I'M JOE MARCOTRIGIANO.
I AM THE CHAIR OF THIS YEAR'S
SALZMAN COMMITTEE.
ON BEHALF OF THE COMMITTEE AND
FOUNDATION FOR THE NIH AND
MRS. SALZMAN WELCOME TO THE
NORMAN P SALZMAN ANNUAL MEMORIAL
SIM PODIUM AND AWARDS IN BASIC
AND CLINICAL VIROLOGY.
IT'S A MILESTONE FOR US THIS
YEAR.
ANOTHER MAJOR MILESTONE IS WE'RE
BACK IN PERSON FIRST TIME IN
THREE YEARS.
SO I THINK THAT IS A MAJOR
ACHIEVEMENT FOR OUR FIELD
WORLDWIDE THAT WE'VE BATTLED THE
PANDEMIC AND WE'RE SOMEWHAT BACK
TO NORMAL AGAIN.
SO, IT'S GREAT TO BE BACK AND
GREAT TO BE HERE.
WE HAVE A VERY EXCITING DAY
PLANNED AND A KIND OF WANT TO
KICK IT OFF.
FIRST UP IS DR. JULIE LOUISE
GERBERDING THE PRESIDENT AND
CHIEF EXECUTIVE OFFICER OF THE
FNIH.
>> THANK YOU FOR YOUR ONGOING
SUPPORT OF THIS VERY IMPORTANT
OCCASION.
WHEN I WAS THINKING ABOUT
DR. SALZMAN I WAS WONDERING IF
HE WERE HERE TODAY AND COMING
OUT OF THE CORONAVIRUS PANDEMIC
WOULD HE BE SURPRISED.
AND I THINK I WOULD HAVE TO SAY
PROBABLY NOT BECAUSE HE SPENT A
GREAT DEAL OF TIME IN HIS CAREER
STUDYING PATHOGENS LIKE POLIO
AND SMALLPOX AND AIDS.
I THOUGHT HE WOULD HAVE BEEN
PARTICULARLY SURPRISED TO SEE
THAT WE WERE PLAGUED WITH YET
ANOTHER RESPIRATORY DISEASE THAT
CAUSED GLOBAL CALAMITY.
I WAS WONDERING WOULD HE BE
SURPRISED ABOUT THE mRNA
VACCINES AND I REALIZED PROBABLY
NOT BECAUSE HE WAS ONE OF THE
PEOPLE WHO FIRST DEFINED BIO
CHEMISTRY OF MESSENGER RNA AND
NO DOUBT UNDERSTOOD AS IT
EVOLVED HOW IMPORTANT THAT
ULTIMATELY COULD BE FOR HUMAN
HEALTH.
BUT I THINK HE'D BE PLEASED HIS
WIFE, FAMILY AND NIH INTEREST
GROUP IN THE AREA AND ORGANIZING
COMMITTEE COMMITTED TO
MAINTAINING THE FOCUS ON EARLY
CAREER SCIENTISTS AND THEIR
MENTORS AND THAT LEGACY IS
CONTINUING.
THANK YOU SO MUCH FOR ENSURING
THAT CONTINUES TO HAPPEN.
THANK YOU ALL WORKING IN THE
FIELD OF VIROLOGY FOR YOUR
CONTRIBUTIONS.
I'M AN INFECTIOUS DISEASE DOCTOR
AND A HARDLY THINK THEY COULD BE
MORE IMPORTANT THAN THEY'VE BEEN
THE LAST THREE YEARS AND
HOPEFULLY THE EYES OF THE WORLD
ARE OPENED UP AND PEOPLE CAN
RECOGNIZE WHAT AN EXCITING TIME
THIS IS IN SCIENCE AND ALSO HOW
CRITICALLY IMPORTANT AND THE
PIPELINE OF THE BEST AND
BRIGHTEST SCIENTISTS AND SUSTAIN
OUR INTRAMURAL WORK IN THE
INTERESTS OF THE HONOR WE'RE
BESTOWING TODAY.
THANK YOU SO MUCH AND THANK YOU
JOE AND I LOOK FORWARD TO
HEARING THE REMARKS THAT FOLLOW.
THANK YOU.
CHARLES RICE IS FROM THE
ROCKEFELLER UNIVERSITY AND
SCIENTIFIC AND EXECUTIVE
DIRECTOR NOR CENTER FOR THE
STUDY OF HEPATITIS C.
CHARLIE STARTED HI GRADUATE
STUDIES AT KAL TECH AND GYM AND
HIS WIFE ELLEN ARE PIONEERS IN
THE IMPLEMENTATION OF MOLECULAR
BIOLOGY TECHNIQUES TO STUDY RNA
VIRUS BIOLOGY SPECIFICALLY IN
ALPHA VIRUSES PIONEERS IN
GENERAL.
CHARLIE DID HEROIC WORK AT THE
TIME THEY SEEM SIMPLE NOW WITH
LOTS OF SEQUENCING AND CLOTH OF
THE STRUCTURAL PROTEINS AND
STAYED ON AS A POSTDOC AND DID
WORK ON VIRUSES SPECIFICALLY
YELLOW FEVER AND 17B STRAIN THE
VACCINE STRAIN FOR YELLOW FEVER.
IT'S CONSIDERED TO BE ONE OF THE
SAFEST AND MOST EFFECTIVE
VACCINES EVER PRODUCED AND MOVED
TO WASH-U IN ST. LOUIS MAKING
THE FIRST CLONES FOR YELLOW
FEVER 17D AND ENABLED THE
PRODUCTION OF mRNA WHICH THEN
CAN BE PUT INTO CELLS.
IT'S A REALLY IMPORTANT
MILESTONE.
HERE AT THE CLINICAL CENTER,
HARVEY ALTER THE STUDY
TRANSFUSIONS PEOPLE WHO
DEVELOPED HEPATITIS POST
TRANSFUSION AND THEY HAD BEEN
SCREENED FOR HEPATITIS A AND B
AND WORKING WITH BOB PURCELL AT
NIAD AND WE USED AN EXPRESSION
CLONING METHOD IDENTIFY A
PATHOGEN AND BOOT STRAP TO GET
MOST THE GENOME AND TURNED OUT
TO BE A NEW VIRUS AT THE TIME.
THAT'S WHEN CHARLIE GOT BACK
INVOLVED THROUGH THE
INTERACTIONS WITH STEVE
FINESTONE WHO I THINK IS HERE
AND STEVE GOT CHARLIE BACK INTO
WORKING ON HEPATITIS C VIRUS.
CHARLIE AND HIS LAB FIGURED OUT
THE POLY PROTEIN PROCESSING OF
THE VIRUS AND THE PRIME END
WHICH THEN ALLOWED FOR IN VITRO
METHODS TO EVOLVE AND MAKE THE
CLONE IN LABORATORY AND THEN
MAKE AN mRNA AND PUT THAT INTO A
CHIMPANZEE'S LEVER AND CAME OUT
AS HEPATITIS C VIRUS.
CHARLIE CONTINUED WITH REPLICONS
AND INFECTIOUS CLONES IN THE
LATE '90s AND EARLY 2000s.
I CONTACTED CHARLIE AROUND 2000
ABOUT BECOMING A POSTDOC IN HIS
LAB AND I WAS CONVINCED HE
WOULDN'T RESPOND TO MY E-MAIL
BECAUSE WHY WOULD HE WANT A
STRUCTURAL PERSON IN HIS LAB AND
I WAS VERY WRONG.
HE CONTACTED ME BACK AND INVITED
ME TO COME AND CHARLIE AT THAT
TIME WAS ASSEMBLING A VERY
DIVERSE GROUP OF PEOPLE TO BRING
DIFFERENT TECHNIQUES TO WORK ON
HEPATITIS C VIRUS.
AFTER ABOUT A YEAR LAST DOWN IN
CHARLIE'S OFFICE AND SAID
THERE'S ONLY 10 PROTEINS IN HCV.
THE DRUG COMPANIES FIGURED OUT
THE POLYMERASE AND PROTEASE AND
WE HAVE TO DO MORE AND WE SET
ABOUT SOLVING STRUCTURES IN NS2
AND CHARLIE WAS RIGHT.
ONCE WE HAD REPLICON SYSTEMS AND
HIGH THROUGHPUT SCREENING
METHODS AND COMPOUNDS WERE
POTENT AND FOUND IN THE
LABORATORY THAT INHIBITED HCV AT
PI
PICOMOLE CONCENTRATIONS AND IT
WAS A NEW CLASS OF ANTIVIRALS
AND NOW NS5A INHIBITORS ARE
CURING GREATER THAN 95% OF THE
CHRONIC INFECTIONS OF HEPATITIS
C VIRUS AND CHARLIE SORT OF LAID
THE FOUNDATION FOR US TO DO THAT
WORK STRUCTURALLY AND HELPED IN
THE MECHANISM AND DESIGN OF THE
COMPOUNDS.
NOW WE'RE CURING MILLIONS OF
PEOPLE YEAR TO DATE AND LOTS OF
PEOPLE HAVE BEEN CURED OF HCV TO
THIS DATE AND CHARLIE'S BEEN AN
EXCELLENT MENTOR FOR YEARS.
HE'S ALWAYS VERY SUPPORT
SUPPORTIVE.
HE ANSWERS MY PHONE CALLS WHICH
I'M SURPRISED HE DOES AND
ACCEPTS MY INVITATION TO COME
AND SPEAK TODAY.
WITH MORE, CHARLIE, PLEASE.
>> WELL, I CAN ZIP THROUGH THE
FIRST PART OF MY TALK.
IT'S A REAL PLEASURE AND HONOR
TO BE HERE.
I THINK THIS SYMPOSIUM IN HONOR
OF NORMAN SORT OF EMPHASIZES
SORT OF THE MENTORS AND MENTEES
AND IS A WONDERFUL LEGACY
BECAUSE I KNOW FROM READING
ABOUT NORMAN AND HAVING MET HIM,
THIS WAS A VERY IMPORTANT ASPECT
OF HOW SCIENTISTS CAN CHANGE THE
FUTURE AND HELPING THE NEXT
GENERATION OF YOUNG SCIENTISTS
AS JOE SAID WE DO HAVE A CURE
FOR HEPATITIS C AND THAT LEAVES
A MOLECULAR VIROLOGIST WITH THE
QUESTION WHAT TO DO NEXT.
AT LEAST IN TERMS OF MY LAB WE
HAVE STARTED TO WORK IN
DIFFERENT AREAS AND I'LL MENTION
A FEW AT THEN OF THE TALK BUT
THERE'S STILL A LOT OF
CHALLENGES AHEAD BOTH IN
HEPATITIS C AND OTHER INFECTIOUS
DISEASES WE'VE SEEN.
SO THE SUCCESS STORY, LET'S SEE
IF THIS IS GOING TO WORK.
SO I THINK AS JOE SAID THIS
MYSTERIOUS AGING OF
POST-TRANSFUSION HEPATITIS
NOTICED BY HARVEY ALTAR AND
OTHERS NOT DUE TO HEPATITIS A OR
B HAPPENED IN THE 1970s.
THIS REALLY WAS QUITE A MYSTERY.
THE INFECTION COULD BE TREATED
WITH INTERFERON BUT VERY POORLY
IS THAT GAVE RISE TO AN
INCREDIBLE EFFORT TO TRY TO
FIGURE OUT WHAT THE AGENT WAS.
AS JOE SAID, THAT WAS
ACCOMPLISHED BY MICHAEL HOUTMAN
AND HIS TEAM AT CHIREON IN 1989
AND LOOKED VERY MUCH LIKE YELLOW
FEVER A VIRUS I HAD BEEN WORKING
ON.
AND THE EARLY STUDIES SHOWED THE
ASSOCIATION OF THE AGENT WITH
LIVER DISEASE, SEMINOLE WORK BY
BOB PURCELL AND UNDERSTANDING
THE PROPERTIES OF THE AGENT OF
THE CHIMPANZEE MODEL AND
CHALLENGING AND TERRIFIC WORK.
IN 1989 WITH THE USE OF A PANEL
OF PEDIGREE EXAMPLES HARVEY
COLLECTED IN MOLECULAR BIOLOGY
IN THE CHIRON ROUTE AND NON A
AND B WAS UNVEILED AND
DIAGNOSTICS SHORTLY THEREAFTER A
LOT OF MOLECULAR VIROLOGISTS
LIKE MY LAB GOT INVOLVED AND IN
2011 THE FIRST AGENTS THAT
DIRECTLY WERE TARGETING THESE
DIRECT-ACTING ANTIVIRALS
PROTEASE INHIBITORS WERE ADDED
TO THE CURRENT TREATMENT OF PEG
INTERFERON AND THEN IN 2014,
'15, IT BECAME POSSIBLE TO GO
WITH AN ALL-ORAL TREATMENT FOR
HEPATITIS C WHICH COULD CURE
MORE THAN 95% OF THOSE THAT WERE
INFECTED.
IN FACT THAT NUMBER IS PROBABLY
99.9% OF THOSE INFECTED.
SO I HAD TO SORT OF SHOW THIS
SLIDE AND JOE HAS ALL RIGHT
INTRODUCED IT BECAUSE THERE YOU
SEE THE THREE POSTER CHILDREN
THE VIRAL TARGETS THAT BECAME
PART OF THE TARGETS AND THE
PROTEIN I HAVE TO SAY WAS A
LUCKY DAY FOR ME WHEN JOE DID
SEND THE E-MAIL AND ASKED ABOUT
BECOMING A POSTDOC IN THE LAB
BECAUSE IT ALLOWED US TO REALLY
EXPAND OUR ATTACK ON THIS USING
STATE OF THE ART STRUCTURAL
VIROLOGY APPROACHES.
WE DO HAVE THESE REMARKABLE
TREATMENTS AND THIS SHOWS YOU
SOME OF THEM, TWO TO THREE
MONTHS OF TREATMENT THE VIRUS IS
GONE.
SO IT HAS -- I THINK ONE OF THE
THINGS WE WERE ALL PUZZLING OVER
WAS SINCE ACD IS SO DIVERSE, HOW
COULD YOU COME UP WITH A DRUG
REGIMENT THAT WOULD ACTUALLY
COVER THAT INCREDIBLY HUGE VIRAL
SEQUENCE SPACE.
BUT THESE COMBINATIONS DO.
AND SO RESISTANCE IS REALLY NOT
A GREAT PROBLEM.
SO, WHERE WE ARE NOW AND I THINK
FRANCIS COLLINS HAS TAKEN UP THE
CHARGE TO GET CLOSER TO THIS
ERADICATION BUT THERE ARE SOME
THINGS WE FACE AND THEY'RE
REALLY IN THE ARENA OF ECONOMICS
AND PUBLIC HEALTH.
ALSO, SINCE THIS IS A RELATIVELY
ASYSTEMATIC -- ASYMPTOMATIC
TREATMENT AND IT USED TO BE THE
OLD TREATMENTS THERE WERE GROUPS
LIKE HIV AND CO-INFECTED
PATIENTS HARDER TO TREAT.
THAT'S NOT REALLY THE CASE NOW
FOR THE DIRECT-ACTING ANTIVIRAL
COCKTAILS.
VIRUS RESISTANCE IS NOT A HUGE
PROBLEM AND WE STILL SORT OF
LACK SOME ACTION.
THERE'S STILL A LONG WAYS TO GO
IN THE UNITED STATES.
AND IN EGYPT THEY MANAGED TO
CURE OVER 90% OF THEIR
POPULATION BY HAVING A REALLY
TIGHT GOVERNMENT PROGRAM AND
COMING UP WITH A WAY OF MAKING
SHE'S DRUGS AFFORDABLE.
WE'VE STILL GOT A WAYS TO GO.
THERE IS SOME UNFINISHED
BUSINESS FOR HEPATITIS C.
I'LL JUST SORT OF POINT OUT THAT
TO SOME EXTENT BECAUSE OF THE
CHALLENGES WITH THIS PARTICULAR
CHRONIC VIRAL INFECTION AND HOW
MANY FEEL A VACCINE IS NEEDED TO
ACHIEVE GLOBAL VIRAL CONTROL OF
THIS VIRUS.
AND I THINK GIVEN THE SUCCESS OF
THE COVID-19 VACCINE THIS REALLY
HAS RESURRECTED MORE ACTIVITY IN
THE HIV SPACE TO DEVELOP A
VACCINE AND JAKE LIANG IN THE
AUDIENCE HERE REALLY I THINK
HELPING TO FACILITATE THAT BY
CALLING FOR US TO CONSIDER THE
POSSIBILITY OF DOING CONTROLLED
HUMAN INFECTION MODELS FOR
TESTING OUT VACCINE REGIMENTS
AND I THINK THAT WOULD REALLY
GREATLY ACCESS -- ACCELERATE
WITH A SMALL CLINICAL PROGRAM
WHAT KINDS OF VACCINE MODALITIES
WILL WORK BEFORE YOU GO INTO
MORE EXPENSIVE TRIALS.
SO IN TERMS OF DISEASE
PROGRESSION, I THINK WE HAVE A
LOT OF DESCRIPTIVE DATA FROM
ACTUALLY ANALYZING LIVER SAMPLES
FROM HIV-INFECTED PEOPLE AND
REALLY WHAT DRIVES THE
PROGRESSION TO CIRRHOSIS AND THE
DEVELOPMENT OF WHAT DRIVES LIVER
CANCER IS NOT COMPLETELY CLEAR.
AND THE GOOD NEWS IS SHOWN HERE
ON THE LEFT THIS IS A STUDY THAT
CAME OUT ABOUT A YEAR AGO WHICH
SHOWS YOU THE DIFFERENCE IN THE
PATTOCELLULAR CARCINOMA INCI
INCIDENTS IN VIRAL CURE
POPULATION AND YOU CAN SEE IT'S
BETTER IF YOU'RE TREATED BUT IF
YOU LOOK AT THAT THERE'S ALSO
SOME BAD NEWS AND THAT IS IF
YOU'RE CHRONICALLY INFECTED WITH
HEPATITIS C AND B IS NOT GOOD
BUT EVEN IF YOU'RE CURED OF
HEPATITIS C OR HAVE SUPPRESSED
HPV YOU'RE STILL IN THE HIGHER
RISK GROUP OF DEVELOPING LIVER
CANCER.
THOUGH WE GET RID OF THE VIRUS
PRESUMABLY IN THE CASE OF HCV
THE FACTOR ARE GIVEN RISE TO AN
ONCOGENIC EVENT HAVE ALREADY
OCCURRED IN SOME PEOPLE.
THOUGH THE VIRUS IS GONE THEY
DEVELOP CANCER.
SOW WE'VE BEEN TRYING FOR
LONGER -- SO WE'VE BEEN TRYING
FOR LONGER THAN I'D LIKE TO
ADMIT FOR DEVELOPING THIS AND
THIS OCCUPIED TIME AND EFFORT.
SOME OF THAT JOE IS IN THE LAB
SO HE PROBABLY REMEMBERS SOME OF
THIS AND PEOPLE HAVE TRIED TO
ADAPT IT TO MOUSE CELLS AND
PEOPLE ARE STILL TRYING TO DO
THAT AND CREATE AN INFECTION
MODEL THAT MIMICS WHAT WE SEE IN
PEOPLE.
AND TRYING TO IDENTIFY FACTORS
THAT WERE HUMAN SPECIFIC
DIFFERENT IN MICE AND POSSIBLY
WE COULD THEN ENGINEER ANIMALS.
THAT DID WORK AND WE MANAGED TO
MAKE TRANSGENIC MICE FOR ATP
REPRODUCTION AND ENTRY BUT IT
DIDN'T ESTABLISH A CHRONIC
INFECTION.
SO IT WAS VERY LOW-LEVEL
REPLICATION.
SO THE OTHER THING WHICH I THINK
IS ALSO BEEN USEFUL IN THE
HEPATITIS C WORLD WITH DUCK
HEPATITIS IS LOOKING FOR A
RELATIVE OF HEPATITIS C THAT
NATURALLY INFECTED SMALL ANIMAL
LIKE THE LABORATORY MOUSE SO WE
GOT INVOLVED IN THIS AND SORT OF
WITH THE ADVENT OF NEXT
GENERATION SEQUENCING, PEOPLE
WERE BEGINNING TO FIND HEPATITIS
C RELATIVES ALL OVER THE PLACE
IN NATURE.
THIS UNFOLDED I WAS AT THE
SCHOOL OF PUBLIC HEALTH AND IAN
IS WHAT YOU WOULD CALL A VIRUS
HUNTER AND THEY WOULD SEQUENCE
ANIMAL SAMPLES AND LOOKING TO
SEE IF THEY FOUND ANY
INTERESTING INFECTIOUS AGENT.
THE FIRST THEY FOUND WAS ONE
THAT ACTUALLY INFECTED THE FIRST
REPORT WAS DOGS BUT THAT TURNED
OUT NOT TO BE RIGHT.
IT ACTUALLY INFECTS HORSES.
SOW AT LEAST ON THE ROCKEFELLER
CAMPUS HORSES ARE NOT ALLOWED
BUT NONETHELESS WE STARTED
WORKING ON THIS IN COLLABORATION
WITH A GROUP IN CORNELL IN
ITHACA IT'S NOT THE SMALL ANIMAL
MODEL WE WANTED.
AND THE NEXT CALL FROM IAN CAME
FROM AND HE'D IDENTIFIED A NEW
VIRUS IN DEER MICE THE THOUGHT
THIS IS GREAT.
WE HAVE A VIRUS AND TRIED TO
INFECT LABORATORY MICE AND THAT
FAILED AND EVEN IF WE HAD
SUCCEEDED EVEN USING THIS MODEL
IN ITS NATURAL HOST IS NOT A
GOOD IDEA BECAUSE THE DEER MICE
ARE FAR DISTANT GENETICALLY AND
WHAT WE HAVE FOR DOING
LABORATORY STUDIES IN MICE WOULD
HAVE TO BE REINVENTED FOR THAT
MODEL.
SO OTHERS HAVE FOUND VIRUSES IN
BATS WHICH HAVE SORT OF A
POPULAR RESERVOIR FOR MANY KINDS
OF VIRUSES.
.
AGAIN, NOT BE AN EASY LABORATORY
MODEL AND IN GIBBONS NOT A SMALL
ANIMAL MODEL AND NON-HUMAN
PRIMATE MODEL AND SO IT TURNED
OUT IAN TEAMED UP WITH A COUPLE
PEST CONTROL GROUPS IN THE NEW
YORK CITY AREA AND TRAPPED A
BUNCH OF RATS AND SLICED AND
DICED AND SEQUENCED THESE GUYS
AND FOUND TWO TYPES OF NORWAY
RAT HEPATOVIRUS AND TELL YOU
WHAT WE FOUND USING THE MODEL
AND TRY TO BE SHORT SO I DON'T
RUN OVER TOO MUCH TIME BUT IT IS
KIND OF IRONIC THE POTENTIAL
ANIMAL MODEL WE'VE BEEN
SEARCHING FOR DECADES WAS
ACTUALLY SCURRYING AROUND THE
STREETS AND DINING ON NEW YORK
CITY GARBAGE BEFORE IT GOT
PICKED UP AND SO ON.
WE GOT THESE VIRUSES FROM IAN
AND WE HAVE A VARIANT THAT
DIFFERS BY THREE OR FOUR.
I'LL SHOW YOU SOME PROPERTIES.
ONE OF THE THINGS THE VIRUS DOES
IS REPLICATE AS YOU CAN SEE IN
THE UPPER LEFT HAND PANEL TO
HIGH LEVELS QUICKLY.
IT'S CLEARED USUALLY IN TWO TO
THREE WEEKS IN THE C57 BLACK 6
MICE IS A POPULAR STRAIN OF
LABORATORY MICE.
THIS INCREASE IN VIREMIA LATER
ON, NOT AT THE BEGINNING, IS
ASSOCIATED WITH A RISE IN AL
ALOMINO TRANSFERASE AND IN K
CELLS THEY COME TO THE LIVER
QUICKLY AFTER OF THE INFECTION
AND ACCUMULATE THERE.
BUT WE KNOW THEY'RE NOT
IMPORTANT FOR VIRAL CLEARANCE
BECAUSE IF YOU GET RID OF THEM
THE VIRUS IS STILL CLEARED WITH
APPROXIMATELY THE SAME KINETICS.
.
YOU LOOK AT CELLS INVOLVED IN
THE CD4T CELLS YOU CAN SEE A
PEAK THAT SHOW UP IN THE LIVER
SORT OF AFTER SEVERAL DAYS AFTER
INFECTION.
AND A VA WENT ON TO SHOW CD4s
WERE IMPORTANT BUT IT WAS
PROBABLY CD8s ANTIGEN SPECIFIC
THAT WERE IMPORTANT SORT OF IN
CLEARANCE.
SO THIS IS GREAT.
AND IT'S A POTENTIAL MODEL FOR
LOOKING AT THINGS YOU CAN LOOK
AT WITH AN ACUTE INFECTION MODEL
LIKE THIS.
BUT IT'S NOT LIKE HEPATITIS C IN
HUMANS WHERE 70% OF HUMANS
BECOME CHRONICALLY INFECTED AND
OFTEN DON'T KNOW THEY'RE
INFECTED.
IT TURNS OUT IF YOU DEPLETE CD8
AS YOU CAN SEE HERE IN THE GRAY
DOTS IT SLOWS CLEARANCE AND IF
YOU DEPLETE CD4 CELLS BEFORE
VIRUS INFECTION AND IN
ADDITIONAL TREATMENTS AFTER
VIRUS INFECTION THE ANIMALS
BECOME CHRONICALLY INFECTED.
SO THIS GIVES US BASICALLY A
CHRONIC INFECTION CAPACITY VIRUS
MODEL WHERE IT WORKS IN
LABORATORY MICE.
AND THE CD4s COME BACK AND ONCE
THE ANTIBODY USED TO DEPLETE IT
BUT IT DOESN'T INFLUENCE THE
NATURE OF THIS CHRONIC
INFECTION.
MOST THE ANIMALS STILL REMAIN
CHRONICALLY INFECTED.
SO SOME OF THE THINGS WE'VE
LEARNED FROM THIS ARE THAT WE
BELIEVE THIS INFECTION IS
EXCLUSIVELY FOR HEPATOCYTES IN
THE LIVER AND BECAUSE A SMALL
MICRO RNA FOUND IN HEPATOCYTES
IS REQUIRED FOR THE REPLICATION
OF THE VIRUS AS IT IS FOR
HEPATITIS C.
