND FOR TRUE
MORTALITIES, MINOR ANTAGONISM.
THAT SAME SET OF WEIGHTS, WE'VE
NOW TAKEN UNDER 20%, 25%, 30%
WEIGHT LOSS, AND LOOK AT THE
DIFFERENCE.
AT 20% IS INSIGNIFICANT SYNERGY,
25% MODERN ANTAGONISM, 30%
WEIGHT LOSS, STRONG SYNERGY.
THIS IS A CONCERN TO ME, THAT
BASED UPON OUR WEIGHT LOSS,
WE'RE SEEING DIFFERENT EFFECTS.
TO ME, THAT'S A HUGE CONCERN.
AND I'M WAY OVER TIME.
SURE.
CAN I SHOW ONE MORE SLIDE?
IT'S THE CLINCHER.
OKAY?
I THOUGHT THIS WAS GOING TO BE
REALLY QUICK.
OKAY.
SO I SHOW YOU THIS GRAPH OF
INFLUENZA, HISTO COMES UP, WHAT
WAS ASTONISHING TO ME WAS IN
THESE HAMSTERS, TITERS GO UP,
THEY COME DOWN, CLINICAL SIGNS
CONTINUED, HISTOPATHOLOGY
CONTINUED.
I HAVE NEVER SEEN THIS BEFORE.
I'VE WORKED ON 30 DIFFERENT
VIRUSES, 15 DIFFERENT MODELS.
THIS WAS ASTONISHING TO ME.
THE VIRUS WAS DOWN, IS VIRUS
STILL THERE, IS IT PERSISTENT,
IS IT CHRONIC?
IS THERE SOMETHING -- KIND OF AN
OBSERVATION HYPOTHESIS, IS THE
VIRUS ACTUALLY LEADING TO A DOWN
REGULATION OF GENES THAT ARE
ALLOWING HEALING TO OCCUR?
A LOT OF THINGS GOING ON.
I THOUGHT THAT WAS KIND OF
INTERESTING.
OKAY.
SORRY.
[APPLAUSE]
>> I TOTALLY APPRECIATE -- ANY
ONE BURNING QUESTION BEFORE WE
MAYBE SAVE THE DISCUSSION FOR
THE PANEL M TIME?
ALL RIGHT.
WE'LL DEFINITELY COME BACK TO
YOU.
THANK YOU SO MUCH OUR NEXT
SPEAKER IS ZONDA WANG.
>> HELLO, EVERYONE.
I'M VERY HAPPY TO BE HERE,
BECAUSE I'M NEW BIOLOGIST BUT --
TODAY I'M GOING TO SHARE WITH
YOU OUR EFFORT IN DEVELOPING
UNCONVENTIONAL ANIMAL MODELS,
UNCONVENTIONAL SPECIES MODELS.
THE DEFINITION OF UNCONVENTIONAL
IS VERY LOOSE.
BASICALLY FOR ME, IT CAN MEAN
ANYTHING NOT THE -- MOUSE OR
RAT, BUT STACEY GAVE ME A VERY
GOOD DEFINITION, FANCY MODEL, SO
I'LL USE THAT FANCY MODEL FROM
NOW ON.
SO MY EXPERTISE IS IN -- AS I
SAID I'M NOT AWARE OF IT, SO MY
EXPERTISE IS SO SPECIALIZED THAT
THAT -- ON THE ANIMAL GENOME IS
THE ONLY THING I KNOW, NOTHING
ELSE.
SO I COLLABORATE WITH -- FIRST I
THANK -- I DON'T KNOW WHY IT'S
NOT GOING FORWARD.
SO I HAVE A LOT OF PEOPLE TO
THANK BUT THESE ARE SOME OF TH
THEM.
INSTEAD OF READING THEIR NIAMS,
I WOULD LIKE YOU TO SEE BILL
WOLD AND -- AMONG THE FIRST TO
USE THE -- MODEL AND MANY
VIROLOGISTS REACHED OUT TO ME OR
I REACHED OUT TO THEM TO
COLLABORATE, AND SOME OF THEM
ARE IN THE AUDIENCE.
MY GOOD FRIEND, AND PETER AND
RYAN.
SO FUNNY THING IS THAT THIS
ISN'T MY FIRST TIME TO MEET
THEM.
WE HAVE BEEN COLLABORATING, HAVE
WORKED TOGETHER, PUBLISHED
TOGETHER FOR TIMES, BUT OF
COURSE BART IS MY COLLEAGUE,
I'VE MET HIM BEFORE.
SO THE PEOPLE ON THIS PANEL ARE
ONES THAT WORK WITH ME ON --
HAMSTER MODELS AND THEN I
STARTED WORK WITH TWO
INDIVIDUAL, ALISTAIR MCGREGOR
AND DR. ANICE LOWEN, AND
RECENTLY WE ESTABLISHED A COLONY
AND ACTUALLY WE CAN HANDLE --
PRETTY WELL, THAT'S IT, NOT
TOO -- I WOULD LIKE TO
ESPECIALLY THANK STANLEY FOR
PROVIDING ME -- ABOUT 10 YEARS
AGO, SO THAT'S HOW WE MADE --
AND STEFAN FROM HAWAII, I HAVE
NOT MET HIM YET BUT HE PROVIDED
THE PIGGY BACK VECTOR.
AND THEN TONY SCHOUNZ SENT ME
MORE THAN 100 DEER MICE SO THANK
THEM AND I HAVE A -- A LOT OF
MONEY AT THE BEGINNING FROM THE
UNIVERSITY OF UTAH SCIENCE --
RESEARCH, THAT'S THE PROGRAM --
MOST MODEL I DEVELOP THROUGH
THAT -- ALL RIGHT.
SO MICE, WE'RE GOOD, WE'RE
POWERFUL, AND WHY I'M INTERESTED
IN THE FANCY MODELS.
AND I THINK -- IN THIS CASE I
THINK FAN SATISFY -- I SAID THAT
YESTERDAY I HAVE A ZOO AND I
WILL SHOW YOU AT THE END OF MY
TALK, I WANT TO MAKE A LIST BUT
TODAY I JUST ONLY SHOW YOU THE
HAMSTER.
WE TALK ABOUT HAMSTER -- AND
DRAWBACKS AND SO FORTH.
I JUST WANTED TO MENTION A
FEW -- WE HAVE NOT TOUCHED.
FIRST THE HUMAN ADENOVIRUS, THE
HAMSTER IS THE ONLY RODENT FOUND
TO BE PERMISSIVE FOR HUMAN
SPECIES, THE PARENTAL STRAIN FOR
MOST OF THE VECTORS USED TODAY.
SO IF YOU WANT AN -- VIRUS TO
CURE CHILDREN, YOU NEED TO USE
HAMSTER AS MODEL.
AND IF YOU WANT TO HAVE A --
COMPETENT -- VECTOR -- HAMSTER
MODEL, BECAUSE IN MOUSE, IT JUST
DO NOT REPLICATE.
AND ANOTHER VIRUS -- IT'S
CONSIDERED AS THE ONLY ANIMAL
MODEL THAT RESEMBLES THE
HANTAVIRUS PULMONARY SYNDROME,
ESTABLISHED AS A MODEL FOR
HORIZONTAL TRANSMISSION OF THE
AND VIRUS.
MY COLLEAGUE JOHN MORREY --
COLORADO STATE, THEY USE HAMSTER
FOR YEARS FOR WEST NILE, YELLOW
FEVER, AND TH RECENTLY EQUINE --
VIRUS, ZIKA -- IN THE HAMSTER.
THE LAST TWO VIRUS, WE TALK A
LOT, AND BASICALLY AMONG MANY
THINGS, HAMSTERS -- STRAIN OF
INFLUENZA AND ALSO TO FEW
STRAINS OF EVEN -- AND AMONG THE
GOOD FEATURES ABOUT USING THE
HAMSTER AS A MODEL FOR SARS 2,
WE ALREADY TALK A LOT BUT
BASICALLY WHAT I WANT -- HERE IS
THAT TWO -- HUMAN CONTACT --
DISTANCE -- TRANSMISSION,
PETER -- BUT THE ORIGINAL
STRAIN -- ALL RIGHT.
I WOULD LIKE TO MENTION TWO
KNOWN INFECTIOUS DISEASE THAT
HAMSTERS HAVE BEEN USED VERY
READILY.
ONE IS METABOLIC SYNDROME,
ANOTHER IS CIRCADIAN RHYTHM
DISORDER.
THE REASON I MENTION THESE TWO
DISORDERS, BECAUSE THEY ARE
HIGHLY RELEVANT TO -- INFECTIONS
BECAUSE FOR EXAMPLE FOR THE
METABOLIC SYNDROME, AND HAMST
HAMSTERS -- METABOLISM MIMIC --
AND FEW THINGS.
NUMBER ONE, CETP, CLS TROLL
ESTER TRANSFER PROTEIN, WHICH IS
VERY KEY, VERY IMPORTANT PROTEIN
IN -- HUMAN HAS IT, HAMSTER HAS
IT, BUT NO MICE AND NO RAT HAVE
IT.
SO TOTALLY DIFFERENT, AND ALSO
HEPA -- AND IN MOUSE IS A B-48.
SO IF YOU WANT TO USE -- WANT TO
STUDY METABOLIC SYNDROME AS A
CO-MORBIDITY IN VARIABLE
INFECTION I THINK IT COMES AS A
VERY GOOD MODEL.
CIRCADIAN RHYTHM DISORDER,
HAMSTERS ARE AMAZING.
THEY HAVE VERY PRECISE CIRCADIAN
RHYTHM.
IF YOU HAVE HAMSTER FEMALE TODAY
AS CIRCLE ONE, 400 DAYS LATER --
THEY ONLY RESPOND TO LIGHT, THEY
DO NOT RESPOND TO HORMONE.
VERY STRAIGHT, VERY NICE.
AND THEN I LIST -- OF THE 100
TOP SETTING -- DRUGS -- AND THEN
ABOUT 58% OF DRUGS ONLY HAVE A
HALF-LIFE SIX HOURS.
SO THIS BEGS THE QUESTION
WHEN -- WHEN YOU TEST DRUGS AND
ANIMAL MODELS -- AND ALSO
TRANSMISSION I WAS THINKING THIS
MORNING, WILL TRANSMISSION OCCUR
DIFFERENTLY AT A DIFFERENT --
SIR CADECIRCADIAN CYCLE.
SO NO ONE HAS STUDIED THIS YET.
ALL RIGHT.
FINALLY, FUN FACT ABOUT HAMSTER,
TO BE USED AS AN ANIMAL MODEL,
YOU LIKE IT A LOT.
NUMBER ONE, THEY HAVE THE
SHORTEST GESTATION AMONG ALL
THE -- MAMMALS, 16 DAYS, AND
LARGE -- SIZE.
18, 20 PUPS PER LITTER.
FEMALE HAS SOME DIFFICULT RESULT
OF THEM, AND THEN THEY'RE VERY
EASY TO HANDLE.
IN THE PAST, MORE THAN 10 YEARS,
I THINK ONLY TWO INCIDENTS GET
BITTEN BY A HAMSTER.
I DON'T KNOW WHAT THEY DO, I
KEEP TELLING THEM THAT THEY DO
NOT BITE, YOU BITE THEM, THEY DO
NOT BITE YOU.
ALL RIGHT.
ANOTHER REASON, I DON'T HAVE
TIME TO TALK IN DETAIL IS THAT
CERTAIN HUMAN SET OF HANDS
WORKING THE -- IN A MOUSE.
THE REASON I MENTION THAT IS
BECAUSE WE HAVE AN ONGOING
EFFORT TO HUMANIZE THE --
SYSTEM.
WE KNOW THAT HUMANIZED MOUSE
MODEL REALLY NOT WORKING.
THERE ARE NO -- NO GERMINAL
CENTER FORMATION AND THEN
SCIENTISTS HAVE BEEN PUTTING SO
MANY HUMAN GENES INTO THAT
MOUSE, IT'S STILL NOT WORKING
WELL.
AND THEN THESE FEW OF THESE
GENES I LISTED HERE ARE AMONG
THE GENES WE HAVE BEEN --
SCIENTISTS HAVE BEEN TRYING TO
KNOCK INTO THE MOUSE GENOME.
HAMSTER LITERALLY HAVE THEM.
ALL RIGHT.
SO THERE ARE MANY GOOD REASONS
TO -- HAMS SISTERS, BUT AT THAT
TIME, THERE ARE NO GENERAL
MODIFICATION TECHNIQUES
AVAILABLE.
SO FIRST BUSINESS FOR US IS TO
ESTABLISH -- TECHNIQUES AND
LUCKILY CRISPR CAME ALONG AND WE
QUICKLY JUMPED ON TO IT AND I
THINK ALL OF YOU HAVE HEARD A
LOT AND KNOW A LOT ABOUT CRISPR
BUT I JUST SIMPLY SAVED THAT.
YOU CA DESIGN -- TARGET ANY GENE
YOU WANT AND BRING THE CASE LINE
INTO THE GENERAL LOCUS, YOU WANT
TO TARGET WE WILL INTRODUCE --
DNA BREAKS AND THE CELL WILL
NORMALLY USE -- JOINING TO
REPAIR THE GENOME WHICH IS
ERROR-PRONE SO INTRODUCE THE
INSERTION THE DELETIONS -- AND
IF YOU WANT TO KNOW -- -- AND --
SO WHEN WE --
>> SO AT THAT TIME WHEN WE
STARTED WITH HAMSTERS AND CRISPR
ONLY WAS DEMONSTRATED TO BE
WORKING IN HUMAN AND THE MOUSE
CELLS, NOT OTHER SPECIES.
SO WE WILL NO IDEA, NOBODY HAD
NO IDEA, AND HE SAID OH, YOU
WANT TO TRY IN HAMSTER?
I DONE KNOW, MAY WORK, MAY NOT.
SO WE JUST USE KIDNEY HAMSTER
CELLS AND WE DESIGNED MULTIP
MULTIPLE -- AND DIFFERENT
DOMAINS, AND OTHER GENES AND
THEN WE BASICALLY TRANSVECT THE
PLASMA CARRYING THE -- THE NK
CELLS AND THEN WE DID THE SO
CALLED -- TERM BUT BASICALLY
WHAT WE DID IS THAT IF GENE
TARGETING HAPPENED, IN THIS CASE
FOR EXAMPLE -- WILL BE ABOLISHED
SO IT WILL BE PARTIALLY
RESISTANT TO THE -- THIS IS --
IN THIS CASE, YOU HAVE SOME DCH
RESISTANT, BASED ON THAT --
EFFICIENCY, SO WE WERE VERY,
VERY EXCITED BUT THERE'S A HUGE
PROBLEM BECAUSE WORKING WITH
HAMSTER IS DIFFERENT FROM OTHER
SPECIES.
