WELCOME TO EVERYONE.
AND THANK YOU FOR JOINING THE
ACHIEVEMENTS OF MARJORIE
ROBERT-GUROFF WHOM WE'RE HERE TO
HONOR TODAY.
I FIRST MET MARJORIE NOT AS A
FELLOW SCIENTIST BUT AS A FELLOW
PARENT WHEN MARJORIE'S DAUGHTER
SARAH AND MY SON ALEX WERE
CLASSMATES AT WINGATE ELEMENTARY
SCHOOL.
THAT WAS LONG BEFORE WE KNEW
EACH OTHER IN SCIENTIFIC REALMS.
I'VE WORKED WITH MARJORIE NOW
FOR NEARLY 18 YEARS SINCE THE
FORMATION OF THE VACCINE BRANCH
AND MARJORIE'S BEEN A GREAT
CONTRIBUTOR OVER THAT TIME.
YOU'LL HEAR FROM OTHER SPEAKERS
ABOUT HER ACCOMPLISHMENTS OVER
THE YEARS.
ONE MAJOR THEME SINCE I'VE KNOWN
MARJORIE HAS BOTH THE PIONEERING
USE OF REPLICATION ADENOVIRUSES
AS VACCINE VECTORS.
YOU'LL HEAR MORE FROM A NUMBER
OF SPEAKERS.
THEN IN RECENT YEARS I WANT TO
MENTION TWO I THINK VERY
IMPORTANT DISCOVERIES SORT OF
AHEAD OF THE FIELD MARJORIE'S
MADE.
ONE IS THE IMPORTANCE OF
NON-NEUTRALIZING ANTIBODIES IN
PROTECTION AGAINST VIRUS
INFECTIONS IN PARTICULAR SIV AND
HIV MARJORIE MADE IN HER MACAQUE
STUDIES, IF MY MEMORY IS CORRECT
PRECEDE THE OBSERVATION OF SUCH
THINGS IN HUMAN TRIALS IN THE
PHASE 3 TRIAL.
THAT WAS PRESCIENT AND THE
SECOND WAS THE ROLE OF GENDER OR
SEX DIFFERENCES IN IMMUNE
RESPONSE TO VACCINES.
AND WE'VE BEEN SEEING MORE AND
MORE EXAMPLES OF THAT.
IT'S TRUE WITH COVID AND WITH A
LOT OF THINGS AS WELL.
YOU'LL HEAR MORE FROM MANY
SPEAKERS WHO WORKED WITH
MARJORIE THOUGH WE'VE BEEN
TOGETHER IN THE VACCINE BRANCH
I'VE NEVER HAD THE PLEASURE OF
BEING IN HER LAB.
I'LL LEAVE IT TO THE OTHER
SPEAKERS WHO WORKED MORE
CLOSELY.
I WANT TO INTRODUCE OUR KEY NOTE
SPEAKER BOB GALLO WHO I THINK
NEEDS NO INTRODUCTION TO THE
AUDIENCE BUT NEVERTHELESS I'LL
GIVE A BRIEF INTRODUCTION.
HE'S THE FOUNDER AND DIRECTOR OF
THE INSTITUTE OF HUMAN VIROLOGY
AT THE UNIVERSITY OF MARYLAND
BALTIMORE BUT ALSO ALUMNUS OF
THE NATIONAL CANCER INSTITUTE.
WHERE A LOT OF DISCOVERIES WERE
MADE AND ALSO THE FOUNDER OF THE
GLOBAL VACCINE NETWORK AND I
WANT TO MENTION AT LEAST THREE
THINGS, ANY OF WHICH WOULD BE A
LIFE TIME ACHIEVEMENT ONE OF THE
DISCOVERY OF INTERLEUKIN 2 THAT
LOU THE GROWTH OF T LYMPHOCYTES
THAT'S OPENED UP THE WORLD OF
THE IMMUNOLOGY AND THE DISCOVERY
OF THE HTLV-1 WE'LL TALK ABOUT
TODAY AND ADULT T CELL LEUKEMIA
AND THE OTHER IS THE DISCOVERY
OF THE AIDS VIRUS NOW KNOWN AS
HIV AND WE'LL BE HEARING MORE
ABOUT THAT AS WELL.
SO IT'S INCREDIBLE FOR ANYONE TO
MAKE SO MANY LANDMARK
DISCOVERIES IN ONE LIFE TIME AND
IT'S A GREAT PRIVILEGE TO HAVE
BOB HERE AND HE'LL TELL US ABOUT
MARJORIE'S CONTRIBUTIONS IN THE
EARLY DAYS.
BOB, THANK YOU.
>> I WANT TO SAY THEY SAW ME
LIMPING AND I SAID I HAD KNEE
SURGERY.
THIS IS A MARJORIE TALK SO THERE
WERE CLINICAL ASSOCIATES AND IN
1971 PEOPLE THINK WE HAD A LOT
OF HELP I CAN TELL YOU IT DIDN'T
START THAT WAY.
I HAD ZERO -- I CAME IN '65 AND
PERSONALLY WAS SUPPOSED TO WORK
WITH WENDELL ROSS AND WENT TO
WORK AND IN NIH'S WISDOM THEY
SENT ME TO THE CHILDHOOD
LEUKEMIA WARD AND I WENT TO THE
LAB AND WAS ALONE FOR YEARS AND
BE THE WE CONVERTED AMY GRIMES
AN EDUCATOR AT THE FOURTH GRADE
AND BECAME MY TECHNICIAN.
THERE WERE MANY LONELY YEARS BUT
THIS IS THE BEGINNING OF HAVING
SOME HELP.
AND MY MAIN INTEREST WAS
LEUKEMIA AND IN ANIMAL THERE'S A
COMMON CAUSE AND THERE WERE RNA
TUMOR VIRUSES SO I THOUGHT AND
THEN THE PROVIRUS THEORY WAS
THOUGHT TO BE CORRECT AND THERE
WAS A DISCOVERY BY KEVIN IN
BALTIMORE FOR REVERSE TRANSCRIPT
TRANSCRIPTASE I WAS COMPARING
THEM WORKING ON PHA STIMULATED
LYMPH SITES.
WE DIDN'T KNOW -- LYMPHOCYTES
AND THEY WERE BEING DESCRIBED IN
THE BEGINNING AND THEY DESCRIBED
THIS PLANT PROTEIN COULD MAKE
LYMPHOCYTES.
WE DIDN'T KNOW WHICH ONES BECAME
CELLS AND THEY LOOKED LIKE ACUTE
LYMPHOCYTIC CELLS.
ONE OF THOSE WAS DNA POLYMERASE
AND WITH BALTIMORE AND OTHERS WE
PUBLISHED IN SCIENCE WE KNEW
THREE AND CALLED THEM ALPHA,
BETA, GAMMA.
MARJORIE WAS ALREADY TRAINED IN
BIOCHEMISTRY.
SO MARJORIE HAD HISTORY AND THE
STAGE WAS SET AND CURTAIN BOWS
WERE RATHER FAR AWAY.
THE TIMING WAS BAD FOR ALL OF
US.
THIS IS NOW THE 1970.
AND WE MOVE FROM THE '60s TO THE
'70s.
AND THERE WAS A HEALTHY RESPECT
FOR PANDEMICS BUT IN THE '70s IT
WAS THOUGHT TO BE A WASTE AND
THE CANCER CENTER WAS SHUT DOWN
AND THERE WAS A MEETING AND
THERE'S NO EVIDENCE FOR ANY
INFECTIOUS INVOLVEMENT OF HUMAN
CANCER IT WAS ALL CHEMICALS AT
THE TIME THOUGHT VERY WRONGLY
AND THE DEPARTMENTS AND SOME
MEDICAL SCHOOLS WERE TERMINATED.
HARD TO BELIEVE.
THIS IS AROUND THE MID 1970s.
RETROVIRUSES DON'T INFECT HUMANS
AND THEY THOUGHT YOU DON'T NEED
TO FUND IT.
THE ARGUMENT WELL OF WAS ALL
MONEY SHOULD GO TO DEGENERATIVE
DISEASES AND MOLECULAR BIOLOGY
AND GENETICS AND SOME OF THOSE
DISEASES WERE INVOLVING VIRUSES.
SO I SAID THE LAST ONE NO
VIRUSES WERE INVOLVED IN HUMAN
CANCER.
THESE BIASES WERE FINISHED BY
THE BEGINNING OF THE 1980s AND
WERE SHOWN TO BE INVOLVED IN THE
CAUSE OF ABOUT 20% HUMAN CANCERS
AND THERE DOESN'T SEEM TO BE
ANYTHING ELSE REQUIRED SO THE
RETROVIRUSES WERE FOUND IN
HUMANS AND FOUND FATAL
NEUROLOGIC DISEASE AND EVEN SOME
AUTOIMMUNITY.
AND WE LOOKED AT HOW THE
DEVELOPMENTS OCCURRED.
SO OUR APPROACH WAS METHODS
USING GROWTH FACTORS.
I FELT WE HAD TO HAVE PRIMARY
CELLS AND WE NEEDED GROWTH
FACTORS.
THEY WERE JUST DISCOVERED.
THEY WERE DISCOVERED BY
VANDERBILT WITH AN EPITHELIAL
GROWTH FACTOR.
THEY WERE NOT LIKE CYTOKINES.
IT WAS AUTOCRINE AND THE CONCEPT
HAD NOT YET BEEN DEVELOPED AND
WE THOUGHT WE NEEDED SPECIFIC
ASSAYS FOR FINDING A RETROVIRUS.
WE KNOW THERE'S A MECHANISM FOR
THE PROVIRUS AND A TOOL THAT
IMPACTED MOLECULAR BIOLOGY.
IT WAS 1,000 TIMES MORE
SENSITIVE THAN MICROSCOPY AND IT
BECAME A SURROGATE MARK AND
ALLOWS FOR FREQUENT SAMPLING
IMPORTANT FOR VIRUSES INVOLVED
IN PERIODIC BURST.
I CAN'T EMPHASIZE THE FIRST A IS
OFFICE BUT B MAY BE MORE
IMPORTANT.
HTLV-1 I CAN'T SAY ABOUT HTLV-2
SO MUCH BUT IN HIV TEND TO COME
OUT IN PERIODIC BURSTS
ESPECIALLY WITH HTLV-1.
NO WAY YOU'LL FIND THE VIRUS BY
ELECTRON MICROSCOPY EASILY
BECAUSE YOU CAN'T SAMPLE EVERY
15 MINUTES OR STIMULATE A
CULTURE REGULARLY ALL THE TIME
BUT YOU CAN WITH THE REVERSE
TRANSCRIPTASE ASSAY.
IT'S NOT A HISTORY THAT'S REALLY
BEEN DESCRIBED.
THERE'S NO PLACE WE EVER KIND OF
GATHERED THIS TOGETHER AND SAID
THIS IS HOW IT HAPPENED.
WE TALK ABOUT IT BUT IT'S NOT
REALLY WRITTEN.
THERE WERE STRONG VIEWS AGAINST
THE POSSIBILITY OF HUMAN ANTI
ANTIVIEANT
ANTIVIRUSES AND ANIMAL MODELS
WERE SELECTED FOR WHICH ARE
OBVIOUS AND THE SAME WITH MOUSE
LEUKEMIA VIRUS.
THE ONES REALLY KNOWN AT THE
TIME.
THERE WAS LITTLE EVIDENCE
CAUSING AIDS UNTIL THE '70s
SHOWN BY A GROUP AT DAVIS
CALIFORNIA AND THEY ONLY TESTED
SMALL ANIMAL RETROVIRUSES.
AND HUMANS WERE PROTECTED AND
LASTLY AND WHAT YOU WON'T
APPRECIATE IN TODAY'S WORLD IF
YOU'RE YOUNG, HUMAN RESEARCH WAS
NOT APPRECIATED.
CANCER IS CATCHING LIKE AN
INFECTION SEEMED TO BE A
PRIMITIVE VIRUS AND SLOW VIRUS
NOT REALLY THOUGHT ABOUT AND
THOUGHT HUMAN RESEARCH WAS, IF
YOU PARDON ME THE PITS.
THE LAST SOPHISTICATED.
IT WAS A POORMAN'S WAY OF DOING
RESEARCH.
A FEW MAJOR KEY PAPERS IN THAT
PERIOD FROM OUR GROUP.
MARJORIE ET AL AND JACK GABRIEL
WITH MARJORIE AND OTHERS AND
POLYMERASES AND POLYMERASES TO
DEPOLYMERASE GAMMA AND OUR
FRIEND, NEOPLASTIC CELLS AFTER
THOROUGH REVIEW.
MARJORIE ET AL.
GAMMA IS CONFUSED WITH
TRANSCRIPTASE AND LYMPHO BLASTS.
AND THERE WERE TALKS ON HUMAN
BLOOD CELLS.
SHE DIDN'T WORK IN BLOOD CELL
STORIES.
WE CULTURED HUMAN MYELOID CELLS
AND WE HAD AN EMBRYONIC GROWTH
FACTOR AND ONE BECAME THE FIRST
MYELOID CELL LINE HH60 WHICH
BECAME THE THERAPY FOR
PROMILOCYTIC LEUKEMIA WON A
PRIZE FOR CLINICAL USE OF RETINO
RETINOACID AND SOME CALLED IT
THE FIRST RECOGNIZED CYTOKINE.
WE CULTURED HUMAN T CELLS IN
1967 THEN THIS POINTED US
TOWARDS T CELLS BY NECESSITY.
WE HAD THE ABILITY TO GROW HUMAN
T CELLS AND COULD SHOW T CELLS
COULD BE GROWN BEYOND A SINGLE
MITOSIS.
IN 1978 WE ISOLATED A STRAIN OF
LEUKEMIA VIRUS AND CAUSED
CHRONIC MYELOID LEUKEMIA AND
FOUND WHAT HAD T CELL LEUKEMIA
AND THESE TWO THINGS PUSHED US
TOWARDS T CELL MALIGNANCIES.
THIS IS THE FIRST COLLECTED
ELECTRON MICROGRAPH THE THIRST
HUMAN RETROVIRUS AND THAT'S THE
LEUKEMIC PAERPZ
PAPERS WE ISOLATED BIOPSIES OF
THE LESIONS THAT WERE
INFILTRATED WITH MAG IG NANT
CELLS AND AN AGGRESSIVE DISEASE
AND RECOGNIZED IN JAPAN.
IN JAPAN THEY KNEW THE DISEASE
BUT DIDN'T FOE THE VIRUS FIRST.
WE FOUND IT IN SPORADIC CASES.
ONE WONDERS IF WE WERE LIVING IN
JAPAN IF IT WOULD HAVE OCCURRED
EARLIER BECAUSE IT WAS DESCRIBED
AS AN EPIDEMIC IN SOUTHWEST
JAPAN AND THROUGHOUT BUT LESION
IS CHARACTERIZED BY THE
INFILTRATED LEUKEMIC CELLS AND
ABOUT 20% IS CALLED SMOLDERING
OR CHRONIC THOSE ARE MORE MILDER
DISEASES BUT WERE RAPIDLY FATAL
WHICH HAPPENED IN THIS YOUNG MAN
WITH NO PAST HISTORY, PERSONAL,
SEXUAL, NOTHING.