SO WE BELIEVE IT'S PROBABLY AN
EXCLUSIVE HEPATOTROPIC INFECTION
UNLIKE OTHER MODELS AND HE WAS
ABLE TO SHOW YOU GENERATE A
STRONG IMMUNE RESPONSE AND THERE
IS IMMUNITY OF T CELL DEPENDENT
CLEARANCE AND THESE ANIMALS ARE
SUSCEPTIBLE TO REINFECTION
THOUGH IT'S CLEARED MORE QUICKLY
AND WITH THE INCREASE INFECTION
AND I THINK THIS IS A REALLY
INTERESTED MODEL TO STUDY BASIC
ANTIVIRAL IMMUNOLOGY FROM HIRES
HOST INTERACTIONS AND IT IS
THOUGH THE DETAILS IN TERMS OF
THE ACTUAL ANTIGENS ARE GOING TO
BE DIFFERENT THAN HEPATITIS C IT
CAN BE USED AS A PLATFORM TO TRY
AND TEST DIFFERENT IMMUNIZATION
STRATEGIES.
THE QUESTION WAS IS THIS A MODEL
FOR ATV RELATED LIVER DISEASE
LONGER TERM WHICH IS WHAT WE'RE
TRYING TO UNDERSTAND IN THE CASE
OF CHRONIC HEPATITIS C
INFECTION.
THIS IS ONE OF THOSE THINGS THAT
SORT OF POPS UP BY CHANCE IN
SCIENCE AND WE HAD A COUPLE OF
THESE ANIMALS THESE MICE
CHRONICALLY INFECTIONS WITH NRHV
THAT SOMEHOW IT ESCAPED
SACRIFICE IN THE ANIMAL FACILITY
AND SOMEHOW THESE TWO MICE WERE
APPROACHES TWO YEARS OLD HAD
MANAGED TO HIDE OUT -- THIS WAS
LIKE RETIREMENT COMMUNITY FOR
LABORATORY MICE.
ANYWAY, WHEN THEY WERE
SACRIFICED WE FIND THAT THERE
WAS PRETTY STRIKING PATHOLOGY IN
THE LIVER ALMOST KIND OF IN A
CIRRHOTIC AND YOU DON'T REALLY
KNOW IF THIS IS JUST A PHENOTYPE
OF OLD MICE.
IT'S NOT LIKE WE HAD UNINFECTED
CONTROLS.
WE ONLY HAD A FEW ANIMALS.
BUT IT GOT US INTEREST IN MAYBE
TRYING SOME LONGER TERM
INFECTION MODELS AND SEEING WHAT
HAPPENS.
DO THEY DEVELOP ADVANCED
PATHOLOGY AND THAT'S WHAT I'M
GOING TO FOCUS ON FOR THE REST
OF THE TALK.
THIS IS WORK THAT WAS REALLY
CARRIED OUT BY MARIANNA BATISTA
AND SCOTT CHENG AT KETTERING AND
A GRADUATE STUDENT AND WE HAVE
KIND OF AN INTERNATIONAL
COLLABORATION WITH VARIOUS LABS
TO WORK IN THIS SPACE BUT BRAD
ROSENBERG AND OTHERS ON THE
IMMUNOLOGY SIDE AND TIM SHEEHAN
ON THE MOUSE GENETICS SIDE AND
SCOTT LOWE ON THE CANCER SIDE.
WE SET UP THIS RATHER LARGE
PILOT EXPERIENCE WITH 95C57
BLACK 6 MICE AND A MIXTURE OF
MALES AND FEMALES SO IF YOU LOOK
AT THIS SCHEME HERE CD4s WERE
DEPLETED AT DAY MINUS 3 AND THEN
ON DAY ZERO THEY WERE EITHER
SORT OF SHAM INOCULATED WITH PBS
OR WITH VIRUS AND THIS IS A VERY
HOT VIRUS.
IT'S 10 TO THE FOURTH GENOME
EQUIVALENCE AND THERE WAS
ANOTHER CD4 DEPLETION IN 10
WEEKS AND WE'RE GOING TO TAKE
MONTHLY BLEEDS AND LOOK AT
THINGS IN THE PERIPHERY BUT HAVE
TERMINAL TIME POINTS AND COLLECT
SAMPLES TO DO ALL KINDS OF
THINGS AT 6, 12 AND 18 MONTHS.
THAT WAS ALL WELL AND GOOD AND
INITIATED.
AT THE SIX-MONTH HARVEST TIME
COVID-19 SHOWED UP.SO A LOT OF
THE FACILITIES WE WERE PLANNING
ON USING TO AND EVEN GETTING IN
THE ANIMAL FACILITY FOR THINGS
THAT WERE NOT CORONAVIRUS
FOCUSSED WAS VERY CHALLENGING.
.
AND MANY OF THESE SAMPLES ARE
STILL SORT OF UNDER ANALYSIS.
I WILL SHOW YOU A LITTLE BIT
ABOUT WHAT HAPPENED.
IF YOU LOOK AT THIS SLIDE IN THE
UPPER LEFT THIS SHOWS VIREMIA
OVER TIME AND SEE THIS HIGH
LEVEL OF VIREMIA AND SORT OF
THIS PERSISTS FOR A WHILE AND
THEN BASICALLY IT LEVELS OUT AND
PLATEAUS BY ABOUT SIX MONTHS.
THERE DOESN'T REALLY SEEM TO BE
ANY DIFFERENCE BETWEEN THE MALE
OR THE FEMALE MICE.
AND THE OTHER THING THAT'S
INTERESTING IS THAT WE DO SEE
LATE CLEARANCE IN SOME OF THESE
ANIMALS AND AT LEAST IN THIS
PILOT EXPERIMENT THE NUMBERS ARE
SMALL.
THE FEMALE MICE SEEM TO DO A
LITTLE BIT BETTER IN TERMS OF
CLEARING THE VIRUS BUT ACTUALLY
NEITHER SEX IS ALL THAT GOOD AT
CLEARING THE VIRUS.
THAT SHOWS YOU A LITTLE BIT AND
SUGGESTS THERE MIGHT BE SOME
SORT OF SEX-DEPENDENT
DIFFERENCES WHICH IS SOMETHING
WE SEE IN HEPATITIS C
PATHOGENESIS AS WELL.
THAT'S SOMETHING WE'RE KEEPING
OUR EYE ON.
SO WE FROM THESE TERMINAL TIME
POINTS WE COLLECTED LIVER
SAMPLES AND ASKED A LIVER
PATHOLOGIST AT NYU WE WORKED
WITH FOR MANY YEARS TO TAKE A
LOOK AT THESE LIVERS AND WHAT HE
SAW IS SHOWN AND THIS IS A GRAPH
THAT HAS BASICALLY INFLAMMATION
ON THE Y AXIS AND FIBROSIS ON
THE X AXIS AND YOU CAN SEE AT
SEVEN MONTHS POST INFECTION
THESE INFECTED ANIMALS, MALES
AND FEMALES WERE SHOWING
EVIDENCE OF LIVER DAMAGE.
THAT SORT OF GETS WORSE AT 12
MONTHS POSTINFECTION AND THEN AT
18 MONTHS THE TREND IS STILL
THERE BUT AGAIN THERE'S MORE
HETEROGENEITY IN THE OLDER MICE
IN TERMS OF THE LIVER PATHOLOGY
THAT THEY SHOW.
EVEN IN THE ONES IN THE INFECTED
AND IF YOU LOOK AT THE LEFT AT
THE HISTOLOGY YOU SEE
DIFFERENCES IN TWO THINGS THAT
WE SEE IN HUMAN HEPATITIS C
FIBROSIS IN THE LOWER LEFT PANEL
AND INFLAMMATION AND SIMILARITY
TO HCV AND STARTING AT 12 MONTHS
AND BECOMING EVIDENT BY 18
MONTHS IF YOU LOOK AT THE MOCK
LIVERS THEY LOOK LIKE NICE
HEALTHY LIVERS BUT IF YOU LOOK
AT THE ONES BY THE VIRUS FOR 18
MONTHS, BASICALLY WE SAW THE
LIVER TUMORS THAT WERE POPPING
UP AND THIS WAS INTERESTING IN
THE SENSE THAT IN MALES THE
FREQUENCY WAS SURVIVORS OF 18
MONTHS WITH BASICALLY 100% SO
THIS IS REALLY A REMARKABLE
NATURAL INFECTION MODEL THAT
LEADS TO HIGH INCIDENTS OF LIVER
CANCER.
THERE'S BEEN SUGGESTIONS BY
GRANT REVIEWERS THAT WELL, THIS
ISN'T WHAT HAPPENS IN HEPATITIS
C.
NOT EVERYONE GETS CHRONICALLY
INFECTED GETS INFECTED WITH
LIVER CANCER BUT IN THIS
UNIFORMITY WE SEE IS AN ASSET
NOT A LIABILITY.
IN THIS PILOT WE SAW A LOWER
INCIDENTS OF TUMOR FORMATION IN
FEMALE MICE.
IF YOU LOOK AT TUMOR VOLUME
AGAIN THERE SEEMED TO BE A TREND
THOUGH I THINK IT'S NOT HOLDING
UP IN A SECOND EXPERIMENT WE'VE
DONE TO FOLLOW THIS UP IN TUMOR
VOLUME BETWEEN THE FEMALE MICE
AND THE MALE MICE.
IF YOU LOOK AT THE HISTOLOGY AND
THE INFLAMMATION WHICH IS
VARIABLE FROM MOUSE TO MOUSE AND
THE PRESENCE OF THESE LYMPHOID
AGGREGATES WHICH IS COMMONLY
FOUND IN HUMAN HCV BUT WE DON'T
KNOW WHAT THEY'RE UP TO.
THERE'S A CONVERGENCE INFECTED
VERSUS NORMAL AND THERE'S A HIGH
CONVERGENCE OF THINGS SHOWN IN
RED OR DOWN IN BLUE.
AND WE BEGAN TO LOOK AT THE
GENETICS OF THESE TUMORS AND
AGAIN WE FIND THEIR QUITE
HETEROGENOUS AND LOCI ASSOCIATED
WITH CANCERS AND YOU CAN SEE IF
YOUR SAMPLE SHOWS ANY OF THESE
AND FIND A SIGNATURE IN SOME OF
THESE TUMORS.
THIS IS SEVEN TUMORS LOOKED AT
IN SOME DETAIL AND THREE APPEAR
TO HAVE SUSPICIOUS K RAS
MUTATIONS THAT COULD BE DRIVING
THIS AND IF YOU LOOK AT THE
TRANSCRIPTOME AND KRAS TUMORS
WITH PROTEIN AND OTHER THINGS
SEEN AS TRANSCRIPTIONAL CHANGES
IN ATV.
I'M GOING A LITTLE TOO LONG BUT
I THINK IT'S INTERESTING THAT
THIS MODEL SORT OF DOES LEAD TO
HEPATITIS FIBROSIS IN CANCER.
THERE ARE POSSIBLE DIFFERENCES
BETWEEN MALES AND FEMALES WHICH
WE'RE GOING TO LOOK AT MORE
CAREFULLY.
THEY'RE HETEROGENOUS.
THERE ARE SOME COMMON HUMAN ATC
MARKERS UPREGULATED AND MANY K
RAS IS ACTUALLY AN INFREQUENT
DRIVER OF ATV ASSOCIATED LIVER
CANCER BUT STILL EARLY DAYS I
DON'T KNOW WHAT OTHER THINGS
WE'RE GOING TO FIND.
AND SO I THINK WE FINALLY HAVE A
PHYSIOLOGICALLY RELEVANT MOUSE
MODEL TO DISCOVER NEW INSTANCES
AND I WANT TO HAVE A FOOT NOTE
HERE I DON'T KNOW HOW MANY OF
YOU ARE FAMILIAR WITH THE
COLLABORATIVE CROSS GENETIC
MOUSE RESOURCE.
AND WE'VE BEEN EXAMINING THIS TO
SEE IF WE SEE ANY DIFFERENCE IN
THE BEHAVIOR IN THE DIFFERENT
COLLABORATIVE CROSSES AND THIS
IS WORK TIM SHEEHAN HAS BEEN
DOING AT THE UNIVERSITY OF
CALIFORNIA OR UNIVERSITY OF
NORTH CAROLINA.
THE COLLABORATIVE CROSS INCLUDES
A COLLECTION OF PARENTAL MOUSE
STRAINS GENETICALLY VERY DIVERSE
AND GENETICALLY INBRED BUT
DISTINCT FROM EACH OTHER YOU CAN
TAKE A LOOK AT THE STRAINS WITH
WHATEVER YOU'RE INTERESTED IN
LOOKING AT AND BASICALLY SEE IF
YOU SEE AN INTERESTING
PHENOTYPIC DIFFERENCE WITH YOUR
PERTURBATION.
SO AND WHAT TIM HAS DISCOVERED
IS THAT IF YOU AND THIS SHOWS
BASICALLY HOW THE VIRUS BEHAVES
IN THE BLACK 6 MICE AND 71 AND
80 AND POINT OUT CCO71 BECAUSE
THIS MOUSE STRAIN ACTUALLY
ALLOWS INFECTION AND CHRONIC
INFECTION WITHOUT CD84
DEPLETION.
THIS IS A NATURAL MOUSE GENETIC
BACKGROUND THAT ALLOWS THIS
VIRUS TO ESTABLISH A FOOTHOLD.
SO WE'RE ACTUALLY VERY EXCITED
ABOUT THAT.
WE DO SEE IN THE CC071 MODEL
FIBROSIS OVER TIME.
IT'S AN INTERESTING MODEL.
WE'RE WAITING ON THE ANIMALS TO
SEE IF THEY DEVELOP CANCER AFTER
LONGER INFECTIONS BUT THERE'S
SOME ENCOURAGING INDICATIONS IN
THAT DIRECTION.
SO WE'RE REALLY EXCITED ABOUT
THIS MODEL.
IT'S BEEN A CHANCE DIVERSION IN
THE AGING MICE BUT I THINK IT
DOES PROVIDE A PLATFORM TO
REALLY BEGIN TO UNDERSTAND HOW
CHRONIC INFECTION CAN LEAD TO
CANCER AND WE'LL BEGIN LOOKING
AT SEX AND COMORBIDITIES LIKE
ALCOHOL AND DIET.
THE ROLE OF FIBROSIS.
ONE OF THE NICE THINGS THAT
SCOTT LOWE'S LAB IS DEVELOPED IS
A CAR T APPROACH FOR SENESCENT
CELLS THAT CAN REVERSE THE
ABILITY OF THE INSULTS TO
GENERATE FIBROSIS.
THE QUESTION IS IF WE TRY THAT
INTERVENTION IS IT GOING TO
PREVENT THE ANIMALS FROM CANCER.
THE IMPACT OF CURE AND I ALLUDED
TO THAT PEOPLE WITH MORE
ADVANCED LIVER DISEASE EVEN WHEN
THE VIRUS IS GONE ARE STILL AT
RISK OF DEVELOPING CANCER.
SO WE CAN ACTUALLY USE THIS
MODEL TO FIND OUT WHAT'S THE
TIMING OF THAT.
IF YOU GET OF THE INFECTION AND
WE BELIEVE WE CAN DO THAT QUITE
EFFECTIVELY HOW LONG DO THEY
HAVE TO BE CHRONICALLY INFECTED
WHERE IT DOESN'T MAKE A
DIFFERENCE IF YOU GET RID OF THE
VIRUS.
VERY INTERESTED IN USING THIS AS
A MODEL FOR LOOKING AT NOT ONLY
IMMUNE FUNCTION AND DYSFUNCTION
AND ALSO TESTING
IMMUNOTHERAPIES.
AND AS I'VE MENTIONED THE ROLE
OF HOST GENETICS AND INFECTION
AND OUTCOME.
WITH THAT I ACKNOWLEDGE THE
PEOPLE INVOLVED IN THIS.
I'VE POINTED OUT THE PEOPLE THAT
HAVE REALLY BEEN INVOLVED IN THE
PILOT EXPERIMENTS.
THE OTHER REPRESENT GROUPS THAT
WERE COLLABORATING WITH AROUND
THE WORLD AND THERE'S A PICTURE
OF OUR LAB THAT MEETS EVERY
WEDNESDAY WHICH INCLUDES LABS
FROM OTHER FOLKS IN THE NEW YORK
AREA.
YOU CAN SEE THE THREE THAT HANG
OUT IN THE OFFICE AND PROVIDE
SOME ENTERTAINMENT.
I WON'T HAVE TIME TO TALK ABOUT
IT TODAY BUT MAYBE I'LL ZIP
THROUGH THESE BECAUSE I THINK
THEY'RE TOPICS THAT PLAY BE FUND
TO DISCUSS IN THE AFTERNOON.
WE'RE DOING A LOT OF WORK IN
HEPATITIS B AS MANY FOLKS
SHIFTED THEIR ATTENTION FROM HCV
CURE TO A FUNCTIONAL CURE FOR
HBV AND THE VACCINE STRAIN THAT
HAS ONE SHOT LIFE LONG
PROTECTION BUT THERE ARE
UNFORTUNATE VACCINEES OF YELLOW
FEVER THAT DEVELOP THIS ATROPIC
DISEASE.
IT TURNS OUT AGAIN I WON'T SHOW
YOU THE DATA THE DIFFERENCE
BETWEEN THE VACCINE STRAIN AND
THE VIRULENT STRAIN HAS A LOT OF
DO WITH INTERFERON.
IF AN INDIVIDUAL WHO GETS THE
VACCINE IS DEFECTIVE IN TYPE 1
INTERFERON SIGNALLING THEY'RE AT
RISK AT DEVELOPING THE DISEASE
AND IT CAN GO WILD AND CAUSE
PROBLEMS.
VIRUS-HOST INTERACTION.
WE GOT INTERESTED IN INTERFERON
BEING USED TO TREAT HEPATITIS C
AND INFORMATION HOW IT MAY BLOCK
IT IT MAY PROVIDE INCLUDE HOW TO
MOVE FORWARD TO IN MOVE
TREATMENT OVER SYSTEMIC
INTERFERON AND EVEN THOUGH THAT
DIDN'T PAN OUT AND THE DAA TOOK
OVER THE PLAYING FIELD IN TERMS
OF TREATMENT WE DID GET HOOKED
ON TRYING TO UNDERSTAND HOW
INTERFERON WORKS AND IT'S
FASCINATING.
MORE LATELY WE'VE BECOME
INTERESTED IN A PHENOMENON
PEOPLE CALL ANTIGEN INDEPENDENT
INNATE IMMUNITY.
THIS HAS TO DO WITH A MODEL THAT
A LAB HAS BEEN WORKING ON WHERE
IF YOU INFECT A MOUSE WITH SARS
COV2 AND SEE A TRANSIENT WEIGHT
LOSS AS A RESULT WHAT HAPPENS IF
YOU CHALLENGE THAT ANIMAL WITH A
RESPIRATORY PATHOGEN NOT RELATED
TO SARS COV2 AND WHAT ALEX HAS
SHOWN AND FIGURED OUT SOME OF
THE MECHANISMS INVOLVED HERE IS
THAT THESE ANIMALS AT LEAST FOR
A PERIOD OF TIME ARE HIGHLY
RESISTANT TO A CHALLENGE OF
INFLUENZA.
THE INITIAL INFECTION OF SARS
COV2 CHANGED THE BEHAVIOR AND
ALVEOLAR MACRO PHAGES SO
INFLUENZA IS RESOLVED RATHER
THAN LETHAL.
WITH THAT I'LL READ A QUOTE FROM
LOUIS THOMAS WHO WROTE THIS TO
COMMEMORATE THE 40 YEARS OF THE
LASKAR PRIZES MOST THE
CELEBRATED ACHIEVEMENT HAVE
THEIR ORIGINS AND MOMENTS OF
SURPRISE IN THE LABORATORY
UNPLANNED FOR, UNANTICIPATED,
UNPREDICTED.
NO COMMITTEE TO SURVEY THE
FUTURE PROSPECT FOR BIO MEDICAL
RESEARCH CAN POSSIBLY GUESS AT
THE THINGS THAT LAY AHEAD.
NO BRANCH OF GOVERNMENT COULD
HAVE LAID DETAILED PLANS NOR
SCIENTIFIC NEEDS OF THE FUTURE
BEYOND ASSERTING WHAT THE
COUNTRY NEEDED WAS MORE
FUNDAMENTAL KNOWLEDGE ABOUT
HUMAN FORM AND FUNCTION, ABOUT
THE AGENTS AND INFLUENCE OF
RESPONSIBLE DISEASE AND IN
NATURE ITSELF.
SCIENTIFIC RESEARCH WORKS AND
IT'S THE ONLY WAY TO GET AT THE
UNDERLYING MECHANISMS OF DISEASE
AND THE ONLY WAY TO LEARN WHAT
TO DO ABOUT THEY WILL.
AND I THINK THAT IS AS TRUE
TODAY AS IT WAS WHEN LOUIS WROTE
THIS AND I'LL CLOSE BY THINKING
BACK TO MY OWN ORIGINS WHERE
CURIOSITY DRIVEN RESEARCH IN THE
OLD DAYS THAT JOE MENTIONED WITH
THE STRAUSS LAB WE WERE SETTING
AN ENVELOPE STRAND RNA VIRUS
THAT'S NOT A BIG DISEASE THREAT.
CHIKUNGUNYA TURNS OUT TO BE ONE
AND I THINK THE NIH AND NIAID IN
PARTICULAR TO THE SUPPORT OF
THIS WORK AT CAL TECH AND
THROUGHOUT MY CAREER HAS HAD THE
FORESIGHT TO FUND THINGS
OBVIOUSLY TRANSLATIONAL
POTENTIAL AND ALSO THINGS THAT
ENRICH OUR KNOWLEDGE ABOUT
BIOLOGY AND REGARDLESS OF REALLY
TRYING TO BE ABLE TO PREDICT
WHAT THE IMPACT THAT'S GOING TO
BE IN THE FUTURE AND I THINK
IT'S VERY IMPORTANT THAT WE
MAINTAIN THE RIGHT BALANCE
THERE.
SO THANK YOU.
SORRY.
QUESTIONS?
>> I THINK THERE'S ALSO A LOT OF
PEOPLE ONLINE DON'T KNOW.
>> CHARLIE, IMPRESSIVE AND
CUTTING EDGE RESEARCH EVEN ON
THE VIRUS MANY PEOPLE THINK WE
SHOULDN'T AND DON'T NEED TO
STUDY ANYMORE OBVIOUSLY YOU
PROVED THEM WRONG.
TWO QUICK QUESTIONS, IN TYPE 1
INTERFERON INDUCED OR INVOLVED
IN THE HV INFECTION IN VIVO?
THE SECOND QUESTION WOULD BE IS
IT SENSITIVE TO THE DAAs?
HAVE YOU TRIED THAT IN YOUR
MODEL?
>> WE HAVE LOOKED AT THE
BEHAVIOR OF THE VIRUS IN
DIFFERENT KNOCKOUT MICE.
WE DO SEE INTERESTING
DIFFERENCES DEPENDING ON WHETHER
OR NOT THEY'RE DEFECTIVE IN
INNATE IMMUNITY OR NOT.
BUT I DON'T THINK INNATE
IMMUNITY IN AND OF ITSELF IS
SUFFICIENT TO CONTROL INFECTION.
IF FOR INSTANCE IF YOU LOOK AT
THE BEHAVIOR OF THIS AND IN OUR
GENE MOUSE THAT LACKS T AND B
CELLS AND STILL HAS INNATE
IMMUNITY THOSE GET INFECTED TO
HIGH LEVELS AND STAY THAT WAY.
SO FAR WE HAVEN'T SEEN -- WE
HAVEN'T DONE A CONTROLLED
EXPERIMENT AND HAVEN'T SEEN ANY
EVIDENCE THE UKRAINECALLY
INFECTED IMMUNODEFICIENT ANIMAL
DEVELOPED PATHOLOGY.
AND -- AND THE SECOND
QUESTION -- WHETHER
>> WHETHER THE VIRUSES ARE
SUSCEPTIBLE --
>> TIM HAS BEEN LOOKING AT THAT
LOOKS LIKE WE'LL COME UP WITH
CONDITIONS TO CURE.
WE'RE THOUGHT THERE YET BUT
THAT'S SOMETHING WE WANT TO BE
ABLE TO DO TO STUDY THE IMPACT
OF THAT ON PATHOGENESIS.
>> THANKS.
>> OH, NO.
>> YOU MAKE ME WANT TO UNRETIRE.
ONE OF THE QUESTIONS WE'VE HAD
ALONG IS HOW IMPORTANT IS
CIRRHOSIS IN THE DEVELOPMENT OF
CANCER?
AND THE DIFFERENCES BETWEEN THE
U.S. AND JAPAN WHERE HERE WE
MOSTLY HAVE CIRRHOSIS AS A
PRECEDING EVENT, JAPAN, THEY
DON'T ALWAYS HAVE THAT.
SOMETHING TO LOOK AT IN THIS
MODEL AND WONDERING OF THE
ANIMALS THAT DID DEVELOP CANCER,
HOW MANY HAVE PRE-EXISTING
CIRRHOSIS OR ADVANCED FIBROSIS?
>> GREAT POINT AND GREAT
QUESTION.
SOMETHING I DON'T KNOW THE
ANSWER TO BUT IT SEEMS THAT
THESE MOUSE MODELS IN GENERAL
ARE NOT PRONE TO DEVELOPING
CIRRHOSIS AND SO WE RARELY SEE
THAT IN OUR ANIMALS.
SO I KNOW THAT PEOPLE SORT OF
DRAW THIS LINEAR PROGRESSION OF
FIBROSIS, CIRRHOSIS, CANCER BUT
I'M NOT CONVINCED THAT CIRRHOSIS
IS CAUSAL AS OPPOSED TO BEING A
MARKER FOR BASICALLY MORE SEVERE
LIVER DAMAGE IN PEOPLE.
AT LEAST IN THE MOUSE MODEL IT'S
NOT A NECESSARY PRECURSOR.
THERE'S NO PERFECT MODEL AS YOU
KNOW BUT IT'S INTERESTING AND
I'D BE INTERESTED TO HEAR YOUR
THOUGHTS IS SCIRRHOSIS A MARKER
AND CORRELATES WITH ACC BUT
ZVENLT
BUT WE'VE ALWAYS THOUGHT IN
GENERAL IT WAS THE REGENERATION
THAT IN THE PROCESS OF
REGENERATING THE LIVER TO
OVERCOME CIRRHOTIC EVENTS OF THE
MUTATIONS OCCUR INSIDE OF CANCER
BUT NOBODY'S IN VIVO REEL PROVEN
THAT.
>> THE IMMUNE SURVEILLANCE THAT
CAN'T GET RID OF THE VIRUS WILL
BE TRIGGERING A LOT OF
REGENERATIVE ACTIVITY IN THE
LIVER.
WE HAVEN'T DONE A CAREFUL JOB OF
LOOKING AT THAT OVER TIME.
BUT YEAH.
>> THANK YOU, JOHN.
>> IT'S AN INTERESTING MODEL TO
STUDY CANCER AND TO STUDY LIVER
FIBROSIS AND YESTERDAY IT'S A
VIRUS.