NUMBER ONE, HAMSTERS -- ARE
EXTREMELY SENSITIVE TO
ENVIRONMENT EXPOSURE,
PARTICULARLY TO LIGHT.
THIS LIGHT WILL KEEP HAMSTER
EMBRYOS LIKE FAMILY, THEY STOP
DEVELOPING.
IT'S CRAZY.
AND NO COMMERCIAL EMBRYO
CULTURAL MEDIA WORKS FOR HAMS
HAMSTER.
SHEEP, GOAT, MOUSE, EQUINE, YOU
NAME IT, NONE OF THE MEDIA WORK
FOR HAMSTER EMBRYOS.
AND THEN THERE ARE NO --
INJECTION OR -- INJECTION
TECHNIQUE AVAILABLE.
BECAUSE WE NEED THESE TECHNIQUES
TO INTRODUCE CRISPR INTO HAMSTER
EMBRY YOES.
SO BIG CHALLENGE, BUT WE WORKED
ON IT AND THIS ONE TABLE
SUMMARIZES ABOUT ONE YEAR'S
EFFORT.
WE DID -- WHEN WE SHOULD COLLECT
THE EMBRY YOES, WHEN TO DO THE
INJECTION, HOW MUCH TIME WE CAN
KEEP ON THIS -- BY THE WAY WE
WERE -- HOW MUCH WE INJECT IS
VERY, VERY CHALLENGING, BUT WE
GOT IT DONE.
SO WE HAVE THE TECHNOLOGY
AVAILABLE AND THEN WE JUST
IMMEDIATELY APPLY TECHNOLOGY TO
HAMSTER EMBRYOS, AND THIS JUST
SHOWS YOU THAT ONE LITER OF THE
PUPS BORN AND THE YELLOW --
INJECT EMBRY YOES, THE BLACK --
MOTHER WHO ARE -- PRODUCED
BECAUSE HAMSTERS ARE TOO
SENSITIVE TO ANY MANIPULATION.
IF YOU USE A SO YOU DOUGH
PREGNANT FEMALES, WHEN YOU
CREATE EMBRY YOE WILL NOT BE
ABLE TO INITIATE PREGNANCY.
AT THAT TIME WE IMPROVED
CONDITION, NOW WE CAN USE A --
SOME OF THE PREGNANT FEMALES.
SO I JUST WANT TO SHOW YOU, THIS
ONE LITTER AND THESE TWO PUPS,
C5, C6 -- WE KNOW THEY CARRY I
IN -- REMEMBER IT'S JULY 10TH,
2013, ONLY A FEW -- YOU KNOW, A
FEW MONTHS AFTER THE CRISPR WAS
REPORTED.
THEN WE USE C6 FEMALE BECAUSE OF
CARRIER -- WHICH GENERATE A
KNOCKOUT -- ALSO KNOCKOUT SO
THEN WE BRED FEMALE WITH -- WE
FOUND ALL THE ALLELES CARRIED
ALLELE -- GERMLINE, WERE GOOD,
AND THEN WE JUST USE A RATHER --
SEPARATING THAT PRODUCE -- AND
THE COLOR HAS BEEN USED A LOT.
ALL RIGHT.
SO IN THE NEXT FEW SLIDES, I
JUST WANT TO SHOW YOU WHAT WE
CAN DO IN THE HAMSTER.
NOW I SHOW KNOCKOUT.
AS AN EXAMPLE, THIS EXAMPLE IS
THAT YOU'RE KNOCKING IN LARGE
PIECE OF DNA INTO THE JEA NOME.
IN THIS CASE LO LUCIFERASE, INTO
PEER -- WE JUST SAID -- IN
THE -- GENE RIGHT BEFORE THE
LAST STOP CODON, PEPTIDE -- AS
VIROLOGISTS YOU -- SO THE --
2 GENE AND REPORTER GENE WILL BE
EXPRESSED AS A SINGLE MRA DND SO
ANY GENE OF INTEREST CAN BE
ALMOST ANY LOCI IN THE GENOME,
AND 2A PEP TAID OR ERRORS
SEQUENCE, CAN YOU MAKE THE TRANS
GENE AND TOTAL CONTROL OF THE --
SEQUENCE.
SO WE MADE THIS AND WE MADE A
FEW OTHERS TOO.
IN A MOMENT I'M GOING TO SHOW
YOU -- HUMAN RECEPTOR INTO THE
HAMSTER.
AND THEN GENE EDITING, THERE WAS
A VERY FAMOUS TALL HAMSTER
DISCOVERED IN 1988.
ONE OF THE FIRST MAMMALIAN
MUTATION GENE MUTATION FOUND TO
BE INVOLVED IN CIRCADIAN RHYTHM.
VERY FAMOUS HAMSTER.
BUT WE LOST IT.
NOBODY COULD FIND IT.
SO THEN -- WE WORKED TOGETHER
AND SUBMITTED A PROPOSAL --
REGENERATED.
SO GENE EDITING, AND THEN WE
ALSO USE PIGGY BACK -- TRANS
GENESIS AND PRODUCE THE HACE 2.
PIGGY BACK -- IT CAN INTEGRATE
THE TRANS GENE WITH A LARGE
SLICE, MUCH LARGER THAN THE
REGULAR KNOCKINS, AND IF YOU
DON'T LIKE THE TRANS GENE FOR
SOME REASON, YOU WANT TO TAKE IT
OUT LATER, YOU CAN TRANSPOSE TO
REMOVE IT AND LEAVING NO SCARS,
SO IT'S GREAT.
ANOTHER THING ABOUT PIGGYBAC, I
WAS USING -- IN THE EARLY YEA
YEARS, -- THROUGH LIKE A
CRASHING, YOU KNOW, WITHOUT
KNOWING HOW TO GET INTO IT.
MOST CASES THE TRANS GENE COULD
BE DISRUPTED, AND YOU ALSO --
STANLEY SHOWED YOU HAVE THAT --
EIGHT COPIES, AND WE HAVE EIGHT
COPIES SOMETIMES CAUSE GENOMIC
INSTABILITY.
SO IF WE BREATHE ANIMALS, SOME
ANIMAL MAY LOSE SOME -- SO CAL
CALLED EPIGENOMIC SILENCING, BUT
MAMMALIAN CELLS AND THEY WOULD
BE -- SILENCED, BUT THE PIG
PIGGYBAC HAS THE SO-CALLED
GENOMIC INSULATOR FOR WHEN THE
TRANS GENE FORM -- AND ALSO
PREVENTED THAT SO CALLED
POSITIONAL EECT, SO WE LIKE PIG
PIGGYBAC A LOT.
SO THESE TECHNIQUES, THESE
HAMSTER MODELS WE PRODUCE.
BUT THIS IS ONLY HALF OF THEM.
A LOT.
ALL RIGHT.
THEN I JUST WANT TO USE THREE OR
FOUR EXAMPLES TO SHOW THE USE
USEFULNESS OF THE ENGINEERED
HAMSTER MODELS.
THIS ONE WE USED FOR SARS 2 AND
1 INTERESTING THING IS THAT THIS
IS VERY DRAMATIC, WE COMPARE T
THE -- VERSUS WILD TYPE
HAMSTERS, THEY'RE VERY LOW,
THOUSAND TIMES IN THIS CASE
100,000 TIMES.
SO THIS TELLS YOU THAT STAT 2 IS
VERY IMPORTANT.
BUT SURPRISINGLY, IF YOU HAVE SO
MUCH -- HAMSTER, YOU DON'T HAVE
PATHOLOGY.
SO IT'S A DOUBLE EDGED SWORD.
-- BY -- BY THE WAY I SEND THE
HAMSTER ALL OVER THE WORLD.
BELGIUM AND OTHER COUNTRIES TOO.
SO ANOTHER EXAMPLE, THEY USED TO
USE -- AS IMMUNOSUPPRESSANT
WHICH IS -- SO THEY REACH OUT TO
ME ASKING WHETHER WE CAN -- AND
WE SEND THE RAG 2 HAMSTER -- AND
AT THAT TIME WE DID NOT HAVE THE
HACE2 HAMSTER YET.
-- MORE LETHAL BUT -- SO WE
DEVELOPED A DIFFERENT MODEL AND
ALSO I WOULD LIKE TO POINT OUT
THAT IT'S NOT HERE BUT THEY
FOUND NEURAL INFECTIONS,
CARDIOVASCULAR INFECTIONS AND AT
THAT TIME WE THOUGHT MAYBE IT'S
ARTIFACT BUT LATER ON WE'RE
REALIZING THAT A LOT OF PATIENTS
GOT -- INFECTIONS AND HEART
INFORECASTS.
PETER SHOWED THIS, THE --
KNOCKOUT HAMSTER ABSENT OF NKK
CELLS AND B CELLS T CELLS
COMPROMISE.
THEY BEHAVE FROM SURVEYS JUST
LIKE WILD HAMSTERS, MEANING THEY
LOST WEIGHT AND THEY'RE JUST
SITTING THERE.
SO THIS WAS DONE IN THE ROCKY
MOUNTAIN LION, THEY THOUGHT OK
OKAY, THIS -- THE HAMSTER THERE
AND -- EUTHANIZE THE HAMSTER
AND -- VIRUS STILL THERE.
CRAZY.
AND RECENTLY, PETER DID AN
EXPERIMENT AS HE SHOW THAT UP TO
100 DAY, VIRUS STILL THERE, AND
ANIMAL CAN GAIN WEIGHT -- IN
THAT CASE, SO HEALTHY.
CRAZY.
SO NK CELLS, VERY IMPORTANT FOR
CLEARING THE VIRUS.
THEN THIS IS A RESPIRATORY
VIRUS, JUNIN VIRUS, USE TRF1 AS
RECEPTOR AND -- JUST CANNOT BE
AFFECTED.
SO WE KNOCKED IN THROUGH THIS
KNOCK-IN CRISPR MEDIAN KNOCK-IN,
WE PUT THE HUMAN -- SO -- IT
GOOD, AND SO THAT WHEN WE INFECT
THE HAMSTER YOU SEE MODEL 33%
AND MIMICKING HUMAN MORTALITY,
AND ALSO WE SEE DIFFERENT --
NEUROPATHOLOGY IS ON THE -- AND
ALSO TREMORS.
I HAVE REALLY WHAT I -- BECAUSE
I WAS AFRAID IT WON'T WORK.
ALL RIGHT.
ANOTHER EXAMPLE OF A JEAN -- I'M
SORRY.
THIS IS A -- FOR HAMSTER -- FO
FOR -- LAB AND HIS LAB -- ALL
ABOUT CHEMISTRY, PCPHY AS
RECEPTOR, CONFIRM -- AND HAMSTER
IS ONLY A RODENT FOUND TO BE
SUSCEPTIBLE TO HANTAVIRUS.
AND WE WORKED TOGETHER, NUMBER
ONE, WE DELETE A DOMAIN THEY
BELIEVE IS -- CHEMISTRY THAT
THIS DOMAIN IS IMPORTANT FOR
HANTAVIRUS ATTACHMENT TO THE
CELL.
WHEN WE LOOKED AT THIS DOMAIN,
BOOM, TURN OUT THAT THE HAMSTER
100% PROTECTED FOR HANTAVIRUS
INFECTION AND THEY ALSO NARROWED
DOWN THE TWO IMMUNOASSAY --
BOOM, AND HEMISPHERES WERE
LARGELY PROTECTED.
MEANING THAT THESE TWO -- FOR
HANTAVIRUS INFECTION.
ALL RIGHT, HACE2 HAMSTERS --
ALSO -- I JUST WANTED TO SHOW
THAT.
TURN OUT TO BE A LETHAL MODEL.
BUT -- PUBLICLY INFORMED OVER --
A FEW MONTHS LATER.
ALL RIGHT.
SO WITH UNDUE -- ONE OF GENES,
WHAT ARE WE UP TO.
SO I THINK THE FIRST THING --
KNOCKOUT -- GENES, WHEN WE --
GENES, -- IMPORTANT FOR -- AND
ALSO GENE ESPECIALLY FOR CERT
CERTAIN -- CELL, LYMPHOID -- I
SHOULD -- AND OTHER GENE
IMPORTANT FOR USING HAMSTER AS A
HOST FOR HUMAN HEMATOPOIETIC --
BIG PROJECT ONGOING.
AND OTHER TYPES OF GENOME
MODIFICATION IN ADDITION TO
KNOCK IN, KNOCKOUT EDITING, WE
WANT TO -- SPECIFIC AND
SEX-SPECIFIC, I THINK HUMAN
GENOME IS ABOUT 5, 6% GENES
SEX-BIAS AND -- ALSO REPORTER --
AND THEN WOULD LIKE TO PRODUCE
ANTIBODS, THIS IS A WISH LIST.
I SURVEYED -- BUYOLOGIST AND WE
CAME UP WITH A LONG LIST OF THE
ANTIBODY GENES AND WE SUBMIT A
PROPOSAL BUT IT WAS REJECTED
BECAUSE THE -- DON'T THINK
HAMSTER -- IS THAT CRITICAL
BECAUSE -- MICE FANTASTIC.
SO ANYWAY, WE HAVE NOT DONE THAT
YET.
BUT I CAN SAY THAT I HOPE IN A
YEAR OR TWO EVEN, WE SHOULD NOT
TALK ABOUT -- MENTION REAGENT IS
A LIMITATION BECAUSE IT'S NOT A
BIG DEAL.
-- SUPPORT THE RESEARCH, I
THINK A COUPLE OF R24, PROBLEM
SOLVED.
WHEN I DID MY PH.D., I MADE A
HUNDRED -- AND I MANUALLY
SEQUENCED -- I DON'T KNOW HOW
MANY SEQUENCES, RIGHT, BIG FIGHT
TO INCLUDE.
SO ALL RIGHT.
I SAID I HAVE A ZOO, AND I WANT
TO SHOW YOU THE OTHERS.
-- DMI, WHITE FOOT MICE,
PRODUCE -- MODELS AND HERE'S THE
LIST.
SOME OF THEM HAVE BEEN USED.
AND THEN THE OTHER SPECIES I'M
TRYING TO WORK ON.
NO, I'M WORKING ON, SORRY.
URBAN I HAVE ABOUT 300 NAKED
MOLE RATS IN MY LAB.
VERY CUTE ANIMALS.
ACTUALLY THEY CAN BE UP TO
30-SOME YEARS.
THEY NEVER DEVELOP -- THAT'S
VERY INTERESTING.
NO OXYGEN, MICE, 32ND, GET
KILLED.
BUT THIS -- AMAZING HOW THEY DO
THAT.