WE FOUND ANTIBODIES AND
DEVELOPED ASSAYS BY MARJORIE, I
CAN'T RECALL AND SOME FAMILY
MEMBERS.
WIN TIME IT BECAME CLEAR IT WAS
NOT TRANSMITTED BY VIRUS BUT
INFECTED CELLS.
THE PROVIRUS IS WHAT IS
INFECTIOUS WHICH WAS AMAZING AND
THAT WAS DIFFERENT IN RETRO
VIROLOGY AND ENCODED IN GENES
FOR THESE REGULATORY PROTEINS.
THIS WOULD PREDICT ALL THE
THINGS THAT HAPPENED WITH HIV,
CD4 TROPISM AND SMALLER CORE
PROTEIN P34 INSTEAD OF P30.
THOUGH THE GENES ARE NOT SIMILAR
BY SEQUENCE IS ALMOST LIKE A
CONVERGENT BETWEEN THE TWO
FAMILIES OF HUMAN RETROVIRUSES.
AND LOOK AT NUMBER 3, IT'S IT'S
LIKELY THE MOST ONCOGENIC KNOWN
AGENT AND IT'S THE ONLY LEUKEMIA
VIRUS THE FIRST RETROVIRUS AND
LINKED AND IF YOU USE THE
PROVIRUS OR CERTAIN GENES FROM
IT YOU CAN CREATE ADULT T CELL
LEUKEMIA IN MOUSE MODELS.
IT'S AN IMMORTALIZING
TRANSFORMING VIRUS AND LINKED
EPIDEMIOLOGICALLY TO ALL THE IN
VITRO RESULTS YOU HAVE ARGUE
FORCEFULLY IT'S A DIRECT CAUSE
OF THIS CANCER.
AND WE PUBLISHED THIS PAPER
TOGETHER AND WE CAME TO THE
CONCLUSION THAT HTLV-1 IS THE
MOST ONCOGENIC AND IT'S
PARTICULARLY TRUE IF YOU LOOK AT
THE PEOPLE THAT WERE INFECTED AT
BIRTH.
IT WAS REPORTED AROUND 20% TO
25% OF CHILDREN BORN WITH
HTLV-1, NOT BORN WITH IT BUT
INFECTED SOON AFTER IN INFANCY
WILL DEVELOP T CELL LEUKEMIA.
THAT'S A VERY HIGH RATE.
FORTUNATELY, IT'S SO POORLY
INFECTIOUS SO IT BECOMES PUBLIC
HEALTH WISE NOT THAT IMPORTANT
THAT WHO VIRTUALLY FORGOT ABOUT
IT UNTIL THERE WAS PROTEST IN
AUSTRALIA WHERE HALF THE
ABORIGINAL NATIVE PEOPLE
DEVELOPED A SEVERE DISEASE FROM
HTLV AND THEY'RE FOLLOWING IT
MUCH CLOSER AND I HOPE THERE'LL
BE SOMETHING SAID ABOUT IT.
MARJORIE'S PLAYS A KEY ROLE IN
THE ROLE OF HTLV TO LEUKEMIA.
I DON'T KNOW IF I NEED TO SAY
THEM ALL.
IF YOU LOOK YOU'LL SEE
IMMUNOLOGICAL LINKAGE.
AND ESTABLISHING THE LINK WAS
HER MORE THAN ANYBODY ON EARTH.
AND JAPAN PICKED IT UP STRONGLY
TOO BECAUSE THEY HAD MANY MORE
PATIENTS AND THIS IS THE
EARLIEST AND STRONGEST OF THIGHS
KINDS OF PAPERS.
SO IN MANY MANY MORE PAPERS
LINKING HTLV-1 TO VARIOUS
REGIONS.
MARJORIE, I DIDN'T GO THROUGH
EVERYWHERE PAPER YOU PUBLISHED.
THERE'S MORE THAN I'M LISTING IN
THE SLIDE.
THE DISCOVERY AND SOME FEATURES
OF THE HUMAN RETROVIRUS IS
HTLV-2.
THE PAPER IS COMING AROUND IN A
POST-DOC WORKING TOGETHER WITH
US.
IT'S A NEW SUBTYPE WHICH I CAN
SAY I PREDICTED IT WOULD BE A
NEW RETROVIRUS RELATED TO HTLV-1
AND CAME FROM T CELL LEUKEMIA
PUBLISHED IN SCIENCE IN 1982.
A RARE DISEASE LIKELY SOMETIMES
CALLED BY HTLV-2.
IT'S UNIQUE TO THE PATIENT.
IT WAS A YOUNG WHITE MALE BUT
HAD A GIRLFRIEND THAT HE WAS
INFECTED BY A NATIVE ALASKAN
NATIVE AND IT IMMORTALIZED THE
STARTER CELLS LIKE HTLV-1 BUT
STUDIES SHOW IT'S A RARE VIRUS
AND DISEASE AND HELPFUL IN
REASONING ABOUT THE MOLECULAR
PATHOGENESIS OF HTLV-1.
AND IT'S IMPORTANT TO WHAT COMES
NEXT, HIV.
WHY'S IT IMPORTANT FOR HIV?
THERE'S ONE AND TWO AND THERE'LL
BE MORE.
WE CERTAINLY MADE THE FRUITFUL
SUGGESTION THAT IT WOULD BE
HUMAN RESOURCE BECAUSE IT'S
TARGETING THE T CELLS BUT WAS
NOT EXPECTED OUTSIDE THE
EXOGENOUS INFECTIOUS FAMILY
MEMBERS.
NOT TRUE WITH MACAQUE, RAT,
MOUSE, CHICKEN, COW.
MARJORIE PLAYED A KEY ROLE IN
THE PAPER OF HTLV-2 DISCOVERY
AND COULD HAVE BEEN THE FIRST
AUTHOR.
WE SAID, WELL, SINCE IN THE
IMMUNOLOGICAL PAPER MAYBE REMMIN
SHOULD BE THE FIRST AUTHOR AND
IT'S A MORE QUOTED PAPER.
THE FIRST HUMAN RETROVIRUS LED
TO THE DISCOVERY OF HIV.
I WANT TO BE PROVOCATIVE SO FOR
THOSE WHO DON'T KNOW OR BELIEVE
IT, HOW WOULD YOU THINK OF THAT?
WHAT WAS OUR THINKING?
WHAT WE WERE TOLD ABOUT AIDS.
THE CLINICIANS TOLD US IT WAS A
CD4 T CELL DISEASE.
THEY WERE DECLINING.
WHAT DOES HTLV-1 AND HTLV-2 DO?
TARGET CD4 T CELLS AND BLOOD SEX
TO MOTHER AND INFANT SOUNDED
LIKE HTLV-1.
THE LONG LATENCY DISEASE OBVIOUS
FROM THE HISTORY OF AIDS
PATIENTS AND WE KNEW IT WAS TRUE
WITH HUMAN RETROVIRUSES AND
IMPAIRMENTS.
WE KNOW HTLV-1 CAUSED LEUKEMIA
AND IMPAIRMENT.
THEY SHOWED THAT IN PEOPLE WHO
DIDN'T HAVE MALIGNANCY AND
PREVALENCE IN HAITI AND AFRICA.
WHAT WE WERE LEARNING ABOUT
HAITI?
IT HAD A LOT OF AIDS.
WHAT DID WE THINK OF AFRICA?
PEOPLE WERE THINKING THE
EARLIEST CASES THIS WAS AN OLD
VIRUS IN THE RAIN FOREST.
SO THAT SEEMED TO FIT AS WELL.
TECHNICALLY, METHODS FOR GROWTH
AND CULTURE OF HUMAN T CELLS
BASED ON THE 1976 STUDIES
APPLIED TO HTLV-1 AND 2 AND
TESTS FOR RETROVIRUSES BASED ON
REVERSE TRANSCRIPTASE IN THE
'70s AND FELT IT WAS HELPFUL.
IN 1982 THERE WAS MORE FROM THE
FDA AND HUMAN PHILIACS IN WERE
NOT INVOLVED AND THERE WAS
DISCUSSED AS THE MOST LIKELY
THEORY AND YET THINK ABOUT IT
AGAIN.
WHAT ABOUT HEMOPHILICS AND
TRANSFUSED PEOPLE.
THE IDEA WAS YOU GET INFECTION
FROM ROUGH SEX YOU THINK DIDN'T
WOMEN GET EXPOSED TO ROUGH SEX
EARLIER THAN AIDS AND WHAT ABOUT
BABIES AND WHAT ABOUT THIS.
IT WASN'T A GOOD IDEA AND THERE
WAS INFECTIOUS CAUSES AND
ADENOVIRUSES FROM ALBERT
EINSTEIN THERE WAS A UNIQUE TYPE
BUT IT WAS IN GAY MEN AND THERE
ARE IDEAS OF HERPES RELATED
CAUSES AND AN UNNECESSARY
CO-FACTOR AND THEN WE POSTULATED
A NEW T LYMPHOTROPIC RETROVIRUS.
IF YOU KNEW ABOUT THE HISTORY OF
THE FIELD AND YOU KNEW THIS, YOU
CERTAINLY COME TO THAT.
THEN ONE WAS CREATED BY THE
UNITED STATES THE RUSSIAN KGB
THROUGH EASTERN GERMANY.
WE WON'T HAVE TIME TO GO THROUGH
THESE THINGS BUT IT WAS
INTERESTING BEING ATTACKED BY
ALL THESE THINGS THAT YOU EITHER
MADE IT IN THE LAB OR MADE IT UP
AND AIDS PRESENTED UNIQUE
CHALLENGES.
THIS IS A SLIDE SHOWN
REPEATEDLY.
THE CLINICAL LATENCY OF SEVEN
YEARS.
YOU DON'T ASK WHAT THEY DID
SEVEN YEARS AGO BUT WHAT THEY
DID LAST YEAR OR LAST MONTH AND
THE MICROBIAL INFECTIONS THAT A
PERSON WITH AIDS HAD NONE WERE
THE EVIDENT CAUSE AND THAT
CREATED A LOT OF PROBLEMS.
WE HAD HELP FROM CLINICAL
COLLEAGUES.
THEY'RE ALL LISTED HERE AND WELL
KNOWN PEOPLE IN THE FIELD AND
HELP WITH DISCUSSIONS AND HELP
PROVIDING TOO MANY SAMPLES.
WE WERE OVERWHELMED IN THE
LABORATORY AS THOSE WHO WERE
THERE AT THE TIME CAN CERTAINLY
TESTIFY.
WE HAD CULTURES COMING OUT OF
OUR EARS.
NOW, THIS IS AN INTERESTING
SLIDE.
AND WE NEVER PUBLISHED THIS.
THIS HELPED AND SET ME BACK FOR
MONTHS.
I WAS THINKING THINGS AS WE'RE
PREDICTING THINGS ARE COMING
OUT.
WE'RE GETTING MORE IN RETROSPECT
THERE'S HUMAN LESSONS IN THIS
AND WE'RE CONFIDENT WE'RE RIGHT
ABOUT IT SEEMS ALL OF IT.
WHY?
LOOK AT THE SLIDE CAREFULLY AND
YOU SEE IT'S LABELLING HIV AND
HTLV-1 IN THE SAME CELL AND
INDIVIDUAL.
UNFORTUNATELY HERE IN RETRO
VIROLOGY YOU SAY THERE'S VARIETY
INTERFERENCE AND DISMISS THIS AS
THE INTERPRETATION AND PEOPLE
WOULDN'T BE DOUBLY INFECTED AND
IF THEY WERE THERE WOULDN'T BE
IN THE SAME CELL SO THIS WAS A
SAMPLE GIVEN TO US FROM A YOUNG
MAN AND THIS CAME FROM A MAN WHO
WAS ON HONEYMOON IN HAITI AND
HAD AN ACCIDENT AND GOT INFUSED
AND DIED OF AIDS PRETTY RAPIDLY.
THIS IS AN IMMORTALIZED T CELL
BUT HIV IS KILLING THOSE CELLS.
WE SAY IT'S GOT DIFFERENT
MORPHOLOGY AND MOREOVER WITH
MARJORIE'S MONTANA
MONOCLONAL THOUGHT THE CELLS
WERE STILL GROWING AND
EVERYTHING IS GOING THE WAY
PREDICTED AHEAD OF TIME.
I'M THINKING DOWN THE LINE IT'S
GOING TO BE AN ENVELOPE VARIANT
OF HTLV-1 AND 2.
A FEW OF US DISCUSSED IT.
THEN CAME MARJORIE WITH SEROLOGY
ASSAYS AND FORGOT THIS ONCE WHEN
I SAID IT, SHE KNOWS THE DATA
BUT DIDN'T REMEMBER WHAT IT
INFLUENCED IT SO MUCH.
HER SEROLOGY DATA WAS SHOWING WE
DON'T SEE THIS REACTION VERY
OFTEN IT'S RARE.
IN 95%, 96%, 98% WE SEE
PARTICLES ACTING LIKE A VIRUS
EVEN WITHOUT GROWING THEM
SUBSTANTIALLY THEY'RE NOT
REACTING WITH THE HTLV-1
REAGENTS THEY MUST BE DOUBLE
INFECTIONS IS WHAT SHE SAID AND
MAYBE THERE'S MORE DRUG ADDICTS
WE WERE SEEING POSITIVE AND WE
WERE SEEING POSITIVE, NEGATIVE,
NEGATIVE AND POSITIVE-NEGATIVE
AND HOW ARE WE GOING PUBLISH
THIS?
WE DIDN'T KNOW WHAT WAS GOING ON
THEN IT BECAME CLEAR THEY WERE
TWO VIRUSES AND OUR GROUP WAS
ABLE TO SEPARATE THE TWO OUT.
THIS IS BEFORE WE HAD POWER OF
CLONING AND ALL THIS STUFF.
WE HAD TO GO BACK IN TIME AND
SEPARATING THE VIRUS AND THINGS
BECAME OBVIOUS AND WE SAID OH,
MY GOD, WE'VE BEEN WASTING
MOONS.
SO WE LOST A LOT OF TIME AND WE
DIDN'T PUBLISH JUST ON FINDING
VIRUS BECAUSE WHEN I WE
PUBLISHED WE FELT WE HAD TO SHOW
THE CAUSE AND CAN'T GROW IT VERY
WELL AND CHARACTERIZE IT AND I
DESCRIBED A FRENCH PAPER AND
THAT WAS IT.
WE WEREN'T IN A POSITION TO DO
THAT.
WE HAD TO SHOW IT WAS SOMETHING
IMPORTANT OR NOT.
THAT WAS WHERE THINGS WERE.
THIS IS MARJORIE'S CONTRIBUTION.
THE EXCITING FEBRUARY '83 LED US
TO PROVE THIS WAS A DOUBLE
INFECTION SAME CELL AND NOT
ANTICIPATED AND MARJORIE'S
VIROLOGY SHOWED IT WAS RARE AND
I GAVE YOU THAT HISTORY BEFORE.