DO THE RATS DEVELOP CANCER LATS
THAT LIVE IN SANITARY
ENVIRONMENTS AND NOT IN THE
STREETS OF NEW YORK THAT ARE NOT
EXPOSED TO ANY OTHER TOXIN?
>> I'M NOT SURE THE LIFE SPAN IN
THE STREETS OF NEW YORK.
>> WE'VE LOOKED AT THE RAT MODEL
AND I DON'T THINK THEY'VE SEEN
EVIDENCE OF CANCER IN RATS IN
LABORATORY RATS.
>> AND THEN I WONDER IF WE PUT
THE VIRUS INTO MICE WITH MICE
MICROBIOTA AND A TYPE OF IMMUNE
RESPONSE THEY MAY DEVELOP CANCER
FASTER.
>> I HAVEN'T BEEN ABLE TO
REROUTE OR MICROBIOME FOLKS AND
WE DON'T KNOW HOW THIS BEHAVES
IN GERM-FREE MICE.
IT WOULD BE INTERESTING TO LOOK
AT IT IN OTHER MOUSE MODELS TO
SEE HOW IT THE PATHOLOGY AND
PATHOGENESIS.
>> AND I HAVE A QUESTION.
I WAS WONDERING IF YOU'RE DOING
A MORE BROAD SHOTGUN APPROACH OR
MORE IN THE DIVERSITY OUTBRED
MODEL OR
>> THERE ARE A LOT OF
COLLABORATIVE CROSS STRAINS AND
THIS WAS A PILOT EXPERIMENT
TAKING ADVANTAGE OF THE FOUNDERS
THEY HAVE TIM SHEEHAN HAD ACCESS
TO OTHER THAN IT WOULD BE A
COMPREHENSIVE THING TO DO AND
TIM HAS GONE AHEAD AND DONE THAT
FOR CHRONIC VERSUS ACUTE
RESOLVED INFECTION THE CC071 AND
C57 BLACK 6 MICE.
WE KNOW WHAT'S RESPONSIBLE FOR
THIS AND I WON'T TELL YOU
BECAUSE THAT'S HIS STORY.
>> THANK YOU.
>> OKAY.
>> HI.
MY NAME IS ALAN RYAN AND I'M
GOING TO BRIEFLY INTRODUCE OUR
NEXT SPEAKER, CHANTAL ABERGEL IN
MARSEILLE WHERE SHE LIVES AND
WORKS AND FORMER POSTDOC AT
NIDDK WHO WENT BACK TO MARSEILLE
AND HAS BEEN INVOLVED FROM THE
BEGINNING IN STUDYING WHAT WE
ALL CALL GIANT VIRUSES, VIRUSES
FAR MORE COMPLEX THAN THE
VIRUSES YOU AND I STUDY AND THE
MYSTERY OF WHY THEY'RE SEEMINGLY
REDUNDANT WITH HOST FUNCTIONS.
CHANTAL, AARE YOU THERE?
I'M LOOKING FORWARD TO YOUR
TALK.
OVER TO YOU.
>> THANK YOU.
I'M PLEASED TO BE AT NIH.
I CHANGED A BIT MY FOCUS TO GIVE
FOCUS ON THE VIRUSES.
FIRST THE STUDY OF THE VIRUS.
SO THAT WAS THE FIRST VIRUS
DISCOVERED IN 2004 AND AND THIS
GENOME HAS 70% OF THE GENES AND
TO GIVE PERSPECTIVE IT'S A VIRUS
KNOWN AT THE TIME CD1 AND WE
DECIDE
DECIDED FROM WHAT WE LEARNED
FROM ME VIRUS WE WERE ABLE TO
SEE HOW SUCCESSFUL IT WAS NOT
ONLY MORE AND THERE IS NOW THE
VIRUS AND AND IT INCREASES THE
SIZE AND THERE WHAT IS HAPPENING
SO THIS PARTICLE IS OPEN WHERE
YOU SEE THE TRIANGLE OF THE
CAPSID AND WE HAVE THE
COMPARTMENT NOW WHICH IS
ACCOMPANIES THE GENOME.
THE
AS SOON AS THE TARGET IS OPEN IT
REACHED THE MEMBRANE AND IT'S
CREATED BY THE OPENING AND WE
HAVE THE COMPARTMENT INSIDE THE
TRANSCRIPTION OF THE GENES AND
YOU CAN SEE THE VIRAL FACTORY.
AND EXPANDS AND YOU CAN SEE BY
LIGHT MICROSCOPY WITH THE
NUCLEUS AND THE LATE STAGE OF
THE INFECTION WHERE THE VIRAL
FACTORY TAKING A HUGE AMOUNT OF
THE SPACE AND ALSO IT WILL BE
EXPELLED FROM THE CELL THROUGH
LYSASE OF THE CELL AND THERE WAS
EARLY EXPRESSION RATHER THAN
LATE AND THERE ARE AND FROM THE
TRANSCRIPTION THERE'S A SPECIFIC
STRUCTURE WHICH IS GOING TO BE
RECOGNIZED BY THE ENCODED
MACHINERY THAT WILL RECOGNIZE
AND GET INTO IT AND THEN THE
POLYMERASE OF THE VIRUS IS GOING
TO BE ABLE TO GET TO THE ENZYME
AND HAVE THIS TRANSCRIPTION.
AND NOW I WANT TO PRESENT TO YOU
THE VIRUS.
AND BY THE VIDEO AND INSIDE WE
HAVE THIS COMPARTMENT AND THE
CELL WAS SHOWING SOME STRUCTURE.
SO IN THE LAB WE MANAGED TO
DEVELOP AN OPENING THAT ALLOWED
US FOR THE STRUCTURE INSIDE THE
COMPARTMENT THAT CONTAIN DNA.
WE COLLECTED DETE OF THE
STRUCTURE AND WERE ABLE TO SOLVE
STRUCTURES.
AND THIS CONTAINS THE GENOME AND
THE FORM OF THE CELL STRUCTURE
AND THE DNA AND FINALLY THERE'S
STRANDS OF DNA SO WE HAVE IT
INSIDE THE CAPSID AND THE
CONTENT OF THIS FIBER AND THIS
IS ACTUALLY AN ENZYME.
AND THIS STRUCTURE HAS BEEN
SOLVED AND HERE WE LOOK AT THE
STRUCTURE.
THIS WAS TARGETED IN THE CAPSID.
AND THE DIFFERENCE BETWEEN THE
PROTEINS WAS THIS WAS PRESENTED
IN THE CAPSID AND THE SUB UNIT
OF THE RNA POLYMERASE WERE
PRESENTED WITH THE FIBER.
WE WERE OBSERVE IING OBSERVING D
THE RNA POLYMERASE.
AND LOOK AT HOW IT'S RELAXING
A
AND LOOK AT THE PROMOTER COULD
ACTUALLY BE RELAXED AND THEN
PROCEED AFTER.
SO SO AFTER SIX YEARS OF HARD
WORK AND THANKS TO A POSTDOC IN
OUR LAB WE KNOCKED OUT EACH OF
THE OXIDASE AND WE HAVE WILD
TYPE AND AND WHEN WE KNOCK OUT
THE WILD TYPE WE GET SURPRI
SURPRISINGLY STABLE FIBERS.
WHEN WE KNOCK OUT THE STRUCTURE
THE FIBER AND WE'RE ABLE TO SEE
WHAT WE SUSPECT AND MANY CHANGES
WHERE AND WE APPLY THE DIFFERENT
STRUCTURES DIDN'T WORK AND
COMPATIBLE FOR THE ANALYSIS AND
IF WE COMPARE THE VIRUS THAT HAS
BEEN KNOCKED OUT WE ONLY
INCREASE THE SECTION IN THE
MICE.
BUT WE TAKE THE STABLE
STRUCTURE.
IT'S NOT ONE PROTEIN OR ONE
STRUCTURE BUT AND IMAGINE A
REASON FOR THE TWO EXTREME
STRUCTURES BY DESTROYING AND THE
DNA IS ACTUALLY IN SIX STRAINS
THE SIX STRAINS DO NOT ALL HAVE
THE SAME LENGTHS.
THE FIRST IS GOING TO BE DONE
WITH THE FIRST AND YOU CAN SEE
HERE.
AND I TOLD YOU WE WERE WONDERING
WHY IT WAS PRESENTED AND NOT ON
THE GENOMIC SIDE.
WE DID DO THE FIRST AND REPLACED
TERMINAL WITH THE GFP AND ACTUAL
LY SHOWING THE PARTICLE AND THIS
IS ENRICHED.
WE SHAPE THE PARTICLE AND YOU
SEE THE DEGREES.
I'VE TOLD YOU FROM THE BEGINNING
AND THIS CAN BE TOTALLY
DIFFERENT AND GETS INSIDE THE
GENOME FOCUSSED FROM THE ENZYMES
INVOLVED IN MAKING THIS THIS
WORK WAS DONE AND THE TRUCK
STRUCTURE SHOWED AND THE
POLYSACCHARIDE AND THERE ARE
HUGE POLYSACCHARIDE.
THIS ADD INIMPACT ON THE
PARTICLE AND THE INDICATES WHERE
THE CONTROL VIRUS PRESENTED HERE
YOU CAN SEE THE CAPSIDS.
FINALLY, HAVE THE PROTEINS USED
AND YOU CAN SEE THIS IS THE SAME
PROTEIN AS COLLECTED.
OVER ALL IT APPEARS THE ONE
VIRUS.
THIS IS SHOWED SOME EQUIVALENCY
A
AND SECURE ESSENTIAL FUNCTION
SUCH AS GENOME PROTECTION AND
THE ENVIRONMENT.
I WILL NOW GO TO ANOTHER FAMILY
OF THE NOT SO GIANT VIRUSES WITH
PARTICLES AND GENOMES WHICH ARE
AROUND 400KV AND ONE QUESTION
WAS THE VIRUSES HAVE A
POLYMERASE AND LOOK AT THE
TRANSCRIPTION AND YOU NEED TO
WAIT TO TRANSCRIBE THE INFECTION
AND HERE'S THE PARTICLES THERE
IS NO POLYMERASE AND WE STARTED
FROM THE BEGINNING AND AT THE
END OF THE INFECTION WE WERE
SEEING THIS AND DECIDED TO GO
BACK TO THE TIME OF THE
INFECTION.
WE USED A TRICK PROTEIN AND WE
CAN SEE THE PROTEIN EXITING AND
GOING INTO THE CYTOBLAST BUT
POSTINFECTION WHEN THEY FULLY
MATURE WE CAN SEE THOSE VIRUSES
ARE A MISSING LINK BETWEEN THE
VIRUSES AND AT THE INITIATION OF
THE INFECTION CHANGE IT IN SUCH
A WAY THIS IS AFTER IF WE GO
BACK.
IF THERE IS A FUSION BETWEEN H4
AND H2B AND H2A.
IF WE OVEREXPRESS FROM THE
INFECTION YOU CAN SEE THE YAWN
-- NON INFECTED AND ALONG THE
NUCLEI AND THE NORMAL INFECTION
CYCLE WE CAN SEE THIS IS STILL
INFECTION IN THE CASE OF THE
INFECTION IS TAKING PLACE AND I
GUESS I SHOULD MAKE IT SMALLER.
THIS IS IN THE LAB THEY DID IN
VITRO AND ONE DIFFERENCE IS THAT
THEY WERE UNSTABLE AND WE
EXPRESSED INTO THE CELL.
THOSE MOST LIKELY INVOLVED IN
THE TRANSFER OF THE GENOME
INSIDE THE CAPSID AND THIS
TRANSFER ACTUALLY WILL MAKE IT
AND THEY NEED THE POLYMERASE TO
COME TO THE DNA AND AS SOON AS
THE PARTICLE STARTS OPENING THE
NUCLEOSOME WILL BE EXPOSED AND
START THE TRANSCRIPTION AND HERE
I PRESENT ALL THE FAMILIES IN
THE LAB AND THEY'RE INTERESTING
BECAUSE THEY DON'T HAVE THE SAME
GENOME SIDE AND THEY DO HAVE
GENOMES AND THEY'RE TOTALLY
DIFFERENT.
IN EACH THEY'RE 70% OF THE GENES
FOR WHICH WE HAVE NO IDEA OF
WHAT THEY ARE DOING AND DO NOT
RESEMBLE ANYTHING AND YOU CAN
ACCUMULATE AND THEY CORRESPOND
TO FUNCTION AND WE HAVE NO IDEA
WHAT THEY ARE YET OF THOSE GENES
AND PROTEINS MEANT TO DO THE
SAME THING IN THE FUNCTION
PROTEIN WHICH MEANS THUS WE
EXPECT TO DO DISCOVER AND
UNDERSTANDING THE STRATEGIES
DIFFERENT BUT IN BETWEEN THE
DIFFERENT FAMILIES OF VIRUSES.
HERE ARE THE PEOPLE IN THE LAB
WHO I'VE BEEN WORKING WITH IN
THE FIELD AND THIS IS THEY
DEVELOPED THE GENETICS IN THE
LAB AND WE WORKED ON THE LAB AND
I WANT TO THANK ALL THE PEOPLE
WITH WHICH WE'VE BEEN
COLLABORATING AND THE LAB AND
THE DETERMINATION AND BEING ABLE
TO LOOK AT THE DATA AND THANK
YOU FOR YOUR ATTENTION.
I THINK I WAS VERY FAST.
>> ANY QUESTIONS?
CHANTAL, I WAS WONDERING WHETHER
THE HISTONES ENCODED BY THE
VIRUSES ARE SIMILAR OR DIFFERENT
TO THE HISTONES.
>> VERY SIMILAR.
YOU CAN ALIGN THEM TOGETHER.
>> THANKS.
>> HI, CHANTAL.
>> HI.
>> WHAT ARE YOUR THOUGHTS ON HOW
THESE THINGS EVOLVED?
YOU MADE IT SOUND LIKE IT'S A
SEPARATE BRANCH OF LIFE OR THEY
DON'T HAVE ANY KIND OF -- DO
THEY INTERFACE AT ALL WITH THE
EUKARYOTIC GENOMES, DO THEY
INTEGRATE OR RECOMBINE?
WHAT ARE YOUR THOUGHTS ON THAT?
VERY PHILOSOPHICALLY.
>> THANK YOU FOR THE QUESTION.
WE CAN SEE PIECES OF THE GENOME
AND MANY WITH THE COST AND
THERE'S TWO PROCESSES THAT WORK.
ONE VIRUS AND OBVIOUSLY THERE'S
SOME ACQUISITION OF GENES BUT
ALSO SOME LOSS OF GENES AND THE
VIRUS WILL BE ABLE TO MOVE THE
POLYMERASE FROM THE HOST AND
THIS NEED FOR KEEPING THE
POLYMERASE WILL BE DECREASED.
THERE ARE TWO WAYS.
IN THE CASE OF THE FAMILIES
THEY'RE ABLE TO CREATE GENES AND
I CAN SEE AT SOME POINT THERE'S
TRANSCRIPTION AND TRANSLATION.
AND THIS IS POSSIBLE BECAUSE
THEY'RE PC RICH AND THEY'RE MORE
AWARE OF AP RICH GENOMES AND I
THINK THERE IS A STRONG AND SO
KEEP CHANGING TO WIN THE
COMPETITION AND THAT'S MOST
LIKELY WHY THEY HAVE THE
MACHINERY TO BE ABLE TO ADAPT TO
THE HOST ENVIRONMENT.
I HOPE I ANSWERED YOUR QUESTION.
>> ANYTHING ELSE?
THANK YOU, CHANTAL.
IT WAS FANTASTIC.
>> THANK YOU VERY MUCH.
>> LADIES AND GENTLEMEN, I'M
PLEASED TO ANNOUNCE A COFFEE BREAK.
SO OUR NEXT
SPEAKER NEEDS TO REAL
INTRODUCTION, DR. FAUCI WAS THE
DIRECTOR OF NIAID AND RETIRED
LAST DECEMBER FROM THE POSITION
AND CONTINUES TO HAVE AN
EXTREMELY BUSY SCHEDULE.
HE COULDN'T BE HERE TO PRESENT
THE TRIBUTE TO DR. SALZMAN BUT
AGREED TO HAVE A VIDEO.
>> THE IS TONY FAUCI.
I'M SORRY I CAN'T NOT BE THERE
BUT I HAVE THE HONOR OF PAYING
CONTRIBUTE TO NORMAN P. SALZMAN
THE NAMESAKE OF THE SYMPOSIUM.
IN THE NEXT FEW MINUTES I'LL
BRIEFLY SHARE HIGHLIGHTS OF THIS
EXTRAORDINARY 44-YEAR CAREER OF
NORMAN.
NORMAN WAS BORN IN MANHATTAN IN
AUGUST 1926 GRADUATED FROM THE
CITY COLLEGE OF NEW YORK AND
RECEIVED A MASTER'S IN BIO
CHEMISTRY FROM THE UNIVERSITY OF
MICHIGAN.
HE EARNED HIS DEGREE FROM
UNIVERSITY OF ILLINOIS IN 1953.
DIRECTLY AFTER RECEIVING HIS
Ph.D. HE CAME TO THE NIH WHERE
HE WOULD SPEND THE NEXT 33
YEARS.
HE FIRST JOINED WHAT WAS THEN AT
THE NATIONAL HEART INSTITUTE
WHERE HE WORKED IN THE
LABORATORY OF CHEMICAL
PHARMACOLOGY.
IN 1961 HE MOVED TO MY FORMER
INSTITUTE, THE NATIONAL
INSTITUTE OF ALLERGY AND IN
INFECTIOUS DISEASES TO HEAD THE
CELL BIOLOGY IN THE LABORATORY
OF THE VIRUSES AND SIX YEARS
LATER PROMOTED TO CHIEF OF THAT
LABORATORY.
DURING THE NEARLY TWO DECADES IN
THIS POSITION HE ATTRACTED MANY
INNOVATIVE SCIENTISTS TO WORK
WITH HIM.
WHEN I FIRST ARRIVED AS A
CLINICAL SOCIETY IN INTERNAL
MEDICINE HE WAS AN ICONIC FIGURE
IB VIROLOGY FOR THE WORK HE WAS
DOING AND WAS A GOOD FRIEND OF
MY MENTOR DR. SHELDON WOLF
CLINICAL DIRECTOR OF NIBIB.
IT'S IMPORTANT TO REMEMBER THE
SCIENTISTS ON WHOSE SHOULDERS
THE FIELD OF VIROLOGY STANDS
THOSE WHO LAID DOWN THE
FOUNDATION FOR VIROLOGY RESEARCH
AND SET NIAID ON THE PATH TO
BECOME THE RESEARCH POWERHOUSE
IN INFECTIOUS DISEASES IT IS
TODAY.
NORM IS ONE SUCH GIANT.
WHEN HE CAME TO THE NIH AFTER
DISCOVERING HOW TO GROW VIRUSES
IN THE LABORATORY AND AUTHORITY
IN THE CHEMISTRY OF VIRUSES HE
WAS INTERESTED IN UNDERSTANDING
THE GENETIC BASIS FOR ACUTE AND
VIRAL INFECTIONS AS WELL AS FOR
MALIGNANT FRANCE --
TRANSFORMATION AND CONDUCTED
RESEARCH ON THE VIRUSES OF
VIRU
VIRUSES.
AND TO DETERMINE WHAT STEPS IN
THE LIFE CYCLE OF A VIRUS
SELECTED ANTIVIRONES MAY BE
TARGET AND INVESTIGATED SMALLPOX
VACCINES.
HE WAS AMONG THE FIRST
SCIENTISTS TO CAPTURE ELECTRONIC
MICROSCOPIC IMAGES OF REP
INDICATING VIRAL DNA MOLECULES
AND WORKED ON SD40 AND
INCREASINGLY FOCUSSED ON TUMOR
VIRUSES AND INVESTIGATING THE
MECHANISMS BY WHICH VIRUSES
CAUSE CANCER.
IN ADDITION TO HIS RESEARCH
ACCOMPLISHMENTS, NORM GAINED AN
INTERNATIONAL REPUTATION IN PART
BECAUSE IN 1966 AS SHOWN IN THIS
PHOTO, NORM DR. KOZLOF AND
OTHERS FOUND THE JOURNAL OF
VIROLOGY.
TO THIS DAY IT'S A TOP-RANKED
VIROLOGY JOURNAL.
AS RESULT OF HIS LEADERSHIP AND
THE INTERNATIONAL STATURE THAT
HIS LABORATORY ACHIEVED IN
VIROLOGY AND IN CELL BIOLOGY
RESEARCH FORM WAS SELECTED FOR
THE SUPERIOR SERVICE AWARD IN
1983.
SOON AFTER I BECAME CHIEF THIS
WAS THE FIRST CASE OF HIV/AIDS
OF MORBIDITY AND MORTALITY AND
AFFECTED NORM AND IN MY OWN
CAREER.
THIS PHOTO FROM THE MID 1980s
SHOWS THE NIAID LABORATORY
CHIEFS AT THE TIME WITH NORM
PICTURED SLIGHTLY TO THE RIGHT
OF CENTER NEXT TO BILL, PAUL AND
ME AT THE FAR LEFT.
GOING BACK TO THE PRIOR SLIDE NO
SURPRISE BUT GREAT LOSS TO
NIAAID IN 1986 GORGETOWN
UNIVERSITY SUCCESSFULLY
RECRUITED NORM TO MOVE HEADLONG
TO AIDS RESEARCH BY LEADING THE
MOLECULAR VIRAL LABORATORY THERE
AND HE AND HIS LABORATORY GROUP
FOCUSSED ON EVALUATING POTENTIAL
VACCINES FOR HIV/AIDS.
IN THE EARLY YEARS OF THE HIV
EPIDEMIC WE COLLABORATED WITH
NORM'S LAB AND SHOWN ARE TWO
PUBLICATIONS RESULT FROM THE
COLLABORATIONS.
THE FIRST PAPER THE FOURTH MOST
CITED AIDS RESEARCH PAPER IN
1989 DESCRIBED THE FIRST HIV
RESERVOIR IN THE BLOOD.
IN 1994, NORM WAS RECRUITED TO
HEAD THE LABORATORY OF MOLECULAR
RETRO VIROLOGY AT THE FREDERICK
CANCER RESEARCH CENTER FORT
DETRICK A POSITION HE HELD THE
LAST FOUR YEARS OF HIS CAREER.
HIS POSITIVE INFLUENCE ON THE
FIELD OF VIROLOGY REMAINS STRONG
TO THIS DAY.
AMONG THE LARGE NUMBER OF
SCIENTISTS AND PHYSICIAN
SCIENTISTS HE TRAINED AS LISTED
ON THE SLIDE AMONG THOSE WHO
BENEFIT FROM THE GENEROSITY AND
WISDOM OF HIS MENTORSHIP, MANY
HAVE GONE ON TO DISTINGUISHED
CAREERS AT NIH AND OTHER
RESEARCH INSTITUTIONS.
IN THE UNITED STATES AND ABROAD
IT'S A QUALITY AND STROKE OF HIS
MENTORSHIP ONE MENTEE RECEIVED A
NOBEL PRIZE AND SEVERAL HAVE
BEEN ELECTED TO THE NATIONAL
ACADEMY OF SCIENCES.
BEFORE I CKCONCLUDE I WANT TO
THANK LENNOR SALZMAN FOR SERVING
ON THE ORGANIZING COMMITTEE FOR
THE SIM POEM WAS FOR EACH OF THE
PAST 25 YEARS THAT IT HAS BEEN
HELD.
MY SINCERE THANKS, LINER AND
PLEASE ENJOY THE REST OF THE
MEETING.
THANK YOU VERY MUCH.
>> LENNOR.
>> GOOD MORNING AND WELCOME.
I'M HONORED TODAY TO BE HERE
WITH THIS ILLUSTRIOUS GROUP OF
SCIENTIST TO CELEBRATE THE 25th
ANNIVERSARY OF THE NORMAN P.
SALZMAN MEMORIAL AND SYMPOSIUM
IN BASIC AND CLINICAL VIROLOGY.
THIS IS THE FIRST TIME THE
SYMPOSIUM WILL BE LIVE HERE AT
THE LIPSETT AUDITORIUM.
200 REGISTERED AND EUROPE AND
180 REGISTERED.
THAT IS A BIG NUMBER.
FOR ME PERSONALLY THE EVENT IS A
TESTIMONY TO NORMAN'S DEVOTION
TO HIS SCIENTIFIC AND MENTORING
WORK AND TO OTHERS WHO WORK WITH
HIM TO CONTINUE THE HIGHEST
STANDARDS FOR ORIGINAL AND
MERITORIOUS WORK IN ALL
SCIENTIFIC EXPLORATION.
MORE ABOUT THAT IN A MINUTE.
THIS IS MY TIME TO SAY THANK YOU
TO DR. JULIE GERBERDING WHO IS
SITTING HERE WHEN SHE ARRIVED
EARLIER IN THE SYMPOSIUM.
OUR LONG ASSOCIATION AND SUPPORT
FOR THE SUPPORT FOR THE
FOUNDATION FOR NIH MADE THE
CONTINUATION OF THE NORMAN
SYMPOSIUM AND AWARD POSSIBLE FOR
THE LAST 25 YEARS.
OUR GRATITUDE.
TO THE EVENT STOP JENNEL LEWIS
AND ASHLEY SPOLEL AND TO
DR. ANTHONY FAUCI TO YOUR
CONTRIBUTE TO NORMAN WHEN HE WAS
CHIEF OF THE LABORATORY OF
VIRUSES.
REMEMBERING ALL HIS ACHIEVEMENTS
OVER THE YEARS.
THEY VISED ME AND HUGE THANK TO
THE ORGANIZING COMMITTEE AND
VIROLOGY INTEREST GROUP FOR
THEIR SUPPORT IN MAKE THE
SYMPOSIUM A SUCCESS CINCINNATI
1998.
-- SINCE 1998.
ON DECEMBER 11, 2023 IT WILL BE
26 YEARS SINCE THE DEATH OF
NORMAN SALZMAN.
SIX WEEKS AFTER HE WAS DIAGNOSED
WITH ASYMPTOMATIC PANCREATIC
CANCER.
DURING A BITTER COLD DECEMBER
DAY, NORMAN SAT OUTSIDE IN HIS
BELOVED GARDEN CONSIDERING HIS
LIFE AND CONTRIBUTION TO
VIROLOGY.
NOW, THERE ARE MORE THANK YOUS.
I WANT TO HAVE A SPECIAL THANK
YOU FOR DR. CHARLIE ROSE FOR HIS
TALK THIS MORNING.
I WISH I COULD UNDERSTAND A LOT
OF IT BUT I'M A SPEECH
PATHOLOGIST NOT A SCIENTIST.