IF YOU WANT TO STUDY STROKE, HOW
TO PROTECT A PATIENT FROM
ACCIDENT -- DAMAGE, GOOD ONE.
SO CALIFORNIA MICE, THEY ARE
NATURAL HOSTS FOR HANTAVIRUS AND
ALSO OTHER -- MANY OTHER
ZOONOTIC PATHOGENS.
AND THEY HAVE VERY INTERESTING
SOCIAL PAIR BONDING.
THEY ARE MONOGAMOUS TOGETHER.
YOU CANNOT SEPARATE THEM, THEY
WILL NOT ACCEPT A NEW PARTNER AT
ALL.
COTTON RAT, RECENTLY WE GOT
THEM.
GOOD MODEL FOR RSV, WE ALL HEARD
ABOUT THAT, MEASLES, HERPES
SIMPLEX VIRUS, AND PARTICULARLY
A THING WE WOULD LIKE TO WORK ON
IS THE CONNECTION BETWEEN
DEMENTIA ESPECIALLY, AND VIRAL
INFECTION.
THERE IS SO APPLICATION, WE HAVE
HAD INDICATION, SO WE WOULD LIKE
TO INTRODUCE SOME OF THE HUMAN
ALLELES WHICH MAKE IT SO THE
POPULATION TO DEVELOP -- DISEASE
TO 80 AGE, SOMETHING LIKE THAT.
SO WE ARE WORKING ON THAT.
AGAIN, WE LIKE -- DON'T BE
AGGRESSIVE BUT -- THESE THINK
ABOUT AND ALREADY IN DISCUSSIONS
LIKE -- FROM COLORADO STATE FOR
TWO OR THREE YEARS.
HE HAS -- HE SA SENT TO ME -- AD
WE DID SOME OTHER THINGS.
I ALSO TEAM UP WITH A HUSBAND
AND WIFE TEAM FROM MY
UNIVERSITY, THEY DO FIELD
STUDIES.
THEY ARE VERY EXCITED, THEY ARE
GOING TO HAVE ME TRAP
WHITETAILED ANTELOPE GROUND
SQUIRRELS.
WHAT KIND OF PATHOGEN THEY
CARRY, WE DON'T KNOW YET.
BUT THEY ARE DIURNAL, AS I
MENTIONED CIRCADIAN RHYTHM VERY
IMPORTANT IN TERMS OF VIRAL
INFECTION.
YOU KNOW, MAYBE IT'S GOOD MODEL
FOR VIRAL INFECTION TOO.
JUMPING MICE, THEY ARE
SEROPOSITIVE FOR SNV AND THEY
HIBERNATE.
I TALK WITH -- THEY HAVE THE
ANIMALS, AND ANOTHER SPECIES,
SPECIES X, I HOPE YOU GUYS CAN
GIVE ME SOME CLUE.
SO PRESENT -- THAT WORKING WITH
THTHE FANCY -- THEY DON'T HAVE -
ES OR IPS CELLS, ALSO LACK OF
ANIMAL HOUSING CAPABILITIES,
STANLEY, THIS WAS 10 YEARS AGO,
YOU DON'T HAVE CAPACITY TO HOUSE
HAMSTERS.
AND THEN RESEARCH A LOT OF MICE
AND -- DO NOT WANT TO GET INTO
TOO MUCH -- FOR THE PROPOSAL I
SAW NEW STUDY SECTIONS -- NO.
SO I THINK THAT JUST SHOW THE
FUNDING MECHANISM AS A - BATON O
DIRECT THIS.
SO I HOPE THAT WILL HAPPEN.
ALL RIGHT.
A SUMMARY.
YOU CAN READ IT.
IT'S VERY SIMPLE.
ALL RIGHT.
THAT'S ALL FOR ME.
ALL RIGHT.
>> THAT WAS REALLY GREAT.
MAYBE WE'LL GO STRAIGHT TO THE
PANEL DISCUSSION AND THEN IF
PEOPLE -- BART, IF YOU WANT TO
COME UP, WE'LL JUST DO THE PANEL
DISCUSSION NOW.
SO TO ANSWER BROADER QUESTIONS
IF ANYBODY HAS ONE, PLEASE GO
AHEAD.
>> I JUST HAVE A COMMENT FOR
ZONDA.
YOU HAD ON YOUR LIST FOR GUINEA
BIG REAGENTS FOR TGF BETA.
IF YOU'RE LOOKING AT TGF BETA,
THE ACTIVE FORM OF IT, THE
HUMAN -- AND REAGENTS WILL WORK
TBEUT BUTTEFULLY BECAUSE IT'S
ABOUT 99% CONSERVED ACROSS ALL
THE SPECIES, SO I WOULD GIVE
THEM A TRY, THAT'S WORKED REALLY
WELL FOR US AND EVERYTHING.
>> ANY QUESTIONS? I'M HAPPY TO
START.
I THINK AGAIN, YOU BOTH HAVE
REALLY UNIQUE PERSPECTIVES ON
SMALL ANIMAL MODELS, AND I WAS
WONDERING IF YOU BOTH WANTED TO
SORT OF ADDRESS KIND OF GAPS
THAT YOU ZIEBART IN TERMS OF
WHAT -- AS NEW PATHOGENS EMERGE
AND THE DEVELOPMENT OF ANIMAL
MODELS NEEDS TO HAPPEN IN TERMS
OF THE REGULATORY SPACE IN
GETTING LICENSURE OF VACCINES
AND SMALL DRUG TARGETS, HOW THAT
IS PERCEIVED WHEN YOU DON'T HAVE
A WEALTH OF INFORMATION.
AND THEN ZONDA, WHEN THE NEW
PATHOGEN EMERGES, HOW DO YOU
PICK A MODEL SYSTEM?
IT WAS REALLY GREAT IF YOU CAN
TOUCH ON THOSE TOPICS A LITTLE
BIT.
>> FOR ME TO CHOOSE AN ANIMAL
MODEL, THERE'S A DISEASE AND
CERTAIN VIRUS -- WORKING WITH
THAT SPECIES AND THEY HAVE A
NEED TO KNOW WHAT THE CAUSE
EFFECTS ARE IMPORTANT.
SO THEN WE WORK TOGETHER.
THE GENE AND -- IN TERMS OF
WHICH MODEL TO CHOOSE FOR
RESEARCH VIRUS, I HAVE NO IDEA.
I JUST RELY ON YOU GUYS.
>> MY GOODNESS.
WE'RE JUST LUCKY.
JUST FROM PREVIOUS EXPERIENCE
REALLY JUST LOOKING WHAT'S OUT
THERE.
I KNOW I HAD A GRADUATE STUDENT
PROBABLY THREE YEARS BEFORE WE
HAD FUNDING FROM THE NIH, ON --
ALREADY COMING UP IN THE
LITERATURE, JUST WATCHING SOME
OF THE EMERGING INFECTION --
SOCIAL MEDIA IDENTIFYING WHAT IS
OUT THERE UNDER UNIQUE DISEASES,
DISEASES WE DON'T UNDERSTAND,
AND JUST KIND OF --
>> BART, YOU KIND OF TOUCHED ON
THIS AND IT'S SOMETHING ACTUALLY
WE HAVE NOT TALKED ABOUT IN THIS
WORKSHOP, AND THAT'S THE
IMPORTANCE OF THOROUGHLY AND
PROPERLY DESCRIBING THE
TECHNIQUES THAT YOU USED IN YOUR
PUBLICATION, RIGHT?
SO WOULD YOU JUST EXPAND ON THAT
A LITTLE BIT?
A LOT OF TIMES I THINK PEOPLE
SAY REFER TO PRIOR PAPER OR
SOMETHING ELSE.
AND THE INFORMATION IS NOT
THERE.
SO DO YOU HAVE RECOMMENDATIONS
FOR HOW TO APPROACH THAT FOR
EVERYBODY?
>> THE WHOLE IDEA OF SHOWING THE
COMBINATION DRUG STUDIES AND THE
WEIGH LOSS CUTOFF, I'M NOT
AGAINST ANIMAL WELFARE, REALLY,
I'M FOR ANIMAL WELFARE.
BUT I ALSO THINK WE NEED TO MAKE
SAWYER THAT THSURE THAT THE DATE
GENERATING IS GOOD DATA.
EARLY ON IN THE PANDEMIC, WE HAD
A DISCUSSION, RESPIRATORY
DISEASE BRANCH PROJECT OFFICER,
ST. JEUDZ -- WISCONSIN, AND WE
WERE TALKING ABOUT REALLY
AGREEING ON THE MODEL AND REALLY
JUST A WEIGHT LOSS CUTOFF, AND
EVEN THAT WAS A LITTLE BIT OF A
CHALLENGE, BECAUSE ST. JEUDZ
SAID THEIR -- WOULD NOT LET HIM
GO ANY HIGHER THAN A 20% WEIGHT
LOSS.
SO IT KIND OF A CHALLENGE.
THESE ARE UNREGULATED SPECIES,
THAT DOESN'T MEAN I DON'T CARE
ABOUT THEM.
I DO CARE ABOUT THEM.
I'VE NEVER SEEN ANIMALS
MISTREATED IN THE LAB, THANK
GOODNESS, AND I DON'T THINK WE
SHOULD.
PEOPLE PAT US ON THE BACK THAT
WE GET SO MUCH INFORMATION OUT
OF A STUDY THAT WE RUN, ANY
INFORMATION WE CAN GET FROM AN
ANIMAL STUDY, BUT I THINK THERE
NEEDS TO BE SOME AGREEMENT AS TO
KIND OF A STANDARD STUDY.
I GUESS WHEN I SAY THAT, I DON'T
WANT TO BE THE ONE MAKING THE
RULES BECAUSE I THINK EVERYBODY
NEEDS TO BE ABLE TO HAVE THE
FREEDOM TO TRY SOMETHING
DIFFERENT.
BUT THE PEOPLE OVER IN ICUC
COMMITTEE, ARE THEY TELLING THE
SCIENTISTS HOW THEY SHOULD BE
DOING THE STUDY?
I WOULD DISAGREE WITH THAT.
DOES THAT MAKE SENSE?
IT BEEN A HUGE CONCERN FOR ME
FOR -- ACTUALLY WHEN I FIRST
CAME TO USU FOR 15 YEARS, JUST
THIS IDEA THAT I WILL READ A
PAPER ON INFLUENZA AND IT'S A
DIFFERENT MOUSE STRAIN AND A
DIFFERENCE VIRUS AND THEY'RE
SAYING MORTALITY AND THEY DON'T
DESCRIBE WEIGHT LOSS CUTOFF.
THAT SHOULD BE JUST BASIC
INFORMATION IN THERE.
THEN I CAN IS A YOU'RE USING A
20%.
I UNDERSTAND THAT.
AND SO IT'S ALMOST -- WE CAN'T
FORCE PEOPLE DO THAT KIND OF AN
AGREEMENT.
I GUESS I WAS TRAINED LONG AGO,
BUT IF I'M PUBLISHING A METHOD,
I SHOULD PUBLISH THAT METHOD SO
YOU CAN REPRODUCE WHAT I'VE JUST
DONE, AND I'VE GIVEN YOU ALL THE
DETAILS.
AND IT'S ONE OF THOSE I GUESS
GOES WITHOUT ASKING, AND IF I'M
USING A REAGENT, BY PUBLISHING,
I'M SAYING I WILL PROVIDE THAT
REAGENT TO YOU SO THAT YOU CAN
REPRODUCE WHAT I'VE JUST DONE.
MAYBE THIS NEEDS TO BE REVIEWED
FOR YOUNGER SCIENTISTS.
I DON'T KNOW.
AND THAT CAN BE CHALLENGING.
IF IT'S AN EXPENSIVE MONOCLONAL,
WE CAN'T -- YOU KNOW, BUT DOES
THAT MAKE SENSE?
SOMEONE ELSE PLEASE TALK.
>> SO YOUR COMMENT ABOUT WHEN
YOU PUBLISH THOSE REAGENTS,
YOU'RE SAYING THOSE REAGENTS ARE
AVAILABLE, AND THAT'S PART OF
THAT AGREEMENT, RIGHT?
AND IT DOESN'T ALWAYS HAPPEN.
WE ALL KNOW THAT, RIGHT?
OR YOU REACH OUT AND THE
REAGENTS ARE NOT WHAT THEY SAID
THEY WERE AND I CAN SAY AT LEAST
FROM GENERAL VIROLOGY AND ASM
THAT THE PUBLISHERS ARE REALLY
PUSHING BACK ON THIS.
SO IF WE GET AN EMAIL, WE JUST
GOT ONE FROM A GRADUATE STUDENT
SOMEWHERE THAT WAS TRYING TO GET
ACCESS TO SOMETHING THAT SHOULD
HAVE BEEN AVAILABLE AND THE
AUTHORS JUST COMPLETELY IGNORED
THEM, AND ASM SENT THE ETHICS
TEAM AFTER THEM AND WHEN THEY
WOULDN'T COMPLY, THEY CONTACTED
NIH BECAUSE IT WAS NIH-FUNDED
STUDY.
SO THESE THINGS ARE BEING TAKEN
SERIOUSLY.
WHEN YOU PUBLISH AND YOU SAY
THAT OPEN ACCESS AND OPEN
RESOURCES, THEY'RE GOING TO HOLD
YOU TO IT, WHICH I THINK IS
GOOD.
>> RYAN?
>> ZONDA, GREAT TALK.
CAN I ASK A QUESTION ABOUT --
THESE MODELS ARE BEAUTIFUL AND I
THINK THEY'RE SUPER IMPORTANT.
A LOT OF THESE ARE OUTBRED AND
THEY'RE WILD ANIMALS.
WHAT KIND OF ADDITIONAL TOOLS
AND STANDARDIZATIONS WOULD YOU
LIKE TO SEE FOR ALL OF THESE?
I MEAN, THEY'RE PROBABLY NOT
INBRED ALL OF THESE, LIKE YOUR
WESTERN JUMPING MICE WON'T BE
INBRED RIGHT AWAY.
ARE THERE HIGH QUALITY GENOMES
FOR THESE THINGS AVAILABLE?
DO WE NEED TO EVALUATE
MICROBIOME CHANGES THAT EXIST
ACROSS THESE DIFFERENT ANIMALS
AND SORT OF WHAT HELP DO YOU
NEED DO THAT OR WHAT THINGS DO
YOU THINK ARE NECESSARY AS
YOU'RE DEVELOPING THESE MODELS
TO STUDY A DISEASE?
>> GREAT QUESTION.
I THINK A LOT NEEDS TO BE DONE.
AT THIS MOMENT, WHEN WE GET
ABOUT -- CAUGHT ANIMAL, WE JUST
DO A PANEL OF PATHOGENS, WHICH
ARE THE CONCERN OF THE VET, NOT
MY CONCERN ACTUALLY.