IT WAS PART OF OUR DECISION TO
PUBLISH WITH OTHER COLLEAGUES
FIVE PAPERS SHOWING THE CAUSE OF
AIDS.
FOUR IN SCIENCE IN THE SAME
ISSUE MAY 4 AND ONE IN LANCET IN
JUNE AND THINGS BECAME CLEAR.
MARJORIE PUBLISHED HTLV-1
ANTIBODIES AND OTHERS IN LANCET
IN '84 SHOWING WHAT I JUST SAID
THE CONTRIBUTION AND NOT WHEN
YOU IDENTIFIED SHOWING THE CAUSE
OF AIDS OR ONE ANYONE TALKED
ABOUT BUT BEFORE THERE WAS
CLARITY AND THIS IS HOW CLARITY
BEGAN TO OCCUR, SLOWLY, STEADILY
AND DEFINITIVELY.
SO IN THE FREQUENCY DETECTION OR
ISOLATION OF OUR VIRUS BY OUR
GROUP BETWEEN '83 AND '84.
IF YOU LOOK AT THE PAPER IT HAS
MY NAME FIRST AND THERE'S 48
DIFFERENT SAMPLES AND 48
DIFFERENT PATIENTS.
THAT WAS OUR FIRST PUBLICATION
AND IN JANUARY OF '85, 105
ISOLETS AND IN OUR COUNTRY AND
MOST TODAY YOU NEED TO BE
VERIFIED.
WOULD ANYBODY ELSE BE ISOLATING
THE VIRUS?
HOW MANY LABS IN VIROLOGY CAN
USE IL2 AND GROW T CELLS?
NOT MANY AND WE WERE GETTING
BLOOD NOT LYMPH NODES.
THINK OF AIDS PATIENTS, SEVERE
AIDS HOW MANY T CELLS DO THEY
HAVE SO THE NEGATIVES ARE
OBVIOUS.
WE COULDN'T DETECT ANYTHING.
SO WE FELT THAT EVEN WITH
ISOLATES IT WOULD BE INEFFICIENT
TO GO OUT ON A LIMB AND --
INSUFFICIENT AND GO OUT ON A
LIMB AND SAY THIS IS THE CAUSE
OF AIDS AND THE SPECIMENS WERE
LIMITED AND NOT EVEN ALLOWED IN
SOME INSTITUTES AND AIDS HAD TO
BE CLINICALLY RELEVANT AND THE
CONSEQUENCE WAS HARDLY ANYBODY
WAS INVOLVED.
AND IN THE FALL OF '83 THE
CONTINUANCE CELL LINE CULTURE
AND A TOTAL OF 6 AND IT MADE
SEVERAL THINGS POSSIBLE THE
BLOOD TREST AND DRUG SCREEN AND
PROTEINS AND THE KEY COLLEAGUE
WORKS HERE IN NCI AS ONE OF THE
PROJECT OFFICERS WHOSE HUSBAND
AT THE TIME DIED OF AIDS FROM A
BLOOD TRANSFUSION AND THESE WERE
MY TECHNICIANS AND THIS LED TO
THE BLOOD TEST AND IT WAS SAFE,
SENSITIVE, ACCURATE AND
VERIFICATION BECAME ALMOST
INSTANT AND IT REMOVED DOUBTS
FOR US TO PUBLIC AND WE PILED UP
PAPER AND PUN ESTABLISHED.
IT WAS A KEY ADVANCE NOT ONLY
BECAUSE OF PUBLIC HEALTH REASONS
FOR PROTECTING THE BLOOD SUPPLY
BUT ALSO ALLOWED THE EPIDEMIC TO
BE RESOLVED AND ALLOWED THE
FIRST SCREENING FOR HIV DRUGS
AND WHEN THERAPY BECAME
AVAILABLE WE COULD DETERMINE WHO
TO TREAT AS WELL AS BLOCKING
TRANSMISSION AND CRITICALLY FOR
US VERIFIED HIV AS THE CAUSE OF
AIDS.
I'M MENTIONING ONLY WHAT WAS
DONE WITH DIRECTLY IN OUR GROUP
WITH MARJORIE OR IMPACTED OUR
WORK AT HIV IN BALTIMORE.
OF COURSE NOT ALL MARJORIE'S HIV
WORK IS NOTED AND MARJORIE
DEVELOPED THE ASSAY AND MOST THE
IHD RESEARCH AND YOU SEE
GEORGE'S REPORT LATER IN THE
MEETING WHERE EEL GO INTO THAT
AND THOUGH WE HAVEN'T DISCUSSED
IT IN MUCH MORE DETAIL.
I'LL CONCLUDE BY SAYING I HOPE
YOU FEEL THE PRESENTATION IN
SOME WAYS IS MY PERSONAL TRIBUTE
TO YOU.
TO MAKE IT CERTAIN YOU DO WISH
TO END THE TALK WITH THIS,
YOU'VE BEEN ONE OF THE MOST
INTELLIGENT, DEDICATED HUMBLE
COLLEAGUES ANYONE COULD WISH
FOR.
AMONG THE FEW OF A LARGE
TALENTED GROUP WITHOUT WHICH I
WOULD NOT HAVE SURVIVED AND I
KNOW THAT WELL.
I CONSIDER YOU ONE OF THE MOST
VALUABLE PERSONS I'VE EVER
ASSOCIATED WITH LIKE ALL OTHERS
HERE WE WISH YOU SAFETY, HEALTH
AND MUCH DESERVED HAPPINESS.
THANK YOU FOR EVERYTHING YOU DID
FOR US IN THE OLD LABORATORY OF
TUMOR CELL BIOLOGY AND FOR ME
PERSONALLY.
CONGRATULATIONS ON A GREAT
CAREER.
>> THANK YOU VERY MUCH.
>> THANK YOU.
I FEEL LIKE YOU'VE MADE ME
EMOTIONAL.
I APOLOGIZE FOR THAT.
YOU'VE BEEN A GREAT MENTOR.
TOOK A CHANCE ON SOMEONE WHO HAD
NO KNOWLEDGE OF VIROLOGY THE
WORK I DID WAS STUDYING
ADAPTATION TO COLD TEMPERATURES
MAYBE SHOULD BE WARM
TEMPERATURES GIVEN WHAT WE KNOW
TODAY BUT YOU PUT KNOW WORK ON
REVERSE TRANSCRIPTASE AND HAD TO
COMPARE TO POLYMERASE GAMMA.
IT WAS VERY DIFFICULT.
IMMUNE OWE LOGICALLY IT WAS
EASIER.
YOU PROVIDED ME THE RESOURCES TO
DO THAT AND SENT ME TO LEARN HOW
TO MAKE HYBRID ILLNESS AND HOW
TO MAKE MONOCLONAL ANTIBODIES
AND CONTACTED RESEARCHERS
WORLDWIDE TO LOOK AT SEROLOGY
AND LOOK AT CELLS FOR THE
PRESENCE OF HTLV AND ULTIMATELY
HLV.
YOU GAVE ME EXPOSURE TO THE
WORLD OF SCIENCE AND ONE OF THE
LAST THINGS YOU DID BEFORE YOU
LEFT WAS TO START THE WORK WITH
ADENOVIRUS VACCINE.
I GUESS I WOULD CONCLUDE BY
SAYING MUCH OF WHAT I'VE
ACCOMPLISHED IS DUE TO YOUR
MENTORSHIP.
SOW I THANK YOU AND THANK YOU
FOR BEING HERE AND THANK YOU FOR
YOUR KIND REMARKS.
>> IT'S A GREAT PLEASURE TO
INTRODUCE THE NEXT SPEAKER THIS
MORNING JOINING YOU FROM THE
UNIVERSITY OF LOUISIANA AND AS
HIGH AS GOING TO TALK ABOUT SOME
OF HER WORK WITH MARJORIE.
PLEASE.
>> THANK YOU FOR YOUR KIND
INTRODUCTION AND MARJORIE FOR
ALL THE COLLABORATIONS OVER THE
YEARS.
I WANTED TO GIVE YOU A FOLLOW-UP
WITH THE COLLABORATIONS.
SOME WERE CLIFF HANGERS AT THE
TIME WE PUBLISHED THEM.
HERE'S THE FIRST ONE.
CLASSIC IMMUNIZATION OF MACAQUE
S.
THE INITIAL PLAN WAS SIMPLE WHAT
TITE PREPARED -- PREVENTS MUSE
-- MUCOSAL SHIFT.
WE PLANNED TO DO MUCOSAL
CHALLENGES WITH A SHIFT TO
REFLECT ACUTELY TRANSMITTED
STRAINS IN HUMANS AND WE WERE
GOING TO GIVE IT EVERY TWO
WEEKS.
THE IDEA WAS TO USE THE IGG FROM
THE SAME SPECIES.
WE ISOLATED IT FROM RHESUS
MONKEYS AND RELATED TO THE
PANELS THE CLADS WE MADE.
WE SCREENED FOR IN VITRO
NEUTRALIZATION WITH A SHIFT C
THE STRAIN NAME THAT.
THAT WAS THE INTENT.
WE KNOW PEOPLE WHO GET
VACCINATED AGAINST HIV HAVE A
SNOWFLAKE'S CHANCE IN LOUISIANA
TO EVER MAKE AN EXACTLY MATCHED
STRAIN.
SO WE WANTED TO HAVE UP FRONT
HETE
HETEROLOGOUS CHANCES AND WE
WANTED TO TEST DIFFERENT DOSES.
DO TITERS PREDICT IN VIVO
PROTECTION?
COMPLICATED ANSWER.
I TOOK IT FROM THE ABSTRACT AND
YOU CAN START READING HERE.
THIS PREPARATION FAILED TO
PREVENT ACQUISITION OF THE
REPEATED CHALLENGES BUT THE DOSE
OF 400MGs LOWERED THIS AND WE
WERE IN FOR THE SURPRISE.
UNEXPECTEDLY SINGLE GENOME
ANALYSIS REVEALED A HIGHER
NUMBER OF TRANSMITTED VARIANTS
AT THE LOW DOSE IMPLYING
INCREASED ACQUISITION OF VIRUS
AT LOW SHIVIG LEVELS AT
CONCENTRATIONS OBSERVED IN
RHESUS MONKEYS TREATED.
THE CONCLUSION WAS THAT MAYBE
IGG IS TWO PHASED AND DEPENDING
ON THE PLASMA LEVELS IT ADDED
PARTIAL PROTECTION OR POSSIBLY
ENHANCEMENT.
AND IMPORTANT QUESTION THAT
REQUIRES AN ANSWER.
AND WE PUBLISHED THE ANSWER IN
DECEMBER OF LAST YEAR.
THIS IS THE SUMMARY OF THE
SINGLE GENOME ANALYSIS.
THE LOW DOSE CLEARLY HIGHER THAN
THE UNTREATED CONTROLS AND AT
THE HIGHEST DOSE OF 675
MILLIGRAMS PER KILOGRAM ONLY ONE
WAS TRANSMITTED.
HERE ARE THE IN VITRO ASSAYS FOR
C PRIME ADE.
YOUR LOOKING AT NEGATIVE
NEUTRALIZATION IN THE RED CURVE
AND SOMETIMES IN MINUS 300% WITH
THE ONE TEST VIRUS AND IN THE
OTHER CASE TO MINUS ROUGHLY
1600% NEGATIVE NEUTRALIZATION IV
ENHANCE MANY.
THE ENHANCEMENT GOES AWAY IF YOU
HEAT INACTIVATE THE COMPLIMENT
AND SO OBVIOUSLY THIS BEGS THE
QUESTION IS THIS REAL AND THE
SHIVIG NO PREVENTION OF
ACQUISITION IN ANIMALS AFTER
REPEATED DOSE CHALLENGES.
400 MILLIGRAM PER KILOGRAM CLOSE
IS SIGNIFICANTLY LOWER AND THERE
COULD BE ENHANCEMENT.
AND THE ENHANCEMENT WAS LINKED
IN VITRO SO THE CONCLUSION OF
THE PAPER WAS POLY CLONAL SHIVIG
DEPEND ON LEVELS OF THE VIRUS
AND OBVIOUSLY IF YOU VACCINATE
AGAINST HIV THE ANTIBODY LEVELS
DON'T STAY STEADY.
THEY DECLINE AND THE QUESTION IS
WOULD YOU ACTUALLY GET INTO A
DANGER ZONE.
IN OTHER WORDS, THUMBS UP OR
THUMBS DOWN FOR IGG.
AND THE REAL WAS IN VIVO.
IN VITRO YOU CAN GET DIFFERENT
PHENOMENA AND THE IDEA WAS TO
GIVE POLY CLONAL IGG THE BEST
CHANCE TO SCORE IN VIVO AND TO
SETTLE THE QUESTION ONCE AND FOR
ALL IT ENHANCES TO THE BEST OF
CIRCUMSTANCES TO DO IT OR IT
DOESN'T AND THEN QUESTIONS.
AND OPPOSITE OF WHAT WE DID TO
ACTUATE SHIV IG.
HERE WE LOOKED AT ENHANCING IGG.
AND WE SCREENED WHAT I JUST
SHOWED YOU FOR NEGATIVE
NEUTRALIZATION.
WE PULLED ANIMALS WHO HAVE THE
MOST NEGATIVE NEUTRALIZATION AND
MADE THE SHIVIG.
THIS TIME IT WAS AN INFECTIOUS
MOLECULAR CLONE.
THE INFECTIOUS MOLECULAR CLONE
IS HETEROLOGOUS TO THE SHIVIG
DONORS AND THIS YIELDS A UNIFORM
CHALLENGE FOR IN VIVO TESTING AN
R5 VIRUS AND IT'S A DIFFERENT
STRAIN FROM THE VIRUS HARVEST
THE DONOR ANIMALS.
THAT WAS THE SYSTEM WE USED TO
ADDRESS THE KEY QUESTION.
DOES PRE-TREATMENT OF SHIVIG
LOWER THE VIRUS DOSE NEEDED TO
ACHIEVE SYSTEMIC INFECTION.
THIS IS A PARAMETER WE HAVE USED
THROUGHOUT TO NOT GET FOOLED BY
LOW LEVELS OF INFECTION.
THE ANIMALS NEEDED TO HAVE
PERSISTENT SYSTEMIC INFECTION WE
DEFINED AS A VIRAL LOAD
EXCEEDING 10,000 COPIES PER ML
AND I'M NOT GOING TO SHOW YOU
THE DATA.
HERE AND HERE'S IS IN VITRO AND
THE BEST AND THE IGG ISOLATE
FROM THE VARIOUS TIME POINTS
POSTINOCULATION AND I'M SHOWING
THE DOTTED ARROW.
WHEN WE DID THE CONTROL THE HEAT
INACTIVATING SYSTEM THE
ENHANCEMENT WENT AWAY AND THAT
IS SHOWN HERE.
AND WE DID EXPERIMENTS BLOCKING
THE COMPLIMENT RECEPTOR 2 AND
ALSO KNOWN AS CD21 AND BLOCKING
THE RECEPTOR ABOLISHED C PROGRAM
80E.