FOR ALL THE TALKS TO COME, I
THANK YOU ALL FOR BEING HERE AND
MAKING THIS IS SYMPOSIUM ANOTHER
SUCCESS THIS YEAR.
26 YEARS LATER IT'S MY HONOR AND
PLEASURE TO PRESENT THE NORMAN
SALZMAN POST-DOCTORAL MEMORIAL
AWARD TO ALBERTO D. LOPEZ MUÑOZ,
Ph.D. AND HIS MENTOR,
DR. JONATHAN UDEL.
DO WE HAVE A SLIDE?
OH, GOOD.
THANK YOU.
WITH THIS CHECK COMES A CHECKS
OF $2,500 AND A PLAQUE FOR
DR. MUÑOZ AS WELL AS A PLAQUE
FOR DR. UDEL.
THE CHECK WILL BE SENT FROM THE
SALZMAN FUND FROM THE FOUNDATION
AT NIH AND I WAS TOLD THAT THE
PLAQUES WERE HERE.
THE FIRST SLIDE I'D LIKE TO HAVE
IS FOR DR. JONATHAN UDEL.
DO WE HAVE THAT SLIDE?
CHIEF CELLULAR BIOLOGY SECTION,
LABORATORY OF VIRAL DISEASES,
NATIONAL INSTITUTE OF ALLERGY
AND IN FEN HOUSE DISEASES,
NATIONAL INSTITUTES OF HEALTH --
INFECTIOUS DISEASES AND NATIONAL
INSTITUTES OF HEALTH, DR. UDEL
M.D., Ph.D. MENTOR TO ALBE ALBE
MUNOZ FOR CAPSID PROTEIN
EXPRESSION A CORONAVIRUS VIRUS
FOR IMMUNOAND I'D LIKE TO
PRESENT THE PLAQUE TO DR
DR. YEWDELL.
I WAS GOING TO ASK YOU TO COME
TO THE PODIUM.
>> THANK YOU SO MUCH.
I'LL READ THIS AND PLEASE COME
TO THE PODIUM THE WINNER OF THE
SALZMAN AWARD, ALBERTO D. LOPEZ
MUÑOZ IN RECOGNITION FOR HIS
COULD, ENTITLED SURFACED CAPSID
NUCLEODILATION MENTORED BY
DR. YEWDELL AND CHIEF CELLULAR
BIOLOGY SECTION AND LABORATORY
OF VIRAL DISEASES AND NATIONAL
INSTITUTE OF INFECTIOUS
DISEASES, NIH.
HERE'S THIS PLAQUE.
CONGRATULATIONS.
WE HAVE A LOT OF BIO GRAPHICAL
DATA IN THE PROGRAM THAT'S
PRESENTED TO YOU TODAY SO I
WON'T GO INTO THAT EXCEPT TO SAY
THAT DR. ANTONIO LOPEZ MUNOZ IS
SERVING IN THE ASSOCIATION OF
SPANISH SCIENTISTS IN THE UNITED
STATES AND DR. MUÑOZ TOLD ME HIS
INSPIRATION TO BECOME A DOCTOR
CAME FROM THE NAME OF THE STREET
HE LIVED ON AS A YOUNG BOY.
THE STREET WAS NAMED DR. FLEMING
STREET.
IT'S MY PLEASURE TO PRESENT
DR. ANTONIO LOPEZ MUNOZ TO
PRESENT HIS LECTURE AND WE HAVE
THAT ON THE PLAQUE.
THANK YOU.
>> ALL RIGHT THEN.
THIRST, MRS. SALZMAN THANK YOU.
MY NAME IS ALBERTO LOPEZ AND
IT'S A PRIVILEGE TO ACKNOWLEDGE
THE MEMORY OF DR. SALZMAN BY
SHARING THE AMAZING SCIENCE FROM
THE LABORATORY OF DISEASES.
I'LL TALK ABOUT THE MODULATORY
ROLE OF THIS HAS FASCINATED ME
AS A STUDENT BACK IN SPAIN.
LARGE DNA VIRUSES AND HERPES
VIRUSES HAVE INVESTED A HUGE
DEAL IN DEVELOPING THE CYTOKINE
RECEPTORS AND KECHEMOKINES AND
INVADING THE FIRST STAGE OF THE
HUMAN RESPONSE WITH THIS IN MIND
I WONDERED IF THERE WAS A
CYTOKINE BINDING PROCESS
DESCRIBED TO STUDY THESE PROTEIN
AND SARS COV2 VIRUS.
AFTER EXTENSIVE RESEARCH I FOUND
THERE WAS NOT THESE KINDS OF
PROTEINS BUT IT WAS USED BY BOTH
HUMAN AND ANIMAL CORONAVIRUSES.
THE PROTEINS IS USUALLY
INTERINTRINT
INTERCELLULAR PATHWAYS BLOCKING
AT DIFFERENT LEVELS.
I WOULD LIKE TO STATE WHAT WE
KNOW ABOUT THE INFECTION AND THE
MOST ABUNDANTLY EXPRESSED DURING
THE INFECTION AND PROTEIN BINDS
TO THE DNA AND IN TERMS OF
IMMUNE RESPONSE IS A MAJOR
TARGET FOR ANTIBODY RESPONSES
AND CONTAINS NUMEROUS EPITOPES.
AND LATER THE VACCINATION
CONTRIBUTES TO THE N PROTEIN TO
THE IMMUNITY AND CLEAR UP THE
INFECTION FOR A BETTER
PROGNOSIS.
WITH THIS IN MIND WE STARTED
DOING BASIC EXPERIMENTS AND
STAINING WITH IN SITU VIRUSES
AND WE SHOW THE CELL SURFACE
SPIKES IN RED FOR VIRUSES AS
WELL AS IN BOTH CASES AND
CONSISTENTLY WITH ALL THE OTHER
CELL WE TESTED WE FOUND A
DISTRIBUTION PATTERN FOR LIKE
THE NUCLEIC ACID OR PROTEIN IN
GREEN.
WE CONFIRMED BY FORMING THE SAME
EXPERIMENT BY FLOW CYTOMETRY AND
WE POSTINFECTION WE STAIN WITH
MONOCLONAL ANTIBODIES AND FOUND
THE HISTO GRAM WE FOND THE SPIKE
AND THE NUCLEIC ACID AND
CONFIRMED WHEN WE QUANTIFY THE
FLUORESCENT INTENSITY AGAINST
LIKE A MOCK INFECTION CONDITION.
WHERE HE USED THE EPITHELIAL
CELLS AND INFECTED THEM AND
THREE DAYS POST INFECTION BOTH
FOR SARS WE FOUND THE SPIKE ON
THE SURFACE OF SOME OF THESE
EPITHELIAL CELLS FOR A SPIKE AS
WELL AS FOR THE NUCLEIC ACID
PROTEIN AND MORE DISCREET IN THE
INFECTION STILL SIGNIFICANT.
WE UNDERSTOOD THIS COULD BE
HAPPENING IN A BREAK-THROUGH
INFECTION IN HUMANS.
WE ALSO WANTED TO UNDERSTAND HOW
THE END IS BEING SECRETED AND WE
CLONED THE M PROTEIN AND
DETERMINE IF OTHER FACTORS WERE
REQUIRED LIKE FOR THE M PROTEIN
BEING EXPRESSED OUTSIDE.
AND POST TRANSFECTION I STAINED
WITH THE ANTIBODIES AND YOU CAN
SEE WE OBSERVE FOR THE M PROTEIN
COMPARED TO A CONTROL
TRANSFECTION NO OTHER VIRAL
FACTORS ARE REQUIRED FOR HUMAN
CORONAVIRUS AND VIRUS
EXPRESSION.
WE CAN UNDERSTAND THERE IS
EXPRESSION IN THE CELL SURFACE
AND HOW IT'S BIND IS IS THE NEXT
QUESTION I WONDER.
THESE ARE WELL KNOWN FOR BEING
USED AS PRECEPTORS FOR THE
CORECEPTOR OR ATTACHMENT
FACTORS.
THIS ALLOWS US TO PERFORM
INTERACTION ASSAYS AND HERE I
EMBEDDED THE BIO CENSOR --
SENSOR WITH DIFFERENT GLYCANS
AND THE HEPARIN WAS FOUND AND
THIS IS AND I COMPLETED THE
TREATMENT AND WHEN WE INCUBATE
THE CELLS UNTREATED WITH THE
PROTEIN THIS PROTEIN STICKS TO
THE CELL SURFACE AND HERE WE
REMOVED THE CELLS AND THESE
TREATED LOST THE ABILITY TO BIND
AND RETAIN THE PROTEIN ON THE
CELL SURFACE.
FOR THOSE THAT LIKE FLOW
CYTOMETRY YOU'LL NOTICE THE
INTERESTING OBSERVATION WE
OBSERV
OBSERVED ACROSS ALL CELL LINES
AND INFECT THE POPULATION OF
CELLS EXPRESSING ONLY M PROTEIN
BUT NOT SPIKE PROTEIN.
I RECALL THE CONVERSATION AND WE
THOUGHT PERHAPS THIS M PROTEIN
COULD BE TRANSFERRED TO THE
SURROUNDING CELLS THAT WERE NOT
YET INFECTED.
AND ASSIGNED IT AND WELL TURE
WITH RECIPIENT CELL AND THE
INEFFECTIVE CELLS WOULD BE
SUSCEPTIBLE FOR INSECTION AND
THIS RECIPIENT CELL POPULATION
TO ALLOW FLOW CYTOMETRY
SEPARATION.
WE INFECTED THE CELLS AND 24
HOURS LATER WE ASKED IF THE
HYPOTHESIS WAS HAPPENING.
WE HAVE THE DONOR CELLS AND
HAVING THE M PROTEIN ON THE
SURFACE AND IF IT COULD BE
TRANSFERRED TO THE RECIPIENT
CELLS.
I'M SHOWING THE BASELINE WE SEE
NO SURFACE SIGNATURE FOR A SPIKE
AND CAPSID PROTEIN AND YOU CAN
SEE THE DONOR CELL SUSCEPTIBLE
FOR INFECTION AND GAINED
POSITIVE FOR SURFACE M PROTEIN
HOWEVER, THEY REMAIN NEGATIVE
AND HAVE PROTEIN ON THE SURFACE
TELLING US THE PRESENCE OF
GLYCANS ALLOWED THEM TO CATCH,
BIND AND RETAIN THE PROTEIN
AVAILABLE FROM THOSE DONOR
CELLS.
WE WONDERED IF THIS WAS TO
RECONFIRM THE GLYC AN GLYCAN ME
THE CELL AND WE REPLACED THE
RECIPIENT CELLS BY THE CELL
LINES AND YOU CAN SEE NOW THE
RECIPIENT CELL THE BLUE GUYS
REMAIN NEGATIVE FOR THE SPIKE
AND NUCLEIC ACID PROTEIN.
IN THE END THE ABSENCE OF
GLYCANS IN THE RECIPIENT CELLS
IS WHAT PREVENTED OR ALLOWED THE
CELLS TO CAPTURE, BIND OR RETAIN
WHAT WAS ON THE SURFACE OF THE
DONOR CELLS.
WE LOOK AT THE SURFACE OUTSIDE
THE CELL AND THE QUESTION I RAN
THE PROTEIN INTERACTION I USED
THE SARS COV2 STRUCTURE AND
ACCESSORY PROTEIN AGAINST THE 24
HUMAN CYTOKINES AND ETCETERA AND
YOU CAB SEE AMONG ALL ONLY THE M
PROTEIN WAS ABLE TO BIND AT THE
SPECIFIC SUBSET OF CHEMOKINES
AND WHERE I FOUND THE PARTICULAR
SET OF SIX CHEMOKINES HERE IN
YELLOW AND THE QUESTION IS THE
RELEVANCE BETWEEN PROTEIN AND
CHEMOKINES.
I CAME BACK TO THE ASSAY I
LEARNED TO USE IN GRAD SCHOOL
AND WE HAD A FIXED CONCENTRATION
OF CHEMOKINE AND WE HAVE
DIFFERENT CELLS FOR THE GIVEN
CHEMOKINE AND A POSITIVE BINDER
AND WE CAN PREINCUBATE THE
CHEMOKINE WITH A RECOMBINANT
PROTEIN AND SEAS IF THERE'S AN
ENHANCEMENT OR INHIBITION TO THE
LOWER CHAMBERS AND QUANTIFY HOW
MANY CELLS HAVE MY GRA-- MIGRAT
AND THERE'S THE INDUCTION OF THE
CHEMOKINE AND WHEN I PREINCUBATE
THE FIXED CONCENTRATION WITH
INCREASING CONCENTRATION OF THE
SARS COV2 PROTEIN I HAVE
MIGRATION FOR THE CELL LINES
EVEN FOR HUMAN PBMCCs.
IT WAS WORTHY TO BE REACHED OUT
AND THEY HAD THE INTERACTION
BETWEEN MUCK THE MONEY MERS AND
THEY OBSERVED THESE BETWEEN THE
SARS COV2 PROTEIN AND IT WAS
INTERESTING THE INTERACTION.
WE ALSO OBSERVED THE PHENOMENON
FOR THE PROTEIN AND IT'S HAD
GRATION AND TO HAVE THE
RECOMBINANT PROTEIN AND DURING
THE CORONAVIRUS AND THE OTHER
CORONAVIRUSES WHICH I BELIEVE IT
SUGGESTS THE MECHANISM IS
SOMETHING CONSERVED BETWEEN
HUMAN CORONAVIRUS DESPITE THE
SEQUENCE.
FINALLY, I WANT TO SHOW YOU WE
ARE ALSO WONDER ABOUT IF THE M
PROTEIN COULD BE A TARGET AND WE
OPTIMIZED IN THE LAB THE ADDC
BIO REPORT ASSAY AND REAL QUICK
WE INFECTED CELLS AND KNEW 24
HOURS LATER WE'D HAVE PROTEIN ON
THE SURFACE OF THE INFECTED
CELLS AND INCUBATED OVERNIGHT
WITH MICE IMMUNIZED WITH THE
EFFECTER CELLS PART WITH AND CAN
GO THIS IS THE CLEAR ACTIVATION
WE OBSERVED WHEN WE USED AN
ANANTIBIOTIC -- ANTIBODY AND
HERE WE USED THE AND SAW
ACTIVATION WHICH WAS HIGHER AND
THIS SUMMARY I WOULD LIKE TO
TAKEAWAY THE PROTEINS AND IT
COMBINED THROUGH THIS AND INFECT
TO NEIGHBORING CELLS AND
INHIBITS THE PROPERTIES WHICH WE
BELIEVE IT'S A TARGET FOR
ANTIBODY BASED IMMUNITY.
AND THE FUTURE DIRECTION RATHER
THAN SHOWING A LIST OF
EXPERIMENT I WANTED TO GIVE
CREDIT TO JOHN TO MY SUPERVISOR
AND CELL SURFACE EXPRESSION
SEEMS TO BE THE RULE AND SHOWN
THE VIRUS AN RNA BINDING PROTEIN
IS USED TO MODULATE THE IMMUNE
RESPONSE IN THE EARLY STAGE OF
INFECTION.
WITH THAT I WOULD LIKE TO THANK
THE INSTITUTE AND NIAID AND THE
LAB AND MY MENTOR AND I'M DEEPLY
GRATEFUL FOR THE OPPORTUNITY I
GOT FOR COMING HERE I'VE ENJOYED
EVERY DAY IN THE WILL BE AND
HAVING FUN AND LEARNING AND
MATURING AS A SCIENTIST.
THANK YOU EVERYBODY FOR YOUR
ATTENTION.
>> SO HOW DOES M GET OUT OF THE
CELL?
>> THAT'S THE MILLION-DOLLAR
QUESTION WE KNOW IT DOESN'T HAVE
A SINGLE PEPTIDE OR GO THROUGH
THE PATH WE AND HAS TO GO
OUTSIDE THROUGH THE PATHWAY AND
HAVE PLASTIC ENCODING FOR THE
PROTEIN AND WE BLOCKED THE
PATHWAY AND PROTEIN KEPT GOING
OUTSIDE.
>> IT SEEMS LIKE A COOL PATHWAY
TO FIGURE OUT.
AND IT'S BEEN ABLE TO REACH THE
CELL SURFACE BY INTERACTING WITH
GLYCANS INTERNALLY AND EXPOSED
OUTSIDE THE PROTEIN WILL BE
USING IN THAT PARTICULAR
SCENARIO.
I'M TRYING TO EXPLORE THAT WITH
DIFFERENT CELL LINES TO SEE THE
PROTEIN THAT STOPPED COMING OUT
BECAUSE THERE'S NO GLYCAN
EXPRESSION IN THE CELLS.
>> IN TERMS OF THE CHEMOKINE
BINDING SPECIFICITY IS THERE A
MUTATION TO KNOCK THAT OUT
CAPABILITY THAT WILL STILL ALLOW
FUNCTION IN TERMS OF REPLICATION
AND VIRUS PROTECTION?
>> TO THE CHARACTERIZATION OF
THE DOMAINS WE'RE WORKING ON
THAT AND USING ANTIBODIES IN THE
mRNA BINDING DOMAINS AND WHEN WE
BLOCK ONE OR THE OTHER WE LOSE
ABOUT 50% OF THE PROTEIN TO BIND
AND WHEN WE BLOCK BOTH WE NEARLY
LOST THE ABILITY OF THE PROTEIN
TO INTERACTION.
WE KNOW SEE THE RNA BINDING
DOMAINS WE MAIN WELCOME ACROSS
THE CONCERNS.
WE UNDERSTAND THIS NOT VERY
PERMISSIVE THE PERMUTATION BUT
FOR THE FINE TUBE -- TUNE WE
WOULD HAVE TO DO THIS GENESIS.
>> FASCINATING WORK.
CONGRATULATIONS.
>> DOES ANYONE HAVE ANY MORE
QUESTIONS.
>> ONE MORE QUESTION FOR YOU.
I ASSUME -- YOU ANSWERED MY
INITIAL QUESTION HOW
APPROACHABLE IT IS FOR OTHER
VIRUSES.
DID YOU TEST THE CAPSID PROTEIN
IF THEY BIND TO CYTOKINES?
>> BACK IN THE '80s INFLUENZA
PROTEINS AND I HAVEN'T RUN THE
WHOLE ASSAY WITH THE CYTOKINES
BUT I DO SEE THE 12B IN THE
ASSAY AND WITH THAT PARTICULAR
CHEMOKINE I SEE THE FLU M
PROTEIN AND WOULD LIKE TO KEEP
EXPLORING THAT.
>> WITH THAT, CONGRATULATIONS
AGAIN OR YOUR AWARD AND WE'RE
LOOKING FORWARD TO YOUR NEXT
RESEARCH.
NOW, I'M GOING TO INTRODUCE ANIS
LOWEN FROM EMORY UNIVERSITY
SCHOOL OF MEDICINE AND GOING NOT
BE ABLE TO BE IN PERSON BUT GIVE
HER PRESENTATION BY ZOOM.
THE TALK IS STOCHASTIC LOSS OF
DIVERSITY WITHIN THE NEW HOST IS
A MAJOR COMPONENT OF INFLUENZA A
VIRUS TRANSMISSION BOTTLENECKS.
WE LOOK FORWARD TO HEAR YOUR
PRESENTATION.
>> GOOD MORNING, EVERYONE.
THANK YOU FOR THE INVITATION TO
PARTICIPATE IN THE EXCELLENT
SYMPOSIUM.
I'M EXCITED FOR THE AFTERNOON.
IT'S BEEN GREAT SO FAR.
I CHANGED MY TITLE LATE TO
BROADEN IT AND TALK ABOUT
BOTTLENECKS AND INFLUENZA A
DYNAMICS.
THE A VIRUSES ARE CONSTANTLY
CHANGING.
THIS IS ILLUSTRATED HERE WITH
THE PHYLOGENETIC TREE SHOWING
THE HUMAN GENE THAT ENCODES THE
GLYCOPROTEIN AND LOOKING AT THE
H3N2 INFLUENZA VIRUSES THAT HAVE
CIRCULATED THE LAST YEARS AND
THIS SHOWS VARIANTS.
YOU CAN SEE A CLEAR AND THE
PATTERN IN WHICH IT'S REPLACED
BY A NOVEL CLAD IN THIS
SUCCESSIVE FASHION.
THE PROBLEM THIS PRESENTS IS
ILLUSTRATED ON THE TREE WITH THE
Xs.
SO THESE ARE IDENTIFYING STRAINS
CHOSEN TO BE INCLUDED IN THE
SEASONAL INFLUENZA VACCINE.
BECAUSE THE VIRUS CHANGES YEAR
ON YEAR, THE VACCINE NEEDS TO BE
UPDATED FROM ONE YEAR TO THE
NEXT.
AND THIS IS A MAJOR EFFORT FOR
BROAD SURVEILLANCE AND IN DEPTH
CHARACTERIZATION THAT FEEDS INTO
THAT STRAIN SELECTION PROCESS.
DESPITE THAT EFFORT THERE'S
GUESSWORK INVOLVED IN STRAIN
SELECTION BECAUSE THE VACCINE TO
BE PRODUCED OR THE STRAINS NEED
TO BE CHOSEN NINE MONTHS PRIOR
TO THE INFLUENZA SEASON.
IN OTHER WORDS IT REQUIRES
PREDICTING EVOLUTION.
THERE'S A STRONG PUBLIC HEALTH
MOTATION TO BETTER UNDERSTAND
VIRAL EVOLUTION.
WHEN THE FIELD DUG INTO THAT
PROBLEM AND WE SEE AT THE GLOBAL
POPULATION LEVEL EACH CLAD IS
REPLACED BY A NOVEL ONE OVER A
PERIOD OF A FEW YEARS.
AND IT'S A FASCINATING
EVOLUTIONARY DYNAMIC AND ONE
BEST DESCRIBED EXAMPLES OF
PUBLIC SELECTION IN BIOLOGY.
NONETHELESS, A SIMILAR DYNAMIC
IS NOT SEEING OCCURRING WITHIN
INDIVIDUALS.
SO WITHIN THE INFECTED PERSON
TENDS TO BE LOW DIVERSITY AND
RELATIVELY STATIC AND THEY
TYPICALLY REMAIN MINOR OVER THE
COURSE OF INFECTION.
WE'RE NOT SEEING A SIGNATURE OF
POSITIVE SELECTION OF ANTIGENIC
VARIANTS.
WHAT'S SEEN WITH INDIVIDUALS IS
A NEGATIVE SELECTION.
A PURGING OF MUTATION OBSERVED
AS WELL AS GENETIC DRIFT.
RANDOM CHANGES AND FREQUENCIES
ARE OBSERVED WITHIN INDIVIDUALS.
DESPITE PRE EXISTING IMMUNITY
AND MOST INFLUENZA CASES WE DID
NOT SEE ANTIGENICALLY INVOLVE
CASES SWEEP.
THIS CREATES AN INTERESTING
PUZZLE BECAUSE FOR A VARIANT TO
CIRCULATE GLOBALLY IT MUST ARISE
WITHIN INDIVIDUALS.
LEAVING US WITH THE QUESTION HOW
MUTANTS ARISE BEFORE GOING ON TO
CIRCULATE GLOBALLY.
THIS PRESENTS AN APPEALING MODEL
AND THE PROJECTS AND WE LOOK AT
THE INFLUENZA A VIRUS AND USING
THE VIRUS POPULATION.
THIS LEVERAGES LOOKING AT
SEASONAL INFLUENZA A VIRUS AND
IT'S A TEAM EFFORT AND WE HAVE
EXCELLENT WORKING TOGETHER ON
THIS.
OUR PARTICIPANTS ARE A MINIMAL
SEROLOGICAL RESPONSE AND THE
INOCULUM IS A SEASONAL STRAIN.
AND WE'RE USING THIS ATOM IZIZE.
WE CHALLENGED 18 PARTICIPANTS EN
THIS MANNER OF 18, 15 WERE
PRODUCTIVELY INFECTED.
WE SEE THE VIRAL LOAD IN THE
NASAL PHARYNGEAL SITE.
THE Y AXIS IS 45 MINUS THE CT
VALUE FROM THE PCR AND THE X
AXIS IS THE TIME OF INOCULATION
INDICATED HERE WITH THE YELLOW
ARROW.
AND THE DYNAMICS WITH INDIVIDUAL
I'LL SHOW DATA FROM THE ONGOING
EFFORT AND WE DON'T HAVE TO
UPDATE IT FROM EVERYONE JUST
YET.
THE FIRST THING WE SEQUENCED WAS
THE VIRAL INOCULUM.
I'M SHOWING THE DIVERSITY WITH
THE FREQUENCY OF SINGLE
NUCLEOTIDE VARIANTS WITH THE LOG
SCALE AND THE VIRAL GENOME ON
THE X AXIS.
ABOVE A THRESHOLD VEHICLE AND
THIS IS AMONG THE VARIANTS WE
DETECTED FOUR STOOD OUT IN
GREEN.
THE TWO IN THE PA SEGMENT AND
TWO IN NP RELATIVELY HIGH
FREQUENCY AND FOR THOSE WHO TAKE
A CLOSER LOOK AT THE FOUR
MUTATIONS, AND ASKED DO THEY
OCCUR IN NATURE?
ARE THEY SEEN IN CIRCULATING
H3N2 VIRUSES.
TO CUT TO THE CHASE A MAJOR
VARIANT IN THE INOCULUM ARE NOT
OCCURRING IN NATURE AND LIKELY
FROM PATIENTS.
THAT'S SHOWN DOWN HERE.
WHAT'S PLOTTED IN THE GRAPHS ARE
THE HAPLOTYPES OF THE PA AND NP
SEEN AMONG CIRCULATING VIRUSES
SINCE 1980.
THEY'RE VERY WELL CONSERVED AND
NONE MATCH THE MAJOR VARIANTS
SEEN IN THE STOCK.
THE MINOR VARIANTS IN OUR STOCK
ARE THE WILD TYPE.
THE MINOR PA IS THE MN AND THE
OTHER WAS CIRCULATED IN 2009.
THE MAJOR VARIANTS ARE WE
PRESUME ARE ADAPTATIONS.
I'LL SHOW THE DYNAMICS OF THE
HAPLOTYPES IN THE INOCULATED
INDIVIDUALS AND THE F026 AND
STARTING WITH THIS BECAUSE THE
DYNAMICS ARE SIMPLICISIMPLEST.
THE FREQUENCY ON THE Y AXIS OF
THE WILD TYPE HAPLOTYPES OVER
TIME WITH PA ON THE LEFT AND P
ON THE RIGHT.
YOU CAN SEE THE WILD TYPE
HAPLOTYPES STARTED AT LOW
FREQUENCY IN THE INDIVIDUAL THEN
INCREASED IN FREQUENCY.