AND THEN WE SAY OKAY, WE IMPORT
THEM, BUT IN TERMS OF THEIR
GENOME, I THINK THE -- RIGHT NOW
IS OF A RARE GENE, WE GOT THAT
DOWN PRETTY QUICKLY.
ALSO WHETHER WE WANT TO KEEP
INBRED OR OUTBRED.
SO FOR US WHAT DO WE TRY TO DO
THAT, EVEN THE HAMSTER.
WHEN YOU PRODUCE A FOUNDER LINE,
YOU JUST USE THAT FOUNDER LINE,
IT WILL BECOME INBRED.
THIS COULD BE A GOOD THING OR A
BAD THING, WE DON'T KNOW.
SO WE TEND TO HAVE MODEL
FOUNDERS AND THEN PERIODICALLY
BREED WITH FRESH -- RIGHT NOW WE
ARE TRYING TO KEEP THEM AS -- AS
POSSIBLE, WE HAVE NOT TRIED TO
PRODUCE AN --
>> JUST QUICKLY TO THE GENOME.
SO WHEN YOU HAVE A TOTALLY NEW
ONE LIKE THE WESTERN JUMPING
MICE OR ANTELOPE SQUIRRELS, DO
YOU AN TATE THE GENOME --
>> NO.
FOR US, WE JUST DESIGN --
PARAMETERS.
FIRST WE JUST LOOK AT -- GENE --
FOR EXAMPLE AND LOOK AT --
WHATEVER YOU HAVE IT, WE COMPARE
IT.
IF -- SEQUENCE -- WE SEQUENCE
IT, MAKE SURE THAT THERE ARE NO
POLYMORPHISM, SO TO GET OUR --
WE DESIGN TARGET THE SEQUENCE,
AND -- WE JUST -- AMPLIFY THE
SEQUENCE LIKE THAT.
>> YOU KNOW, BECAUSE THERE'S A
FEW OF THESE CONSORTIA THAT ARE
OUT THERE LIKE THE VERTEBRA GENE
PROJECT AND SONOMIA, I WONDER IF
YOU KNOW IF NID OR NIH OR A
FUNDING AGENCY WOULD BE WILLING
TO HELP COVER THE COST OF
DEVELOPING ALL THESE MODELS AND
GETTING JOE NOMES DONE.
I THINK THE SEQUENCING IS NOT
TERRIBLY EXPENSIVE BUT THE
BIOINFORMATICS IS QUITE
TIME-CONSUMING ESPECIALLY FOR
TRULY NOVEL GENOME.
SO I THINK HAVING A PIPE LOIN TO
DO THIS, IT WOULD BE GREAT --
THEY CHARGE YOU FOR THEM, IT'S -
>> ABSOLUTELY.
YOU'RE LUCKY --
>> FANTASTIC.
I'LL ALLOW IT.
>> SO I HAVE A QUESTION FOR
ZONDA AS WELL.
I THINK YOU BRIEFLY TOUCHED UPON
HOW SENSITIVE, THE SYRIAN
HAMSTERS ARE TOO LIGHT.
MOST OF US PERFORM EXPERIMENTS
DURING DAY TIME, AND I'M KIND OF
CURIOUS HOW DO YOU THINK THAT
MAY POTENTIALLY AFFECT THE
OUTCOME OF THE EXPERIMENT?
>> THE EMBRYOS ARE SENSITIVE.
THE ANIMALS ARE NOT.
BUT THEY ARE SENSITIVE TO PHOTO
PERIOD, MEANING THAT THEIR SIR
CADDIAN RHYTHM -- WE HAVE FEW
OCCASION -- SOMEHOW THE LIGHT
JUST DOES NOT TURN OFF OR TURN
ON AND THEN PULL -- THE EXTRA
CYCLE AND TAKE AT LEAST ABOUT A
MONTH FOR FEMALE TO RECOVER.
SPO FSO FOR ADULT ANIMALS -- BUI
THINK THE CIRCADIAN RHYTHM WOULD
REALLY PROVIDE A VERY UNIQUE WAY
FOR US TO TETHER VACCINE -- AT
WHAT STATE OF CIRCADIAN RHYTHM
IS IMPORTANT OR NOT IMPORTANT AT
ALL.
>> ARE THERE ANY QUESTIONS
ONLINE OR IN THE CHAT ROOM?
GO AHEAD, MICHELLE.
>> I HAVE TWO HAMSTER QUESTIONS.
THE FIRST IS IF WE'RE DEVELOPING
REAGENTS AGAINST SYRIAN
HAMSTERS, IS THERE ANY
EXPECTATION THAT THOSE WILL BE
CROSS-REACTIVE IN OTHER HAMSTER
SPECIES?
>> NOT EXACTLY.
BECAUSE THEY ARE QUITE
DIFFERENT, HAMSTER AND -- MANY
YEARS.
WE HAVE TO TEST --
>> WHEN WE WERE WORKING ON THE
SARS 2 MODEL IN THE HA
HACE2 HAMSTER I THINK WE
EVALUATED EVERY CYTOKINE KIT
THAT WAS AVAILABLE ON THE MARKET
FOR HAMSTER TO SEE IF IT WOULD
WORK.
NONE OF THEM DID.
>> THAT'S POSH INFORMATION.
THAT'S IMPORTANT INFORMATION.
MY SECOND QUESTION IS,
THROUGHOUT THE DAY TODAY, I'VE
HEARD SOME PEOPLE SAY THAT
HAMSTERS ARE RELATIVELY CHEAP
AND SOME PEOPLE SAY THAT THEY'RE
EXPENSIVE MODELS.
SO I KNOW THIS IS PROBABLY
SUBJECTIVE DEPENDING ON THE
PERSON, BUT SO HOW DO WE SORT OF
QUANTIFY THAT AND DECIDE, YOU
KNOW, THE EXPENSE?
>> WHEN YOU BUY A HAMSTER FROM
A -- THEY CHARGE MORE THAN MICE.
BUT HAMSTER REPRODUCE SO FAST, I
ANTICIPATE IF YOU HAVE A PAIR OF
HAMSTER, MALE/FEMALE AND A PAIR
OF MICE IN THE SAME ROOM, WITHIN
TWO YEARS, MORE THAN 95% OF THE
ANIMAL WILL BE HAMSTERS.
THEY REPRODUCE AT LIKE CRAZY,
RIGHT?
GESTATION TIME -- AND WHY
HAMSTER -- WE USE -- LARGER
CAGING, JUST -- IF I WANT TO
SELL PETS TO MAKE A LIVING, I
WOULD LIKE TO WORK WITH HAMSTER
THAN WORK WITH MICE.
SO IT SHOULD NOT BE THAT
EXPENSIVE.
IT'S THE MO NOP LI MONOPOLY, I .
BUT WE TRIED A LOT FOR HAMST
HAMSTER -- SO THAT'S WHY THE
HAMSTER I PRODUCE ARE
EXPENSIVE -- BUT IT'S OKAY.
>> THIS IS FOR BART.
I GO BACK TO IN MY PREVIOUS LIKE
I USED TO BE A STRUCTURE BASED
DRUG DESIGN PERSON SO I ASK YOU
ABOUT DRUGS.
SO ONE PROBLEM IN GOING FROM
ANIMALS TO HUMANS IN THE
PHARMACOKINETICS AND DYNAMICS
LIKE FOR SOME CANCER DRUGS,
HUMAN CELLS PUMPED THEM OUT,
WHERE THEY WAY WORK IN MICE.
SO IN YOUR FIELD OF LET'S SAY
ANTIVIRUS, IS THERE ANY LESSONS
LEARNED FROM THOSE THAT WORK IN
ANIMALS, WORK IN HUMANS, OR
DON'T WORK IN HUMANS BUT WORK IN
ANIMALS?
IS THERE SOME TYPE OF
PHARMOKINETIC DYNAMIC MAP, DOES
THIS PARTICULAR STRUCTURE -- IT
WILL BE GOOD BECAUSE THIS MOUSE
HAS THESE TRANSPORTERS OR THESE
SERUM PROTEINS.
IS THERE ANY WAY TO KIND OF
PREDICT AND PUT IN A COMPUTER IF
WE KNOW HOW DRUGS ARE
TRANSPORTED IN OR OUT OF SALES
OR TAKEN UP IN THE MUCOSA OR
THINGS LIKE THAT?
>> GOOD QUESTION.
THE TWO THINGS I SEE ARE GOING
TO KILL A DRU DRUG IN DEVELOPME,
TOXICITY, SOLUBILITY.
IS IT GOING TO BE TOXIC IN THE
HOST, AND IS IT SOLUBLE.
I THINK PROBABLY HALF OF THE
ANTIVIRALS THAT WE EVALUATE,
SPONSORS THROUGH THE ANTIVIRAL
TESTING PROGRAMMING, HAVE
SOLUBILITY ISSUES, AND WE HAVE
LITTLE TRICKS, DID I SAY -- DI
SAY THAT CORRECTLY?
HALF THE STUDIES FAILED.
AND I SAY THAT, ARE PROBABLY
SOLUBILITY ISSUES.
A LOT.
IN FACT, MY LAST --
COMMUNICATION, WE HAVE A LOT OF
LIKE I SAID TIPS AND TRICKS.
SO A DRUG THAT IS NOT VERY
SOLUBLE, YOU CAN -- ORGANIC
SOLVENT AND TAKE THAT -- GO 10%
DMSO, SIZE THE DRUG, THEN GO
INTO -- I CAN TO THAT RAW TEE
TEENLY.
SONICATION HEAT, THESE ARE ALL
THINGS WE MIGHT -- CAN USE ON A
SMALL SCALE BUT NOW FOR
PHARMACEUTICAL DEVELOPMENT,
MANUFACTURING AND LICENSING.
SO SOLUBILITY IS JUST TOP.
GOING FROM AN ANIMAL TO HUMAN
DOSING, IT SOUNDS REALLY WEIRD.
THERE ACTUALLY ARE SOME METRICS
THAT ARE USED, SOME CONVERSIONS,
THEY GO BY SURFACE AREA.
SO THE SURFACE AREA -- THE SKIN
OF A MOUSE COMPARED TO THE SKIN
OF A HUMAN, AND THEY'VE KIND O
OF -- A LITTLE BIT OF NONSENSE
TO ME BUT THAT'S WHAT IT USED,
IS IT ACCURATE?
NOT ALWAYS.
I WILL HAVE A -- OR IT DID WORK
IN VITRO, WHY DIDN'T IT WORK IN
THE ANIMAL?
OFTEN -- HOW SOLUBLE IS IT, HOW
MANY MEMBRANES DOES IT NEED TO
CROSS?
BECAUSE THAT'S WHERE A DRUG WILL
NOT BE ABLE TO CAUSE THAT
MEMBRANE -- IT WILL JUST STOP,
OF ACTIVITY.
THINK ABOUT TAKING A PILL
THROUGH THE -- OF THE STOMACH,
GETS IN THE INTESTINE, HAS TO GO
THROUGH THE INTESTINAL WALL INTO
THE BLOOD, NOW I WANT TO GET TO
THE SURFACE OF THE LUNG, THAT'S
TOUGH.
NOW IT GOT TO COME BACK NA THE E
LUNG THROUGH THE LUNG-BLOOD
BARRIER, AND IN THE CASE OF
RESPIRATORY DISEASE WHERE
THERE'S INFLAMMATION, THERE'S
CONSOLIDATION, HOW DO IS THAT DG
GOING TO GET TO THE SURFACE WHEN
EVERYTHING IN THE LUNGS IS
FIGHTING AGAINST IT?
YOU CAN SAY NEBULIZER, BUT IF
YOU HAVE NEBULIZER, YOU'RE NOT
GOING TO GET IT EITHER.
SO THE LUNG IS A CHALLENGING
LOCATION TO TRY TO GET A DRUG --
DID THAT MAKE SENSE?
>> SO QUESTION FROM ONLINE, THIS
IS FOR ZONDA.
HAVE YOU SEEN WHETHER THE ESTRUS
CYCLE IMPACT VIRAL INFECTION IN
YOUR MODELS LIKE THE INFLUENZA
VIRUS IN GUINEA PIGS?
>> THAT'S A GREAT QUESTION.
I HAVEN'T -- ANY ANIMALS, SO MY
COLLABORATORS, I HAVE IN
DISCUSSION WITH SOME OF THEM,
BUT THIS HAS NOT BEEN DONE YET.
I THINK VERY -- EFFECT AS CYCLE
ALSO CIRCADIAN RHYTHM WOULD BE
VERY IMPORTANT AND I HAVE A
REPORTER HAMSTER THAT WILL MAKE
THAT -- THE MODEL ALIGN VERY
CONVENIENT TO MONITOR THE
CIRCADIAN RHYTHM CHANGE.
GOOD QUESTION.
>> I WANTED TO ASK ONE MORE
QUESTION TOO ABOUT THAT DELIVERY
WERE YOU JUST MENTIONING AND I
THINK THIS IS REALLY
INTERESTING.
SO HOW DO YOU ENVISION OR HOW --
WHAT HAVE YOU LEARNED MAYBE
LESSONS LEARNED ABOUT DIFFERENT
DELIVERY MODALITIES?
OBVIOUSLY NEB LIIZATION OF
STAGED THERAPY IS A BIG THING
NOW TO GET THEM TO THE LUNG, SO
TALKING ABOUT DELIVERY TO
MUCOSAL SITES.
THEN I THINK THIS DOVETAILS
MAYBE TO SORT OF -- ABOUT
MUCOSAL IMMUNITY, TARGETING --
AND -- DRUGS TO MUCOSAL SITES?
>> IN ANIMALS, IN PEOPLE AS
WELL, THEY REALLY NEED TO BE
ADMINISTERED WITHIN 48 HOURS.
HOW MANY PEOPLE ARE GOING TO
KNOW THEY'RE INFECTED WITH FLU
WITHIN 48 HOURS OF BEING
INFECTED?
I'M GOING TO WAIT UNTIL I'M
REALLY SICK BUT I'M NOT GOING TO
GO TO THE DOCTOR.
SO IT'S A CHALLENGE WITH THE
LUNG.
IT'S ALMOST GOT TO BE THERE
EARLY.
SO IS THAT PROPHYLACTICALLY OR
IS IT VACCINATION?
I MEAN, THE VACCINE INDUSTRY,
THERE USED TO BE A HUGE -- I
REMEMBER IT WAS BLUE, A BOOK OF
MUCOSAL IMMUNOLOGY.
IT WAS PROBABLY THIS THICK.
THIS IS THE FUTURE!
YOU KNOW, THIS -- AND IT JUST --
THE THINGS THAT WE'VE WORKED ON
FOR ABOUT 20 YEARS, IT REALLY
HASN'T WORKED OUT VERY WELL.