WE MADE THE SHIVIG POOL AND THEN
WE ASKED END POINT IN VIVO VIRUS
TITRATION WITH THE INFECTIOUS
MOLECULAR CLONE WHETHER OR NOT
WE WOULD SEE A DIFFERENCE IN THE
REQUIREMENT OF VIRUS FOR
OBTAINING A PERSISTENT SYSTEMIC
INFECTION OF CHALLENGE ONE TIME
COMPARING NAIVE UNTREATED
ANIMALS SHOWN AT THE TOP OF THE
TIME LINE VERSUS ANIMALS GIVING
THE SHIVIG 24 HOURS BEFORE THE
VIRUS EXPOSURE.
SO WHAT WE DID WAS WE TOOK A
1:10 DILUTION AND WANTED TO MAKE
SURE THEY WERE SYSTEMIC POSITIVE
AND ONCE WE KNEW THE NEGATIVE
DILUTION WE DID INTERMEDIATE
DILUTIONS SOMETIMES TWO ANIMALS
AT THE SAME TIME AND ANIMALS
THAT WERE CLEARLY NEGATIVE AFTER
THE VIRUS EXPOSURE WERE RECYCLED
AND GIVEN A HIGH DOSE OF THE
VIRUS.
TO GET SOME ADDITIONAL
EXPERIMENT.
AND THEN WE STARTED TO DO THE
SAME DILUTION WITH THE ENSHIVIG
PRETREATED ANIMALS.
WE ALREADY KNEW A 1:1,000
SOLUTION WAS BORDERLINE SO TO
SAVE ANIMALS WE STARTED WITH
1,000 AND WENT ON TO USE FURTHER
DILUTIONS.
HERE'S THE SUMMARY.
WE USED THE CALCULATION TO
ESTIMATE TO EXTRAPOLATE I SHOULD
SAY THE AID50 AND IT TURNED OUT
THAT THE ENSHIVIG PRETREATED
ANIMALS REQUIRED A STATISTICALLY
SIGNIFICANTLY LOWER AMOUNT OF
VIRUS.
SO ROUGHLY THREEFOLD LOWER.
THAT WAS PUBLISHED AS A SAID IN
DECEMBER LAST YEAR.
SO THAT WAS OBVIOUSLY A DONOR
AND SOMETIMES WHEN YOU -- AFTER
DECADES OF TRYING YOU DON'T HAVE
AN AIDS VACCINE YOU NEED TO LOOK
AT THE ELEPHANT IN THE ROOM.
I THINK THE FACT THAT ENHANCING
ANTIBODIES ARE REAL IS AN
IMPORTANT ASPECT OF AIDS VACCINE
DEVELOPMENT TO BE TAKEN INTO
ACCOUNT FOR FURTHER
CONSIDERATION.
I'M JUST SAYING IT RIGHT NOW BUT
YOU DON'T LIKE TO END IT WITH
THAT SO THE OTHER PAPER MARJORIE
CONTRIBUTED AGAIN HER EXPERTISE
IN ANTIBODY IGG EFFECTIVE
FUNCTION INCLUDES AN INTERESTING
CASE AND THIS ANIMAL WAS
VACCINAT
VACCINATED 2003 AND REMAINED
VIREMIC AND HERE IS THIS ANIMAL
VACCINATED IN 2003 WITH PROTEIN
RECOMBINANT PARTICLES GP160 OF
OTHER HIV STRAIN AGAINST
HETEROLOGOUS AND IT WAS A TRIMER
AND HIV TRACT.
THE FIRST CHALLENGE WAS A VIRUS.
NO VIREMIA BUT RESPONSE AND WE
SID ALREADY, THE ANIMAL MADE
NEUTRALIZING ANTIBODIES AGAINST
THIS TIER 1 VIRUS.
LET'S TAKE THE TIER 2 VERSION
WHERE NO NEUTRALIZING ANTIBODY
MADE AND SAME OUTCOME AND THE
ANIMAL REMAINED FLAT LINE AS FAR
AS VIREMIA GOES FOR YEARS.
THIS ANIMAL STARTED TO MAKE SOME
MEDIATED IMMUNE RESPONSES
AGAINST VIRAL TARGETS NOT
PRESENT IN THE INITIAL VACCINE
AND NOT PRESENT IN PUBLISHED
DATA WITH ME FOR YEARS AGAIN AND
THIS WE DID A CASE REPORT IN
'22.
THE CONCLUSION IS THE ANIMAL IS
THE FIRST EXAMPLE OF A
VIRUS-EXPOSED PERSISTENTLY
AVIREMIC ANIMAL THAT HAS MADE
LONG-TERM HIGH-LEVEL CELLULAR
ANTIVIRAL IMMUNITY IN THE
ABSENCE OF AN IDENTIFIABLE
CRYPTIC RESERVOIR THAT WAS
PUBLISHED IN 2012.
AS YOU KNOW I HAVE MOVED AND NOW
THE RESERVOIR IS NO LONGER
CRYPTIC AS YOU WILL SEE.
THIS WAS SUCH A PROVOCATIVE
FINDING THAT AFTER ALL THE YEARS
THE STUDIES WERE MAINTAINED AND
HOW LONG DOES AIDS VACCINE
PROTECTION LAST WITHOUT BOOSTS?
THIS ANIMAL WAS NEVER BOOSTED.
JUST HAD THE VACCINE IN 2003.
WE RECRUITED OTHER ANIMALS WHEN
MARJORIE WORKED WITH US AND WE
DECIDED TO DO A TIER 2R5 SHIFT
AND CHALLENGE MORE THAN 10 YEARS
AFTER THE INITIAL VACCINE
PROTECTION.
SO THIS WAS A TWO-PART
EXPERIMENT AND HERE, MARJORIE,
THANK YOU AGAIN THIS WAS
COLLABORATION WHERE WE TRIED
DIFFERENT WAYS OF INDUCING
SIMULTANEOUS ANTIBODY RESPONSES
AND CELL-MEDIATED RESPONSE.
I HAVE BEEN A BELIEVER FROM THE
GET GO THAT HIV IS SUCH A TOUGH
ENEMY YOU HAVE TO ATTACK IT WITH
MULTIPLE BRANCHES OF THE
MILITARY.
YOU CAN'T JUST HAVE ONE
MODALITY.
WE CALL THIS DEFENSE IN DEPTH.
WE ALWAYS USED THAN ONE
APPROACH.
HERE'S AGAIN ONE OF THOSE
APPROACHES AND WE HAD ANIMALS,
FOUR ADDITIONAL ANIMALS THAT
WERE PROTECTED.
SOME I'M SHOWING HERE.
SOME OF THEM HAD NO VIREMIA
EVER.
THIS ONE INTEROPT 12 AND SOME
HAD LOW LEVELS OF VIREMIA
BEGINNING OR HAVE INTERMITTENT
BUT NEVER REACHING PSI
POSITIVITY.
AND NOW THAT I'VE TOLD YOU ABOUT
THE IMMUNOGENS ARE LISTED HERE
AS I SAID THEY ALL GOT PROTEIN
AND VARIOUS APPROACHES THAT WE
TRIED OUT WITH OVERLAPPING
SYNTHETIC PEPTIDES AGAINST
VARIOUS TARGETS THAT COULD
INDUCE CMI.
THAT'S NOT IMPORTANT.
WHAT IS IMPORTANT IS THE TIME
LINE.
THE TWO ANIMALS IN GREEN AND
THAT WERE FLAT LINERS.
NO VIREMIA WE DISCOUNTED THAT SO
MUCH LATER.
THE EXPERIMENTS STARTED IN 2011
AND THEN ONE WAS FLIPPING IN THE
BEGINNING DIDN'T REACH THE
10,000 LEVEL AND THE PURPLE
ANIMAL WAS LOW-LEVEL
OCCASIONALLY BUT NEVER HIGH
ENOUGH.
I MOVED TO THE RESEARCH CENTER
WHERE WE ESTABLISHED PET CT
SCANNING.
WE WERE ABLE TO LABEL THIS VIRUS
PRODUCING CELLS FROM THE PG2121
AND WAS LABELLED WITH RADIO
ACTIVE COPPER AND THE ANIMAL WAS
SCANNED OVER TIME TO SEE UPTAKE.
AND UPTAKE WE SAW.
AND THERE'S SEVERAL OTHER SPOTS
THAT WERE NOT EXPIRATION OF THE
RADIO ACTIVE LABEL.
IN FACT THIS ONE LYMPH NODE WAS
ACCESSIBLE.
AND WE EXCISED AND SENT THE
LYMPH NODE DA DNA TO UCLA TO
IDENTIFY VARL SEQUEN-- VIRAL SEQUENCES
AND NOW WE HAVE PROOF THE ANIMAL
HAS INDEED A TISSUE RESERVOIR
AND THE DISTRIBUTION OF THE
LABEL IS DIFFERENT FROM ANIMALS
THAT ARE VIREMIC AND HAVE VIRE
SPREAD OF THE SYSTEMIC
CIRCULATION.
THIS IS WHAT I JUST TOLD YOU AND
LET ME GO ON.
WE HAVE TO DO IT IN TWO SESSIONS
FOR LOGISTICAL REASONS.
WE HAVE TWO SETS OF CONTROL.
ONE EXPERIMENT INVOLVED FOUR
INTERACTIVE CHALLENGES AND THE
OTHER ONE THREE AND THOSE ARE
SHOWN HERE WITH THE ARROWS PLUS
OR MINUS THE GRAY.
THE BOTTOM LINE IS ALL OF THESE
CONTROLS BECAME VEREMIC.
THE CONTROLS WERE ALSO ANIMALS
THAT HAD SEEN LIVE VIRUS UNDER
THE PROTECTION OF PASSIVELY
ADMINISTERED ANTIBODIES CLASS
AND THEY BECAME CLEARLY VIREMIC
AND NO PROBLEM THERE.
SO RAT 9 RESISTED LATE
RECHALLENGE SOMETHING LIKE 18
YEARS AFTER THE INITIAL
VACCINATION.
NO VIREMIA.
THE VIRUS WAS THE TIER 2
INFECTIOUS MOLECULAR CLONE.
THE EXTRA ANIMALS HAD AN
INTERESTING AND PROVOCATIVE
OUTCOME.
TWO HAD HIGH PEAK VIREMIA AND
THEY WERE PROTECTED IN THE
BEGINNING.
THEY HAD STERILIZATION IMMUNITY
AND DID NOT EXTEND TO THE
CHALLENGE AND THE OTHERS WERE
ALSO REMAINED FLAT LINED.
THE RED ANIMAL THAT WAS FLAT
LINED AFTER THE INITIAL ACUTE
VIREMIA AND THEY KEPT DOING THE
SAME THING THERE WERE LITTLE
BLIPS BUT NOT MAKING COPIES SO
THE BOTTOM LINE HERE IS THAT AND
ONLY RHESUS MONKEYS WITH
PROTECTION EXISTED LATE AND
THOSE STERILIZING IMMUNITY
BECAME VEREMIC AND THE
CONCLUSION IS AND THIS IS AN
IMPORTANT MESSAGE FOR VACCINE
DEVELOPMENT AND ANTIGEN
PERSISTENCE MAY BE NEEDED FOR
LONG-TERM PROTECTION BUT DOING
IT ONE TIME AND THEN ACHIEVING
PROTECTION IS ANOTHER THING.
AND WE COINED THIS A TERM TO
MAKE IT CLEAR THAT WE NEED
ANTIBODY DEPENDENT ENHANCEMENT
OF VIRUS ACQUISITION.
THE INITIAL ENCOUNTER WAS TO
HAVE NOT CHANGE DISEASE PATTERN
OR ANYTHING.
AND THERE'S COLLABORATION FROM
THE PAPER PAPER AND I THINK WE
NAILED IT.
THIS IS REAL AND IS PASSIVE
IMMUNIZATION.
PASSIVE IMMUNIZATION EXPERIMENT
CAUSE AND EFFECT AND AGAIN THE
TROUBLING OBSERVATION OVER TIME
THERE'S INDIRECT EVIDENCE IN
HUMAN VACCINEES GIVEN THE
MONOMER THIS MAY ALSO MAY BE
TRUE AND IN SUMMARY NUMBER 2,
THE LONG-TERM PROTECTION I'VE
SHOWN YOU THE STORY OF RAT 9 AND
THE BOTTOM LINE IS THE ANIMAL IS
A CASE OF ARREST OF VIRUS
DISSEMINATION OVER MANY YEARS
AND ABILITY TO RESIST LATE
RECHALLENGE MORE THAN 15 YEARS
AFTER THE LAST VIRUS EXPOSURE.
THE ADDITIONAL FOUR ANIMALS
SHOWED THAT ANTIGEN PERSISTENCE
OR ANTIGEN BOOSTING IS REQUIRED
FOR LONG-TERM PROTECTION.
YOU DON'T GET IT IF YOU JUST
VACCINATE AND THIS IS WHAT A
JUST SAID.
AND I'D LIKE TO THANK THE
COLLABORATORS THAT MADE THE
INTERDISCIPLINARY WORK POSSIBLE
AND FIRST AND FOREMOST I'D LIKE
TO THANK MARJORIE AND HER LAB
MEMBERS WHO HAVE CONTRIBUTED AND
ACTUALLY STIMULATED THESE
QUESTIONS FOR THE FOLLOW-UP
STUDIES.
MARJORIE, MANY MANY THANKS.
[APPLAUSE]
THE QUESTION WAS WHETHER OR NOT
WE KNOW WHICH REGIONS OF THE
ANIMAL ARE INVOLVED IN THE
ENHANCEMENT, CORRECT?
WE DID MAPPING AND COMPARED THE
SHIVIG WITH CROSS-NEUTRALIZING
ANTIBODIES AND THE SHIVIG WAS
ISOLATED IN THE LATE STAGE OF
SHIV INFECTION OF RHESUS MONKEYS
AND THE END SHIVIG WAS ISOLATED
EARLY BEFORE THE ANIMALS MADE
NEUTRALIZING ANTIBODIES.
WE DID CROSS-COMPARE PEPTIDE
MAPPING BETWEEN SHIVIG AND END
SHIVIG.
THE SHIVIG, AS EXPECTED, HAD
GOOD REACTIVITY AND THE END
SHIVIG WAS MOSTLY AGAINST GP41
PRACTICALLY NOTHING AGAINST THE
V3 LOOP.
WE ACTUALLY IN THE PAPER WE
DEPLETED THE END SHIVIG OF THE
REACTIVITY AND THERE WAS NO
DIFFERENCE.
SO THE IMPLICATION WAS IN THE
END SHIVIG THE EPITOPE
SPECIFICITY WAS LIKELY SHIVIG 41
BUT THE ANIMALS THAT HAD DUAL
OUTWITH THE SHIVIG, THEY HAD
REACTIVITY AGAINST GP120.
AND THERE IS A PAPER OUT IN PNS.
WE WILL TALK WITH YOU LATER
ABOUT IT WHERE WE SHOWED
ENHANCEMENT WITH MONOCLONAL
ANTIBODIES AGAINST GP120.
SO IT'S NOT ONLY THAT GP41 IS A
SUSPECT.