THIS IS A VERY CLEAR SIGNATURE
OF POSITIVE SELECTION LEADING TO
ENRICHMENT OF THE WILD TYPES
WITHIN THE HOST AND SURPRISE
GIVEN THE VIRUS.
THE PATTERNS SEEN FOR PA AND NP
ARE SIMILAR.
THAT SUGGESTS THEIR LINKED
GENETICALLY AND THE REASSORTMENT
IS NOT SEPARATING THE WILD TYPE
GENE SEGMENTS BUT MOVING
IMPORTANTLY.
NOW LOCK AT ALL SIX INDIVIDUALS
THE ONE IN THE LOWER LEFT AND
THE OTHER PARTICIPANTS ARE
LABELLED HERE WITH THE RELEVANT
PLOTS.
YOU SEE THE DYNAMICS IN THE
OTHER FIVE INDIVIDUALS IS MORE
COMPLEX.
WE HAVE PA AND THEY'RE MOVING IN
DIFFERENT DIRECTIONS.
THE DYNAMICS ARE DISCORDANT
TELLING US ABOUT THE
REASSORTMENT.
IT WOULD ALLOW THE TWO GENOTYPES
PRESENT TO BECOME FOUR POSSIBLE
GENOTYPES AND THEREFORE YOU CAN
HAVE A SITUATION WHERE PA MAY BE
DECLINING IN FREQUENCY WHILE NP
IS INCREASING IN FREQUENCY.
CLEAR EVIDENCE OF REASSORTMENT
IN THE MAJORITY OF INDIVIDUALS.
SECONDLY, YOU HAVE A MESSIER
SITUATION OF THE OTHER FIVE
INDIVIDUALS AND THE FREQUENCY
CHANGES OF THE HAPLOTYPES IS NOT
AS DIRECTIONAL FOR F026 AND
RELEVANT TO NOTE IN EVERY CASE
AT LEAST ONE OF THE PA OR NP
HAPLOTYPE REACHES A FREQUENCY BY
THE END OF THE COURSE AND GIVING
A STRONG POSITIVE SELECTION
ACTING ACROSS THESE INDEPENDENT
INDIVIDUALS.
WE'LL COME BACK IN A MINUTE TO
WHY WE THINK IT HAPPENS IN THIS
CONTEXT AND ONE FURTHER PIECE OF
DATA FROM THE HUMAN.
I'VE SHOWN THE DATA FROM THE
NAVAL -- NASAL PHARYNGEAL SWABS
AND COLLECTING FROM
PARTICIPANTS.
THE NASAL PHARYNGEAL AND WE SEE
WHAT IS DETECTED AT THE
DIFFERENT SITES.
THE RESULTS ARE FROM ONE
PARTICIPANT AT 048.
WE PICKED THIS INDIVIDUAL
BECAUSE IT'S A PERSON WHO HAD
APPRECIABLE VIRAL LOADS AT ALL
FOUR SITES AND THE DYNAMICS ARE
SHOWN HERE IN PINK.
YOU CAN SEE DIFFERENT DYNAMICS
WITHIN THE ONE PARTICIPANT.
SO VERY STATIC SITUATION HERE
FOR EXAMPLE AND THE SWABS WE
HAVE SITUATIONS WHERE THE WILD
TYPE HAPLOTYPES ARE INCREASING
IN FREQUENCY IN THE NASAL
PHARYNGEAL LOCATION.
IT'S AN INTERESTING PICTURE AND
SUGGESTS THERE'S AT LEAST
PARTIALLY DIFFERENT
SUBPOPULATIONS AT THE DIFFERENT
LOCATIONS WITHIN A SINGLE
INDIVIDUAL.
THERE COULD BE MIGRATION BETWEEN
THEM HOWEVER THEY'RE NOT LEVEL
MIXED AND AN IMPORTANT
IMPLICATION IS WE COULD HAVE
DISTINCT NICHES FOR VIRAL
EVOLUTION AND DEEPER
TRANSMISSION WITHIN THE UPPER
ARE RESPIRATORY TRACT OF A
SINGLE INDIVIDUAL.
TO SUMMARIZE THAT PART WE'VE
SEEN IN THE CONTEXT OF
EXPERIMENTAL CHALLENGES POSITIVE
SELECTION IS ACTIVE DURING THE
BRIEF TIME COURSE OF ACUTE
INFECTION.
AND WHY DO WE THINK THIS IS
DIFFERENT FROM NATURAL ACUTE
INFECTIONS, I THINK THERE'S TWO
DISTINCT POSSIBILITIES.
ONE IS THE INITIAL POPULATION
SIZE IS QUITE LARGE AT 3.6 TEN
TIMES TO THE FIVE TIMES 50 IN
THE INOCULUM AND COULD GO TO THE
INITIAL POPULATION SIZE DIFFERS
FROM THE INOCULUM DOSE AND THE
MAGNITUDE OF EFFECTS BE
DIFFERENT.
IT COULD BE THE WILD TYPE AMINO
ACIDS ARE MORE FIT THAN THE
ALTERNATIVES IN HUMANS AND WOULD
DRIVE MORE EFFICIENT SELECTION.
AND THEY COMPRISE MULTIPLE
DISTINCT SUBPOPULATIONS.
WE KNOW FOR INFLUENZA A VIRUSES
TRANSMISSION ASSOCIATED WITH A
TIGHT BOTTLENECK.
THAT MEANS REGARDLESS OF THE
NUMBER OF VARIANTS PRESENT IN
THE DONOR INDIVIDUAL WE SEE THE
VERY FEW VARIANTS ESTABLISHED
INFECT
INFECTION IN AS FEW AS ONE
ESTABLISHED IN THE INDIVIDUAL AS
SEEN IN ANIMAL MODEL SYSTEMS AND
NATURALLY INFECTED HUMANS.
WE WANTED TO BETTER UNDERSTAND
THE POTENTIAL FOR SELECTION TO
ACT DURING TRANSMISSION.
WE QUESTION AT WHAT POINT DOES
THE BOTTLENECK ACT.
I'LL WALK YOU THROUGH
POSSIBILITIES.
IT COULD BE IN THE DONOR HOST
SUCH THAT FEW ARE SAID IN THE
ENVIRONMENT.
THE BOTTLENECK COULD OCCUR IN
THE ENVIRONMENT WHERE YOU HAVE
DILUTION OF VIRUS RELEASED IN
THE AREA OR ACTIVATION OF VIRUS
SUCH THAT IS NO LONGER
INFECTIOUS.
WE WOULD HAVE IT DURING THE
INITIAL INFECTION WHERE IT MEETS
MUCOSAL BARRIERS AND MAY NOT BE
ABLE TO PENETRATE EFFICIENTLY.
OR DURING WITHIN THE RECIPIENT
HOST AND DO VARIANTS INFECT IN
THE RECIPIENT BUT DURING THE
INITIAL EXPANSION OF THE VIRAL
POPULATIONS MANY LINEAGES ARE
LOST.
THE DISTINGUISHED THE
POSSIBILITIES WE DEVELOPED A
BAR-COD
BAR-CODED INFLUENZA VIRUS AND
WE'VE IDENTIFIED A REGION IN THE
GENOMES THE SITE OF DIVERSITY A
NATURAL OCCURRING BAR CODE AND
THIS IS ANOTHER SEASONAL
INFLUENZA VIRUS AND DIDN'T WANT
TO ATTENUATE THE VIRUS BUT
RATHER BUILD IN THE DIVERSITY
SEEN AT THE POPULATION LEVEL
INTO ONE VIRAL POPULATION.
WE FOUND 12 SITES OF VARIATION
AND ALLOWED EACH TO BE ONE OF
TWO NUCLEOTIDE IDENTITIES.
SINCE TWO TO THE 12 IS 4,096 THE
SYSTEM CREATES A LIBRARY WHERE
THERE ARE UP TO THOSE PRESENT
AND DETECTS CHANGES IN THE
GENETIC MAKEUP OF A VIRAL
POPULATION.
WE WORKED HARD TO MIKE SURE THE
BAR CODE WOULD BE FITNESS
NEUTRAL AND WILL DETECT RANDOM
CHANGES IN THE COMPOSITION.
IN OTHER WORDS, DETECT GENETIC
THROUGH AND WE USED THIS WITH
OUR FAVORITE ANIMAL MODEL WHICH
IS A GUINEA PIG.
WE USE THEM FOR INFLUENZA A
VIRUS FOR TRANSMISSION VIRUS
BECAUSE THEY'RE HIGHLY
SUSCEPTIBLE TO INFECTION AND WE
SEE TRANSMISSION FROM THE GUINEA
PIGS AND HAVE TWO WAYS TO MODEL
TRANSMISSION.
WE HAVE INFECTED AND EXPOSED
ANIMALS TOGETHER WITHIN THE SAME
CAGE SO ALL MODES CAN OCCUR AND
HERE THE AEROSOL WITH ANIMALS IN
THE SAME CAGE BUT SEPARATE A
DIVIDER SUCH VIRUS CAN ONLY MOVE
THROUGH THE AIR TO THE EXPOSED
ANIMAL.
WE HAVE FOUR CONTACT
TRANSMISSION PAIRS AND THIS IS
SHOWING THE VIRAL LOAD DYNAMICS
IN THE ANIMALS AND THE
INOCULATED ANIMALS IN BLUE AND
EXPOSED ANIMALS ARE IN RED OR
YELLOW AND EVALUATE THE DYNAMICS
AND HAVE TAKEN THE NASAL WASH
SAMPLES AND APPLIED NEXT
GENERATION SEQUENCING TO THE BAR
CODE OF THE VIRUS.
LOOKING FIRST AT THE DIRECTLY
INOCULATED ANIMALS HERE WE HAVE
REPRESENTATIVE ANIMALS SHOWN AND
THE INOCULATION DOSE IS FIVE
TIMES TEN TO THE FOUR
PLATFORMING UNITS.
AND THE Y AXIS IS SHOWING THE
TITER AND NASAL WASHES ACROSS
TIME.
THE COLORS ARE SHOWING UNIQUE
BAR CODES.
EACH BAR CODE IS REPRESENTED AND
THE HEIGHT INDICATE THE
FREQUENCY IN THE POPULATION AND
YOU CAN SEE A LOT OF BAR CODES
WERE MAINTAINED OVER TIME.
IT'S A HIGH DIVERSITY SYSTEM.
THAT COULD BE SEEN MORE CLEARLY
HERE WITH A QUANTITATIVE
ANALYSIS OF DIVERSITY.
IT TAKES INTO ACCOUNT THE NUMBER
OF BAR COZE -- CODES DETECTED
AND SEE IN MOST ANIMALS A
GRADUAL DECLINE OVER TIME AND
YOU CAN SEE RIGHT AWAY THERE'S A
DIFFERENT DYNAMIC PLAYING OUT
AND FEW BAR CODES ESTABLISHED IN
THE EXPOSED ANIMALS.
THIS REPLICATES WELL WHAT HAS
BEEN SEEN PREVIOUSLY AS
DESCRIBED FROM BOTH ANIMAL
MODELS THERE'S A TIGHT
TRANSMISSION BOTTLENECK AND ONE
OR TWO FOR THESE AND IF YOU LOOK
AT THE TIME POINT POSITIVE IN
THE NASAL WATCHES WE HAVE A LOT
OF DETECTED.
THE FIRST IS HIGH DIVERSITY AND
IT'S IN THE ENSUING 24 HOURS THE
DIVERSITY COLLAPSES AND A COUPLE
LINEAGES PREVAIL AND IT'S
OBVIOUS WHEN WE LOOK AT THE
QUANTITATIVE ANALYSIS OF
DIVERSITY THE EXPOSED ANIMALS
ARE PLOTTED IN RED.
YOU CAN SEE THE SHARP DECLINE
EARLY ON IN THE DIVERSITY AND IT
APPEARS THERE'S A LOT OF BAR
CODES BEING TRANSFERRED FROM THE
ENVIRONMENT TO THE RECIPIENT
ANIMALS AND FEW THAT SUCCEED
WITHIN THE RECIPIENT HOST MOST
ARE LOST DESPITE THE BAR CODES
ARE FITNESS NULT NEUTRAL AND
THEY'RE LOST BECAUSE OF A
STOCHASTIC EVENT.
THESE THREE PAIRS WERE
INOCULATED AND EXPOSED PAIRS
SHOWN ON TOP AND BOTTOM.
AGAIN, VERY DIVERSE ON THE FIRST
POSITIVE DAY AND LOW DIVERSITY
ON SUBSEQUENT DAYS.
WE LOOK AT THESE TWO DAYS AFTER
THE POSITIVE SAMPLE.
SUCH THAT FEW LINEAGES ARE
SUSTAINED IN THE RECIPIENT OVER
THE COURSE OF INFECTION.
AND TO THINK ON THE IMPLICATIONS
OF THOSE RESULTS IT APPEARS THE
LOSS OF DIVERSITY IS PRIMARILY
DRIVEN BY EVENTS THAT OCCURRED
IN THE EXPANSION OF INFECTION
AND NOT AS WE MIGHT HAVE GUESSED
DURING THE DONOR RECIPIENT
TRANSFER ITSELF WE HAVE A
SITUATION AND IT'S HIGH
DIVERSITY OF EVENT INITIALLY AND
THE EXPANSION IS LOST DURING
POPULATION EXPANSION.
WE HAVE A HIGH DIVERSITY TIME
POINT WITHIN RECEPTING
INDIVIDUAL SO WE HAVE AND WE
HAVE INDIVIDUAL FOLKS WHO HAVE
BEEN EXAMINED.
TO FINISH I WANT TO ACKNOWLEDGE
THE GROUP WHO PERFORM THE WORK
AND THOSE WHO HAVE DONE THE
SEQUENCING ON THE DIRECTLY
CHALLENGED HUMANS AND IT'S A
GREAT TEAM EFFORT WITH THE
EXCELLENT COLLEAGUES AT EMORY
AND VIRGINIA TECH.
THE HUMAN CHALLENGE WORK GETS
FUNDED BY THE FOUNDATION FLU
LAB.
THE BAR CODE RELATED PUBLIC
HEALTH IS FUNDED BY NIH.
THANK YOU FOR YOUR ATTENTION.
>> GREAT TALK.
I WAS WONDERING IF THE
BOTTLENECK IS HAPPENING DURING
THE EXPANSION OF THE VIRUS WHY
ONLY IN THE RECIPIENT AND NOT
THE ORIGINAL INOCULATED ANIMALS?
>> GREAT POINT.
WE'RE VERY INTERESTED IN THE
DIFFERENCE WE SEE BETWEEN THE
NATURALLY EXPOSED ANIMALS AND
THE DYNAMIC.
THE KEY DIFFERENCE IN MY FIND IS
IS THE POPULATION SCIENCE
INITIALLY INTRODUCED AND WE KNOW
WE PUT FIVE TIMES TEN TO THE
FOUR INTRANASALLY IN THE
INOCULATED AND THE LARGE
POPULATION SIZE OVERCOMES THE
STOCHASTIC EFFECTS TO ALLOW
LINEAGES TO EXPAND AND TAKE HOLD
IN THE HOST AND THE SITE OF THE
INITIAL DEPOSITION FOR EXAMPLE
BUT I THINK THAT'S THE MOST
LIKELY EXPLANATION.
>> THANK YOU.
>> THANK YOU, NICE TALK AND
INTERESTING TOPIC.
THE FIRST PART OF YOUR TALK YOU
POINTED OUT THERE'S DISTINCT
NICHES IN PARTS OF THE
RESPIRATORY.
WHAT DO YOU THINK COULD BE
CONTRIBUTING TO THOSE.
IS IT IT IMMUNE INVOLVED?
I'M CURIOUS.
>> I AGREE.
IT'S INTERESTING WE HAVE SEEN
SPATIAL STRUCTURE AND IN THOSE
SYSTEMS WE'RE ABLE TO SAMPLE THE
LOWER RESPIRATORY TRACT.
SO WE SEE THESE VIRAL
SUBPOPULATIONS SET UP IN
DISTINCT POPULATION AND DON'T
NECESSARILY MIX WELL.
THE SIMPLE ANSWER IS VIRUSES ARE
MORE LIKELY TO SPREAD IN A
LOCALIZED MANNER TO INFECT
NEIGHBORING CELLS THAN TO
DISPERSE ALONG DISTANCE AND
CREATE STRUCTURE RATHER THAN A
PERFECTLY MIXED SYSTEM.
THE HUMAN RESPIRATORY TRACT,
EVERYTHING'S PRETTY CLOSE
TOGETHER.
WE WERE SURPRISED TO SEE A
SIMILAR SPATIAL STRUCTURE IN
THAT ANATOMICAL LOCATION.
IN THE MOUSE WE DON'T KNOW IT'S
REPLICATING THERE.
IT'S DETECTED THERE BUT COULD BE
MOVING INTO THE MOUSE FROM WHERE
IT'S REPLICATING AND IT'S
FEEDING INTO THE ORAL SITE NOT
JUST ANYWHERE AND NEEDS FURTHER
INVESTIGATION.
RIGHT NOW IT'S SIMPLY AN
OBSERVATION.
>> THANK YOU.
I HAVE A QUESTION ON MY OWN.
CAN YOU COMMENT ON THE HIGHLY
VARIABLE VIRUSES COULD BE
DETECTED PARTICLES?
>> THAT'S A GOOD POINT.
ONE POSSIBILITY WE SEE THEM LOST
EARLIER IS THE TRANSMITTED
GENOMES WHAT WE DETECT COULD BE
AC PACKAGED WITH A GENOME WITH
A DELETION OR NOT ALL THE GENE
SEGMENTS.
SUCH A VIRUS COULD INFECT CELLS
AND COULD START REPLICATION TO
SOME EXTENT BUT WOULDN'T MAKE
ANY PROGENY.
SO IT'S A POSSIBILITY I DON'T
HAVE EVIDENCE TO EXCLUDE IT AND
THIS IS LOOKING AT THE GEN EITY
AND IT COULD BE 12 OR 24 HOURS
BEFORE ANY PROGENY ARE MADE.
THEY WOULD HAVE A STRONG
TEMPORAL ADVANTAGE AND WOULD
TAKE OVER DUE TO THE
EXPERIMENTAL NATURE FOLLOWING
FROM THE FIRST GENERATION.
WE CANNOT EXCLUDE THE
POSSIBILITY OF INFECTED GENOMES.
>> OUT OF CURIOSITY, DO YOU HAVE
A NUMBER OF ALL THE BAR CODES
TRANSMITS?
OUT THEIR 4,000 IB AVERAGE IN
THOSE EXPERIMENTS, HOW MANY WERE
TRANSMITTED IN GENERAL?
>> I DON'T HAVE IT AT MY FIN
FINGER -- TIPS BUT LOOKS LIKE A
LOT.
>> WELL, THANK YOU SO MUCH.
>> WE DON'T HAVE ANY QUESTION
ONLINE SO ANICE THANK YOU SO
MUCH AND HOPEFULLY WE'LL GET YOU
TO THE D.C. AREA NEXT TIME WE
HAVE A MEETING HERE.
>> THANK YOU.
>> OKAY.
WE LITERALLY HAVE -- I'LL
FOLLOW-UP WITH THE QUESTION
ONLINE.
I HAVE A GREAT ANNOUNCEMENT NOW,
WHICH IS THE LUNCH BREAK.
FOR THOSE THAT DO NOT KNOW WHERE
THE FAS TERRACE IS, FOLLOW THE
CROWD.
TRUST ME, THE PEOPLE WILL GO
THERE.
WE'RE SUPPOSED TO COME BACK AT
1:00 SHARP, PLEASE.
GOOD AFTERNOON TO EVERYONE I
AM PATRICIA AND WELCOME TO THE
AFTERNOON SESSION.
IT'S A GREAT PLEASURE FOR ME TO
INTRODUCE THE FIRST SPEAKER OF
THE SESSION, DR. DANIEL CHERTOW.
HE IS THE HEAD OF THE EMERGING
PATHOGEN SECTION IN THE CRITICAL
CARE MEDICINE DEPARTMENT AT NIH
CLINICAL CENTER.
HIS RESEARCH INCLUDES
UNDERSTANDING THE PATHOGEN SINCE
OF SEVERE, EMERGING OR
RE-EMERGING INFECTIOUS DISEASES
INCLUDING EBOLA VIRUS, EBOLA
VIRUS WITH COVID-19 AND CARED
FOR PATIENTS WITH EBOLA IN WEST
AFRICA DURING THE OUTBREAK IN
2015 TO 2016 AND MA AS IT MADE
UNDERSTANDING OF THE PATHOGENIC
DISEASE AND HIS RESEARCH EFFORT
IS TOWARDS UNDERSTANDING THE
HOST PATHOGEN INTERACTION DURING
THE SEVERE COVID-19 AND
ESTABLISH A LANDMARK IN NATURE
DEMONSTRATING SARS COV2 IN AN
TOP CAL SITES INCLUDING THE
BRAIN ANATOMICAL SITES AND THE
PRESENTATION JUST TO SUMMARIZE
THE MOST IMPORTANT STUDIES ON
SEVERE COVID-19 AND THE TITLE IS
SARS COV2 INFECTION AND PER
SISSANCE IN THE HUMAN BODY AND
BRAIN -- PERSISTENCE IN THE
HUMAN BODY AND BRAIN.
THANK YOU PHOR THE INVITATION TO
THE ORGANIZERS AND MRS. SALZMAN
FOR CONTINUING THE AWARDS AND
I'LL STUDY THE FIRST YEAR OF THE
PANDEMIC IN THE NIH CLINICAL
CENTER AND I HAVE NO
DISCLOSURES.
AT THE ONSET NOT MUCH WAS GIVEN
OUTSIDE THE RESPIRATORY TRACT
AND MY GROUP DECIDED TO EMBARK
ON FATAL COVID-19 FIRST TO
DESCRIBE THE BURDEN OF COV2 AND
OUTSIDE THE RESPIRATORY TRACT
AND THE SECOND WAS TO DEFINE THE
CELLULAR TROPISM ACROSS THE
HUMAN BODY AND IMPORTANTLY THE
BRAIN.
THE THIRD IS TO DETERMINE IF
SARS COV2 PERSISTS IN WHAT
TISSUES AND INTERVALS AND TO DO
GOOD VIROLOGY AND DETERMINE THAT
SARS COV2 REPLICATES AND IS
THERE EVIDENCE AS THE EVOLVING
IN ANATOMIC COMPARTMENTS WE
COLLABORATED WITH A NUMBER OF
INVESTIGATORS AND VERY MUCH
REGIONAL HOSPITAL AND EASTERN
SHORE REGIONS OF MARYLAND AND
RECRUITMENT WAS INITIATED WITH
OUR UPSTREAM CLINICAL PARTNERS
AND WE HAD CONNECTION WITH THE
CRITICAL CARE DOCKS -- DOCTORS
WHO ALERTED US TO COVID-19
DEATHS AND THE REMAINS WERE
TRANSFERRED AND AUTOPSY WAS
PERFORMED IN COLLABORATION WITH
A POSTMORTEM INTERVAL OF LESS
THAN 24 HOURS TO PRESERVE TISSUE
AND PROTEIN AND NUCLEIC ACID
WITHIN TISSUE.
WE PERFORMED A TOTAL OF 44
AUTOPSIES BETWEEN MARCH AND 2020
AND MARCH 2021.
EACH AUTOPSY THAT WAS PERFORMED
IS DENOTED HERE ON THE MARYLAND
EPIDEMIOLOGY CURVE OF CASES
OUR CASES WERE WELL DISTRIBUTED
OVER THE FIRST YEAR OF THE
PANDEMIC.
IMPORTANTLY, NONE OF THESE
INDIVIDUALS WERE VACCINATED THIS
IS BEFORE VACCINATES WERE
AVAILABLE AND ALL INFECTED WITH
WILD TYPE AND THE VARIANT OF
CONCERNS HAS IMPLICATIONS FOR
THE GENERALIZABILITY OF OUR
FINDINGS.
WE PROCURED TISSUES FROM ALL
MAJOR ORGANS IN 44 CASE WITH
EXTENSIVE BRAIN SAMPLING IN A
SUBSET OF 11 CASES.
WE PRESERVED OVER 10,000
SPECIMENS ACROSS 85 ANATOMIC
SITES AND BODY FLUIDS IN THREE
WAYS.
FRESH TISSUE AND BODY FLUIDSES
-- FLUIDS FOR DROPLET PCR FOR
SARS COV2 AND PRESERVED ADJACENT
TISSUE FOR FLASH FROZEN SAMPLES
SO IN A SUBSET OF TISSUES WE CAN
PERFORM CULTURE AND LOOK FOR
REPLICATION IN OUR TISSUES AND
TISSUES WERE FIXED FOR
SUBSEQUENT HISTOLOGICAL AND
CHEMISTRY.
AND OUR COHORT WAS DIVERSE --
[TECHNICAL DIFFICULTIES]
OF THE 11 ASSAYS ACROSS ALL
TISSUE GROUPS SAMPLED WITH THE
HIGHEST LEVELS DEPICTED IN RED,
DETECTED IN RESPIRATORY AND
CARDIOVASCULAR TISSUES EARLY IN
INFECTION AND AMONG THE LATE
CASES SARS RNA PERSISTED IN LOW
LEVELS DEPICTED IN THE BLUE
COLOR INCLUDING IN PATIENT 42
DENOTED AT THE TOP OF THE COLUMN
WHO DIED 230 DAYS AFTER INITIAL
COVID-19 ONSET THE TIME OF DEATH
RELATIVE TO ILLNESS ON SET IS AT
THE BOTTOM, EXCUSE ME, 230 DAYS
AFTER ILLNESS ONSET AND THEY
DIED OF CAUTIONS UNRELATED TO
THEIR INITIAL INFECTION.
SO DISPLAYED HERE ARE PCR
RESULTS FOR THE SAME 11 PATIENTS
ACROSS 10 ANATOMIC SITES IN THE
NERVOUS SYSTEM AND DETECTED IT
IN CNS TISSUES AND DETECTED
RESIDUAL RNA IN LATE CASES BOXED
THERE.
WE DEFINED LATE AS SOMEBODY THAT
DIED GREATER THAN OR EQUAL TO 31
DAYS AFTER THE INITIAL ILLNESS
ONSET AND IN THOSE LATE CASES WE
DETECTED RNA IN EACH OF AND
EVERY ONE OF THOSE IN AT LEAST
ONE REGION OF BRAIN AND
INCLUDING PATIENT NUMBER 42 WHO
DIED 230 DAYS AFTER ILLNESS
ONSET.