INTERESTING, THOUGH, ON VIRUS
SPREADING, ON THE VETERINARY
SIDE, WE WORKED ON A
BORDETELLA -- VACCINE FOR
PUPPIES, SO THERE'S THE
RESPIRATORY BACTERIAL DISEASE IN
DOGS, AND WE ACTUALLY DEVELOPED
A LICENSURE SO IT WAS ONSET OF
IMMUNITY IT WOULD BE LOOKED AT.
IF I KNOW THAT I HAVE TO TAKE --
I'M GOING ON VACATION, I'M GOING
TO TAKE MY DOG TO THE KENNEL, IF
I VACCINATE IT AT 48 HOURS
BEFORE TAKING TO THE KENNEL, WE
SHOWED THAT THE VACCINE WAS
EFFICACIOUS IN PROTECTING THE
ANIMAL FROM GETTING THAT
INFECTION.
THINKING ABOUT THESE
TRANSMISSION STUDYS THAT HAVE
BEEN TALKED ABOUT, THAT'S KIND
OF COOL, BUT THERE WAS AN ONSET
OF IMMUNITY, WE HAD THE CLAIM,
48 HOURS.
SO THERE IS MUCOSAL ANTIBODY
COMING UP VERY RAPIDLY.
THAT'S IN A DOG.
I DON'T KNOW IF THEY'RE MAGIC.
I THINK WE COULD -- THAT'S
PROBABLY THE DIRECTION I WOULD
GO FOR HUMAN VACCINES.
KIND OF ONE MORE LITTLE TRIVIA
HERE, SO THINKING ABOUT OKAY,
THE FLUMIST VACCINE.
I WILL OFTEN TEACH WHEN I'M
TALKING ABOUT VACCINES, SAFETY,
EFFICACY, SAFETY, EFFICACY.
A KILL VACCINE, VERY SAFE.
-- ADMINISTERED, DEVELOPS
SOMETIMES LIFELONG SYSTEMIC
IMMUNITY, NOTHING AT THE MUCOSAL
SURFACE.
FLUMIST VACCINE, ADMINISTERED MU
KOMUCOSALLY, ADMINISTERS VERY GD
IGA TYPE MUCOSAL SURFACES, BUT
THE MUCOSAL IMMUNE SYSTEM IS
SHORT-LIVED.
YOU KNOW, SIX MONTHS A YEAR.
SO I'D HAVE TO KEEP GETTING
VACCINATED OVER AND OVER AGAIN
TO HAVE THAT MUCOSAL ANTIBODY
THERE.
SO WE GET SAFETY, EFFICACY.
I TEACH IN THE VET SCHOOL, I
TELL THE VETS, YOU NEED TO
TALK -- WHAT IS YOUR PATIENT
GOING TO NEED, SAFETY AND
EFFICACY, YOUNG ANIMALS, OLDER
ANIMALS.
I DON'T KNOW IF THAT ANSWERED
THE QUESTION, BUT --
>> ANY OTHER QUESTIONS?
I THINK WE THEN ARE AT PERFECT
TIMING FOR A BREAK.
WAS IT 10 MINUTES, 20 MINUTES?
2:30.
IS THAT RIGHT?
OKAY, GREAT.
THANK YOU BOTH TO THE SPEAKERS.
THANK YOU.
[APPLAUSE]
>> WE'RE GOING TO GO AHEAD AND
GET STARTED ON OUR LAST SESSION
OR LAST PART OF THIS SESSION FOR
THE SMALL ANIMAL MODELS
WORKSHOP.
SO WE HAVE A PANEL DISCUSSION,
AND I'M GOING TO ASK EVERY
MEMBER OF THE PANEL TO FIRST
INTRODUCE THEMSELVES AND THEN I
KNOW A NUMBER OF THEM HAVE
SLIDES.
SO WE'LL GO DOWN THE ROW,
INTRODUCE, AND WE'LL START WITH
PRESENTATIONS.
THANK YOU.
>> GOOD AFTERNOON, EVERYONE.
MY NAME IS JOY LIU.
I'M A PROGRAM OFFICER FROM BASIC
IMMUNOLOGY BRANCH IN THE
DIVISION OF ALLERGY, IMMUNOLOGY
AND TRANSPLANTATION OF NIAID.
>> MY NAME IS MICHELLE ARNOLD.
I'M A PROGRAM OFFICER IN THE
DIVISION OF MICROBIOLOGY AND
INFECTIOUS DISEASES AT NIAID.
>> I'M ERIC STEMMY, I'M ALSO IN
THE DIVISION OF MICROBIOLOGY AND
INFECTIOUS DISEASES IN THE
RESPIRATORY DISEASES BRANCH, AND
I'M THE TEAM LEAD FOR THE
CENTERS OF EXCELLENCE FOR
INFLUENZA RESEARCH AND RESPONSE.
>> STACEY SCHULTZ-CHERRY, I AM
AT ST. JUDE CHILDREN'S RESEARCH
HOSPITAL.
>> SO FIRST I WOULD LIKE TO
INTRODUCE OUR SBIR REGION
DEVELOPMENT PROGRAM.
IF YOU CAN SEE THE SLIDES, IT'S
OVER THERE.
SO THIS PROGRAM ACTUALLY STARTED
IN 2018.
THE GOAL IS TO DEVELOP AND
VALIDATE RELIABLE ANTIBODIES FOR
TRACKING IMMUNE RESPONSE AND
IDENTIFY IMMUNE CELLS IN
UNDERREPRESENTED MAMMALIAN
MODELS AND NON-MAMMALIAN MODELS.
SO WHEN WE TALK ABOUT
NON-MAMMALIAN MODELS, WE MEAN
ARTHROPOD, AMPHIBIAN, FISH,
NEMATODE, MARINE ECHINOID, AND
BY UNDERREPRESENTED MAMMALIAN
MODELS WE MEAN GUINEA PIG,
FERRET, BAT, MINK, HAMSTER,
BIRD, COTTON RAT, CAT, RABBIT
AND MARMOSETS.
THIS PROGRAM HAS BEEN SUPPORTED
BY THE DIVISION OF ALLERGY,
IMMUNOLOGY AND TRANSPLANTATION,
THE NIAID, AND WE USE SBIR AS A
CONTRACT MECHANISM.
SO OVER THE YEARS WE HAVE FUNDED
220 PROJECTS.
YOU CAN SEE THE FUNDING
PROPOSALS ACCORDING TO THE
TARGETED SPECIES.
OVER THE YEARS WE HAVE HAD SIX
GUINEA PIG PROJECTS, ACTUALLY
THREE COMPLETED AND THREE
ONGOING, ANTIBODIES PRODUCED IN
THE RABBIT AND THE MOUSE AND
PENDING APPLICATIONS, ACTUALLY
IT'S A FLOW CYTOMETRY AND ALSO
YOU CAN ALSO SEE HAMSTER AND THE
FERRET ARE POPULAR SPECIES IN
OUR PROGRAM.
WE HAVE TWO COMPLETED PROJECTS
AND TWO ONGOING PROJECTS, THE
ANTIBODIES ALSO PRODUCE IN THE
IMMUNIZED MICE AND ALSO RABBITS.
WE ACTUALLY PLAN TO USE THESE
ANTIBODIES TO SUPPORT FLOW
CYTOMETRY EVEN THOUGH
FLUORESCENCE -- AND ALSO FOR THE
FERRETS PROJECTS, WE HAVE THREE
COMPLETED, ONCE WE USE THEM IN
THE SAME APPLICATIONS.
ADDITIONALLY, WE ALSO HAVE SOME
MIXED PROGRAMS, PROJECTS, ET
CETERA, FOR FERRET AND GUINEA
PIG.
WE ALSO HAVE ONE LIKE
NON-SPECIES IN THIS PROJECT --
NINE SPECIES IN THIS PROJECT,
PEOPLE TRIED TO GENERATE --
ANTIBODIES TO RECOGNIZE IMMUNE
CELLS IN DIFFERENT SPECIES.
YOU CAN ALSO SEE WE HAVE SOME
ANTIBODIES LIKE RABBIT, SWINE,
AND ZEBRAFISH OTHER SPECIES TOO.
SO THIS SLIDE SHOWS COMPLETED
ANTIBODIES FOR 26 FERRET
ANTIGENS.
WE HAVE CD3, CD11C, CD19, CD56.
THESE ANTIBODIES PRODUCED BY
SILVER LAKE RESEARCH CORPORATION
AND THE ANTIBODIES HAVE BEEN --
BY FLOW CYTOMETRY AND
IMMUNOFLUORESCENCE USING THE
FERRET PBMCs.
WE ALSO HAVE LIKE CD ANTIBODIES
AGAINST CD56, CD64, CD86, AND
THEY ALSO HAVE BEEN TESTED BY
USING THE FERRET PBMC CELLS AND
THE COMPANY PRODUCING THE
ANTIBODIES IS THE AMPERSAND
BIOSCIENCES.
I ALREADY PUT THE NAMES OF THE
CONTACT PERSONS SO YOU CAN -- IF
YOU NEED, YOU CAN CONTACT THEM
DIRECTLY.
IN ADDITION TO THE SURFACE
MARKERS, WE ALSO HAVE 11
CYTOKINES PRODUCED -- THE
ANTIBODY AGAINST CYTOKINES BY
AMPERSAND.
SO THESE ANTIBODIES HAVE BEEN
TESTED IN -- USING THE -- FROM
STIMULATED FERRET PBMCs OR
WHOLE BLOOD CELLS.
IN THE PARALLEL AM P AMPERSAND
ALSO -- DIFFERENT CYTOKINE -- TO
DIFFERENT PANELS PRODUCED THEM
IN -- CAGE AND ALREADY TESTED
THEM USING THE NASAL -- FERRETS.
ADDITIONALLY WE ALSO HAVE
AVANGGEN, ANOTHER CONTRACTOR WE
ACTUALLY HAVE UNDER OUR PROGRAM
AND THEY ARE PRODUCING
ADDITIONAL ANTIBODIES FOR
CYTOKINES SUCH AS L1 BETA, L6,
L8, AND IT HAS BEEN TESTED USING
THE SUPER NATANT FROM STIMULATED
FERRET PBMCs.
AND AVANTGEN ALSO PRODUCED TWO
RECEPTOR ANTIBODIES FOR
L1 RECEPTOR AND THE TPNF AND
TESTED BY FL FLOW CYTOMETRY.
THEN WE ALSO HAVE FOUR COMPLETED
HAMSTER ANTIBODIES AGAINST CD16,
CD64, CD69 AND CD115.
AND THESE ANTIBODIES PRODUCED BY
SILVER LAKE RESEARCH
CORPORATION.
THEY TESTED THE FINDING OF THE
ANTIBODIES TO HAMSTER PBMCs.
WE HAVE ADDITIONAL ANTIBODIES IN
THE PIPELINE.
YOU CAN SEE WE HAVE ONE FERRET
ANTIGEN ANTIBODIES AGAINST
CD11C.
IT HAS BEEN TESTED BY USING THE
FLOW CYTOMETRY AND FLUORESCENCE
STAINING USING FERRET PBMCs
AND THE SILVER LAKE RESEARCH
CORPORATION IS PRODUCER, AND WE
HAVE 29 HAMSTER ANTIGEN
ANTIBODIES IN THE PIPELINE.
AND THE FOURTH ONE, FOURTH SET
ANTIBODIES INCLUDING --
CYTOKINE -- GENES AND ALSO
FOUR -- MARKERS AND THESE
ANTIBODIES HAVE BEEN TESTED
USING LEUM NEXT, ELISA OR FLOW
CYTOMETRY IMMUNOFLUORESCENCE
STAINING VALIDATED BY USING THE
TRANSFECTED AND NON-TRANSFECTED
CELL LINES OR HAMSTER SERUM,
CELL AND TISSUE SAMPLES AFTER
STIMULATION OR INFECTION.
THESE ANTIBODIES ARE PRODUCED BY
AMPERSAND BIOSCIENCES.
AND WE HAVE ONE ADDITIONAL
ANTIBODIES PRODUCED BY SILVER
LAKE THAT'S CD32 AND HAS BEEN
VALIDATED BY IMMUNOFLUORESCENT
STAINING.
WE HAVE ANOTHER GROUP MA MAKING
SOME ADDITIONAL ANTIBO KEYS F AR
HAMSTERS SUCH AS CD8 ALPHA,
CD3E, IL-17, ISG15, CCL2, CCL3,
IL-6, IFNA RECEPTOR, LY6G,
CD169, TMEM119 AND CD16 AND
P2RY12, CX3CR1, AND THEY HAVE
TESTED THE FINDING OF THE
ANTIBODIES TO THE HAMSTER PBMC
TISSUE SERUM/PLASMA WITH OR
WITHOUT THE INFECTION BY USING
FLOW CYTOMETRY, OR ELISA OR IHC,
WESTERN BLOT.
THIS COMPANY IS CALLED QOOLABS.
I HAVE INCLUDED THEIR
INFORMATION THERE FOR YOU TO
CONTACT.
SO THAT'S ALL FOR ME.
I'D LIKE TO MENTION THAT WE
ACTUALLY GREATLY APPRECIATE IF
YOU COULD LET US KNOW WHAT KIND
OF ADDITIONAL TARGETS AND
SPECIES YOU NEED TO ADVANCE THE
FIELD.
IF YOU HAVE ANY QUESTIONS OR ANY
COMMENTS OR ANY SUGGESTIONS,
PLEASE CONTACT ME.
I LEAVE MY EMAIL THERE.
WITH THAT, THANK YOU SO MUCH.
>> THANK YOU.
SHOULD WE JUST GO THROUGH AND DO
ALL THE PRESENTATIONS FIRST?
MICHELLE, OR --
>> YEAH.
SO I JUST WANTED TO MENTION THAT
THE DIVISION OF MICROBIOLOGY AND
IP FORECASTTIOUS DISEASES HAS
ITS OWN PROGRAM FOR GENERATING
REAGENTS FOR PRE-CLINICAL MODELS
OF INFECTIOUS DISEASES.
SO WE'RE USING A CONTRACT
MECHANISM TO DEVELOP MONOCLONAL
KNOCKOUT ANTIBODIES, AND I'VE
JUST MADE A LIST OF THE ONES
THAT WE HAVE ALREADY MADE THAT
VA BEEN DEPOSITED INTO BEI, SOME
OF WHICH ARE PENDING, WHICH
MEANS THE PAPERWORK HAS BEEN
SUBMITTED.
THEY WILL BE AVAILABLE SOON.