THE ENTIRE ENVELOPE CAN BE
CAUSING ENHANCEMENT.
>> OKAY.
IF THERE ARE NO FURTHER
QUESTIONS, THANK YOU VERY MUCH,
RUTH.
THIS WAS A FANTASTIC
PRESENTATION AND WITH THIS WE
HAVE A LITTLE BREAK
>> I WANT TO TALK ABOUT OUR WORK
ON THE IMPACT OF IMMUNOGEN
DESIGN AND REPLICATION ON
PRECLINICAL AND CLINICAL
IMMUNO-GENESIMMUNO
GENESSE -- IMMUNOGENICITY.
MARJORIE HAS MADE TREMENDOUS
CONTRIBUTIONS NOT ONLY SPECIFIC
TO HIV BUT ALSO IN THE VACCINE
AND ADENOVIRUS RECOMBINANT AND
THEY WERE ISOLATED HERE AT NIH
IN THE EARLY '50s AND THEN
THROUGH SOME WORK WITH BOB
CHANNIC'S LAB VIRUSES WERE
LICENSED IN 1980 AND THEN IN
1997, MARJORIE ALONG WITH A
GROUP AT YF PUBLISHED -- THIS
PAPER LUBEK ET AL. AND
DISCOVERED CHIMPANZEES WERE
PROTECTED AGAINST A VIRUS.
IN THE LATE '90s I WAS ASKED TO
COLLABORATE WITH MARJORIE AND I
WORKED WITH ADENOVIRUS
RECOMBINANTS AND SHE HAD
RECLINARECLIN
RECOMBINANTS AND IF YOU DON'T
HAVE A CORPORATE PARTNER IT'S
TOUGH SLEDDING AT THAT TIME AND
DURING THAT PHASE MARJORIE HAD
PUBLISHED MORE THAN 60 PAPERS ON
AD RECOMBINANTS AND
EXTRAORDINARY CONTRIBUTIONS THAT
HIGHLIGHT LEAD IT ASPECTS OF THE
ADVANTAGES OF REPLICATION
CONFIDENT ADENOVIRUS AND THE
REPLICATION OF THE VIRUS HAD
ADVANTAGES IN TERMS OF THE
MAGNITUDE OF THE IMMUNE RESPONSE
AND SOME EXTENT THE DURABILITY.
IN ADDITION, THERE WAS INDUCTION
OF MUCOSAL IMMUNITY AND THE
IMPORTANCE OF A PROTEIN BOOST
AND GETTING THAT RESPONSE TO THE
NEXT LEVEL.
AND SO WE INITIATED PHASE 1 OF
HIV RECOMBINANTS IN 2011 AND A
COUPLE YEARS BEFORE US OF H5
RECOMBINANTS IN HUMANS THAT
PAVED THE WAY TO STUDY DIFFERENT
ASPECTS OF USING THE VIRUS IN
HUMANS.
TREMENDOUS CONTRIBUTIONS FROM
MARJORIE THAT LAID THE
FOUNDATION FOR A LOT OF THE WORK
I'LL DESCRIBE TODAY.
THERE IS A REALLY IMPORTANT
IMPACT OF VACCINE PLATFORM ON
IMMUNOGENICITY.
A LOT OF THESE HAVE VARYING
AMOUNTS OF ATTENTION OVER THE
YEARS FROM THE HIV FIELD BUT
THEY CONTRIBUTE TO THE RESPONSE,
MAGNITUDE AND VARIABILITY.
IT'S NOT A SIMPLE STICKING IN A
120 PROTEIN.
THERE ARE ALL THESE OTHER FACTOR
GO INTO OUR RESPONSE TO VIRUSES
AND THOSE INCLUDE ENVELOPE
CONFIRMATION THE VALENCY AND THE
LOW AFFINITY INTERACTION, THE
REPLICATION OF THE VIRUS THEY
COME TOGETHER TO GIVE US A
DURABLE RESPONSE THAT IS OF HIGH
MAGNITUDE.
WE HAVE BEGUN STUDYING THESE
FACTORS IN SOME ISOLATION
THROUGH THE USE OF VIRUS-LIKE
PARTICLES.
THIS IS THE WORK OF OUR LAB IN
WHICH HE IS STUDIES THE IMPACT
OF VALENCIES AND THE SPIKE AND
DOSE REPLICATION IN SMALL
ANIMALS AND RABBITS AND
ANTOGENIC AND WE'RE ABLE TO
MODULATE THE DIFFERENT FACTORS
IN ISOLATION AND TRY TO
UNDERSTAND BETTER WHAT ARE THE
MOST IMPORTANT FACTORS FOR
VACCINE IMMUNOGENICITY IN HIV
AND TWO HAVE GOTTEN ATTENTION
THE ANTIGENICITY AND MAINTAINING
THE SPIKE AND THOSE ARE
CONSERVES HAVE BEEN RESOLVED BY
THE WORK OF JOHN MOORE AND
STABLIZING HIV SPIKE AND WE VA
HIGH VALENCY PART KALTZ AND
THESE ARE PARTICLES WHERE WE'RE
ABLE TO MAKE BEAUTIFUL PRIMERS
THAT COAT THE SURFACE SO HIGH
VALENCY, ANTIGENICALLY THEY LOOK
FANTASTIC BY PROBING WITH
MONOCLONAL ANTIBODIES OR BY EM
AND MAKE THE STRUCTURAL
BIOLOGISTS JEALOUS AND YOU WOULD
ANTICIPATE IF VALENCY WERE THE
ONLY CRITICAL FACTORS THIS WOULD
BE AN APPROACH AND HIV IS NOT AN
EASY TARGET.
IF YOU DO THE EXPERIMENT WITH
VLP THAT HAS IT FROM BIRD FLU
AND IF YOU DO THE SAME
EXPERIMENT WITH VLP WITH THE
PRI
TRIMERS I HEAD TO YOU DO NOT SEE
A RESPONSE AND THERE'S AN
INHERENT LACK OF IMMUNOGENICITY
THAT HAS OTHER FACTORS TO
GENERATE AN IMMUNE RESPONSE.
SO VERY TOUGH TARGET.
AND IT WILL BE IMPORTANT FOR THE
NEXT GENERATION OF VACCINES TO
INCORPORATE MANY OF THE
ADVANTAGES OR THE WAYS I SHOWED
YOU IN THE SECOND SLIDE HOW WE
SENSED VIRUSES INTO THE VACCINE
AND TAKE ADVANTAGE OF
REPLICATING VECTORS, FOR
EXAMPLE, THAT HAVE A LOT OF
THOSE ATTRIBUTES.
THERE'S A KNOWN HIERARCHY OF
IMMUNOGENICITY FOR ANY GIVEN
TARGET ANTIGEN AND THAT IS BY
THERE'S ATTENUATED
AND REPLICATING VECTORS HAVE A
NUMBER OF ADVANTAGES YOU CAN SEE
AND THAT IS THAT THEY MAINTAIN
THE APPROPRIATE CONFIRMATION
BECAUSE THEY'RE PRODUCING EN
ENVELOPE IN VIVO AND CONTINUE TO
REPLICATE IN THE HOST UNTIL THE
IMMUNE SYSTEM SAYS STOP.
THEY HAVE TLR AGONISTS AND SIGHT
CYTOKINES AND FOR ALL THESE
REASONS WE HAVE FOCUSSED ON
REPLICATING VECTORS IN OUR
ANIMAL AND HUMAN WORK.
AND THERE'S EXPRESSION OF GLYCO
GLYCOPROTEINS AND THIS WAS A
NATURAL VIRUS TO START THE WORK
WITH BECAUSE IT'S BEEN GIVEN TO
MORE THAN 10 MILLION PEOPLE IN
THE MILITARY WITH A 99.3%
EFFICACY RATE AND EXTRAORDINARY
SAFETY RECORD.
IT'S NOT ASSOCIATED WITH
INCREASED ACQUISITION RISK AND
WE'VE NOW EITHER STARTED OR
COMPLETED MULTIPLE CLINICAL
TRIALS TO EVALUATE THE SAFETY,
DOSE, ROOT IMMUNOGENICITY AND
STARTED WITH WORK TO ESTABLISH A
DOSE AT ROOT AND I'LL SHOW A
LITTLE BIT OF THAT WORK TODAY.
WE THEN ALONG WITH MARJORIE
STARTED THIS EARLY TRIAL OF AD 4
WITH GAG OR A C150 A 1086
ISOLATE THERE AND WE'RE IN THE
MIDDLE OF THE TRIAL THE AD 4
THAT ENCODES THE 1086
CONFIRMATIONALLY NATIVE LIKE OR
A STABILIZED NFL FROM RICH WYATT
IN CLINIC.
THE EARLY WORK SHOWED IT TO BE A
PROMISING PLATFORM PUBLISHED
FROM OUR GROUP IN 2019 AND 2021.
WE EITHER GAVE IT BY AN ORAL
CAPSULE SIMILAR TO THE MILITARY
OR PAINTED IT ON THE TONSILS OF
PARTICIPANTS AND BY AN
INTRANASAL SPRAY AND THERE'S TWO
FORMS OF ATTENUATION.
ONE IS ON THE G.I. TRACT AND THE
SECOND IS AN INSERT THAT TURNS
OUT TO BE OVERLY ATTENUATED.
YOU GET A NICE BUMP EARLY BUT
YOU DON'T GET THE DURABILITY IF
YOU HAVE MORE LONG-LASTING
DURABILITY AND YOU PUT IT ON THE
TONSILS AND HAVE BETTER
REPLICATION AND GOT DURABLE
RESPONSES.
THIS IS AT SIX MONTHS.
NO DIFFERENT FROM THE RESPONSE
OF 28 DAYS.
WE ALSO SAW THIS NICE INCREASE
IN THE MEMORY COMPARTMENT OF
MONOCLONAL ANTIBODIES WE
DERIVED.
A SINGLE IMMUNIZATION DROVE SIX
MONTHS TO A YEAR OF SOMATIC
MUTATION AND THEY BECOME MORE
POTENT AND THERE'S B CELL
EXPANSIONS THAT PARALLEL THIS
OVER A LONG PERIOD OF TIME.
PARTICIPANTS IN THE TRIAL WOULD
SHED NUCLEIC ACID AND INFECTIOUS
VIRUS WAS SHED 4 TO 7 DAYS AND
THE MEDIAN DURATION OF SHEDDING
WAS ONE DAY WHEN YOU LOOK AT THE
INDIVIDUAL LEVEL.
VERY PROMISING DATA.
WE HAD NO TRANSMISSION BETWEEN
PARTICIPANTS AND INTIMATE
CONTACTS AND THIS LOOKED LIKE
QUITE A GOOD IMMUNOGENIC.
WE DID NOT SET OUT THIS APPROACH
FOR SARS COV2 IN MARCH OF 2020
WHEN I LOOKED AT THE LIST OF
VACCINES AND SAW THERE WERE NOT
ONLY REVIEW COMPETENT APPROACHES
AND NO MUCOSAL APPROACH AND THIS
WAS OUR FOURTH TRIAL AND KNEW IT
WAS AN IMMUNOGENIC PLATFORM.
WE DEVELOPED A PARTNERSHIP WITH
A COMPANY IN BELGIUM AND THEY
ARE VERY GOOD FIT FOR US AND THE
SMALLER MACHINES CAN FUNCTION AS
A FACTORY FOR VACCINES AND WE'RE
PARTNERED WITH OTHERS AT DRUMMER
HITCHCOCK AND THEY'RE REMARKABLY
ONE OF THE FEW CENTERS THAT HAVE
DEVELOPED AN EXTRAORDINARY TOOLS
FOR MEASURING MUCOSAL IMMUNITY.
IT'S QUITE SOMETHING YOU HAVE A
VIRUS LIKE SARS COV2 AND FEW
PEOPLE ARE FOCUSSED ON MUCOSAL
IMMUNITY BUT THAT GROUP HAS A
GREAT SUITE OF TOOLS.
IT'S A NICE COLLABORATION
BETWEEN THE GROUPS.
SO I'M GOING TO GO QUICKLY
THROUGH GO THROUGH THE
DURABILITY OF RESPONSE TO
VACCINE AND PRE-CLINICAL RESULTS
AND OUR PLANS FOR PHASE 1 AND 2.
FIRST INDUCTION OF MUCOSAL
IMMUNITY.
AND THERE'S NO BETTER EXAMPLE
THAN POLIO AND IT'S BEEN DONE
MANY WAYS, MANY TIMES.
IF YOU HAVE EXPERIENCE WITH TYPE
1 AND TYPE 3 POLIO BY AN ORAL
ROUTE SO THIS IS A TYPE 2
CHALLENGE OR YOU GOT TYPE 1,
TYPE 3 ORALLY AND GOT TYPE 2 BY
IPV INTRAMUSCULARLY YOU
REPRESENTED IT FULLY BUT IF YOU
HAVE AN ORAL POLIO VACCINE THAT
INCLUDE THE TYPE 2, THEN THE
TYPE 2 CHALLENGE IS RESTRICTED
IN THESE PEOPLE.
AN INTRAMUSCULAR VACCINE DOES
NOT INFER THE ABILITY TO
INTERRUPT TRANSMISSION.
IT'S A VERY IMPORTANT LESSON IN
POLIO.
AND THE IDEA FOR SARS COV2 IS A
MUCOSAL RESPONSE WOULD CURTAIL
VIRUS SHEDDING.
IT'S A HIGHLY INFECTIOUS VIRUS.
IT MAY BE WE'RE UNABLE TO
COMPLETELY PROTECT AGAINST
INFECTION.
YOU MAY RAISE THE BAR SOMEWHAT
BUT AT A MINIMUM YOU WOULD LIMIT
THE ABILITY AND THE REASON IS IF
YOU GIVE AN INTRAMUSCULAR
VACCINATION IT MAY LEAK AND GET
A LOWER RESPIRATORY TRACT
PROTECTION BUT HAVE LIMITED
ABILITY TO STOP THE SHEDDING OF
THE VIRUS.
IF YOU GET A LOCAL MUCOSAL
INFECTION YOU GET B CELLS AND
THEY PRODUCE DIME ERIC IGA AND
CAN CAPTURE VIRUS AT THE EARLY
STAGE AND PRESENTS TO THE IMMUNE
SYSTEM OR STOPS INFECTION AND SO
THIS IS A VERY POTENT ABILITY TO
BLOCK TRANSMISSION POTENTIALLY.
AND THIS IS ACTUALLY QUITE
APPLICABLE TO RESPIRATORY
VIRUSES NOT JUST A POLIO
PHENOMENON SHOWN IN THE PAST FOR
INFLUENZA VIRUS CHALLENGE
STUDIES BY BRIAN MURPHY FOR
USING FLUMIST.