FROM OUR FROZEN SAMPLES WE HAD A
SUBSET OF TISSUES AND GREW VIRUS
FROM 25 FOCUSSING ON TISSUES
OUTSIDE THE RESPIRATORY TRACT
AND USING A VIROCELL CULTURE
THAT OVEREXPRESSED AND
SUCCESSFULLY GREW SARS COV2 FROM
NON-RESPIRATORY TISSUES THE E
EYEEY
EYEEYE,
DIFFERENT ORGANS AND ON THIS
FIGURE LOG TRANSFORMED RNA
LEVELS ARE SHOWN ON THE Y AXIS
AND ON THE LOG AXIS, LOG
TRANSFORM DAY OF ILLNESS ON THE
X AXIS LONG TRANSFORMED DAY OF
ILLNESS IS SHOWN AND SARS RNA
LEVELS WERE HIGHER IN
RESPIRATORY TISSUES SHOWN IN RED
COMPARED TO NON-RESPIRATORY IN
BLUE FOR MONTHS FOLLOWING
ILLNESS ONSET AND AYER RATE OF
DECLINE INVOLVED RNA LEVELS WAS
NOTED IN RESPIRATORY COMPARED
WITH NON-RESPIRATORY TISSUES
SUGGESTING A MORE RAPID
CLEARANCE FROM RESPIRATORY
COMPARED WITH NON-RESPIRATORY
TISSUES.
FINALLY WE EVALUATE THE
PROPORTION OF SARS COV2 SPIKE
GENE VARIANT POPULATIONS ACROSS
DIFFERENT TISSUES WITH THE SAME
PATIENT.
AND HAD BRAIN TISSUE BRACKET IN
RED NOT PRESENT IN THE LUNG
SAMPLE FROM THE SAME PATIENT.
THESE SUGGEST COMPARTMENTIZING
AND WHERE WE SEQUENCED VIRUS THE
PREDOMINANT VARIANT AND WE WERE
ABLE TO DETECT MINOR VARIANTS IN
ONE ANATOMIC DEPARTMENT NOT
DETECTED IN THE RESPIRATORY
TRACT.
A SUBSET INCLUDE THYROID AND
ESOPHAGUS AND YOU CAN SEE THE
BROWN STAINING AND IN TOTAL WE
DETECTED SARS COV2 WITHIN MORE
THAN 30 CELL TYPES ACROSS 35
TISSUES.
SHOWN HERE IN THE BRAIN AND
FINAL CORD ON THE RIGHT AND WE
OBSERVED STRAINING WITH THE
MAGENTA AND THESE OBSERVATIONS
WERE ORTHOGONALLY VALIDATED
USING TWO DISTINCT PROBES ONE
WITH SPIKE AND CAPSID AND CHROME
GENIC IHC AND IN SUMMARY WE
OBSERVED SARS COV2 BURDEN IS
HIGHEST IN RESPIRATORY TISSUES
BUT WIDELY DISSEMINATED IN THE
BODY AND BRAIN AND RNA CAN EXIST
IN ANATOMICAL COMPARTMENTS OVER
SEVEN MONTHS AND THE VIRUS
INFECTS SEVERAL TYPE AND ABLE OF
EVOLVING IN MULTIPLE TISSUES
INCLUDING BRAIN AND WE OBSERVED
A PAUCITY OF INFLAMMATION AND
EFFECT OUTSIDE THE LUNGS IN
EARLY CASES AND AS LATE AS OUR
LATE CASES IF YOU LOOK AT
NON-RESPIRATORY ORGANS, THE
LIVER, KIDNEY, ETCETERA THERE
WAS AN ABSENCE OF HISTO PATH
LOGIC FINDINGS AND THERE WAS A
PREDOMINANTLY ABSENCE OF IMMUNE
CELL INFILTRATE IN THE TISSUES
WE CONFIRMED WERE INFECTED.
AND SO THIS WORK WAS TOOK A
YEOMAN'S EFFORT AND WAS TEAM
SCIENCE AND I WANT TO HIGHLIGHT
NAMES AND THE PRIMARY
COLLABORATORS AT THE UNIVERSITY
OF MARYLAND WERE ALLISON
GRAZIOLI A CRITICAL CARE AND
NEPHROLOGY DOCTOR WHO HELPED
FACILITATE THE BUILDING OF OUR
NETWORK AS WELL AS CAPPEL AND
OUR PHYSICIANS AT MARYLAND THAT
HAVE PARTICIPATED IN BOTH THESE
STUDIES AND ONGOING DOWN STREAM
WORK WE'RE DOING ON THE TISSUES.
SIDNEY STEIN AND MY GROUP WAS
THE POSTDOC THAT LED THE STUDY
AND THE FIRST AUTHOR ON THE
PAPER.
SIDNEY DID MORE OR LESS ALL THE
DROPLET DIGITAL PCR ACROSS
THOUSANDS OF THESE TISSUES AND
PARTICIPATED IN A GOOD MAJORITY
OF THE AUTOPSIES THEMSELVES AND
AMY DEWITT HELPED WITH THE
VIROLOGY WORK IN THE CONTAINMENT
LABS AND KAREN PETERSON AND
CLAYTON WINKLER DID THE
IMMUNOFLUORESCENCE IN THE BRAIN
AND ELI AND HIS GROUP DID THE
HIGH-SEQUENCING SINGLE CELL HIGH
THROUGHPUT SEQUENCING OF OUR
VARIANTS ACROSS THE DIFFERENT
TISSUES AND THEN OF COURSE OUR
NCI PARTNERS.
THIS WAS DONE AND IT'S
INTERESTING TO HISTORY TO NOTE
DR. KLINER WHO HEADS THE
ANATOMIC PATHOLOGY LAB HAS BEEN
HERE SINCE THE BEGINNING OF THE
AIDS EPIDEMIC AND WHEN THIS
STARSTAR
ED -- STARTED THEY HAD
FAMILIARITY WITH A NEW OUTBREAK
THOUGHT TO BE HIGHLY CONTAGIOUS
AND SHOWED GREAT FORTITUDE AND
SCIENTIFIC CURIOSITY TO DO THESE
STUDIES AND HAVE DONE THESE HAND
IN HAND WITH THEM AND THEY WOULD
GET CALLS FROM THE ADMISSIONS
TEAM SAYING WE NEED TO
FACILITATE A POSTMORTEM INTERVAL
OF 22 HOURS POST DEATH WHICH IS
ALMOST UNHEARD OF WITH THESE
TYPES OF STUDIES AND WITH THAT I
WILL CLOSE AND TAKE YOUR
QUESTIONS.
>> QUESTION?
CHARLIE?
>> AMAZING STUDY.
CONGRATULATIONS.
AND I WAS WONDERING, DO YOU HAVE
ANY INFORMATION ON PEOPLE THAT
HAVE HAD A MILDER NON-LETHAL
OUTCOME WHERE YOU KNOW SOMETHING
ABOUT THEIR HISTORY AND THEN
THEY SORT OF RAN INTO A WALL OR
SOMETHING LIKE THAT?
>> SO THIS WAS AN IMPORTANT
QUESTION.
THANK YOU FOR YOUR QUESTION,
DR. RICE AND FOR YOUR COMMENT OF
THE DAY.
THIS WAS A QUESTION THAT CAME UP
RIGHT FROM THE BEGINNING ABOUT
REALLY THE GENERALIZABILITY OF
OUR FINDINGS AND I MADE THE
COMMENTS ABOUT VACCINATIONS AND
THERE WERE PATIENTS THAT GAVE US
INSIGHT INTO PERHAPS THE ANSWER
TO YOUR QUESTION.
NOW, TRULY THE ANSWER TO YOUR
QUESTION REQUIRES MORE WORK AND
FIGURED WE'D SET THE EXAMPLE AND
OUR WORK WOULD BE DONE.
I THINK PERHAPS WE'LL PICK UP
THE GAUNTLET AND TRY TO ANSWER
THOSE QUESTIONS BUT I WILL SAY
THAT I ALLUDED TO THIS, THERE
WAS A 6-YEAR-OLD WHO CAME IN
WITH SHE HAD AN UNDERLYING
SEIZURE DISORDER AND WAS ON MEDS
AND DURING COVID THERE WERE
ISSUES AND WAS OFF HER MED AND
SHOWED UP IN THE HOSPITAL AND
WAS EPILEPTIC AND DOESN'T KNOW
SHE HAD COVID AND INCIDENTALLY
TESTED POSITIVE BUT AT THAT TIME
EVERYONE WAS BEING TESTED AND
HAD A COMPLICATION AND
UNFORTUNATELY DIED WITH
COMPLICATION FROM HER
NEUROLOGICAL ISSUES WITHIN 48
HOURS OF HOSPITALIZATION AND
BECAUSE SHE WAS COVID POSITIVE
SHE WAS PREFERRED TO US AND
FOUND VIRAL RNA IN VARIOUS
LEVELS THROUGHOUT HER BODY.
THIS IS A YOUNG HEALTHY BODY
THAT TYPICALLY DOESN'T HAVE
SEVERE ILLNESS AT LEAST NOT
ACUTE ILLNESS FROM COVID AND WE
FOUND VIRAL RNA IN MULTIPLE
ANATOMIC COMPARTMENTS.
>> FANTASTIC STUDY.
AS YOU KNOW, SYSTEMIC
MICROVASCULAR THROMBOSIS IS
DESCRIBED WIDELY IN POSTMORTEM
COVID-19 AND IT MAY BE ONE OF
THE UNDERLYING COMMON PATH LOGIC
MECHANISMS.
HOW FREQUENTLY DID YOU OBSERVE
IT AND WAS IT RELATED TO VIRAL
ACTIVITY?
>> SO WE DO DO SPECIAL STAINS
LOOKING FOR PRESENCE OF PLATELET
RICH THROMBINE AND IN THE LUNGS
IN THIS COHORT IT WAS EASILY
DETECTABLE AND PRETTY
WIDESPREAD.
IN OTHER ORGANS IT WAS PRESENT
BUT NOT AS PREDOMINANT AS MIGHT
THINK.
AND IT GETS BACK TO THE COMMENT
ABOUT OUR HISTO PATHOLOGIC
FINDINGS.
WHEN YOU LOOKED AT HISTO
PATHOLOGIC SAMPLES YOU SAW AN
APAUCITY -- AND IT'S NOT SAYING
IT'S NOT CONTRIBUTING TO
PATHOGENESIS.
AND THERE ARE WAYS TO BIAS YOUR
VIEW.
AND THERE'S THE CONCEPT OF WIDE
SPREAD ENDOTHELIAL ITIS AND SAW
A COBBLESTONING OF ENDOTHELIAL
CELLS AND ABSENT BOTH IN THE
LUNGS AND OUTSIDE THE LUNGS.
I THINK THERE'S MORE TO DO ON
THE HOST SIDE TO UNDERSTAND WHAT
THE DRIVERS OF THE ORGAN INJURY
AND DYSFUNCTION.
>> CONGRATULATIONS.
AMAZING JOB.
I'M WONDERING IF DID YOU OBSERVE
ANY CYTOTROPIC EFFECT OUTSIDE
THE RESPIRATORY TRACT OR DO YOU
THINK IT'S JUST A RESERVOIR FOR
REACTIVATION FOR THE VIRUS IN
THE FUTURE.
>> WE LOOKED CAREFULLY AND DID
NOT FIND EVIDENCE OF CYTOPATHIC
EFFECT OUTSIDE THE RESPIRATORY
TRACT.
IN PARTICULAR AND OBVIOUSLY
WE'RE WIDE INTERESTED IN THE
BRAIN AND PURSUING ADDITIONAL
STUDIES IN THE BRAIN.
AND AN ABSENCE OF PATHOLOGY IN
THE BRAIN AND AS YOU LOOK IN THE
VIRUS YOU MAY TAKE A VIRUS LIKE
RABIES.
RABIES CAN GET IN THE BRAIN BUT
THEY HAVE THESE NEURONS IN THE
BRAIN BUT ABSENCE OF THAT THERE
ISN'T REALLY CYTOPATHIC EFFECT
OF THE NEURONS BUT THE
TRANSCRIPTION FILE IS ALTERED.
BECAUSE YOU DON'T SEE HISTO
PATHOLOGIC INJURY DOESN'T MEAN
THERE'S NOT DYSFUNCTION.
>> CONGRATULATIONS.
AND THE VARIANTS YOU SEQUENCE ON
THE TISSUES PERSISTENTLY
AFFECTED CAN YOU COMMENT WHETHER
ANY OF THOSE MUTATIONS ARE FROM
VARIANTS YET TO BE EMERGING 2021
OR SO.
>> THE ANSWER IS NO.
MOST OFTEN THE PREDOMINANT
PATTERN IN THE INDIVIDUALS WHERE
WE SEQUENCE VIRUS ACROSS SITES
THE PREDOMINANT PATTERN IS THE
MAJOR VARIANT PREDOMINATED
ACROSS MOST TISSUES BUT THERE
WERE REPEATED EXAMPLES WHERE
MINOR VARIANTS WERE PRESENT
OUTSIDE THE LUNGS THAT WERE NOT
DETECTED IN THE LUNGS.
AND CLEARLY FROM SOME OF THE
WASTE WATER STUDIES THAT HAVE
BEEN DONE, THERE ARE SUGGESTIONS
THAT WE KNOW AND
IMMUNOCOMPROMISED INDIVIDUAL
WHERE'S THEY REPLICATE FOR A
LONG PERIOD OF TIME PERHAPS IN
THAT POPULATION THAT'S WHERE
SOME OF THE VIRAL EVOLUTION IS
TAKING PLACE.
>> CLEAR PRESENTATION.
VERY NICE.
I WAS WONDERING HOW YOU
EXTRAPOLATE ALL THIS INFORMATION
TO LONG COVID?
DOES THE VIRUS HAVE TO BE THERE
FOR LONG COVID?
>> THANK YOU FOR YOUR QUESTION
AND COMPLIMENT.
AS YOU KNOW AND AS WE DESCRIBED,
WE DID OBSERVATIONAL SCIENCE.
IN GETTING TO MECHANISM OF LONG
COVID OR EVEN UNDERLYING FACTORS
WITH LONG COVID IS CHALLENGING
AND WERE AIR FULL NOT TO
EXTRAPOLATE THE STUDY OF ACUTE
COVID FOLKS TO LONG COVID.
HAVING SAID THAT IT PUT A STAKE
IN THE GROUND AND HIGHLIGHT THE
POTENTIAL FOR VIRAL PERSISTENCE
AND THE POTENTIAL CONTRIBUTING
PERHAPS TO LONG COVID AS AN
UNDERLYING HYPOTHESIS THAT NEEDS
TO BE EVALUATED AN TED TESTED
FURTHER.
I WON'T COMMENT ON CAUSALITY BUT
THE FIELD HAS MOVED AND THE
PIECE OF LONG COVID HAS TO BE
EVALUATED.
>> ALSO, IN RELATIONSHIP TO THE
QUESTION THAT DR. ALTER RAISED,
DO YOU THINK AND THE TITER OF
THE VIRUS SINCE YOU HAVE
ANTIBIOTICS AFTER THE INITIAL
INFECTION.
LIKE A COLD INFECTION BUT STILL
OR SARS AFTER BECAUSE THE DATA
SHOWS ALMOST ALL ORGANS HAVE
SIGN OF REPLICATION OF THE
VIRUS.
WE DON'T KNOW ONCE THE
ANTIBODIES APPEAR MAYBE YOU HAVE
LOW LEVEL IN SOME ORGAN.
>> I THINK THE PREDOMINANT
HYPOTHESES AROUND LONG COVID IS
IS THERE VIRAL PERSISTENCE AND
THE HOST RESPONSE TO PERSISTENT
VIRUSES CONTRIBUTING TO SIGNS
AND SYMPTOMS.
THE PROTHROMBOTIC UNDNATURE, IS
THAT CONTRIBUTING TO CELL
DYSFUNCTION AND OTHERWISE HE'S
ARE ALL BEING EVALUATED BUT
CHALLENGING.
ONE OF THE STUDIES AND INCLUDING
DR. KNOPF OR BRINGING A COHORT
OF INDIVIDUALS CHARACTERIZED
WITH LONG COVID AND DO A
SAMPLING OF A SUBSET OF TISSUES
YOU CAN ACCESS AND TO SEE IF
THERE'S AN ASSOCIATION TO BEGIN
WITH BETWEEN VIRAL PERSISTENCE
AND GET CLOSER THERE TO TARGET
ASSOCIATION.
>> THANK YOU, DANIEL.
NOW LET'S MOVE TO THE NEXT
SPEAKER, AND IT'S A GREAT
PLEASURE FOR ME TO INTRODUCE
DR. URSULA BUCHOLZ ALSO KNOWN AS
ULA.
THE CHIEF OF THE -- NO?
THERE'S ANOTHER SLIDE?
NO?
OKAY.
LET ME GIVE YOU THE TITLE THEN.
ULA IS THE CHIEF OF THE RNA
VIRUS SECTION NOTICE LABORATORY
OF INFECTIOUS DISEASE AT THE
NIAID.
SHE COMPLETED HER Ph.D. IN
VIROLOGY AT THE UNIVERSITY OF
BERLIN IN GERMANY WHERE SHE
STUDIED ON HUMAN RESPIRATORY
STITIAL VIRUS AND FOLLOWED THE
LEAD FOCUSES ON THE SYNCYTIAL
VIRUS AND BECAME THE CHIEF OF
THE SECTION AND PRIMARY RESEARCH
INTEREST IS TO DEVELOP A LIVE
ATTENUATED PEDIATRIC VACCINE
AGAINST THE RCV WHICH CAUSES A
PRIMARY DISEASE IN INFANTS, THE
ELDERLY AND THE
IMMUNOCOMPROMISED INDIVIDUALS.
THE TITLE OF HER PRESENTATION
TODAY IS DEVELOPMENT OF LIVE
INTRANASAL VACCINES FOR ACTIVE
IMMUNIZATION AGAINST YOUNG
CHILDREN IN RESPIRATORY
SYNCYTIAL VIRUS DISEASE.
>> THANK YOU FOR THE KIND
INTRODUCTION AND THANK YOU TO
THE ORGANIZERS AND MRS. SALZMAN
FOR LETTING US TALK ABOUT OUR
WORK.
THIS IS GOING TO BE A
PRESENTATION ABOUT OUR CLINICAL
STUDY PROGRAMS WITH OUR CLINICAL
COLLABORAT
COLLABORATORS THE COMMUNICATION
RESEARCH AND WITH RODCHESTER
VANDERBILT AND LONG STANDING
COLLABORATION WITH THE IMPACT
NETWORK UP TO 18 CLINICAL SITES
AND THE RESPIRATORY SYNCYTIAL
VIRUS HAS AN RNA GENOME
COMPRISING 10 GENES AND 11
PROTEINS AND CAN BE MANIPULATED
BY THE SYSTEM PETER COLLINS SET
UP AT THE NIH.
RSV HAS TWO MAJOR PROTECTIVE
ANTIGENS OF FUSION GLYCOPROTEIN
F AND G AND THE PROTEINS THAT
INHIBIT THE INTERFERON RESPONSE
AND ANOTHER FACTOR OF THE RNA
REGULATORY FACTOR M2T.
RSV IS THE MOST IMPORTANT CAUSE
OF SEVERE LOWER RESPIRATORY
DISEASE IN INFANT DISEASE
CAUSING SEASONAL YEARLY
EPIDEMICS AND MOST THE BURDEN
OCCURS IN CHILDREN ABOVE SIX
MONTHS OF AGE.
THE MAJORITY IN CHILDREN ABOUT
SIX MONTHS OF AGE.
THIS YEAR TWO APPROACHES
RECEIVED APPROVAL IN '23.
ONE IS INDIRECT AND THIS DURING
WEEKS 32 TO 36 OF PREGNANCY AND
HALF LIFE MONOCLONAL SEVEN
MONTHS WHICH IS SUPPOSED TO BE
USED FOR SEASONAL IMMUNIZATION
OF NEONATES AND CHILDREN UNDER 8
MONTHS OF AGE BUT DUE TO
AVAILABILITY ISSUES THE
RECOMMENDATION THIS YEAR IS
UNDER SIX MONTHS OF AGE.
PROVIDING SHORT TERM PASSIVE
PROTECTION LEAVING OLD ER
INFANTS AND CHILDREN WITHOUT
ACTIVE IMMUNITY TO RSV AND A
VACCINE FOR THOSE STILL MUTED IN
THE POPULATION AND WOULD BE A
PART OF THE PEDIATRIC SCHEDULE.
RSV INFECTS THE RESPIRATORY
EPITHELIUM.
INFECTION IS RESTRICTED TO THE
RESPIRATORY TRACT AS FAR AS WE
KNOW AND I TALKED ON THIS SIDE
THE HIGH ATTACK RATE EARLY IN
LIFE AND BASICALLY EVERYBODY HAS
ANTIBODIES BY AGE TWO.
CAUSES THE RESPIRATORY DISEASE
FROM UPPER RESPIRATORY ILLNESS
WITH FREQUENT PERIODS OF
WHEEZING.
PRIMARY INFECTIONS ARE
ASYMPTOMATIC AND LET SEVERE OR
DURING THE SARS COV2 PANDEMIC WE
HAD A BIG CHANGE IN EPIDEMIOLOGY
WITH NO SIGNIFICANT WITH NO
CIRCULATION WITH LOCK DOWN
GUIDANCE IN EFFECT AND THIS LEFT
A HUGE POPULATION OF HIGHER AGE
GROUPS WITH A HIGHER LEVEL OF
SUSCEPTIBILITY TO RSV AND IN '22
WE OBSERVED WORLDWIDE O
HOSPITALIZATIONS IN OLDER
CHILDREN AND WE DO NEED ACTIVE
IMMUNITY TO INDUCE IMMUNITY IN
OLDER INFANTS AND YOUNG
CHILDREN.
SO FOR US THE TARGET POPULATION
STARTING IN FOUR TO SIX MONTHS
OF AGE WE'RE DEVELOPING
INTRANASAL VACCINE FOR
ADMINISTRATION FOR ANTIBODIES
AND RESIDENT T CELLS AND B CELLS
TO INDUCE RAPID PROTECTION EARLY
IN LIFE LIVE VACCINES CAN BE
IMMUNOGENIC IN PASSIVE
ANTIBODIES.
THESE ARE ADJUVANTING AND AND
OUR RSV NON EXPERIENCE THAT
INCLUDES PASSIVE IMMUNITY BUT
NOT ACTIVELY ENCOUNTERED RSV AND
SUBUNIT VACCINES BECOME
CONTRAINDICATED BECAUSE INDUCED
MARKERS OF PRIMING FOR ENHANCED
RESPIRATORY DISEASE.
THE PRIME FOR ENHANCE DISEASE
WITH RSV INFECTION.
THIS IS TRUE EVEN FOR TH1
ADJUVANTS BUT NOT ASSOCIATED
WITH LIVE VACCINES.
THE PROB FOR US IS WE'RE DEALING
WITH AN INFECTION AGENT AND OUR
EFFICACY AND IT'S HARD TO
PROTECT EFFICACY AND PROTECTION
AGAINST RSV ASSOCIATED ILLNESS
WHILE STERILIZING IMMUNITY.
THE BIG CHALLENGES ATTENUATION
AND IMMUNOGENICITY ARE ADVERSELY
CORRELATE.
WE'RE USING REVERSE GENETICS TO
AND AND WE'RE DEALING WITH NOT
HAVING A FULLY PERMISSIVE ANIMAL
MODEL AND FOR THE LAST BIT OF
ATTENUATION WE RELY ON PHASED 1
STUDIES IN RSV NEGATIVE CHILDREN
BECAUSE POSITIVE CHILDREN WOULD
BE NOT AS ACCEPTABLE OR THE
VACCINE AND NEED TO STUDY THE
VACCINE CANDIDATES IN THE
SEASONS FREE FROM COMMUNITY RSV
ILLNESS AND ALSO DEALING WITH
THE INCIDENTS OF RESPIRATORY
ILLNESSES OF ALL CAUSES IN THE
YOUNG PEDIATRIC POPULATION EVERY
PARENT OF YOUNG CHILDREN KNOWS
VERY WELL.
AND THE LIVE VACCINES MAY BE
AFFECTED BY THE PRESENCE OF A
LONG LASTING ANTIBODY.
AND DESIGN THE ATTENUATING
MUTATION WE CAN DELETE CODONS
AND/OR STABILIZE PALPITATIONS
AND STABILIZE THESE AGAINST
ATTENUATION AS OTHERS IN THE LAB
HAVE DONE.
AND THE ANTAGONIST AND WE CAN
COMBINE MUTATIONS AND OTHERS IN
OUR LAB WORK WITH THE
OPTIMIZATION AND CAN OPTIMIZE
OPEN READING TO INCREASE ANTIGEN
EXPRESSION OR REORDER THE
GENOME.
I WANT IT TALK ABOUT THE LEAD
CANDIDATES THAT ARE CURRENTLY
THE MOST PROMISING AND HOPEFULLY
MAKING INTO EFFICACY STUDIES AND
THE LEAD CANDIDATE AND THIS IS
BASED ON A DELETION OF THE
INTERFERON AN TAG GIFT WHICH IS
PATHOGENICITY FACTOR BECAUSE IT
PROMOTES SHEDDING OF BRONCHIAL
EPITHELIAL CELLS AND IS
BENEFICIAL FROM THAT
PERSPECTIVE.
IN ADDITION, THIS CANDIDATE
CONTAINS A DELETION OF ONE COE
ON POLYMERASE PROTEIN THIS IS
GOOD FOR SAFETY IN THE LOWER
RESPIRATORY TRACT AND THE
VACCINE CANDIDATE LOWER THAN THE
UPPER TRACT.
AND IN ADDITION WITH THE
MUTATION TO STABILIZE THE VIRUS.
THIS CANDIDATE HAS BEEN
SEQUENTIALLY EVALUATES FOR
SAFETY AND CERTAINLY NEGATIVE
CHILDREN IN ORDER OF FIVE AND
TEN TO THE SIX DOSE IN 20
VACCINEES.
THE STUDY ESTABLISHED THE SAFETY
THAT WAS NO EXCESS RESPIRATORY
ILLNESS COMPARED TO THE PLACEBO
RECIPIENTS AND TITERS WERE
RELATIVELY LOW WITH LESS THAN
TWO LOGS PER MILLILITER OF THE
VACCINE AND WE AND WE SAW
ANTIBODY RESPONSE WITH 80% OF
THE VACCINES HAVING SERUM
ANTIBODIES WITH IMMUNIZATION AND
THE TITERS IS VERY LOW THIS IS
SIMILAR TO WHAT WILD TYPE IN
INDUCES IN THE FIRST INFECTION
IN YOUNG CHILDREN.
THE VACCINE TAKE OF 100% AND WE
FOLLOWED THE KIDS THROUGH THE
WINTER WE SAW SUBSTANTIAL RISE
IN NEUTRALIZING ANTIBODIES
DURING THE SEASON SO THESE
VACCINE CANDIDATES PRIME FOR A
STRONG RESPONSE AND SAW THE
RISES IN ABSENCE OF THE RSV
DISEASE IN MANY CASES.