SO EACH OF THESE MONOCLONAL
ANTIBODIES HAS BEEN DEVELOPED
WITH A CORRESPONDING KNOCK-IN
AND KNOCK-OUT CELL LINE SO WE
CAN ENSURE THAT THE ANTIBODY IS
RECOGNIZING THE TARGET OF
INTEREST AND NEGATIVE IN A CELL
LINE OF INTEREST AS WELL.
AND THE CERTIFICATE OF ANALYSIS
FOR EACH OF THESE ANTIBODIES AS
WELL AS THE CELL LINES ARE ON
THE BEI WEBSITE OR ARE AVAILABLE
ON THE BEI WEBSITE.
SO WE'RE DOING THIS FOR BOTH
FERRET AND SYRIAN HAMSTER
REAGENTS.
WE ALSO HAVE SEVERAL THAT ARE
STILL UNDERGOING SYNTHESIS.
WE DON'T KNOW THE OUTCOME YET
BECAUSE WE HAVEN'T GONE THROUGH
ALL THE STAGES BUT WE DO HAVE A
COUPLE OF ADDITIONAL ONES IN
DEVELOPMENT, WHICH I'VE LISTED
HERE.
IN THE PAST, WE'VE USED REQUEST
FOR INFORMATION OR RFI TO ASK
FOR INPUT.
OUR RFI IS CLOSED BECAUSE THIS
WAS INITIATED FOUR YEARS AGO.
SO IN THE FUTURE, IF WE CHOOSE
TO DO THIS AGAIN, WE WILL LIKELY
PUT OUR ANOTHER RFI AND TRY TO
MAKE EVERYBODY AWARE OF IT SO
YOU CAN PROVIDE INPUT AS TO THE
MOST NEEDED REAGENTS RELEVANT TO
YOU.
>> SO I JUST WANTED TO PLUG
BRIEFLY THE REAGENTS THAT HAVE
BEEN CREATED THROUGH THE CEIRR
NETWORK AND THE PREVIOUS
GENERATION.
THESE ARE ALL AVAILABLE ON OUR
WEBSITE.
YOU SEE THE LINK THERE.
A VARIETY OF ANTIBODIES, BLOOD
MATERIALS, CELLS, PROTEIN, RES
PIER SAMPLES AND TISSUES AS WELL
AS FERRET REAGENTS AS WELL
MOSTLY PRODUCED BY STACEY THERE
AT ST. JUDE.
THOSE OF YOU THAT HAVE USED THE
WEBSITE, WE ARE GOING TO BE
GETTING READY TO UPDATE AND
REVAMP THE WEBSITE, SO ONE, IF
YOU SEE THINGS MISSING YOU WOULD
WANT TO SEE AND TWO, TO SAY IT
WOULD BE REALLY GREAT IF WE
COULD DO THIS WITH WHAT YOU
HAVE, PLEASE REACH OUT TO ME AND
WE CAN TRY AND GET THAT IN THE
PIPELINE AS WE REVISE THE
WEBSITE A LITTLE BIT.
>> GREAT, THANK YOU.
I THINK STACEY HAD A QUESTION.
>> MY QUESTION WAS FOR JOY.
IN TERMS OF VALIDATION, HOW WERE
THE VALIDATIONS DONE?
WAS IT JUST DOING FLOW CYTOMETRY
WITH FERRET, HAMSTER PBMCs AND
YOU GET A SHIFT SO YOU ASSUME
THAT YOU'RE ACTUALLY RECOGNIZING
THE TARGET OF INTEREST, OR IS IT
LIKE WHAT WE'RE DOING, WHICH IS
HAVING TO USE KNOCK-IN,
KNOCK-OUT CELL LINES TO ENSURE
THAT THE TARGET IS ACCURATE?
>> THAT'S A GREAT QUESTION.
THIS PROGRAM AS I SAID IS NOT
ESTABLISHED TOO LONG AGO, SO AT
VERY BEGINNING, WE HAVE LOW BARS
OF -- TO COME INTO THE PROGRAM.
WITH THE PROGRESS OF THE PROG
PROGRAM, OUR REQUIREMENT FOR
PHASE ONE STUDIES COME MORE AND
MORE STRINGENT.
WE ACTUALLY IN THE PAST, WE DO
ASK THEM TO SHOW THE SPECIFI
SPECIFICITY, BUT WE DID NOT ASK
THEM TO DO LIKE THE AFFINITY
ASSAYS, ESPECIALLY USING LIKE A
BELL SENSOR TO TEST THE
AFFINITY, NOW WE REQUEST THEY
HAVE TO DO THE AFFINITY ASSAYS.
WE ALSO ASK THEM TO DO KNOCK-IN,
KNOCK-OUT CELL LINES.
ADDITIONALLY, WE ASK THEM TO USE
THE COMPUTATION ASSAY USING
THEIR RECOMBINANT PROTEINS TO
BLOCK THE FUNDING OF THE
ANTIBODIES TO THE -- ANTIGENS.
SO WE REALIZE, YOU KNOW, IT
NEEDS TO BE MORE STRINGENT WHEN
WE MAKE SURE THE ANTIBODIES TO
BE MORE SPECIFIC, SO THAT'S WHY
WE HAVE ADDED NEW REQUIREMENTS
INTO THE PROGRAM.
I JUST WANT TO LET EVERYONE
KNOW, I PRINT SOME INFORMATION
HERE IF YOU WANT TO KNOW THE
ANTIBODY TARGETS WE HAVE, PLEASE
COME TO GET THE PAPER AFTER THE
SESSION.
>> THERE'S A COMMENT IN THE CHAT
BOX FROM LAURIE STEPHEN, WE HAVE
DONE WESTERNS ON THE EXPRESSED
PROTEINS AND CROSS REACTIVITY
WITH PROTEINS EXPRESSED IN THE
SAME CELL LINE, ALSO BETA TEST
WITH FERRETS THAT HAD THE FLU.
>> IS THIS RESPONSE TO THE ONE
WITH SBIR?
>> UH-HUH.
>> GO AHEAD.
>> SO ARE THERE ANY CENTRALIZED
EFFORTS TO SEE ANY CROSS
REACTIVITY WITH ANTIBODIES THAT
ARE ALREADY OUT THERE, LIKE
ANTIMOUSE, ANTIHAMSTER, ET
CETERA?
>> SO I CAN ADDRESS THAT.
WE'VE DONE -- AND I THINK
PROBABLY EVERYBODY IN THIS ROOM
HAS DONE A LOT OF THAT, IS ORDER
A COMMERCIAL ANTIBODY THAT'S
SUPPOSEDLY WORKS ACROSS MULTIPLE
SPECIES OR TO CONSERVE TARGET
AND YOU TEST IT.
MOST OF THE TIME IT DOESN'T
WORK.
I DON'T KNOW OF A CENTRALIZED
PLACE AND THAT'S SOMETHING WE
SHOULD THINK ABOUT WHERE YOU
ACTUALLY SHARE THAT DATA BECAUSE
YOU DON'T PUBLISH IT UNLESS
YOU'RE REALLY DOING AN ANTIBODY
VALIDATION PAPER.
BULL IT WOULD SAVE EVERYBODY,
THE COMMUNITY, SO MUCH TIME TO
HAVE ACCESS TO THAT INFORMATION
SO THAT YOU KNOW I TESTED THIS
USING THESE ASSAYS USING THESE
CELLS AND IN OUR HANDS, RIGHT?
BECAUSE THEN COW YOU COULD GO D
DO IT IN YOUR OWN HANDS OR SAY
THIS IS EXACTLY WHAT WE WANTED
TO DO AND THERE'S NO POINT.
TESTING SOME OF THESE ANTIBODIES
SPECIFICALLY DEVELOPING -- WHERE
DEVELOPING FOR FERRET, WE WILL
WANT TO SEE IF THEY CROSS REACT
WITH HAMSTER SO WE KNOW THAT UP
FRONT.
>> SO ONE OF THE THINGS WE'VE
RECENTLY LAUNCHED IS CALLED FLU
HUB.
IT'S INTENDED TO BE A WEBSITE
THAT SORT OF CENTRALIZES ALL OF
NIAID AS INFLUENZA RESOURCES AND
I THINK THIS MIGHT BE A NICE
PLACE FOR SOMETHING LIKE THAT TO
LIVE AS WE HAVE VERSION 1.0 NOW
AND AS WE MOVE ON, I THINK THIS
WOULD BE A GREAT RESOURCE WE CAN
START TO BUILD AND HAVE THAT
CONVERSATION AND BUILD UP A
DATABASE LIKE THAT.
>> I THINK FOR ME, WE WOULD NEED
TO UNDERSTAND ALL OF THE -- LIKE
ALL OF THE QUESTIONS WE WOULD
NEED TO ASK TO SORT OF
CATEGORIZE AND MAKE SURE ALL
THAT INFORMATION IS THERE,
RIGHT, SO THAT IS AS ERIC SAID A
DISCUSSION WE WOULD PROBABLY
NEED TO HAVE IN ORDER TO MAKE
SURE THAT THAT IS AS BROAD AS
POSSIBLE AND EFFECTIVE.
>> FOR ME, MY PROGRAM NOW IN
SOLICITATION WE REQUIRE THEM TO
DO THE MARKET ANALYSIS TO SHOW
THE ANTIBODIES ON THE MARKET
DOES NOT MEET THEIR NEED SO THAT
THAT CAN JUSTIFY THEIR PROPOSED
ANTIGEN.
AND ALSO WE REQUIRE THEM TO DO
THE CROSS REACTIVITY AT LEAST TO
THREE SPECIES IN THEIR PHASE TWO
STUDIES SO THAT WE CAN KNOW
WHETHER THE ANTIBODIES CAN BE
USED FOR OTHER SPECIES IN THE
FUTURE.
WE ALSO HAVE ONE PROJECT AS I
MENTIONED, THEY ARE PRODUCING
ANTIBODIES AGAINST -- LIKE --
REACTIVE AGAINST NINE SPY SHE'S.
SO IN THAT CASE, ONE ANTIBODIES
CAN SERVE A LOT OF STUDY
RESEARCH NEED.
>> THESE ANTIBODIES BEING
GENERATED, IS THE PRIMARY
PURPOSE FOR DETECTION OF
PROTEINS BY, FOR EXAMPLE, ELI I,
LELUMINEX AND FLOW CYTOMETRY,
BECAUSE A LOT WERE IMMUNE CELLS.
BIGGER QUESTION I HAVE IS, DO US
AS INVESTIGATORS HAVE ACCESS TO
EITHER THE HYBRIDOMA OR MONL
CLONAL ANTIBODY SEQUENCE SO THAT
WE CAN ACTUALLY GENERATE IT AND
POTENTIALLY USE IT FOR DEPLETION
STUDIES?
OR BLOCKING STUDIES INA VERY VO?
>> THE ANSWER FOR OUR AGENTS IS
GOING TO BE ABSOLUTELY.
ANYTHING WE MAKE, WE VALIDATE,
WE'RE HAPPY WITH, WE'LL GET IT
SEQUENCED SO THAT WE CAN EVEN
MAKE -- SO IF WE LOSE A
HYBRIDOMA, YOU CAN MAKE
SYNTHETIC ANTIBODIES.
BUT THAT WAS OUR GOAL, WHAT
WOULD BE ACCESSIBLE IS PURIFIED
ANTIBODY, CERTIFICATE OF
ANALYSIS, KNOCK IN, KNOCK OUT
CELLS, AND THEN THE ACTUAL
HYBRIDOMA ITSELF.
BUT TO YOUR POINT, YEAH, A LOT
OF IT IS DETECTION AND
QUANTITATION, BUT WORKING --
WE'RE DOING A LOT OF OUR WORK
WITH PAUL THOMAS, SO HE'S
SPECIFICALLY LOOKING FOR
DEPLETION ANTIBODIES.
AND HE HAS A CD TH CD8 THAT LOOE
A FAIRLY DECENT DEPLETER.
SO THAT'S WHERE I THINK FEEDBACK
FROM THE COMMUNITY IS REALLY
HELPFUL OF IS IT JUST SIMPLY YOU
WANT TO LACK AT THE NUMBER OF
IMMUNE CELLS FILTRATING IN, OR
DO YOU WANT TO KNOW THEIR
FUNCTION, DO YOU WANT TO GET RID
OF THEM, BASICALLY DO YOU WANT A
GIANT MOUSE, RIGHT?
SO THAT'S THE KIND OF
INFORMATION THAT'S REALLY
HELPFUL FROM US.
>> I WAS MORE INTERESTED FOR THE
COMPANIES THAT ARE PRODUCING
THESE, AND THE RELATIONSHIP WITH
THE NIH AN DO WE HAVE ACCESS TO
THE SOURCE MATERIAL TO GENERATE
IT OURSELVES.
TO PRODUCE IT OURSELVES AT A
QUANTITY THAT WOULD BE GREAT FOR
DEPLETION EXPERIMENTS.
OR WHEN YOU'RE DETECTING IMMUNE
CELLS, TO BE ABLE TO CO INSURING
JUGATE DIFFERENT FLUOROPHORES
READILY.
>> THAT'S FOR OUR PHASE TWO
STUDIES, WE ASK THEM NOT ONLY --
FOR WILL VALIDATED ANTIBODIES
BUT ALSO WANT THEM TO MAKE READY
TO USE -- FORMS, OTHER KIND OF
MEASURE SO THAT THEY CAN SCALE
UP FOR PRODUCTION ALSO MARKETING
THE REAGENTS.
HOWEVER, FOR US -- I'M SO SORRY.
FOR US, THE PROGRAM IS FOR THE
SMALL -- I DON'T THINK WE CAN
JUST -- YOU KNOW, BECAUSE THE
PROPERTY STILL BELONGS TO THEM,
SO WE DON'T -- WE DON'T HAVE THE
AUTHORITY TO HAVE THE, YOU KNOW,
HAVE THEM TO PRODUCE ANTIBODIES
LIKE THE BIG QUANTITY.
THEY DON'T HAVE THE CAPACITY TO
DO THAT FOR -- STUDIES, BUT I
THINK IF YOU WANT, YOU CAN
EXPLORE THE POSSIBILITY WITH
THEM.
MOST OF THE ANTIBODIES ARE
RECOMBINANT ANTIBODIES, SO THEY
HAVE THE SEQUENCE ACTUALLY.
SO I THINK YOU CAN DIRECTLY TALK
TO THEM AND SEE WHAT'S THE
POSSIBILITY.
>> IT SOUNDS LIKE THOSE WOULD BE
MORE FOR PURCHASE.
THOSE ARE GOING TO BE ONES THAT
ARE GOING TO BE PURCHASED WHERE
OURS ARE GOING TO BE FREE TO THE
COMMUNITY.