THERE ARE PROTECTIONS FROM
SEASONAL CORONAVIRUS THAT
CORRELATES WITH MUCOSAL IGA
SHOWN IN THE '80s AND NOW
THERE'S PROBABLY MORE THAN 20
PAPERS IN THE LITERATURE SHOWING
THIS PHENOMENON IN EXPERIMENTAL
ANIMALS IF YOU GIVE AN TRA NASAL
VACCINATION YOU'RE ABLE TO
RESTRICT THE CHALLENGE VIRUS AND
YOU SEE LIMIT THE AMOUNTS
CHAIRED TO SHAM IMMUNIZATION OR
INTRAMUSCULAR VACCINE WITH THE
SAME VECTORER SHOWN IN BLUE.
THIS HAS BEEN SHOWN IN HIGH
SUSCEPTIBLE ANIMAL MODELS AND
LOTS OF VACCINES.
IT'S QUITE A ROBUST PHENOMENON
THAT INTRANASAL VACCINE CAN
LIMIT THE ABILITY TO INFECT A
HIGHLY SUSCEPTIBLE ANIMAL AND
LIMIT THE ABILITY TO TRANSMIT.
SECONDLY THE DURABILITY OF THE
RESPONSE TO VACCINE.
IT'S BEEN WELL TELEGRAPHED AND
MOST PEOPLE IN THE ROOM ARE
SIMILAR TO THE ISSUE WITH mRNA
VACCINES THE DURABILITY IS QUITE
PURE BUT IT'S NOT NEW OR
SPECIFIC TO SARS COV2.
IN THE PAST THE MODERNA H7
VACCINES WHEN GIVEN YOU HAD HIGH
LEVELS OF NEUTRALIZING
ANTIBODIES AFTER 28 DAYS OF THE
IMMUNIZATION AND BY DAY 200 YOU
WERE BARELY ABOVE GMT OF 10.
NOT DURABLE.
SIMILAR RESULTS WITH RABIES
VACCINE USING mRNA.
ON THE RIGHT IS IF YOU SPLIT THE
H5 VACCINE IN WHICH IF YOU GIVE
THE VACCINE AND YOU CAN AD ASO1
OR 3 AND JACK UP THE TITERS 200
TO 300 BUT BY 12 MONTHS YOU CAN
BARELY TELL THE PERSON HAS BEEN
VACCINATED.
SO THIS IS A VERY DIFFERENT
PHENOMENON FROM WHAT I'LL SHOW
IN THE NEXT COUPLE SLIDES AND
YOU GIVE A LOT OF VIRUS
INFECTION.
IT'S KNOWN THE HALF LIFE OF IGG
WILD TYPE INFECTION FOR EXAMPLE
WITH MEASLES OR A VACCINIA OR
THE HALF LIFE OF THE IGG IN THE
PERIPHERAL BLOOD IS IN THE ORDER
OF HUNDREDS TO THOUSANDS OF
YEARS ESSENTIALLY MEANING FLAT
BUT IF YOU GIVE A PROTEIN LIKE
DIPHTHERIA, TETANUS, OTHER
APPROACHES, THE ANTIBODY
RESPONSE EVENTUALLY WANES AND
REQUIRES BOOSTING.
THESE ARE THE RESPONSES IN A
COHORT IN THE MILITARY TO
REPLICATION COMPETENT AT 4 THE
MILITARY VACCINE THAT'S GIVEN ON
THE G.I. TRACT.
SO IF YOU'RE NEGATIVE AT THE
OUTSET YOU THEN DEVELOP A
RESPONSE AND THEN BRING THE
PEOPLE BACK FIVE TO SIX YEARS
AND THERE'S NO CHANGE FROM THE
PEAK OF OTHER SPOTS.
IF YOU'RE ZERO POSITIVE
PRE-VACCINATION YOU'RE ABLE TO
BE REINFECTED BECAUSE IT'S A
MUCOSAL INFECTION AND IF YOU
BRING PEOPLE BACK FIVE TO SIX
YEARS LATER THERE'S NO CHANGE IN
NEW TITER.
QUITE DURABLE RESPONSES TO THE
VACCINE GIVEN ORALLY IN THE
MILITARY.
WE SAW A SIMILAR PHENOMENON IN
OUR STUDIES WITH H5 WHEN WE
BROUGHT PEOPLE BACK THREE TO
FIVE YEARS LATER.
THERE WAS NO CHANGE REALLY IN
THEIR H5 TITERS AND AT THREE TO
FIVE YEARS SO DURABLE ANTIBODY
RESPONSES TO THIS VACCINE.
JUST BRIEFLY I OFTEN GET ASKED
ABOUT PRE-EXISTING IMMUNITY.
IT'S NOT A BINARY EVENT.
IF YOU'RE GIVING IT ON A MUCOSAL
SURFACE IT'S NOT THEY'RE ROUTINE
PROTECTED AGAINST VACCINATION.
PEOPLE ZERO POSITIVE CURTAIL
THEIR DAYS OF SHEDDING SHOWN IN
THE LOWER RIGHT BUT TO LESS THAN
SEVEN DAYS BUT THERE'S ONLY SIX
PEOPLE IN THE ENTIRE TRIAL WE
DID NOT INFECT WITH THAT OR THE
RESPONSE ALSO DEPENDS -- THE
ABILITY TO INFECT DEPENDS ON
YOUR PRE-EXISTING TITER WHICH IS
NOT BINARY.
IT'S A CONTINUUM SHOWN IN THE
UPPER RIGHT.
THERE ARE SOME PEOPLE WITH
EXTRAORDINARY HIGH TITERS BUT
PEOPLE HAVE QUITE MODEST TITERS
AND ALSO TWO OTHER FACTORS AND
THAT'S AN EFFECT OF THE
IMMUNOGENICITY OF THE TRANS GENE
PRODUCT.
PEOPLE WE WOULD BE IMMUNIZING
WITH AD 4 AND SARS 2 NOT ONLY
IMMUNIZED WITH AN IMMUNOGENIC
TRANS GENE WHICH WOULD SHIFT
THIS OUT FURTHER BUT HAVE A
MEMORY RESPONSE PREEXIST FROM
PRIOR VACCINATION OR INFECTION.
THIS WOULD BE IN MOST PEOPLE
WOULD BE BOOST.
THEN THE MUCOSAL INFECTION ROUTE
IS KNOWN THAT YOU CAN BYPASS
MOST THE SYSTEMIC RESPONSE BY
MUCOSAL INFECTION.
S THE REACTOGENICITY IS SHOWN
HERE AGAINST THE DATA FOR THE
PFIZER AND MODERNA mRNA
VACCINES AS PART OF THEIR EUA.
AND WE HAVE A CATEGORY EXCEPT
FOR PEOPLE WITH A SORE AND
SCRATCHY THROAT OR COUGH BUT NO
GRADE 3 OR GRADE 4 REACTIONS.
WE HAVE NOT SEEN THAT IN ANY OF
OUR CLINICAL TRIALS AND HALF THE
PEOPLE ARE COMPLETELY
ASYMPTOMATIC TO THIS VIRUS.
SO JUST A LITTLE BIT OF
PRE-CLINICAL RESULTS.
WE'VE NOW BUILT VIRUSES THAT CAN
EXPRESS A NUMBER OF DIFFERENT
VARIANTS OF CONCERN.
WE'RE DOWN TO NOW OMICRON.
THESE ARE ALL STABILIZED SPIKES
THAT WILL HAVE HAMSTER DATA NEXT
WEEK AND WE'RE PLANNING A
CHALLENGE STUDY OF THE BEST TWO
CANDIDATES WE'VE ALREADY PUT
INTO HAMSTERS.
WE DO QUITE WELL WITH REGARD TO
NEUTRALIZING ANTIBODIES.
IF EITHER IF YOU GIVE THE SARS
COV2 WUHAN OR DELTA SHOWN AS
INDIA IN THE UPPER LEFT, WE GET
NEUTRALIZING ANTIBODY RESPONSES
CLOSE TO 1,000 AND IF YOU
IMMUNIZE THEM WITH J&J
INTRAMUSCULARLY AND RESPONSES
ARE SOMEWHAT LOWER TO DELTA AND
EVEN LOWER TO OMICRON.
VERY IMPORTANTLY WE ALSO INDUCE
A MUCOSAL RESPONSE.
THIS IS WORK FROM PETER AND
MARJORIE ACKERMAN'S LAB AND RUTH
CONNOR SHOWING BINDING
ANTIBODIES IN THE UPPER
RESPIRATORY TRACT OF HAMSTERS
AFTER VACCINATION AND YOU CAN
SEE WE INDUCE NASAL IGA AND IGG
AND IGM.
YOU DON'T SEE THIS USING J&J AT
26IM AND THAT VACCINE LOOKS
BARELY ABOVE THAT OF SHAM OR FLU
VACCINE.
THESE DO INDUCE ANTIBODIES IN
THE UPPER RESPIRATORY TRACT TO A
VARIETY OF DIFFERENT PROTEINS
SHOWN HERE.
LASTLY THE PHASE 1 TO 2 PLANS.
THIS IS THE DESIGN OF THE
CLINICAL TRIAL WE BEGIN THIS
SUMMER.
PEOPLE WILL RECEIVE AD 4 SARS
COV2 INTRANASALLY BY OTHER
PROCESSES AND THEY'RE SIMILAR
BUT THE FDA WANTS BOTH PROCESSES
TESTED.
THEN WE'RE HOPING TO GET
INTERNATIONAL PARTNERSHIP TO TRY
TO GET SOME INDIVIDUALS WOULD
ARE SARS COV2 NAIVE.
THAT'S BECOME INCREASINGLY
DIFFICULT TO OBTAIN.
AND IN USING THIS TO A VACCINE
PRODUCT AND HERE I'M SHOWING IT
AGAINST mRNA BUT COULD BE ANY
OTHER VACCINE AND PROTECTION
COULD BE LIFE LONG.
AND THIS IS NOT A CONCERN FOR
MRNA.
PRODUCTION CAPACITY CONSTRAINTS
ARE REALLY NONE.
THERE ARE A LOT OF CAPACITY
CONSTRAINTS FOR mRNA.
THIS IS VERY INEXPENSIVE ABOUT A
PENNY PER DOSE AND HAS MINIMAL
COLD CHAIN REQUIREMENTS AND A
SCALEX MUCH PRODUCE 500 MILLION
DOSES IN FIVE DAYS.
REALLY, YOU CAN PRODUCE HUNDREDS
OF MILLIONS AND BILLIONS OF
DOSES QUITE RAPIDLY WITH THIS.
AND EMMA AND ELEANOR AND BEN
LEECH AND NIAID PERSONNEL AND
INVESTIGATORS AT THE VRC, FRED
HUTCH AND FLORYN KRAMER AND
MARJORIE AND DIVISION OF AIDS
HAVE SUPPORTED A LOT OF WORK AND
PARTICIPANTS.
I'LL STOP THERE AND TAKE
QUESTIONS.
>> VERY NICE STUDY.
I WANTED TO REINFORCE YOUR IDEA
OF INTRANASAL MUCOSAL VACCINE.
WE HAVEN'T USED THE REPLICATING
VIRUS BUT THERE'S BEEN DEVELOPED
A MUCOSAL NANO PARTICLE S1 BASED
PROTEIN VACCINE WITH TLR LIGAND
AND CYTOKINE AGINENTS AND WE
TESTED THIS IN RHESUS MACAQUES
AND WERE ABLE TO GET MORE
CLEARANCE IN THE NOSE AND NASAL
PHARYNX COMPARED TO INTRAMUSS
INTRAMUSCULARLY AND THINK IT'S
IMPORTANT NOT ONLY TO NIP THE
INFECTION IN THE BUD BEFORE IT
BECOMES SYSTEMIC BECAUSE ONCE
IT'S SYSTEMIC IT CAN CAUSE CAN
CAUSE BLOOD CLOTTING AND CAUSE
PROBLEMS AND IF YOU CAN CLEAR IT
YOU MAY REDUCE AND WE'RE SEEING
THESE IN HAMSTERS.
>> HAMSTERS IS A MORE
SUSCEPTIBLE MODEL.
>> WE USED IT IN ALUM AND NOW
REVIEWERS ARE SAYING WHY DIDN'T
YOU PRIME WITH ONE OF THE
EXISTING VACCINE VACCINES --
[OFF MIC]
>> IT MAY HAVE TO GO TO THE
TOTAL DOSE TO ME WAS AN IN
EXPECTED ANSWER.
IT'S NOT THAT SIMPLE.
IF YOU'RE TALKING ABOUT JUST THE
B CELL RESPONSE AND THE HSN, YOU
CAN GET THAT WITH A
NON-REPLICATING VECTOR.
SO THAT HAS BEEN SHOWN FOR FLU
AND ALSO FOR SARS COV2 AND LOOK
AT THE PAPER THE HSM AND THE
MEMORY B CELL COMPARTMENT FOR
PEOPLE WHO GOT mRNA ISN'T
DIFFERENT BUT FOR mRNA THEY'RE
NOT BEING PUSHED IN THE LONG
LIVED PLASMA CELL POOL.
SO THERE'S A CLEAR DIFFERENCE IN
THE BREADTH AND DURABLE OF YOUR
SERUM RESPONSE BETWEEN mRNA
VERSUS INFECTION.
SO THAT I THINK IS THE
IMPORTANCE OF THE REPLICATION
THAT GIVES YOU THAT PUSH THAT
GETS THEM OUT OF THE MEMORY B
CELL AND INTO THE BONE MARROW
AND PRODUCING A LONG LIVED
RESPONSE.
BUT IN RABBITS FOR HIV THE
IMMUNE RESPONSE KEEPS GOING FOR
SIX MONTHS TO A YEAR.
[OFF MIC]
>> YES.
>>
[OFF MIC]
>> I'M HOPING SOMETHING THAT CAN
GET CLOSER TO TAKING ADVANTAGE
OF ALL THOSE PARAMETERS THAT A
LIVE VIRUS DOES FOR YOUR IMMUNE
RESPONSE BUT WE'LL HAVE TO SEE.
ALMOST NO ONE PUBLISHES
DURABILITY DATA AND LIKE SARS
COV2 IN 28 DAYS OF SUBMITTING
YOUR PAPER AND THE
SELF-REPLICATING mRNA I HAVEN'T
SEEN DURABILITY.
>> ON THE DOSE I THINK DO YOU
KNOW WHAT IS THE DOSE -- YOU
MUST KNOW THE DOSE COMPARED TO
THE ADENO 4 VACCINE THAT HAS
BEEN GIVEN?
>> YEAH, BUT IT'S MUCH MUCH
LOWER FOR INTRANASAL I THINK
IT'S 10 TO THE 9.
WHEN WE PUT IT ON THE NOSE THE
DOSE AT WHICH YOU START TO GET
INFECTION IS BELOW 10 SIMILAR TO
SARS COV2.
IMPORTANTLY, IT DOESN'T MATTER
WHAT DOSE YOU GIVE AS LONG AS
YOU GET AN INFECTION.
WE WENT UP FROM 10 TO THE 8 AND
YOU SEE ONCE A PERSON'S INFECTED
YOU SEE NO DIFFERENCE FROM
SOMEBODY WHO GOT 10 TO THE 8
VERSUS 10 TO THE 2.
>> DO YOU EXPECT THE RESISTANCE
IN TERMS OF THE FDA FOR
INTRANASAL ADMINISTRATION.