THE SECOND LINE OF PHARMACY
CANDIDATES IS WITH OPEN READING
THAT'S A SMALL OVERLAPPING IN
THE M2 GENE WHICH EXPRESSES AN
RNA REGULATORY PROTEIN.
IF YOU DELETE TO THE OPEN
READING FRAME THERE'S LESS
REPLICATION AND HIGHER LEVEL OF
GENE TRANSCRIPTION AND THAT OF
COURSE RESULTS IN HIGHER PROTEIN
EXPRESSION AND IN THIS CASE
LOOKING AT BLOTS OF THE
PROTECTIVE ANTIGENS F AND G AND
YOU CAN SEE HOW MUCH MORE F AND
G IS SYNTHESIZED IN A VIRUS WITH
AN M2D DELETION AND HIGHLY
RESTRICTED IN CHIMPANZEES AND
GENE DELETION WAS HIGHLY STABLE.
THIS BLUE LINE RESULTS FROM A
PHASE 1 STUDY IN 6 TO 34 MONTHS
OLD WITH NASAL WASHES COLLECTED
OVER SEVEN SEQUENTIAL TIME
PERIODS OVER 17 DAYS AND SAW
SHEDDING IN OUR EXPERIENCE THAT
IS A LITTLE BIT ON THE HIGH
SIDE.
WE WENT BACK TO THE DRAWING
BOARD AND ADDED ANOTHER MUTATION
POLYMERASE WE KNOW FROM OTHER
VIRUSES AND CANDIDATES INDUCES
TEMPERATURE SENSITIVITY AND THIS
IS ALSO MUTATION THAT HAS BEEN
STABILIZED AGAINST D
DE-ATTENUATION OVER TIME AND
THIS YOU SEE ALL THE REDUCED
LEVEL OF SHEDDING AND ALSO MOVED
THE PEAK OF SHEDDING TO A LATER
TIME POINT WHICH IS ALSO A MARK
ER OF ATTENUATION OF THE VIRUS.
THIS CANDIDATE HAS BEEN
EVALUATED IN TWO STUDIES
MEANWHILE AND THE TEMPERATURE
SENSITIVITY AGAIN IS GOOD
ADDITIONAL SAFETY FEATURE TO
REDUCE AND PROTECT THE LOWER
RESPIRATORY TRACT FROM
REPLICATION.
WE AS YOU SAW IN THE LAST WE
LOOKED FOR SHEDDING BY THE ASSAY
ON THE X AXIS AND THE Y CHASMS
A
-- AXIS AND COMPARING THE
LINEAR CORRELATION SHOWS WE HAVE
ABOUT A TENFOLD HIGHER LEVEL OF
COPIES SO INDEED IN VITRO IT
SEEMS THE M2D DELETION DOES WHAT
WE PREDICTED RESULTING IN MORE
POSITIVE AND NEGATIVE SHED AND
MORE RNA SYNTHESIS PER PFU AND
HOLDS UP FOR THREE COMBINED DATA
FROM MANY STUDIES.
LOOKING WHETHER THIS TRANSLATES
TO BETTER IMMUNOGENICITY AMONG
PRIMARY INFECTION IN BOTH CASES
WE SAW LEVELS FROM PRE TO POST
IMMUNIZATIONS AND ANTIBODIES BUT
NO BIG DIFFERENCE IN THE IMMUNE
RESPONSE IN THE PRIMARY
RESPONSES IN YOUNG CHILDREN.
WE ALSO FOLLOWED THE CHILDREN
OVER THE RSV SEASON TAKING A
PRE-SEASON SAMPLE AND
POST-SAMPLE SEASON AND WE FOLLOW
THE KIDS OVER THE SEASON TO GET
REPORTED ON ATTENDED ACUTE
RESPIRATORY ILLNESS AND TAKE
SAMPLES TO IDENTIFY RSV
ASSOCIATED ILLNESS.
WE USUALLY SEE HERE IN THE GRAPH
THE RISES HERE AND YOU LOOK AT
THE RIGHT GRAPH SHOWING PLACEBO
RECIPIENTS GIVES THE COMPARISON
THIS SB WHAT WE SEE IN THE
WINTER SEASON WHEN SEEING RSV
FOR THE FIRST TIME AND THEN
SIMILAR LEVELS OF IMMUNOGENICITY
OF CANDIDATES AND TO WILD TYPE
RSV.
YOU CAN ALSO SEE SOME OF THE
VACCINEES HAVE THESE SUPER
STRONG RISES IN THE RSV SEASON
AND THE STAR IS THE ONE INCIDENT
WITH A REPORT ABOUT RSV ILLNESS
IN THE SEASON AND THAT WAS THE
PARTICIPANT THAT HAS NOT DONE
CONVERSIONS POST VACCINE AND SAW
TWO REPORTS IN THE RECIPIENTS.
AND PRIMING FOR STRONG RESPONSES
AND THAT IS SOMETHING WE SEE
GENERALLY IN THE STUDIES.
YOU CAN SEE THE SECOND RSV
INFECTION PROVIDES THE
SUBSTANTIAL TITER SIMILAR TO
WHAT THAT LEVEL OF SERUM
ANTIBODIES ARE AND THIS IS
ASOLID PROTECTION AGAINST
ILLNESS.
AND FOR US THE QUESTION IS HOW
CAN WE FOLLOW THE NICE IMMUNE
RESPONSES HOW CAN WE GET KIDS TO
FUL
BE FULLY PROTECTED LATER IN
LIFE.
GOOD VACCINE TAKE SHEDDING BY
CULTURE AND LOWERING THE NS2
CANDIDATE AND THAT WAS THE
BIGGEST ARGUMENT FOR MOVING --
FOR THE GREATER COLLABORATIVES
MOVING THIS CANDIDATE FORWARD AS
THEY WANTED TO GO FORWARD WITH
THE MOST PROMISING SAFETY
PROFILE.
WE HAVE ALSO DONE A SMALL
POSTDOC RECENTLY LOOKING AT THE
MOST PROMISING CANDIDATES
DEVELOPMENT IN PHASE 1 POST
IMMUNIZATION AND WE LOOKED AT
COMBINED EFFICACY AND SHOWING IN
THE BLUE LINES AND ALL VACCINES
FROM THE SEVEN STUDIES ON THE
BLACK LINES AND LOOKING AT THE
MOST PROMISING WE DO SEE SOME
SUGGESTIVE DATA THAT SUGGESTS
EFFICACY WHEN YOU LOOK AT
VACCINE EFFICACY AND LOWER
ILLNESS OF UP TO 88% AND WE THEN
LOOK AT THE SUBSET OF KNOWN
RESPONDERS WE DID NOT SEE LOWER
RESPIRATORY TRACT ILLNESS IN
THAT GROUP AND GIVEN SMALL
NUMBERS AND IT'S JUST POSTDOC
ANALYSIS BUT JUST KIND OF
PROMISING.
LOOKING AT THE FOR THE POSTDOC
ANALYSIS ALL TOGETHER THE FIRST
IS VACCINEES WITH EVIDENCE OF OF
WILD TYPE DURING THE SURVEYED
SEASON AND YOU CAN SEE THE
PRIMING FOR THE STRONG RESPONSES
IF YOU SEE WILD TYPE DURING THE
SEASON WE ALSO SAW THE
DISABILITY TO ANTIBODY TITERS
AND THE POSTDOC ANALYSIS
CONFIRMATION THAT ANTIBODY
TITERS TO VACCINE ARE COMPARABLE
TO THOSE AND THE BIG QUESTION
HOW CAN WE HAVE VACCINES FROM
6.4 TO 10.2?
TO THAT END WE'RE WORKING ON THE
VIRE -- VIRUS F PROTEIN WITH
THE MOST POTENT STABILIZED FOR
EXPRESSION.
THE BOVINE INFLUENZA VIRUS IS
PROVIDE
PROVIDING PATHOGEN AND DOWN TO
TWO YEARS IN A DIFFERENT VERSION
OF THE RSV ANTIGEN AND LIKELY
DIFFERENT SET.
WE IMPROVED THIS VECTOR IN MANY
WAYS AND READY TO.
THE PREVACCINE STUDY HOPEFULLY
NEXT YEAR.
THE DISADVANTAGE IS IT'S A
SINGLE PROTEIN.
WE'RE LOOKING AT THE PRIMARY
IMMUNIZATION AFTER THE IMMUNITY
BY THE LONG LASTING ANTIBODIES
OR WITHOUT PRE F HAS WANED AND
LOOK AT THE PRE F VECTOR.
WE KNOW FROM MANY PRE CLINICAL
AND CLINICAL STUDIES THAT THE
SECOND TAME OF AN RSV VACCINE
VACCINE IS NOT GOING TO WITH THE
VICT
VECTOR INDUCED TO RSV BY THE
FIRST LAB ATTENUATED VACCINE.
WE HAVE TWO PROMISING
CANDIDATES.
THE CANDIDATE IN SANOFI WILL GO
POWERED FOR EFFICACY SCHEDULED
TO BEGIN EARLY IN 2024 AND THIS
IS OF COURSE SUPER EXCITING
ONGOING STUDIES.
WE FURTHER STUDIED LEAD
CANDIDATES INCLUDING THE
CANDIDATE IN SMALL PHASE 1/2
STUDIES AND CHARACTERIZE THE
PEDIATRIC IMMUNE RESPONSE
FOLLOWING INTRANASAL
IMMUNIZATION AND THIS IS AS MUCH
AS WE CAN BY SINGLE CELL
SEQUENCING GETTING FROM A SMALL
NUMBER OF CHILDREN IN THE
STUDIES.
IN ADDITION WE'RE MOVING FOR THE
VICTORY VACCINE CANDIDATES AND
LOOKING AT PRIME RICH FACTORS IN
LAB ATTENUATED RSV.
AND THIS IS WHERE WE ARE THIS
WAS A HUGE COLLABORATION AND
PETER COLLINS AND OUR STAFF
SCIENTISTS WHO CARRIED A BIG
LOAD AND OUR CLINICAL
COLLABORATORS I MENTIONED
EARLIER THE SANOFI TEAM AND ALL
THE STUDY PARTICIPANTS IN THIS
MANY PHASE 1 STUDIES.
THANK YOU VERY MUCH.
>> ANY QUESTIONS?
>> GREAT DATA.
ANY EVIDENCE FOR EMERGENCE OF
VACCINE ESCAPE VARIANTS
ESPECIALLY NOW THAT ELDERLY ARE
BEING RECOMMENDED TO RECEIVE THE
VACCINES AS WELL.
>> THERE ARE HUGE COLLABORATIVE
NETWORKS LOOKING AT THIS.
SO FAR RSV IS GENETICALLY STABLE
AND THERE'S A SUB GROUP A AND B
AND CHANGES BETWEEN SEASONS AND
THE IT'S REQUIRED ADDITIONAL
AMINO ACIDS LIKE 12 OR 18.
THEY'RE NOW FURTHER TEMPLATES
FOR ADDITIONAL MUTATIONS.
IF WE WOULD SEE VARIANTS THE F
IS ALSO FUNCTIONALLY CONSTRAINED
AND CAN'T ESCAPE FAR WITH
EPITOPES.
>> WE THOUGHT OF THE
CORONAVIRUS.
IT DOESN'T MUTATE AS MUCH AS.
>> WHAT DO YOU KNOW ABOUT THE
LOCAL MUCOSAL RESPONSES?
>> THE SARS COV2 PANDEMIC
IRONICALLY HAS BROUGHT OUT
CLOSER TO HAVING BETTER TOOLS IN
OUR GROUP.
THE BIG PROBLEM IS WITH THE
PEDIATRIC STUDIES THE TOOLS
YOUNG TODDLERS DON'T TOLERATE
THEM VERY WELL AND THE CLINICAL
TEAMS ASKED TO US REFRAIN FROM
USING THEM AND BACK TO NASAL
SWABS AND WASHES IT'S A
DIFFICULT SUBSTRATE BUT OUR
TOOLS HAVE IMPROVED A LOT AND
WE'RE RETROSPECTIVELY DOING THE
ANALYSIS AND HOPING TO GET MORE.
WE'RE LOOKING AT PBMCs AND
LOOKING AT THE LINEAGE
DEVELOPMENT.
>> GREAT WORK, ULA.
MODERNA IS DEVELOPING AN mRNA
VACCINE FOR RSV AND DO YOU
BELIEVE THE LIVE ATTENUATED TO
BE SUPERIOR TO mRNA AND WHY?
>> IT STILL REMAINS TO BE SEEN
IF LIKE I KNOW MODERNA AND
OTHERS ARE PLANNING TO GO TO
PEDIATRIC COHORTS WITH mRNA
VACCINES.
I MEAN WE'RE AT LID AND THERE'S
A HISTORY OF VACCINE ENHANCED
DISEASE.
FOR US WE'LL STAY WITH LAB
ATTENUATED VACCINES BECAUSE WE
KNOW THEY WILL PROVIDE A VERY
GOOD BASELINE ALL ANTIGENS OF
RSV AND GOOD MUCOSAL IMMUNITY.
THE HARD THING IS TO PROTECT THE
UPPER RESPIRATORY ATTRACT
AGAINST SUSCEPTIBILITY TO
INFECTION AND I THINK ALSO IN
OUR APPROACHES DURING THE
PANDEMIC WE FEEL MORE STRONGLY
THAN EVER IT'S A GOOD APPROACH
FOR A RESPIRATORY VIRUS, LIVE
ATTENUATED OVER mRNA INJECTABLE.
>> I'M CURIOUS ON THE DECISION
TO GO WITH THE BOOST WITH THE
BPRP AND GETTING THE DESCAB
SOLUBLE.
WHAT'S THE ADVANTAGE THERE?
>> ONE OF THE ADVANTAGES IS
WE'RE AGAIN GOING INTRANASALLY.
WE DO SEE SERUM IMMUNE RESPONSES
BUT WE KNOW EVERYTHING ELSE IN
BETWEEN AND THAT'S THE MUCOSAL
IMMUNE RESPONSE INITIATING FIRST
AND IT'S A BIVALENT VACCINE AND
PROTECT AGAINST ANOTHER MAJOR
PLAYER OF RESPIRATORY ILLNESS IN
CHILDREN AND GETTING THE SUPER
GOOD ANTIGEN.
THAT'S BASICALLY.
WE MIGHT PRE-CLINICALLY LOOK AT
OTHERS BUT THAT WILL BE OUR
AVENUE.
>> FOLLOWING THAT QUESTION, WHY
DO YOU THINK CAN YOU SPECULATE
IF THE BOOST IS AS SUPERIOR AS
THE IMMUNOMODIFICATION?
>> IT'S MADE IT STABLE.
THE F PROTEIN IS FIRST SECRETED
AS A PRE-FUSION AND GETS
TRIGGERED AND FALLS TO POST-F
AND A GROUP HAVE SHOWED IF YOU
ABSORB HUMAN SERA WITH POST F
THE NEUTRALIZATION ACTIVITY OF
THE CELL IS STILL THERE AND
NEUTRALIZING ANTIBODIED ARE NOT
CAPTURED WITH POSTS F AND
BASICALLY WE KNOW THAT THE SITE
0 MOST CRITICAL EPITOPE FOR
NEUTRALIZING ANTIBODY --
>> SO WHY NOT IMMUNIZE FROM THE
VERY BEGINNING WITH PRE F THEN?
>> BECAUSE THERE WAS ONE ANIMAL
MODEL IF YOU DO THAT YOU STILL
TRIGGER MARKERS THAT PREDICT
SUSCEPTIBILITY FOR ENHANCED
RESPIRATORY DISEASE.
THE NEXT TIME YOU WOULD GET
INFECTED WITH WILD TYPE RSV IT
MAY NOT BE THE ENTIRE POPULATION
BUT IT MIGHT BE IN SOME
INDIVIDUALS YOU SEE THE ENHANCED
RESPIRATORY DISEASE THAT WAS
OBSERVED WHEN IT INACTIVATED IN
THE EARLY DAYS OF RSV.
>> LAST QUESTION.
>> THANKS FOR YOUR TALK.
I WAS CURIOUS, WHAT CAUSES THIS
ENHANCED -- I FORGOT WHAT TO
CALL IT.
>> ENHANCED RESPIRATORY DISEASE.
THE WAS OBSERVE ED LIKE TRYING
TO DO WORK THROUGH ACTIVATE AND
THEN IMMUNIZED CHILDREN AND IT
WAS ALL GOOD UNTIL YOU GOT TO
RSV NAIVE POPULATIONS WHERE THE
PRIMARY IMMUNIZATION WAS WITH
LOTS OF SOON YOU ARE IMMUNOGENIC
EPITOPES AND LOOKED AT THE
STRONG NEUTRALIZING EPITOPES AND
STRONG INDUCTION OF ANTIBODIES
BUT NOT NEUTRALIZING AND INDUCED
RESPONSE AND LIKE A HUGE
OVERREACTION ENHANCED
RESPIRATORY DISEASE AGAINST THE
RSV ANTIGEN THAT CAME FROM THE
COMMUNITY SO UNINHIBITED RSV
APPLICATION, ABSENCE OF
NEUTRALIZATION WITH A STRONG
IMMUNE RESPONSE AND IN THE EARLY
STUDY TWO TODDLERS PASSED AWAY.
YOU CAN RING KING ET AL.
>> THANK YOU, ULA FOR YOUR GREAT
PRESENTATION.
>> ALL RIGHT.
THE LAST PRESENTATION OF THE DAY
IS DR. TIJANA IVANOVIC.
SHE WAS BORN IN SERBIA BUT CAME
TO THE UNIVERSITY OF CALIFORNIA,
LOS ANGELES TO DO HER UNDER
GRADUATE DEGREE AND GOT A Ph.D.
FROM HARVARD AND POST DOCKED
WITH STEVE HARRISON AN
OUTSTANDING VIROLOGIST OF VIRAL
AND CELLULAR FACTORS OVER MANY
YEARS.
WHILE POSTDOC IN STEVE'S LAB SHE
DEVELOPED SINGLE PARTICLE
METHODS FOR STUDYING VIRAL
FUSION.
AFTER HER POSTDOC SHE WENT TO
BRANDEIS UNIVERSITY AS AN
ASSISTANT PROFESSOR AND HER
PROGRAM WAS MOSTLY ON VIRUS
ASSEMBLY AND HOST INTERACTION
AND ADAPTATION SHE WAS SUPPORT
THE NIH'S NEW INNOVATOR AWARD
AND G. HERALD AND LEAH MATHERS
CHARITABLE FOUNDATION AND WAS
RECRUITED TO COME TO NIH AND
NIAID IN THE LABORATORY OF VIRAL
DISEASES AND STARTED IN MAY THIS
YEAR.
SO WITH THAT, I WILL INVITE
TIJANA TO COME UP AND TELL US
WHAT SHE'S BEEN DOING.
>> THANK YOU, JOE FOR THE KIND
INTRODUCTION.
THANK YOU SO THE ORGANIZERS AND
MRS. SALZMAN FOR THE INVITATION
AND A NICE DAY OF TALKS
INCLUDING BY TO THE AWARDEE.
I'D LIKE TO SHARE WORK FROM MY
LAB BRANDEIS AND HERE.
WE'RE INTERESTED IN
UNDERSTANDING HOW PHENOTYPIC
PROCESSES SO THOSE NOT ROOTED IN
GENETIC CHANGE CAN INFLUENCE AND
CHANGE VIRAL EVOLUTION.
THIS STORY WILL BE ABOUT VIRUS
SHAPE AND STRUCTURE.
SHAPE IN VIROLOGY CONFERS
IMPORTANT FUNCTION AND UNDER
SELECTIVE PRESSURE AND DEFINING
THE PRESSURES AND PROCESSES IS
DIFFICULT ESPECIALLY WHEN
DEALING WITH ONLY SNAPSHOTS IN
TIME.
SO VIRUSES HERE PRESENT UNIQUE
OPPORTUNITY BECAUSE OF THEIR
RAPID VIRAL REPLICATION, THE
RAPID EVOLUTION AND PRESENT
THEIR OWN CHALLENGES AND THIS IS
THEIR SMALL SIZE.
HOW DO WE STUDY THESE AND THIS
IS WHAT WE EMBRACE IN OUR LAB WE
STARTED WITH THE MOST EXTREME
EXAMPLES AND POLY MORPHIC
VIRUSES AND THEY GENERATE A
MIXTURE OF SHAPES THEY INCLUDE
PARTICLES AND FILAMENTS OF
VARIOUS LENGTHS AND IT
REPRESENTS A SINGLE VIRUS
PARTICLES AND SPHERES AND
NOODLES ENCODE THE GENOME AND
SHAPE IS A PHENOTYPIC TRAIT AND
WE FOCUSSED ON THE INFLUENZA
VIRUS BUT I HAVE EXAMPLES OF SO
MEMBERS FROM DIFFERS FAMILIES
HERE LISTED ON THE LEFT
INCLUDING DANGEROUS PATHOGENS
AND EMERGING VIRUS AND SEEMS
IMPORTANT TO UNDERSTAND.
THIS TALK I'LL LEAD YOU THE OUR
DISCOVERY OF THE FUNCTION OF
VARIANT SHAPE AND GROWING
UNDERSTANDING OF THE REGULATION
OF VARIANT SHAPE.
AND BOTH OF THESE STUDIES WERE
ENABLED BY EMPLOYING SINGLE
VIRION AND THE HAVE THE ABILITY
TO STUDY AND CHARACTERIZE THE
FULL DIVERSITY OF A POPULATION
RATHER THAN MEASURING AN AVERAGE
INHERENT IN THE BULK OF
APPROACHES.
AND SO THE FIRST PART OF MY TALK
WILL BE ON THE FUNCTION OF
VARIANT SHAPE AND THIS WORK WAS
LED BY A FORMER POSTDOC IN THE
LAB.
SOME FUNCTION OF THE SHAPE
DERIVE FROM OBSERVATIONS
DOMINATE IN ANIMAL INFECTIONS
AND A FEW PATHOGENS IN VITRO ARE
FOR SPHERICAL FORMS.
I SUGGESTED SOME UNIQUE IN VIVO
PRESSURE THAT FAVORED FILAMENTS.
THEY ENCODE THE SAME GENOME AND
ALSO THE SAME COPY NUMBER.
SO REALLY THE ONLY THING THEY
DIFFER IN IS WHAT'S DISPLAYED ON
THE SURFACES.
AND THESE PROTEINS ARE GENERALLY
INVOLVING ANTIGENS AND WE
HYPOTHESIZED THEY MAY COME FROM
THE MEDIATING CELL ENTRY.
I'M SHOWING AN ELECTROMICROGRAPH
OF A FLU PARTICLE.
YOU CAN SEE A DENSE ARRAY OF
GLUTAMATE AND THIS BINDS TO
RECEPTORS WHICH COMBINE TO THE
ENTIRE PARTICLE AND THE
ENDOSOMES TRIGGER FUSION BY THE
BASE DOMAIN PORTION OF THE
MOLECULE WHICH UNDER GOES A
SERIES OF FUNCTIONAL REARRA
REARRANGEMENTS AND BRINGS THEM
TOGETHER ALLOWING THEM TO FUSE.
BOTH FUNCTION UNDER THE ANTIBODY
PRESSURE AND SO YOU HAVE
ANTIBIOTICS THAT BIND TO THE
HEAD OF HA AND BLOCK ATTACHMENT
OR THE BASE OF HA AND PREVENT HA
CONFIRMAT
CONFIRMATIONAL CHANGES AND
THOUGHT THEY MAY NEUTRALIZE
ANTIBODIES WITH MORE SPIKE
PROTEINS ON THE SURFACES AND THE
FRACTIONATED VIRUSES HAVE
SPHERES OR FILAMENTS IN TWO
PREPS AND IN THE PRESENCE OF
INCREASING ANTIBODY
CONCENTRATIONS AND WITHOUT
ADDING INHIBITORS BUT WITH
EITHER TYPE OF ANTIBODY
FILAMENTS WERE LESS ATTENUATED
FOR THE SAME ANTIBODY
CONCENTRATION.
HERE WHAT IS PLOTTED IS THE FULL
INHIBITION FOR THE PREPS IN
THEIR SPHERES OR FILAMENTS AND
TAKES THE SAME IN THE CONTEXT OF
FILAMENTS AND SPHERES.
WHAT IS THE MECHANISM THAT LEADS
TO THIS RESISTANCE.
WE STARTED ADDRESSING THE
DISCUSSION IN THE CONTEXT OF
ANTIBODIES THOSE THAT BIND EHA
DO NOT INTERFERE WITH ATTACHMENT
BUT PREVENT CONFIRMATIONAL
CHANGES IN THE ENDOSOME.
WE HAD A NICE ASSAY WE DEVELOPED
DURING MY POSTDOC JOE MENTIONED.
THIS IS HERE WE CAN OBSERVE
VIRUS FUSION AT THE LEVEL OF
SINGLE VIRIONS.
AND YOU'RE SEEINGS FUFG AND
MERGING THEIR ENVELOPES WITH A
SINGLE LAYER ON THE MICROSCOPE
COVER SLIP AND DEPOSITING THE
DYE TO THE TARGET MEMBRANE.
YOU SEE THIS AS FLASHES AND
FLUORESCENCE AND THE DYE
DISSIPATE FROM THE SITE OF
FUSION AND YOU SEE MANY HERE IN
THE MOVIE.
THIS INCLUDES FLUORESCEIN AND
THIS WE EMPLOY MICROSCOPES.
YOU GET THE PROCESS AND WE COULD
DERIVE OR EXTRACT FROM THE
INFORMATION FOR FILAMENT VIRUSES
IN A SORT OF MORE MIXED
POPULATION I DESCRIBED EARLIER.
AND EXTRACT THE DATA FROM THE
EXPERIMENTS PERFORMED WITH THE
MORE SPHERICAL OR MOMENTOUS
PREPS WE CAN GET IDEA FOR LONG
VERSUS SHORT PARTICLES IN THE
FUSION ASSAY AND IN THE
PRESENTATION OF THE BASE BINDING
ANTIBODY WITH CONFIRMATIONAL
CHANGES.
THE PROBABILITY OF THE FUSION OR
FUSION EFFICIENCY OR THE END OF
THE REACTION GOES DOWN WITH
DECREASING ANTIBODY
CONCENTRATION AND THAT EFFECT
OCCURS FOR HIGHER ANTIBODY
CONCENTRATION FOR THE
FILAMENTOUS PARTICLE ARE FASTER
AT FUSING AND BOTH PARTICLE
TYPES HOLD THE SAME VALUE.
WHAT DOES IT TELL US ABOUT THE
UNDERLYING MECHANISM?
SO THIS COMES FROM THE FUSION
AND HERE'S A MORE INTENSE
DEPICTION FROM MY POST DOCTORAL
WORK.