>> YEAH, OUR CONTRACT, THEY
AGREE THAT AS LONG AS THERE'S A
NEED FROM NIAID SUPPORTED
RESEARCH INVESTIGATORS, THEY CAN
GIVE DISCOUNT, OR OTHERWISE YOU
CAN WORK WITH LIKE COLLABORATORS
TO HELP THEM TO VALIDATE THE
ANTIBODIES.
YOU CAN PROVIDE SOME FREE ONES,
FREE ANTIBODIES TO YOU.
>> STANLEY?
>> I WAS GOING TO ASK A QUESTION
ALONG THE SAME LINE.
SO WHEN DATA ARE AVAILABLE SHO
SHOWING HOW THEY VALIDATED, WILL
THE COMPANIES BE ABLE TO PUT THE
SAME AMOUNT OF DATA ON THE
WEBSITES AS THE NIAID ANTIBODIES
ARE GOING TO BE?
SO WE CAN SEE AS WAS MENTIONED
BY COMMERCIAL ANTIBODIES, WE
OFTEN GET THINGS THAT DON'T
WORK, BUT WITH THE COMPANIES,
WILL WE SEE DATA SHOWING THAT
THEY DO WORK?
WILL THAT BE AVAILABLE?
>> WE HAVE THE REPORT.
WE DO REVIEW THE REPORT, YOU
KNOW, IN T IN A TIMELY FASHION,I
DON'T THINK IT'S UP TO THE
COMPANY WHETHER THEY CAN SHARE
THE DATA OR NOT SH BU, BUT I DOK
AS LONG AS YOU ARE INTERESTED IN
THEIR ANTIBODIES, THEY ARE
WILLING TO SHARE THEIR DATA WITH
YOU.
>> SO IS THE DEAL TO DISTINGUISH
THIS FROM A USUAL COMMERCIAL
ANTIBODY, IS THE DIFFERENCE THAT
THE NIH IS PUTTING MORE MONEY
INTO IT OR WOU GETTING A REDUCED
PRICE?
WHAT'S THE ADVANTAGE OF GETTING
IT THROUGH THESE COMPANIES AS
OPPOSED TO A COMMERCIAL
ANTIBODY?
>> THIS IS A MECHANISM FROM HHS.
IT'S TO SUPPORT SMALL BUSINESS.
>> I THINK IF I CAN COMMENT, I
THINK THE NEED IS SO LARGE FROM
A VERY SMALL GROUP OF PEOPLE FOR
THESE REAGENTS, AND THAT THE
COST BENEFIT FOR A COMPANY IS
PROBABLY NOT THERE TO MAKE IT ON
THEIR OWN.
SO I THINK THE NIH IS TRYING TO
HELP FILL THAT GAP WHERE, LIKE,
YOU KNOW, THERE'S ONLY SO MUCH
THAT A COURSE CAN DO TO -- AT
LEAST THAT WAS MY UNDERSTANDING
OF THIS PROGRAM INITIALLY.
>> YEAH, THE -- IS NOT HUGE.
IT'S LIKE 600K FOR TWO YEARS FOR
AN AWARD.
FOR PHASE TWO, WE HAVE UP TO
$1.5 MILLION, BUT YOU'RE REALLY
ONLY -- THE SBIR IS ONLY
SUPPOSED TO BE FOR SMALL
BUSINESS.
>> AND THE LAST QUESTION, IS
THERE OVERLAP BETWEEN THE
ANTIBODIES THAT YOU HAVE AND
THAT NIAID HAD?
I COULDN'T TELL FROM THE LIST.
BUT IS THERE SOME OVERLAP THERE?
>> WE MAY HAVE SON OVER SOME OVP
ONES, YEAH.
>> SORRY.
I'M GOING TO I TAKE MY MODERATOR
POSITION HERE.
CURIOUS, YO ERIK YOU MENTIONED S
THING CALLED FLU HOME FOR
AGGREGAGGREGATING ALL OF THE THS
FLU-RELATED BECAUSE OBVIOUSLY
CEIRR HAS A VARIETY OF RESOURCES
DISTINCT FROM CBI AND IRR AND --
COMPANIES I'M CURIOUS IF FLU
HOME IS GOING TO BE THE SITE
THAT ALLOWS ONE JUST LIKE A
CATALOG OF ALL OF THESE THINGS
THAT ARE COMMERCIAL, LIKE NIAID
FUNDED, CEIRR REAGENTS, AND ALSO
POTENTIALLY HAVE A SYSTEM
WHEREAS SOMEBODY ELSE MENTIONED,
INVESTIGATORS CAN WRITE IN AND
SAY, HEY, I TESTED THAT
ANTIBODY, AND THIS WAS THE FLOW
CHART I GOT FROM MY PBMCs.
LIKE, YOU KNOW WHAT I'M SAYING?
>> MA?MAYBE PART OF THAT IS USER
ERROR, BUT IS THAT A
POSSIBILITY?
>> YEAH, SO IT'S FLU HUB, NOT
HOME, SO SORRY IF IT DIDN'T COME
CLEAR.
SO FLUHUB.
WE'VE GOT A VERY FANCY LOGO AN
EVERYTHING.
SO RIGHT NOW IT MORE OF A LIST
OF INFLUENZA-RELATED PROGRAMS
LIKE CEIRR, LIKE CIVICS, LIKE
THE HIPCs, THINGS LIKE THAT
AND IT'S STARTING TO BUILD UP
THINGS LIKE COMPUTATIONAL
RESOURCES, DATABASES RELATIVE TO
INFLUENZA SORT OF LOOSELY MAPPED
TO OUR UNIVERSAL INFLUENZA
VACCINE STRATEGIC PLAN.
SO AS WE'RE BUILDING IT UP, WE
WANT TO DO THINGS AND FIGURE OUT
WHAT EATION USEFUL FOR THE
COMMUNITY.
SO I THINK LIKE MICHELLE SAID,
IT'S A DISCUSSION WE NEED TO
HAVE RIGHT NOW.
SO WHAT DOES SOMETHING LIKE THIS
LOOK LIKE?
IS IT FLOW POTTS L POTTS, SOME E
THAT'S FILTERABLE, SORT OF STAN
ARDIZED, THE WAYS WE CAN
REPRESENT THE WAY THINGS WERE
DONE AND WHAT WORKED AND DIDN'T
WORK.
WE DO AS THE U.S. GOVERNMENT
NEED TO BE A LITTLE BIT CAREFUL
ABOUT APPEARING TO ENDORSE ONE
PRODUCT OVER ANOTHER PRODUCT, SO
IT'S LESS OF US SAYING YES, THIS
COMPANY'S PRODUCT WORKS AND NO,
THIS COMPANY'S PRODUCT DOESN'T
WORK, SO IT'S A LITTLE BIT OF A
DISCLAIMER THERE, BUT I THINK
THAT'S SOMETHING WE CAN
NAVIGATE.
>> I LIKE THE TABLE FORMAT.
SO WE USE INTERNALLY, JUST
BECAUSE IT'S EASY AND THEN WE
CAN DO POWER VI TO SET UP
DATABASES WITH THAT.
ONE THING THAT WOULD BE HELPFUL
IF YOU'RE REQUESTING GOING TO T
WAY IS TO PUT THE CONTACT OF THE
PERSON, IF THEY AGREE, THAT DID
THE TESTING SO THAT IF YOU
WANTED TO REACH OUT TO ME
DIRECTLY AND SAY WHAT DID YOU
DO, CAN YOU SHARE THE DATA, YOU
CAN AT LEAST HAVE THOSE
CONVERSATIONS.
>> WE CAN LINK TO THE PROTOCOLS,
THINGS LIKE THAT.
WE CAN HAVE THOSE THINGS AND
LINKS THAT CAN BE PULLED UP FOR
PEOPLE TO SHARE AND TRY AND
REPRODUCE TOO.
>> CAN I ADD REALLY BRIEFLY,
IT'S LIKELY THAT, YOU KNOW, THE
RESULTS THAT ONE PERSON HAS IN
THEIR LAB MAY DIFFER FROM
ANOTHER, SO THERE SHOULD BE A
WAY TO CAPTURE THAT INSTEAD OF
JUST ONE PERSON HAVING THE SAY.
SO THESE ARE ALL THINGS THAT
WE'RE TRYING TO CONCEPTUALIZE
AND THAT'S PART OF THE OUTCOME
OF THIS WORKSHOP THAT MIGHT BE
REALLY BENEFICIAL.
>> THIS IS A QUESTION ABOUT THE
SMALL BUSINESS REAGENT PROGRAM.
IT WAS KIND OF UNCLEAR ABOUT THE
ANTIBODY.
SO ARE THEY MAKING THE -- WHEN
THEY HAVE THESE ANTIBODY
REAGENTS, ARE THEY MAKING THE
ANTIBODY SEQUENCES AVAILABLE?
IF THEY ARE NOT, I THINK THEY
SHOULD.
BECAUSE ONCE PEOPLE LIKE ME
START USING THESE ANTIBODIES,
THERE'S AN OPPORTUNITY THAT WE
COULD USE STRUCTURE BASE
ANTIBODY DESIGN TO MAKE THE
ANTIBODIES MORE SPECIFIC OR MAKE
THEM BROADER DEPENDING ON WHAT
THE COMMUNITY NEEDS.
SO I THINK IF THEY DON'T MAKE
THE ANTIBODY SEQUENCING
AVAILABLE, THEN THE NEXT
GENERATION OF IMPROVED
ANTIBODIES PROBABLY CAN'T BE
MADE.
SO THEY SHOULD REALLY BE MAKING
THE ANTIBODY SEQUENCES
AVAILABLE.
>> THANK YOU.
I WILL DISCUSS THIS WITH MY
SUPERVISOR AND MAYBE WE CAN ADD
THAT INTO OUR NEXT SOLICITATION.
>> JUST ANOTHER COMMENT FROM
LAURIE STEPHANS.
SHE JUST MENTIONED THAT ESV
CURRENTLY HAS ANTIBODIES FOR
SALE WITH DATA.
I DON'T KNOW IF THAT INCLUDES
SEQUENCING OR --
>> ARE THEY ONLINE?
WOULD THEY LIKE TO UNMUTE AND
MAYBE JUST SPEAK TO THIS --
MAYBE TO THIS TOPIC ON THE
SEQUENCE AVAILABILITY
SPECIFICALLY BECAUSE IT WAS
BROUGHT UP BY BOTH -- THAT WOULD
BE REALLY NICE.
IS ONE OF THE INDIVIDUALS
INVOLVED IN THE SBIR GRANT
ONLINE THAT WOULD LIKE TO YOU BE
UNMUTE?
IT WON'T LET THEM UNMUTE.
IS THERE A WAY WE CAN FIX THAT,
AV ISSUE?
BECAUSE THIS MIGHT BE THE MOST
DIRECT LINE OF COMMUNICATION.
>> PERHAPS YOU CAN GO TO THE
CHAT.
>> EXACTLY.
SPECIFICALLY ANSWER THE QUESTION
ABOUT WHETHER THE SEQUENCES FOR
THE ANTIBODIES THAT HAVE BEEN
CREATED BY AT LEAST YOUR COMPANY
THAT YOU ARE INVOLVED WITH WOULD
BE MADE AVAILABLE.
PRESUMABLY UPON REQUEST OR AS
PART OF THE DATA CERTIFICATE.
>> APOLOGIES.
THE MICROPHONES ON THE FAR END
THERE ARE UNMUTED.
THEY JUST HAVE TO OPEN THEIR
MICROPHONES BUT THEY ARE
UNMUTED.
SORRY.
>> WHILE I GUESS MAYBE WE WAIT
FOR THAT ANSWER TO COME THROUGH,
IS THERE ANYBODY ELSE ONLINE WHO
WANTED TO ASK A QUESTION?
CAN I COME BACK TO THE FLU HUB?
SO IS THIS -- THIS IS STARTING
TO SOUND A LITTLE BIT
REMINISCENT OF IRR BEFORE IT WAS
INTERNATIONAL REAGENT RESOURCE
AND IT WAS INFLUENZA REAGENT
RESOURCE?
OBVIOUSLY WHEN WE TALK ABOUT THE
VARIETY OF NIAID PROGRAMS AND
CIVICS, WHERE YOU ALSO HAVE NOW
CHIM OR CHALLENGE VIRUSES THERE,
AS PART OF CEIRR, WE'RE
DEVELOPING, YOU KNOW, RESCUE
SYSTEMS AND SERUM ASSAYS AND
LOTS OF THINGS.
SO I GUESS MAYBE CAN YOU --
>> HI.
CAN YOU HEAR ME?
[ECHOING]
OH, MY GOD!
>> APOLOGIES.
>> IS THAT BETTER?
>> PLEASE MUTE YOUR SPEAKERS.
>> CAN YOU HEAR ME AT ALL?
>> THANK YOU.
WE CAN HEAR YOU BETTER NOW.
THANK YOU.
>> ONE MORE TRY, THEN I'LL LOG
OUT.
>> NO, LAURIE, WE CAN HEAR YOU.
>> OH, OKAY.
IT AN ECHO FOR ME SO I WASN'T --
I WAS WORRIED THAT IT WAS ECHO
FOR YOU.
>> IT'S CLEARED UP NOW.
COW PLEASE -- THANK YOU FOR
UNMUTING.
WOULD YOU LIKE TO ANSWER THE
QUESTION ABOUT THE SEQUENCE
AVAILABILITY?
>> YEAH.
SO THE ISSUE WOULD BE THAT A
COMPETITOR COULD THEN JUST MAKE
THE ANTIBODY AND ALL THE EFFORT
THAT WE'VE PUT IN IS LOST.
SO --
>> IS THERE A PATHWAY TO
POTENTIALLY MAKE IT AVAILABLE
FOR RESEARCH PURPOSES TO
SCIENTISTS?
>> I MEAN, WE'D HAVE TO ENTER
SOME KIND OF AGREEMENT
POTENTIALLY, YEAH.
I MEAN, WE HAVE SCALED UP A LOT
OF THE ANTIBODY SO WE CERTAINLY
CAN MAKE -- THOSE ARE AVAILABLE
FOR SALE AND WE CAN MAKE LARGE
QUANTITIES AVAILABLE.
BUT IN ORDER TO SEQUENCE IT AND
GIVE THAT OUT, THAT'S THE ONLY
CONCERN, IS THAT, YOU KNOW, THEN
ALL OF A SUDDEN EVERYBODY COULD
MAKE IT.
>> DID YOU HEAR THAT COMMENT?
>> I UNDERSTAND THAT CONCERN
FROM A SMALL BUSINESS, THAT
EVERYBODY CAN MAKE IT.
BUT TO BE BRUTALLY HONEST, IF
PEOPLE WANT TO MAKE ANTIBODY,
THEY CAN ORDER THE ANTIBODY AND
USE MASS SPEC TO SEQUENCE THE
ANTIBODY IF THEY REALLY WANT TO
MAKE IT.