>> THE INTRANASAL PART WE WORKED
THROUGH WITH THE FDA OVER MANY
YEARS.
WE HAVEN'T HAD TO DO ANY TOX
SINCE THE FIRST TRIAL.
THEY'RE COMPLETELY COMFORTABLE
WITH INTRANASAL.
INITIALLY THERE WERE CONCERNS
SPIKE WAS GOING THROUGH THE
RECEPTOR BUT NO ONE HAS SEEN
THAT AND THERE'S ABOUT A DOZEN
INTRANASAL VACCINES BEING TESTED
IN PHASE 1 NOW.
NO ONE HAS OBSERVED THAT.
I THINK THOSE TWO ISSUES ARE NOT
GREAT ISSUES.
THE ONE LAST ONE TTS WE HAVE TO
WATCH FOR BUT THERE'S NOT GREAT
DATA THAT SAYS IT'S ALL
ADENOVIRUSES.
SO YOU DON'T SEE TTS OR
THROMBOTIC COMPLICATIONS WITH
THE ORAL AND IN THE AD 4 IN THE
MILITARY YOU DON'T SEE WAVES OF
INTRANASAL INFECTION.
IT'S PROBABLY SOMETHING TO DO
WITH 10:11 IM AND WHERE IT'S
EXPRESSING PROTEIN THAT PROBABLY
HAS TO DO --
>> ON THE TTS SPECIFICALLY, CAN
ONE USE THE OLD DATA THE ADENO 4
VACCINE DATA IN THE MILITARY TO
SUPPORT SAFETY CONSIDERATIONS
BECAUSE --
>> IT WILL BE MENTIONED.
THAT WILL WILL BE AN IMPORTANT
ISSUE WITH OTHER ADENOS.
WE KNOW THEY HAVE A PROBLEM WITH
THROMBOSIS BUT WE SAW THAT IN A
MILLION VACCINATIONS.
THE ARGUMENT IS CAN WE LOOK BACK
AT THE MILLION VACCINATIONS WITH
ADENO 4 VACCINE AND BECAUSE I
DON'T KNOW HOW GOOD THE DATA IS
FOR THESE RECRUITS, THE MILITARY
RECRUITS.
>> WITH MORE THAN 10 MILLION
PEOPLE AND YOU WOULD HAVE
EXPECTED MORE THAN THAT NOW YOU
WOULD EXPECT MORE THAN 50 CASES
OF STRONG THROMBOSIS BUT IT'S
NOT BEEN REPORTED.
SOME CRITICS WOULD SAY WELL,
THEY WEREN'T REALLY LOOKING FOR
IT SO MAYBE IN THE MILITARY
YOU'RE DOING ALL KINDS OF CRAZY
THINGS YOU MAY BE DEHYDRATED AND
MAYBE YOU HAD THROMBOSIS AND IT
WAS MISSED.
WHO KNOWS.
BUT IT WILL BE RAISED THAT THERE
ARE THOSE DATA.
WHETHER THE FDA WILL PAY
ATTENTION I DON'T KNOW.
I THINK PROBABLY THEY'LL REQUIRE
IT TO BE SCREENED FOR OVER A
LONGER PERIOD OF TIME AND MAYBE
BY THE TIME THAT COMES AROUND,
THERE'LL BE MORE INFORMATION
ABOUT THE ACTUAL MECHANISM OF
WHAT IS INDUCING PEOPLE
POSTULATING CONTAMINANT AND A
VARIETY OF OTHER ISSUES ON WHAT
INDUCES THAT.
IT'S JUST GOING TO TAKE TIME.
>> FINAL QUESTION, SINCE WE HAVE
YOU HEAR AN OPPORTUNITY TO
LEARN.
WHY DO YOU THINK THE MUCOSAL
ESPECIALLY ADENO VECTOR WHICH IS
A UNIQUE OPPORTUNITY, WHY DIDN'T
IT PROGRESS FASTER THAN THE TWO
YEARS AND A HALF WE ARE NOW?
>> WELL, THAT'S EASY.
SO IT WAS MORE IF YOU TALK TO
PEOPLE THERE'S TWO FACTORS.
ONE IS A LOGISTIC ONE AND
ASPECTS GOT BOUGHT BY EMERGENT
AND IT WAS A CONTRACT
MANUFACTURER FOR OTHER VACCINES
PART OF OPERATION WARP SPEED.
THEY PUT THAT AND WE HAVE A NEW
CORPORATE PARTNER AND THAT FIXED
THAT BUT IF YOU TALK TO PETER
POLASI OR OTHERS PRODUCING
VACCINES, IF YOU WEREN'T PART OF
THAT INITIAL SELECTED VACCINE
YOU GOT SHOVED TO THE SIDE AND I
WAS HARD TO GET FUNDING AND GET
A CONTRACT MANUFACTURER EVEN IF
YOU GOT THE MONEY FROM A FUNDER.
>> CLINICAL TRIALS?
>> FOR US IT'S NOT AN ISSUE.
BUT YES, IT WAS A BRUTAL YEAR TO
GET THIS TOGETHER.
>> ONE MORE QUICK QUESTION, I
DON'T WANT TO RUN OVER TIME.
ONE POTENTIAL CONCERN HAS BEEN
RAISED ABOUT INTRANASAL VACCINES
WAS RAISED RECENTLY BY SOMEONE
AT YALE WITH AN INTRANASAL SPIKE
PROTEIN BEING USED WITHOUT
ADJUVENTS AND HER CONCERN IS
THINGS CAN YOU PUT THROUGH THE
NOSE CAN GO THROUGH THE
OLFACTORY SYSTEM TO THE BRAIN
AND CAUSE POTENTIAL PROBLEMS.
I'M WONDERING IF YOU'VE RAN INTO
THAT CONCERN ADJUVANT VIRUSES IF
THERE EVIDENCE THEY ARE CAN GO
TO THE BRAIN AND IF SO WHAT'S
RESULT.
>> WAS CAUSE IT GOT TO THE
OLFACTORY BULK AND THE DATA FOR
SPREADING TO THE BRAIN WAS
WEAKER IF YOU LOOK AT THE
DETECTION SYSTEM USED IN THE
SLIDES WE WERE ASKED TO DO THE
SAME STUDY AND THERE WAS NO
SPREAD TO THE BRAIN.
WE'VE INTRANASALLY IMMUNIZED
PEOPLE AT THE CLINICAL CENTER
AND I'VE BEEN TOLD SIMILAR DATA
WERE OBTAINED FOR FLUMIST THAT
THIS IS GETTING TO THE OLFACTORY
BULB IS SOMETHING THAT HAPPENS
NOT JUST WITH VIRUS BUT OTHER
PROTEINS.
BUT REALLY SPREAD TO THE BRAIN
IS THE KEY THING.
SO WE HAVE NO EVIDENCE OF ANY
ISSUE.
>> THERE'S OLD LITERATURE THAT
SAYS IF YOU PUT GOLD
MICROPARTICLES IN THE NOSE YOU
CAN SEE THEM IN THE BRAIN BY EM.
>> I DIDN'T KNOW ABOUT THE
BRAIN.
I KNOW THERE ARE SIMILAR DATA
ABOUT THE OLFACTORY BULB.
>> THANK YOU.
FASCINATING.
THANK YOU SO MUCH FOR THAT.
AND I MUST SAY THAT I DON'T KNOW
IF ANYBODY WILL HAVE THE CHANCE
TO MENTION THAT WITH THE ADENO
4.
IT STARTED A LONG TIME AGO WITH
BOB AND WITH WYATT AND THEN WHEN
WYATT DECIDED TO ELIMINATE THE
WHOLE ADENOVIRUS DEPARTMENT, I
WAS THEN THE CONSULTANT FOR
WYATT SO I REMEMBER AND MARJORIE
INSIST
INSISTED AND THAT SAYS SOMETHING
ABOUT STAMINA AND GOOD THINGS
ABOUT NIH AND NCI THAT WE WERE
ABLE TO WE HAD POPULAR PROJECTS
AT TIMES.
ANYWAY, THE NEXT SPEAKER WILL BE
MICHAEL THOMAS OF HOWARD
UNIVERSITY.
I BELIEVE THAT HE LEFT
MARJORIE'S GROUP RECENTLY AND IS
NOW AT HOWARD UNIVERSITY AND HIS
TITLE IS ROELDZ AND FUNCTIONS OF
THE ADENOVIRUS EARLY GENES IN
VACCINE-INDUCED IMMUNE
RESPONSES.
>> THANK YOU.
I FEEL HONORED TO SHARE THE
PODIUM WITH YOU GUYS.
I TELL SOME PEOPLE THE STORY
HERE THEY DON'T SEE A WOW FACTOR
IN IT BUT IN 2007, PART OF MY
F31 WHY WE HAD A TRAVEL AWARD SO
I WENT TO CANADA WHERE THEY WERE
HAVING A HIV MEETING AND ONE OF
THE SPEAKERS THERE WAS
DR. MARJORIE GUROFF AND TALKING
ABOUT REPLICATING THE ADENOVIRUS
AND I WAS DOING WORK WITH
REPLICATING ADENOVIRUS.
I WENT TO HER AFTER HER TALK AND
I WENT TO HER AFTER THE TALK AND
I TOLD HER I WAS ABOUT TO
GRADUATE AND WAS INTERESTED IN A
POST-DOC POSITION IN HER LAB.
AT THAT POINT SHE DIDN'T HAVE
ANY POSITIONS BUT TOLD ME KEEP
IN CONTACT AND IF SOMETHING
COMES UP AND WE'LL SEE AND
SOMETHING CAME UP BUT SHE
DOESN'T REMEMBER THAT.
AND WE LOOKED TO BETTER
UNDERSTAND THE ROLE AND FUNCTION
OF THE ADENOVIRUS EARLY GENES.
OF COURSE YOU KNOW THAT
ADENOVIRUS HAS BEEN DEVELOPED
FOR A WIDE VARIETY OF DIFFERENT
INFECTIOUS DISEASES AND WE'RE
FOCUSSING ON HIV.
AND MOST RECENTLY OUT OF THE
AIDS VACCINES FOR COVID THAT YOU
FIND A ROLE WORLDWIDE OR IN THE
CLINICAL DEVELOPMENT AIDS, SIX
ARE BASED ON ADENOVIRUS.
SO OUR GOAL IS TO DEVELOP A
BETTER UNDERSTANDING OF THE
VIRAL AND CELLULAR FACTORS THAT
INFLUENCE THE IMMUNE RESPONSES
TO IMMUNOGENS EXPRESSED WITHIN
THE ADENOVIRUS.
SO ADENOVIRUS IS A DNA
CONTAINING VIRUS THAT WAS
DISCOVERED IN THE 1950s AND IN
HEALTHY INDIVIDUALS YOU GET MILD
UPPER RESPIRATORY TRACT
DISEASES.
BUT ADENOVIRUS CAN KILL AND WE
HAVE SEEN EVIDENCE OF THAT NOT
TOO LONG AGO IN UNIVERSITY OF
MARYLAND.
IT INFECTS CELLS BY BINDING TO
THE CAR RECEPTOR AND IT'S
INFECTED AND TRAVELED TO THE
LATE ENDOSOME WHERE THE VIRAL
PARTICLE IS DESTABILIZED.
THE PARTICLE TRAVELS TO THE
NUCLEUS.
WE'VE KNOWN FOR A LONG TIME THAT
ADENOVIRUS STRUCTURED GENES THE
LATE AND STRUCTURAL GENES FIBER
AND THOSE PROTEINS ACT LIKE THIS
AND EVERYONE USING ADENOVIRUS
FOR VACCINE STUDIES ARE
UTILIZING THIS THE ADENOVIRUS
STRUCTURAL PROTEINS WILL AS AN
ADJUVANT AND WE THEY'D EARLY
GENE E1A.
SO MANY VIRUSES YOU SEE IN THE
CLINIC NOW DON'T HAVE THIS GENE
AND I'LL TALK TO YOU ABOUT THAT.
ANYHOW THE E1A IS IMPORTANT
BECAUSE IT TRANSACTS ALL THE
OTHER ADENOVIRUS GENES AND THEY
ARE EXPRESSED BEFORE VIRAL DNA
SYNTHESIS AND THOSE LABELLED L
ARE EXPRESSED AFTER ADENOVIRUS
GENO SYNTHESIS.
AND THESE OTHER GENES IN
ADENOVIRUS AND WE HAVE MORE
WE'LL SHOW YOU LATER.
WHAT ABOUT THOSE GENES AND DO
THEY CONTRIBUTE TO THE VACCINE
INDUCED IMMUNE RESPONSES WE SEE?
PEOPLE ASKED THE QUESTION BEFORE
US AND WE WANTED TO LOOK AT IT
DIFFERENT
DIFFERENTLY AND HOW DO THEY
FUNCTION IN IMMUNE RESPONSES?
ONE OF THE THINGS WE NOTICED WAS
THAT AND HERE WHAT WE'RE DOING
IS USING THIS IODINE FOR DNA AND
WE'RE LOOKING AT THE DNA AND
THERE ARE CELLS IN TISSUE
CULTURE AND OTHERS GOING THROUGH
THE CELL CYCLE AND IN CANCER
CELLS YOU CAN FIND DNA CONTENT
BETTER THAN D2M.
SO HERE WE INFECTED BRAIN CANCER
CELLS AND 48 HOURS AFTER WE ARE
LOOKING AT THE DNA CELL CYCLE
PROFILE WHERE WE QUANTIFY THEM
AND WE CAN SEE THAT THERE'S
ENHANCED OR INCREASED LEVEL OF
CELLS WITH DNA CONTENT GREATER
THAN D2M.
THIS IS 48 HOURS.
120 HOURS YOU SEE THE INCREASE
AND WE OVERLAID TO SLOW YOU IT
BETTER.
SO TO UNDERSTAND THIS THING I'VE
BEEN TALKING ABOUT THIS DNA
GREATER THAN 4N AND HOW IT
BECOMES RELEVANT, SO THIS IS
YOUR NORMAL CELL CYCLE.
SO CELLS GO FROM G1 TO S, G2 TO
M AND DAUGHTER CELLS AND GO BACK
TO G1.
BUT THERE'S THIS ALTERNATE CELL
CYCLE IN WHICH CELLS GO FROM G1
TO S, G2, THEY SKIP M ALTOGETHER
AND REENTER THE CELL CYCLE.
THE PROBLEM WITH THIS IS THAT IT
IS PART THIS VICIOUS CYCLE WHERE
YOU HAVE DNA DAMAGE, DNA DAMAGE
SEEMS TO LEAD TO THIS THING I'M
TELLING YOU ABOUT, THE DNA
GREATER THAN 4N AND SEEMS TO
MAKE CELLS MORE TOLERANT TO DNA
DAMAGE AND THAT SEEMED TO LEAD
TO GENOME INSTABILITY AND MORE
GENOME INSTABILITY ENHANCES DNA
DAMAGE WHICH ENHANCES THIS,
ENHANCES THAT AND GETS CONFUSED
LIKE THAT.