HERE YOU CAN SEE THE AGE OF
FUSION IS INDEPENDENT OF EACH
OTHER.
ALL EVENTS TAKE PLACE AT THE
SURFACE OF PARTICLES AND HERE IN
GREEN IS THAT REGION OF THE
VIRION WHICH INTERFACES THE
TARGET BIO LAYER.
AND IT'S TRIGGERED AND THIS
HAPPENS ANYWHERE AT RANDOM IN
THE INTERFACE AND INDIVIDUALS
CANNOT BRING TWO MEMBRANES
TOGETHER.
SOME HAs ARE LOST AS
PARTICIPANTS AND THERE'S
REDUNDANCIES AND YOU'LL HAVE
SOME NEXT TO EACH OTHER AND THEY
CAN COLLAPSE AND BRING THE
MEMBRANES TOGETHER.
THE TRANSITION IS FAST.
AND THE EARLY TRANSITION OF
EXTENSION IS SLOW.
SO THE OVER ALL RATE OF FUSION
IS DETERMINED BY THE RATE OF
EXTENSION AND HUMAN CLUSTERS
WITH THE GROUP THAT CAN COLLAPSE
TOGETHER.
WHICH IS IN TURN RELATED TO THE
ACTIVATION.
AND NOW YOU CAN IMAGINE WHAT
WOULD HAPPEN IF YOU HAD ADDED
ANTIBODIES TO THE MIX.
YOU INCREASE THE FRACTION THAT
CANNOT PARTICIPATE AND RESTRICT
POSSIBLE PROFUSION CLUSTERS
FORMING.
YOU HAVE FEWER WAYS OF FORMING
THE CLUSTERS AND YOU MIGHT
DERIVE IT'S IMPOSSIBLE TO
ASSEMBLE A FUSION CLUSTER.
SO THIS LED TO ME PREDICTION
LARGER PARTICLES MIGHT HAVE MORE
OPPORTUNITIES FOR FUSION CLUSTER
FORMATION FOR THE SAME EXTENT OF
ENACT VASION FOR THE HA FOR THE
SPHERICAL PARTICLES.
THIS WAS TESTED AND WE COULD
MAKE PREDICTIONS ON A COMPUTER.
THESE ARE FROM COMPUTER
SIMULATIONS.
YOU CAN SEE THE LARGER PARTICLES
ARE FASTER AT FUSING AND TAKES
MORE ACTIVATION FOR FLOW OF
FUSION AND ALL PARTICLE SIZES
HERE INCLUDING THE PATCHES ARE
HAs FOR GIVEN PARTICLE TYPE AND
THE RANGE OF PARTICLE SIZES
REPRESENT NATURAL VARIATION OF
FLU PARTICLES.
ALL PARTICLE TYPES HAVE A NEAR
PERFECT CHANCE OF INFUSION BY
THE SMALLEST FUSION OF
ACTIVATION OF HA AND GREATER
THAN 90% IN ACTIVATION FOR ANY
EFFECT OF THE FUSION YIELD TO
SUGGEST THAT FILAMENTS MAY
RESIST THE SLOW DOWN OF FUSION
AND REDUCTION OF FUSION
EFFICIENCY BY ANTIBODIES.
I'M PULLING OUT THE
REPRESENTATIVE DATA FOR THE
PATCHES THAT REPRESENT THE
POPULATIONS THAT WERE DERIVED BY
FRACTIONATION AND HERE I ENABLED
DIRECT COMPARISON OF SIMULATION
AND THEORY AND HIGHLIGHTING THE
PARTS OF THE INACTIVATION CURVES
AND THE SINGLE PARTICLE
EXPERIMENTS SIMULATE THE MODEL
AND SHOWED FILAMENTS CAN RESIST
THE SLOW DOWN OF FUSION PRODUCED
BY ANTIBODIES AND THERE'S ACTION
INFUSION PROBABILITY.
SO WE'RE IN THE PROCESS OF
GENERATING QUANTITATIVE MODELS
WITH ANTIBODIES THAT INHIBIT
ATTACHMENT AND INCLUDE OTHER
TYPES OF ACTIVATION SUCH AS THE
OBSERVATION FOR THE ADVANTAGE OF
FILAMENTS NOT LIMITED TO THE
SOLUBLE INHIBITORS AND FLU AND
IF YOU TAKE PROTEINS FOR OTHER
FILAMENT VIRUSES AND YOU SEE
EBOLA ANTIBODIES AND RE SIST TH
ACTIVATION AND IT'S IMPORTANT
FOR ADAPTATION.
THIS LED TO THIS MODEL.
BY ALWAYS ASSEMBLING A MIXTURE
OF PARTICLE SHAPES AND VIRUSES
HAVE READY PARTICLE TYPES WITH
ROUNDS OF INFECTION UNDER
PRESSURE.
BY ALWAYS HAVING A SUB
POPULATION OF FILAMENTS YOU WILL
BE ABLE TO INITIATE AND
REPLICATE IN THE PRESENCE OF
INHIBITERS AND LEAD TO MUTATIONS
WHEN AND WHERE THEY'RE NEEDED.
AND SPHERES MAY DOMINATE MORE
ECONOMICALLY EFFICIENT IN
ASSEMBLY AND ENTRY.
HOW POTENT WOULD A STRATEGY BE
AND WE ENCOUNTER AN INITIAL
INFECTION STARTING WITH A SINGLE
VIRUS PARTICLE.
AND YOU'RE TALKING ABOUT
PROVIDING ADVANTAGE IN
CONDITIONS OF ATTENUATION.
WE WANTED TO ASK IF THE
POPULATION DISTRIBUTION IS A
FIXED FEATURE OF THE VIRAL
GENOME OR EXPECT TO INCREASE THE
PROPORTION OF BENEFICIAL TYPES
AS NEEDED.
BECAUSE YOU CAN IMAGINE IF YOU
REQUIRE GENETIC CHANGE FOR SUCH
A SHIFT IT WOULDN'T BE SUCH AN
EFFICIENT METHOD UNDER PRESSURE.
AND THE FORMER IS ASSUMED BY THE
LITERATURE.
THE DISTRIBUTIONS ARE FIXED.
BUT TO PROPERLY ADDRESS THE
QUESTION WE NEEDED A NOVEL
METHOD TO STUDY SHAPE.
THIS WORK WAS LED BY A TALENTED
POST DOC IN THE LAB AND JOINED A
YEAR AGO AND I'LL SHARE SOME OF
THE WORK HERE.
THE CHALLENGE WAS TO STUDY SHAPE
AND WE USED MICROSCOPY AND IT'S
INHERENTLY LOW THROUGHPUT.
WE CAN MEASURE SAME BY MEASURING
THE SCATTERED INTENSITY OF THE
VIRUS PARTICLE SMALL PARTICLES
WILL SCATTER LIGHT LESS THAN
LARGER PARTICLES.
THIS IS ILLUSTRATED HERE.
WE VALIDATED THIS APPROACH BY
COMPARING PURIFIED SAMPLES
DERIVED FROM ELECTROMICROSCOPY
AND YOU'LL SEE THE NOISE PEAK
INHERENT IN THE BUFFER AND FOR
SPHERICAL PARTICLES A SINGLE
FIRST HERE.
YOU'LL SEE THE SECOND BUMP UP
HERE IN THE FILAMENTOUS
PARTICLES.
TO INCREASE THE SENSITIVITY OF
THE APPROACH WE INCLUDED A
SECOND DIMENSION.
IF YOU USED FLUORESCENT ANTIBODY
COMBINED TO SURFACE OF VIRUS
PARTICLES YOU CAN PULL THE
PARTICLE POPULATION IN THE
FLUORESCENT DIMENSION AND YOU
CAN SEE THE CLOUDS ON THE RIGHT
SEPARATE FROM THE BUFFER NOISE
AND SO GREATLY EXTENDED THE
DYNAMIC RANGE OF WHAT CAN BE
MEASURED AND INCREASING THE
SENSITIVITY TO HUNDRED THOUSANDS
TIMES OF MICROSCOPY AND WE WERE
IN A GREAT POSITION TO START
ASKING QUESTIONS ABOUT PARTICLE
SHAPE IN ATTENUATED INFECTIONS.
AND THE BEAUTY IS WE CAN MEASURE
MANY SAMPLES AUTOMATICALLY.
IT'S A GREAT OPPORTUNITY FOR
ANALYZING SHAPE IN DYNAMIC
SYSTEMS.
AND WE STARTED WITH A VERY BASIC
EXPERT IN VIROLOGY THE TIME
COURSE OF INFECTION.
AND NOW THERE WAS A NEW LENS TO
INTERROGATE THE DATA WITH AND
THIS IS THE ABILITY TO COUNT
PARTICLES AND MEASURE SHAPE.
AND TO THE INCREASED SENSITIVITY
HE WAS ABLE TO DO IT IN TWO
EXTREMES.
HE INFECTED CELLS AT HIGH
MULTIPLICITY AND LESS THAN 1% OF
CELLS WERE INFECTED AND THE
INFECTED CELLS RECEIVED AT MOST
THE SINGLE INFECTIOUS UNIT.
SO THE YIELD IN THE LATTER CASE
ESPECIALLY WAS INHIBITED VIRUS
PREP AND ONLY LOOKED AT THE
FIRST ROUND AND NOT NEWLY
INFECTED CELLS.
WHAT YOU OBSERVE YOU GET MORE
PARTICLES PRODUCED AND EVEN THE
PARTICLES ARE MEASURABLE AND
COUNTABLE AND WE COULD DERIVE
PARTICLE SHAPE AND THE FIRST
SURPRISE IS SHAPE NOT GOING TO
BE FIXED SO THE VERY SAME VIRUS
HOW MUCH WENT INTO CELLS IN THE
FIRST PLACE PRODUCED PARTICLES
OF DIFFERENT SHAPES AND LESS
VIRUS INPUT PRODUCED MORE
FILAMENT AND PRODUCED MORE
SPHERES.
THE SHAPE IS NOT CONSTANT
THROUGHOUT INFECTION.
THERE'S MORE FILAMENTS INITIALLY
PRODUCED AND MORE SPHERES AND
LATER YOU GET MORE FILAMENTS
AGAIN.
SO SHAPE IS NOT UNIQUELY
SPECIFIED BY THE GENOME AND
SIMPLY CHANGING HOW MUCH YOU PUT
INTO A CELL CAN SHAPE OUTCOMES.
INFECTIONS OF DIFFERENT CELL
TYPES SHOW THERE'S ALSO
CELL-BASED DIFFERENCES THAT CAN
INFLUENCE SHAPE.
BUT WHAT ABOUT THE ANTIBODIES I
TOLD YOU FILAMENTS CAN RESIST
ANTIBODY PRESSURE?
CAN THE PRESENCE OF ANTIBODIES
OR VIRUS REPLICATION BY
ANTIBODIES INFLUENCE THE
RESULTING PARTICLE SHAPE?
THIS WOULD BE IMPORTANT TO HELP
VIRUSES INFECT NEURONS OF CELLS
AND THIS IS SPECIFIC FOR TWO
VIRUS STRAINS AND EXPERIMENTS IN
TWO CELL TYPES.
I'LL ILLUSTRATE RESULT FOR A
SINGLE NEUTRALIZING AND
NON-NEUTRALIZING ANTIBODY AND
THIS IS BEFORE THE VIRUS BOUND
TO CELLS DURING ENTRY AND YOU
SEE REDUCTION IN VIRUS YIELD AND
STRIKINGLY INCREASE PROPORTION
OF FILAMENTS.
IT SEEMS VIRUS CAN RESPOND TO
ANTIBODY PRESSURE BY PRODUCING
MORE FILAMENTS.
NEXT WE ASKED WHERE BASICALLY
HOW DOES THE NEUTRALIZATION OR
WHEN THE ANTIBODY WAS PRESENT
RELATES TO THIS SHAPE OUTCOME.
WE HAD VARIATION OF THE
EXPERIMENT WHERE THE ANTIBODY
WAS PRESENT BEFORE ENTRY AND
DURING THE FIRST FOUR HOURS
ENTRY OR AFTER THEY WERE
COMPLETED.
I SEE MOST ANTIBODIES TO AN
EXTENT INHIBITED ASSEMBLY.
THE EFFECT OF ANTIBODY PRESENCE
WAS MORE POTENT ON SHAPE DESPITE
THE INHIBITION OF VIRUS
PRODUCTION.
YOU STILL SEE INCREASED
FILAMENTS WHEN THE ANTIBODY WAS
PRESENT AND BINDING TO AN
ANTIBODY WITHOUT INFECTING VIRUS
PRODUCTION CAN TRIGGER INFECTION
TO START PRODUCING MORE
FILAMENTS.
NEUTRALIZATION WAS REQUIRED AND
YOU HAD TO INHIBIT THE
NEUTRALIZING.
AND THERE MAY BE TWO MECHANISMS
BY WHICH A VIRUS CAN RESPOND TO
SHAPE BUT INFECTED CELLS CAN
SENSE THE PRESENCE OF
ATTENUATING PRESSURE AND LEADS
TO THE RESPONSE THAT WOULD BE
BENEFICIAL IN THE NEXT CYCLE.
THE RESPONSES TO ANTIBODIES ARE
FAST.
AND YOU START TO SEE INCREASED
IN FILAMENTS WITH NO EFFECT ON
THESE AND THIS CONVINCED US THE
INFECTION IS SENSING THE
PRESENCE OF ANTIBODIES AND DO
NOT REQUIRE GENETIC CHANGE TO
START CHANGING THE ASSEMBLY
CHARACTERISTICS.
AND WE'VE SHOWN TRANSIENT
CHANGES IN SURFACE TENSION CAN
TRIGGER SHAPE CHANGES.
AND THERE'S BEEN A SERIES OF C
ELEGANS AND IT'S SHAPED TO
EXTERNAL CONDITIONS IF IT'S NOT
A FIXED FEATURE OF THE VIRAL
GENOME SO VIRUSES ARE MORE
POWERFUL OPPONENTS THAN
PREVIOUSLY ANTICIPATED.
SIMPLY CHANGING THE VIRUS AMOUNT
AND EXTENT OF REPLICATION AND
CHANGES OF REPLICATION INHI
INHIBITORS AND CHANGES AND SO
THIS LED TO COMBINING TO THIS
MODEL AND INCREASE PROPORTION OF
FILAMENTS BOUND BY ANTIBODIES
CAN STILL MEDIATE ENTRY AND
ALLOW SOME LEVEL OF REPLICATION
EVEN IF SOME COULD BE BENEFICIAL
AND THE VIRUSES CAN ACQUIRE TRUE
RESISTANCE AND INFECT NEURONS OF
CELLS IN OUR SPHERES CAN
DOMINATE AGAIN AND MORE QUICKLY
SPREAD THROUGH POPULATION.
I MENTIONED KEY PLAYERS IN THE
WORK AND IT'S GROUP EFFORT AND
STUDENTS WHO CONTRIBUTED TO THE
WORK ARE STEVEN AND NICK AND
SPECIAL THANKS TO COLLABORATORS
WHO PROVIDED WORK AND I DIDN'T
INCLUDE SOME WORK ON
QUANTITATING VIRUS ATTACHMENT
AND BOSTON COLLEGE.
A LOT OF THE FUND IS EXTRAMURAL
FUNDING LISTED HERE FROM MY TIME
AT BRANDEIS BUT NOW HERE AT THE
DIR AND COLLABORATE IN THIS
ENVIRONMENT.
>> TAKE A PERSPECTIVE ON VIRUSES
WITH REGARD TO SHAPE.
MY QUESTION RELATES TO IN VIVO
VALIDATION.
AND CAN YOU VALIDATE OR HAVE YOU
TRIED TO VALIDATE IT IN A MOUSE
MODEL OF SUB LETHAL INFECTION?
>> GREAT QUESTION.
WE'RE STARTING THAT WORK AND
THIS IS ONE OF THE
COLLABORATIONS WE STARTED AND WE
HAVE SOME MOUSE SAMPLES.
IT WILL WORK IT'S JUST NOT A
TRIVIAL TRANSITION BECAUSE WE
WANT TO BASICALLY HAVE A WAY OF
STUDYING ANALYZING VIRUS SHAPE
WITHOUT MUCH MANIPULATION AND
YOU WANT THE DIRTIEST ANIMAL
SAMPLES AND HAVE A WAY OF THE
SHAPE OUT AND WE'RE MAKING
PROGRESS BUT THE ASSAY REQUIRES
FINE TUNING BUT WE'LL THERE BE.
>> AND FOLLOW ON TO THAT IS IF
THE ASSAY CAN BE FINE TUNED IT
WOULD BE FINE TO COLLABORATE TO
SEE IF THERE'S ANATOMICAL IMPACT
OF EVEN WITHIN THE UPPER
RESPIRATORY TRACT WITH VIRUS
SHAPE AND REAPPLY THAT.
>> THANK YOU.
THAT'S WHERE WE'RE HEADING WITH
THE ANIMAL SAMPLES AND WE DON'T
NEED A LOT AND THAT'S THE POWER
OF THIS METHOD.
I THINK IT'S REALISTIC TO GET
THERE.
I'M HOPEFUL.
DOES IT DIFFER BETWEEN THE TWO
FORMS?
YOU COULD IMAGINE IT WOULD
INFLUENCE THE ABILITY TO INDUCE
NEUTRALIZING ANTIBODIES AT
VARIOUS DOSES.
>> THE DENSITY IS VERY HIGH.
>> INTERESTING TOPIC.
IT'S FASCINATING THE FACT THAT
IN SPHERICAL AND FILAMENTOUS YOU
GET THE SAME COPY OF GENOME BUT
IS IT POSSIBLE TO LOOK THE OTHER
WAY WIN VIRIONS AND COULD THAT
IMPACT THE SHAPE OF THE VIRUS?
>> I SEE.
AND THIS WAS A CONVENIENT SYSTEM
TO ASK THE QUESTIONS OF THE
ADVANTAGE OF EVERYTHING ELSE.
OTHERWISE LIKE RESPIRATORY
SYNCYTIAL VIRUS WOULD HAVE
MULTIPLE COPIES AND WOULD HAVE
THEN ADDITION TO THE ADVANTAGES
THE ADVANTAGE OF DELIVERING
MULTIPLES FROM A SINGLE
INFECTION EVENT.
AND LOOK EIGHT AT IT IN THE
CONTEXT OF NO GENOME.
WOULD IT AFFECT THE SHAPE?
>> THE NO GENOME PARTICLES WOULD
NOT YIELD AN INFECTION.
THAT WOULD NOT BE ADVANTAGEOUS.
FLU IS KNOWN TO GENERATE EMPTY
PARTICLES TOO.
WE DON'T THINK SO BASICALLY IN
OUR EXPERIMENTS WE DON'T SEE
EVIDENCE OF EMPTY PARTICLES
BEING MORE PROMINENT IN EITHER
PARTICLE FORM.
SOO UNDER THE CONDITIONS WHERE
YOU GET MORE SPHERES OR
FILAMENTS THE SAME NUMBER OF
PARTICLES YIELDS THE SAME OUTPUT
AGAIN.
SO IT SEEMS THE NUMBERS DON'T
CHANGE BETWEEN THE TWO PARTICLE
FORMS.
GOOD I'M WONDERING IF THE GENOME
HAS INVOLVEMENT IN THE SHAPE OF
VIRIONS.
>> YEAH.
I DON'T KNOW.
YEAH.
>> THAT WOULD BE REALLY
INTERESTING.
THANK YOU.
>> THANKS.
>> THANK YOU FOR THE
PRESENTATION.
I'M WONDERING IF THE
DISTRIBUTION AND SHAPE IS BINARY
OR ONLY SEE LONGER FILAMENTS OR
IF YOU'VE OBSERVED PARTICLES IN
BETWEEN.
>> IT'S NOT BINARY.
THERE'S A WHOLE DISTRIBUTION OF
SHAPES I THINK I SHOWED EARLIER
IN MICROGRAPHS.
YOU HAVE EVERYTHING TWICE THE
LENGTH OF THE SPHERE TO ALMOST A
THOUSAND TIMES THE LENGTH OF A
SPHERE AND ANYTHING IN BETWEEN
AND SO I THINK WITH MORE
MATERIAL WITH THE LARGER PART OF
MATERIAL YOU HAVE MORE
OPPORTUNITIES OF ENTRY UNDER
EXTENSIVE PRESSURE.
THE DISTRIBUTION OF PARTICLE AND
THE WAY WE PRESSURE THIS WE'RE
NOT MEASURING THE SIZE PRECISELY
BUT YOU DO SEE TWO DISTINCT
PEAKS.
THERE'S A DISTRIBUTION AROUND
EACH.
SO THERE'S SOMETHING BINARY
ABOUT IT.
EITHER THE FIRST OR SECOND
POPULATION BUT WITHIN THE
POPULATIONS THERE'S A HUGE
VARIABILITY.
>> THANK YOU.
>> I HAD A QUESTION.
IN WERE ONE SLIDE YOU HAD A
VIRION WITH A GREEN SURFACE AS
THE CONTACT SURFACE, DOES THAT
CHANGE BETWEEN SINGLE PARTICLES
OR THE ELONGATED PARTICLE?
ARE YOU JUST ENGAGING A LARGER
SURFACE?
>> THAT'S THE ASSUMPTION IN THE
SIMULATION MODEL THAT WOULD BE
THE CONTACT AREA OF THE SMALL
SPHERICAL PARTICLE AND THE
CONTACT AREA WOULD INCREASE IN
LENGTH BECAUSE THE FILAMENTS
HAVE THE SAME DIAMETER OF THE
SPHERE JUST LONGER.
YOU WOULD HAVE MORE --
>> YOU WOULD BE FUSING THROUGH
THE LENGTH OF THE FILAMENT NOT
THE TIP?
>> THAT'S THE ASSUMPTION BUT IT
SEEMS THE DATA QUANTITATIVELY
WITH THE ASSUMPTION.
>> OKAY.
ALSO YOU WOULD EXPECT -- BECAUSE
YOU HAVE THE IF SURFACE AND SO I
CAN'T IMAGINE IT'S A FLIMSY
PARTICLE AND YOU'D HAVE TO
ENGAGE THE LENGTH.
>> LAST QUESTION.
ARE THERE OTHER ANTIBODIES
SPECIFICALLY RECOGNIZE ONE FORM
OR THE OTHER FORM AND HAVE THOSE
BEEN --
>> IT'S THE EXACT SAME HA.
THE SAME GLYCOPROTEINS
DISPLAYED.
>> SORRY.
>> THAT'S THE CHALLENGE THE SAME
HAs AND DENSITY ON THE SURFACE
AND MOST ANTIBODIES AS WE KNOW
THEN BIND TO -- SO THEY WOULD
NOT BE DIFFERENT.
>> BECAUSE NEUTRALIZING BY
DEFINITION MAY PROBABLY WORK BUT
NOTE NEUTRALIZING MAY BE HIDDEN
IN ONE FORM AND NOT ANOTHER OR
VICE VERSA.
>> IT'S THE SAME HAs.
>> I'M JUST TALKING ABOUT
EXPOSURE OF THE EPITOPE?
>> RIGHT.
>> IN THE FILAMENT YOU HAVE THE
ENDS WHICH IS A PORTION OF THE
SPHERE.
SO IF THERE'S ANY DIFFERENCE
YOUR DISPLAYING THE FILAMENT AND
HOW DO YOU TARGET ONE VERSUS THE
OTHER?
WE'RE THINKING OF METHODS THAT
ARE SO A STUDENT IN THE LAB IS
WORKING ON BUILDING THE ORGANIC
CONSTRUCT THAT CAN BE CODED WITH
VIRUS PARTICLES AND ASSEMBLED
INTO SHELLS TO AN INDIVIDUAL
COMPONENT CAN HAVE ANTIBODIES
AGAINST HA AND THEN STIMULATE
THE CELL AND HAVE COOPERATIE IV
ENCLOSURE AND SHAPE AND THIS IS
ONE DIRECTION WE'RE THINKING IN
THOSE TERMS.
HOW DO YOU TARGET NOT ONE VERSUS
THE OTHER BUT HOW DO YOU ENSURE
YOU TARGET ALL THE SHAPES?
RIGHT.
>> I'M WONDERING WHETHER YOU
THINK MORPHOGENESIS HAS AN
INFLUENCE IN THE SHAPE AND HOW
MUCH MATRIX THERE IS OPPOSED TO
OTHER VIRUSES AND THE CURVATURE
OF THE MEMBRANE AND IF YOU THINK
OF THE ABUNDANCE HAS AN
INFLUENCE AND WHETHER YOU CAN
PROBE THAT.
>> WE TRIED TO ASK THAT
PARTICULAR QUESTION AND WE WENT
INTO THE DIRECTION OF THESE
EFFECTS AND ANY PROTEIN
EXPRESSION WILL TAKE TIME AND I
DIDN'T SHOW THE BACK UP SLIDE
BUT YOU CAN TAKE THE INFECTION
AND CHANGE IT INTO HYPERTONIC
AND MEASURE SHAPE OR WHAT'S
PRODUCED AFTER 30 MINUTES TO
INCREASE OR DECREASE TENSION.
AND WITHIN 30 MINUTES HAVE YOU
SPHERES VERSUS FILMIMENTSS --
FILAMENTS AND PROTEINS AND WE
DON'T HAVE ANY EVIDENCE OF THE
ADDITIONAL COMPONENT.
THE A LITTLE BIT WE LOOKED WE
DIDN'T FIND ANYTHING BUT SHOULD
LOOK INTO THAT MORE CAREFULLY.
THANK YOU.
>> CAN YOU FOLLOW UP DURING THE
RECEPTION?
THANK YOU.
WE NEED TO WRAP UP.
WELL, THAT CONCLUDES OUR
MEETING.
FIRST I'D LIKE TO THANK LENNOR
COMING DOWN HERE.
WITHOUT YOU THIS IS IMPOSSIBLE.
ALSO SO TO THE SPEAKERS.
THANK YOU FOR ACCEPTING THE
INVITATION.
THANK YOU SO MUCH.
OF COURSE THE NIH FOR ALL YOUR
SUPPORT AND THEY'RE BEHIND THE
SCENES.
WITHOUT THEM NONE OF THIS WOULD
WORK SO THANK YOU SO MUCH ALL
THE TEAM AND ESPECIALLY ASHLEY
AND JENNELLE.
THANK YOU SO MUCH.
IT'S BEEN A FANTASTIC MEETING
AND LENNOR DECIDED TO HAVE THE
SPECIAL 25 ANNIVERSARY A
RECEPTION FOLLOWING IN THE
FOYER.
PLEASE STICK AROUND OUTSIDE THE
LIPSETT AND WE CAN CONTINUE WITH
SCIENTIFIC CONVERSATIONS OR NOT.
THANK YOU SO MUCH.
SEE YOU NEXT YEAR.