SO I UNDERSTAND THAT ISSUE, BUT
THAT'S NOT REALLY -- SO IF WE
WANT TO MAKE ANTIBODY, WE COULD
ORDER IT FROM THE COMPANY, SPEND
$3,000, MASS SPEC AND REVERSE
ENGINEER IT AND PEOPLE HAVE DONE
THAT IN THE PAST.
I JUST WANT TO BRING THAT AS A
POINT.
>> NO, I UNDERSTAND THAT AND
THAT'S YOUR RIGHT, BUT IF WE PUT
ALL SEQUENCES ONLINE, THEN
OUR -- SYSTEMS COULD COME IN AND
JUST MAKE THEM ALL AND THEN WE
HAVE NOTHING.
AND PART OF THE REASON THAT WE
DID THIS IS YOU KNOW, BECAUSE IT
WAS A GOOD OPPORTUNITY TO GET
INTO SOMETHING NEW, BUT AS JOY
MENTIONED, IT'S NOT -- WE PUT A
LOT OF EXTRA TIME INTO IT.
IT'S NOT JUST ALL FUNDED.
AND YOU KNOW, WE ARE A SMALL
BUSINESS SO WE STILL -- A LOT OF
YOU GUYS ARE GETTING -- YOU'RE
GETTING -- YOUR LABS ARE PAID
FOR AND YOUR SALARIES ARE PAID
FOR.
I STILL HAVE TO PAY FOR MY
STUFF.
SO --
>> THANK YOU, LAURIE.
I THINK WE UNDERSTAND THE
CONCERNS ON THE SMALL BUSINESS
SIDE.
I THINK MAYBE THERE ARE SOME
INTERNAL DISCUSSIONS ON YOUR END
AS WELL AS ON THE NIH SIDE TO
ESSENTIALLY THINK ABOUT PATHWAYS
FORWARD.
OBVIOUSLY MAYBE NOT MAKING THEM
FREELY AVAILABLE ON WEBSITES,
BUT MAYBE PATHWAYS FORWARD SUCH
THAT RESEARCH SCIENTISTS, YOU
KNOW, COULD WORK WITH COMPANIES
TO GET THE SEQUENCES TO DO
ADDITIONAL SCIENTIFIC INQUIRIES.
I THINK THAT'S WHAT WE'RE ASKING
FOR.
SO YEAH, MAYBE OFFLINE QUESTIONS
AND CONVERSATIONS ARE NEEDED
AROUND THAT.
THANK YOU.
ANY OTHER QUESTIONS ONLINE FROM
OUR AUDIENCE?
GO AHEAD, JOY.
>> AS A PROGRAM OFFICER
OVERSEEING THIS PROGRAM,
ACTUALLY WE HAVE EXPERIENCED A
LOT OF DIFFICULTY.
ONE DIFFICULTY IS WE DON'T KNOW
WHAT'S THE ANTIGEN LIST,
WHETHER -- WHICH ONE YOU LIKE,
YOU WANT, WHICH IS THE MOST
IMPORTANT ONES, PRIORITIES WE
WANT TO HAVE THE PRIORITIES FROM
THE -- YOU KNOW, THE FEW, WANT
TO MEET YOUR NEED.
SO IF WE CANNOT HEAR YOUR NEEDS,
WE DON'T KNOW WHICH ANTIBODIES,
YOU KNOW, WE SHOULD DEVELOP.
ANOTHER THING IS WE ACTUALLY
HAVE DIFFICULTIES TO FUND
COLLABORATORS TO VALIDATE OF OUR
ANTIBODIES, BECAUSE THERE'S A
DISCONNECTION BETWEEN THE END
USERS AND PRODUCERS, IT'S KIND
OF A DIFFICULTY FOR US TO MAKE
SURE THE ANTIBODIES ACTUALLY CAN
WORK WELL IN THE REALTIME
INFECTION MODELS OR SAMPLES.
SO IT WILL BE GREAT IF WE CAN
ESTABLISH SOME KIND OF
CONNECTION BETWEEN THE END USERS
AND THE PRODUCERS.
THAT'S MY COMMENT.
>> CAN I RESPOND TO THAT?
I KNOW PREVIOUSLY THERE WAS A
WORKING GROUP, I WAS A PART OF
IT, WE HAD MAYBE TWO CALLS, BUT
THERE WAS A FERRET REAGENT
WORKING GROUP AT LEAST, AND I
THINK WE CAN COME BACK TO THAT,
NOW THAT THERE'S BEEN SO MUCH
PROGRESS MADE.
AND SORT OF LIKE TO LOOK AT
REDUNDANCIES AND SORT OF I
AGREE, I THINK AS INVESTIGATORS,
WE WOULD LOVE TO HAVE A SEAT AT
THE TABLE TO TALK DIRECTLY TO
THOSE MAKING THE RESOURCES TO
FURTHER OUR SCIENCE.
>> AND IT SOUNDS LIKE THAT'S A
FERRET WORK GROUP, BUT MAYBE WE
PULL IN HAMSTERS AS WELL OR A
SEPARATE WORKSHOP FOR --
>> I THINK TWO DIFFERENT WORKING
GROUPS WOULD BE IDEAL, JUST TO
ALSO SEE WHERE PROCESSES ARE.
MICHELLE, I'M GOING TO TURN IT
OVER TO YOU.
>> THANK YOU PANELISTS.
THANK YOU GUYS SO MUCH!
ALL RIGHT.
>> I'M JUST GOING TO MAKE A FEW
CLOSING REMARKS.
OH, I SHOULD PROBABLY BRING -- I
JUST WANTED TO DO A QUICK
SUMMARY TO CLOSE OUT THE
WORKSHOP.
BUT BEFORE I DO THAT, I JUST
WANT TO POINT OUT, I DO
APPRECIATE THE THANKS THAT
EVERYBODY HAS OFFERED IN MY
DIRECTION, BUT THIS IS NOT A
SINGLE PERSON CREATING THIS
MEETING.
IT REALLY TAKES AN ENTIRE TEAM
OF PEOPLE.
SO FIRST, OF COURSE, STEVEN
BLACK WHO IS ONE OF OUR PROJECT
COORDINATORS HAS DONE A
FANTASTIC JOB HELPING OUT AT
EVERY STEP OF THE WAY.
ALONGSIDE STEVEN, THERE HAS BEEN
AN ENTIRE TEAM OF PROGRAM
OFFICERS INCLUDING ERIK STEMMY,
KATE, LAUREN, JENNIFER GORT GORN
AND NITA ALL WHO HAVE CONTRIBUTE
TODAY DEVELOPING THIS PROGRAM,
SO WE ALL HAD EXTENSIVE
CONVERSATIONS ABOUT WHO TO
INVITE, WHAT WE HOPE TO
ACCOMPLISH, AND I THINK ALL
TOGETHER, YOU KNOW, WE PUT OUR
BEST IDEAS FORWARD IN ORDER TO
CREATE THIS WORKSHOP.
SO BEFORE WE CLAP FOR ALL OF
THESE PEOPLE, I JUST WANTED TO
ALSO ACKNOWLEDGE THE EXTERNAL
CONTRIBUTION OF MAHUL AND SEEMA
WHO ALSO MADE SUGGESTIONS FOR
PUTTING THIS WORKSHOP TOGETHER.
FINALLY OUR EXCELLENT MODERATORS
FOR EVERY SINGLE SESSION WHO
HAVE REALLY DONE A GREAT JOB ON
KEEPING EVERYBODY ON TIME AND
ALSO KEEPING THAT CONVERSATION
DRIVING FORWARD.
SO LET'S -- I GUESS WE'LL CLAP
FOR EVERYBODY, RIGHT?
SO JUST A COUPLE OF LESSONS THAT
THIS IS NOT A COMPREHENSIVE LIST
BUT A COUPLE OF LESSONS THAT
I'VE TAKEN AWAY FROM THIS
WORKSHOP.
AND THAT I'VE HEARD MULTIPLE
TIMES.
ALSO HAVING COLLATED YOUR
RESPONSES FROM THE CONFERENCE IO
APP, THE QR CODE QUIZ OR
QUESTIONS.
SO FIRST CHOOSING MODELS BASED
ON THE QUESTION THAT YOU'RE
SEEKING TO ANSWER CAME ACROSS
VERY LOUD AND CLEAR IN THE
WORKSHOP.
I ALSO THINK THAT CONSIDERATION
OF THE DEMOGRAPHICS AND THEN
OTHER HEALTH CONDITIONS REALLY
ADDS A LOT OF COMPLEXITY TO
THESE SYSTEMS.
AND IT IS UNLIKELY THAT WE WOULD
BE ABLE TO SORT OF ADDRESS EVERY
SINGLE CONDITION IN A SINGLE
SYSTEM, SO THEREIN THE DIVERSITY
OF THE ANIMAL MODELS THAT YOU
ARE ALL GENERATING REALLY WILL
BE ABLE TO PROVIDE NEW
INFORMATION, AND ADDITIONAL
EXPERIMENTAL SUPPORT TO
UNDERSTANDING THE PATHOGENESIS,
TRANSMISSION, IMMUNOLOGY AND
OTHER ASPECTS OF RESPIRATORY
VIRUS INFECTIONS.
WE'VE HAD A LOT OF PEOPLE
MENTION THAT THERE SHOULD BE
PROPER QUALIFICATION OR
VALIDATION.
THANK YOU, BART, TODAY FOR SORT
OF DISTINGUISHING THOSE TWO
THINGS.
WE DO USE THAT TERM VALIDATION,
WHEN PERHAPS WE REALLY ARE
TALKING MORE ABOUT QUALIFICATION
OF ALL THESE REAGENTS.
BUT IN ANY -- IN EITHER SENSE,
IT'S ESSENTIAL THAT WE MAKE SURE
THAT THE REAGENTS ARE USABLE.
SO THIS NOT ONLY INCLUDES WHAT
WE JUST DISCUSSED, WHICH ARE THE
COMMERCIAL AND SHARED ANTIBODY,
BUT ALSO SEQUENCING THE VIRUSES
AS YOU AMPLIFY THEM IN YOUR OWN
LABS TO MAKE SURE THAT YOU KNOW
WHAT YOU'RE WORKING WITH.
AND THEN THE WELL ANNOTATED
GENOMES WHICH BECAME MORE
APPARENT DURING THE DISCUSSIONS
TODAY.
THEN OF COURSE JUST TO SORT OF
BRING US BACK TO THE FIRST
SESSION OF THE FIRST DAY IS, YOU
KNOW, PUTTING IN SOME EFFORT
INTO LINKING THE STUDIES IN
SMALL ANIMALS TO STUDIES IN
HUMANS.
AND HOW ARE WE GOING TO BEST DO
THAT MOVING FORWARD.
SO A COUPLE OF THE NEEDS THAT
I'VE IDENTIFIED BASE ON THE
DISCUSSIONS HERE.
WE'VE JUST TALKED ABOUT THIS
COMPREHENSIVE DATABASE.
IT'S SOMETHING THAT HOPEFULLY
WE'LL BE ABLE TO FACILITATE AND
ENGAGE WITH ALL OF YOU IN ORDER
TO BUILD SOMETHING THAT'S USEFUL
FOR THE COMMUNITY.
AS WELL AS THE IMMUNOLOGICAL
REAGENTS AND I'VE SORT OF
CATEGORIZED THESE INTO SORT OF
DETECTION OF ANTIBODY SUBTYPES,
B CELLS, T CELLS AND MUCOSAL
TARGETS.
WHAT I HAVEN'T HEARD ARE ABOUT
NON-IMMUNOLOGICAL TARGETS, SORT
OF UNIDENTIFIED.
EVEN THOUGH A LOT OF THE TARGETS
THAT WE'RE TALKING ABOUT ARE
CALLING FOR NEEDS AND STUDIES OF
IMMUNOLOGY, PLEASE DON'T EXCLUDE
OTHER THINGS IF THOSE ARE THE
KIND OF ANTIBODIES OR THINGS
THAT YOU NEED TO DETECT.
SO I THINK WE ALSO NEED TO HAVE
GREAT ROBUST DISCUSSIONS ABOUT
APPROPRIATE EXPERIMENTAL USE OF
THE MOUSE ADAPTED STRAINS VERSUS
HUMAN STRAINS AND EXPERIMENTAL
SYSTEMS.
AND I'VE HEARD SEVERAL TIMES
ABOUT THE DEVELOPMENT OF UPDATED
MOUSE STRAINS, ESPECIALLY IN THE
CASE OF INFLUENZA, SO THIS IS
SOMETHING ALSO TO CONSIDER.
I'M REALLY GLAD THAT WE GOT TO
HEAR ABOUT THE COLLABORATIVE
CROSS MOUSE MODEL.
AND CERTAINLY THINK THAT THERE'S
A LOT OF REALLY INTERESTING AND
USEFUL POTENTIAL THINGS TO
HAPPEN THERE.
BUT I CAN ALSO SEE A NEED FOR
EXPANDING THAT MOUSE MODEL USE
AS WELL AS THE ASSOCIATED
EXPERTISE WITH THAT.
AND THEN FINALLY, SOME OF THE
BROADER COLLABORATIONS THAT
WE'RE ALWAYS LOOKING TO
ESTABLISH.
WITH ENGINEERS, INFORMATICIANS
AND OTHER UNIQUE VIEWPOINTS THAT
ARE REALLY GOING TO HELP DRIVE
OUR WORK AND YOUR WORK IN SMALL
ANIMAL MODELS FORWARD.
SO THOSE ARE THE TWO SUMMARIES
THAT I REALLY WANTED TO PROVIDE
FOR YOU JUST TO SORT OF WRAP UP
THE WORKSHOP AND BRING THINGS
TOGETHER.
IN THE CONFERENCES IO APP, WE
UNVEILED A LAST QUESTION WHICH
IS TO PROVIDE US WITH ANY
FEEDBACK OR THOUGHTS THAT YOU
MIGHT HAVE ON THE WORKSHOP OR
ANYTHING ELSE.
IF YOU DON'T WANT TO USE THAT
APP, FEEL FREE TO EMAIL ME OR
STEVEN BLACK.
IT HELPS IF YOU KNOW US BOTH, SO
WE CAN KEEP TRACK OF THINGS WITH
YOUR FEEDBACK.
HAVE I FORGOTTEN ANYTHING?
SO I GUESS WITH THAT, I'M GOING
TO SAY THANK YOU ALL VERY MUCH
FOR YOUR ENGAGEMENT AND
PARTICIPATION IN THIS MEETING.
I THINK IT WENT VERY WELL.
AND WE'LL CLOSE THE MEETING.
THANK YOU.
[APPLAUSE]