SO DNA DAMAGE RESPONSE IS A VIEW
BUT YOU CAN HAVE DAMAGE FROM
DIFFERENT CHEMICAL, DIFFERENT
AGENTS.
SO IN RESPONSE TO DOUBLE STRAND
BREAKS YOU HAVE THE MAJOR KINASE
ATM RECRUITED TO DOUBLE STRAND
BREAK AND WHILE THERE IT
PHOSPHORYLATED DOWN STREAM
MEDIATORS AND EFFECTERS THAT CAN
STOP THE CELL IN THE CELL CYCLE
AND IF IT WAS SINGLE STRAND
BREAK THAN ATR WOULD DO THE SAME
THING AND STOP THE CELL IN THE
CELL CYCLE.
SO WE'VE WANTED TO KNOW THEN
WHETHER THIS WAS POSSIBLY AN
ADENOVIRUS INFECTED CELLS
BECAUSE WE SAW A LOT OF EVIDENCE
THAT SUGGESTS THE DNA DAMAGED
AGENT OR DAMAGE ITSELF CAN
STIMULATE THE IMMUNE RESPONSE.
SO ONE OF THE FIRST THINGS WE
DID WAS INFECT CELLS WITH WILD
TYPE ADENOVIRUS AND THE VIRUS IN
WHICH WE DELETED THE E1B55K GENE
THE GENE ITSELF IS PROBABLY
IMPORTANT AND THE IMPORTANCE OF
THIS VIRUS HERE IS IT'S BEEN
USED IN CLINICAL TRIALS IN THE
U.S. AND IS ACTUALLY BEING USED
TO TREAT HEAD AND NECK CANCER IN
CHINA.
YOU CAN SEE WHEN WE INFECT CELLS
WITH THE WILD TYPE VIRUS THIS
STAYS IN THE NUCLEUS AND THIS IS
THE SAME PHOSPHORYLATION YOU'D
FIND IN RESPONSE TO DNA DAMAGE
AND THE SAME YOU FIND ON ATM IN
RESPONSE TO DNA DAMAGE AND YOU
CAN SEE HERE THAT THE WILD TYPE
ADENOVIRUS INDUCED HIGHER LEVELS
OF THE PHOSPHORYLATED PROTEIN OF
THE DNA PHOSPHORYLATED DAMAGED
PROTEIN AND HERE I MEASURED AND
QUANTIFIED BY FLOW.
THESE PROTEINS HERE THIS IS JUST
SHOWING YOU THE RESPONSE TO DNA
DAMAGE, THE MRN COMPLEX IS
RECRUITED TO THE BREAK AND THERE
THEY RECRUIT ATM, ATM
PHOSPHORYLATED H2X AND ATM CAN
ALSO PHOSPHORYLATE DOWN STREAM
MEDIATORS THAT CAN STOP THE CELL
THAT CAN INDUCE DEATH OR THAT
CAN CAUSE REPAIR FOR WHATEVER
THE DAMAGE IS.
SO THESE TWO VIRUSES THEN WE
KNOW THAT IT INDUCES DNA DAMAGE
SIGNALS AND COMPARED THEM AS FAR
AS THEIR ABILITY TO WHEN THEY
INFECT CELLS TO INDUCE INNATE
IMMUNE RESPONSE AND HERE WHAT
WE'RE DOING IS MEASURING
CYTOKINES PRODUCED IN A54 THE
CELLS AND IN HELA CELLS.
IN EACH CASE THE VIRUS WHERE WE
DELETE THE E1G GENE PRODUCED
MORE CYTOKINES OR IT ALLOWED
CELLS TO PRODUCE MORE CYTOKINES
THAN THE WILD TYPE VIRUS.
NOW, WITH THE HELA CELL AND THE
GENES ARE DIFFERENT AND ALLOWED
THEM DIFFERENT AND THEY DON'T
ALL HAVE THE SAME GENES OR THE
SAME SPLICED VARIANTS OF THE
GENES AND THE TUMOR CELLS ARE
WHAT CAUSED THEM TO BECOME TUMOR
CELLS, THAT ITSELF IS DIFFERENT
BECAUSE OF THAT YOU CAN HAVE
DIFFERENT CYTOKINES PRODUCED IN
THE DIFFERENT CELLS.
ANYHOW, WE LOOKED AT A DIFFERENT
FACTOR.
THIS IS LOOKING AT AND THE
LIGAND THIS IS AN ACTIVATING
MARKER YOU FIND IN NKT CELLS AND
GANA DELTA CELLS AND TUMOR
CELLS TEND TO HAVE HIGHER LEVELS
AND YOU SEE THIS BLACK LINE.
THIS IS THE LEVELS ON THE HELA
CELL.
I INFECTED THEM WITH THE WILD
TYPE VIRUS WITH THE DELETION IN
THE E1B AND YOU CAN THEN LOOK
OVER TIME AND YOU CAN SEE OVER
TIME THE LEVELS OF THE INCREASE
WAS IN THE DELETED FILES.
THIS AND WHAT I SHOWED YOU
BEFORE HAVE BEEN INFORMING US
ABOUT E1B55K AND THE POTENTIAL
ROLE IN INFECTED CELLS.
WE WANTED TO EVALUATE THE IDEA
THAT VIRUSES ARE ABLE TO INDUCE
THIS GREATER THAN 4N ARE MORE
IMMUNOGENIC THAN THE VIRUSES
THAT DON'T PRODUCE AS MUCH.
THAT WAS KIND OF REALLY
SIMPLISTIC AND NOW THAT WE THINK
ABOUT IT, IT'S KIND OF LIKE
SILLY BUT IT WAS SOMETHING TO GO
ON BECAUSE WE COULD MEASURE IT
AND THEN WE COULD SEE WHETHER IT
CORRELATES.
SO WHAT WE DID IS FIRST MADE
VIRUSES.
THIS IS SHOWING AGAIN THE
ADENOVIRUS GENOME AND HERE'S
THERE'S MORE BUT THESE ARE THE
TWO MAIN ONES AND HERE IN THE E4
REGION THERE'S SEVERAL OTHER
PRODUCTS.
SO WE MADE VIRUSES IN WHICH WE
DELETED THE E1B GENE AND WE
EXCHANGED THE E3.
AND THE E3 AND ADENOVIRUS IS NOT
NECESSARY FOR OPTIMAL VIRUS
REPLICATION.
IN FACT, WE DON'T REALLY WANT TO
KNOW BECAUSE WHAT IT DOES IS IT
DOWN REGULATES THE MOLECULES.
WE CAN'T HAVE THAT SO WE GOT RID
OF THIS.
AND THIS YOU'VE HEARD OF BEFORE.
THIS IS JUST FULL AND SINGLE
CHAIN HIV LINKED TO THE D1, D2
DOMAIN OF CD4.
THIS IS FIRST DESCRIBED EARLIER
AND WAS SHOWN THAT IT CORRELATED
WITH PROTECTION AND WE SHOWED
THAT WHEN WE MADE A VACCINE IT
CAN GENERATE HIGH NEUTRALIZING
ANTIBODY AND BOOST AGAINST THE
PEPTIDES.
SO WE MADE THE VIRUS, DELETED
E1B AND VERIFIED THAT IT
PRODUCED A TRANS GENE.
AND THIS IS WHAT IS MADE IN THE
CELL SO WE JUST LOOKED IF THE
PROTEIN IS IN THERE YES OR NO
AND THERE'S NOT A LOT IN THE
CELL BUT WHEN YOU LOOK AT IN THE
MEDIUM THIS MAJORITY OF THE
PROTEIN IS SECRETED.
AND RELATIVE TO THE VIRUS WE
DELETED THE E1B, THEN THIS WILD
TYPE VIRUS PRODUCED MORE OF THE
TRANS GENE.
SO WE JUST GOING IN THINK WOW,
IF THE TRANS GENE IS ANYTHING TO
SAY THAT THIS VARIOUS IS AT A
DISADVANTAGE.
KNOWLEDGES WE DECIDED TO
IMMUNIZE MICE.
AND WE COLLECTED BLOOD AND I'M
SHOWING YOU ONE FOR THE SAKE OF
TIME.
WE LOOKED AT LEVELS OF BINDING
ANTIBODIES AGAINST THE VIRUS,
AGAINST THE ADENOVIRUS.
THE LEVELS OF ANTIBODY MADE THE
SAME AND DIDN'T SEE ANY
DIFFERENCE AS FAR AS AGAINST THE
TRANS GENE IN EITHER OF THE
FORMS.
AND THE VIRUSES THAT MAKE
GREATER THAN 4N SHOULD BE MORE
IMMUNOGENIC BUT FOR THE I WAS
EXCITED BECAUSE BECAUSE ONE OF
THE FIRST THINGS WE DID WAS
SHOWED THIS VIRUS THAT IS IN THE
CLINIC BEING USED TO TREAT
CANCER IN CHINA, MADE IT TO
PHASE 3 CLINICAL TRIALS IN THE
U.S., I SHOWED THIS VIRUS CAN
INDUCE IMMUNE RESPONSES SIMILAR
TO VIRUSES BEING USED FOR
PRECLINICAL TRIALS SO I WAS
EXCITED ABOUT THAT.
I WAS ALSO EXCITED ABOUT THIS
BECAUSE I KNOW THIS VIRUS AND
NOW I KNOW HOW I CAN IMPROVE ON
THIS.
THE ADENOVIRUS E1B GENE PRODUCT
CONTROLS LEVELS OF SURFACE OF
INFECTED CELLS AND PROTEINS AND
TRANS GENE AND IT CONTROLS
LEVELS AND TYPES OF CYTOKINES.
SO THAT'S THE E1B.
THERE'S STILL A LOT OF GENES
THAT ARE ADENOVIRUS.
SO WE'VE FOCUSSED ON THE E4
GENES AND WE DID THIS BECAUSE IF
YOU LOOK HERE AT THE ADENOVIRUS,
IT CAN ONLY CARRY THIS WILD TYPE
AND CARRY A TRANS GENE ABOUT
2.5KB.
SO YOU COULDN'T REALLY PUT IN A
HUGE CANCER GENE AND YOU
COULDN'T INVERT MULTIPLE GENES
IN THERE BUT IF WE DELETED THE
E1B WE GET ABOUT 1,000 BASE
PAIRS OF SPACE AND PUT IN 1 TO 4
AND WE GET ANOTHER 1,000 BASE
AND CARRY CAPACITY.
NOT ONLY THAT, WE LEARNED THAT
THESE GENES, SOME OF THESE GENE
PRODUCTS, WELL OTHERS HAD SHOWN
THAT THESE PRODUCTS INHIBIT AND
HERE WE'RE SHOWING WE
CHARACTERIZED THE VIRUS JUST AS
FAR AS HOW MUCH PROGENY IS MADE
AND THIS IS LOOKING AT THE DNA
OF VIRUSES.
THIS IS LOOKING AT THE LATE
VIRAL PROTEINS BY WESTERN BLOCK
AND THIS IS THE TRANS GENE.
THIS IS LOOKING AT A TRANS GENE
PRODUCED BY WESTERN BLOCK.
SO HERE THIS VIRUS WE CREATED BY
DELETING E1B AND E4 HAVE MUCH
EFFECT ON THE DNA.
WE HAVE DONE THIS AND WE DON'T
ALWAYS GET THIS HERE.
THIS IS JUST THIS TIME.
AND THE LATE VIRAL PROTEIN MAKES
QUITE A BIT BUT IT MAKES THE
TRANS GENE WHICH IS THE
IMPORTANT THING.
SO WE FIRST THEN INFECTED CELLS
AND AGAIN WE WANTED TO SEE HOW
THESE VIRUSES, HOW THEY WHETHER
THEY CONTROL THE CYTOKINES IN
THE CELL AND WE RANSOM PCR AND
THIS IS A549 CELLS AND DID
EXPERIMENTS ALSO AND WE NOTED
THAT THE VIRUS WHERE WE DELETED
E1B AND E14, THEY SEEM TO BE A
LOT MORE RED ON THE SIDE THAT IS
ENHANCED.
THIS IS A FOLD COPY AND IN SIDE
THERE'S MORE RED AND NOT ONLY
MORE RED SOME ARE SIGNIFICANTLY
HIGHER THAN BLACK.
WE LOOKED IN GENERAL ALL OF THEM
AND THAT PATTERN REMAINED AND
ALL THOSE ABOVE ZERO ABOVE THE
MARK THIS WAS ALSO SIGNIFICANTLY
HIGHER THAN IN THE WILD TYPE
VIRUS.
WE THEN WENT TO MICE AND
IMMUNIZED MICE AND LOOKED AT THE
IMMUNE RESPONSE.
SO HERE I'M SHOWING YOU BIND
BEING ANTIBODIES AGAINST THE
ADENOVIRUS AND THIS THE LEVELS
OF BINDING IG IS THE SAME
AGAINST IT THE ADENOVIRUS THE
MUTATIONS MADE IN THE VIRUS THEY
DON'T AFFECT THIS.
WELL, THAT'S TELLING ME THAT
THIS, THIS IS AGAINST THAT THING
THAT'S OUTSIDE THE VIRUS AND
THAT'S A RESPONSE AGAINST THIS.
THAT'S SO THEY'RE THE SAME.
BUT NOW WHEN WE LOOK AT THE
GP120 WE NOTE THE VIRUS THAT WE
CREATED CAN INDUCE HIGHER LEVELS
OF BINDING ANTIBODY IN THE CASE
BY SIGNIFICANTLY HIGHER LEVELS
OF BINDING ANTIBODIES.
BINDING ANTIBODIES ON THE WHOLE
ARE NOT THE SAME SO IN HUMAN THE
IG1 IS THE MOST FUNCTION.
IT HAS THE MOST FUNCTION.
AND IT'S EQUIVALENT IN MOUSE IS
THE IG2A.
SO IG2A IS PREDOMINANTLY MADE
AGAINST VIRAL PATHOGENS.
WE LOOKED AT IG2A AND SEE THERE
AGAINST THE ADENOVIRUS THE
LEVELS OF ANTIBODIES MADE ARE
THE SAME.
HOWEVER, AGAINST THE TRANS GENE
THE LEVEL OF ANTIBODIES MADE IN
THE VIRUS THAT WE CREATED AS
THIS ENHANCE.
SO WE REMEMBER I SHOWED YOU
THIS, WHEN WE JUST DELETED E1B
AND THERE WAS NO DIFFERENCE.
BUT IN THESE, WHERE I DELETED IN
ADDITION E41 TO 4, NOW YOU CAN
SEE MARK DIFFERENCES.
SO THIS ALLOWS US TO CONCLUDE
THE E4, 1 TO 4 IT IMPACTED
ANTIBODY THAT ARE REPORTED TO
PLAY A VIRAL PATHOGEN AND THE
VIRUS MADE THIS VACCINE
CANDIDATE INDUCED HIGHER LEVELS
OF RESPONSE THAN THE WILD TYPE.
WE THEN WENT TO NON-HUMAN
PRIMATES AND WE IMMUNIZED ORAL
INTRANASAL TWICE AND BOOSTED FOR
THE C.
THESE ARE THE NUMBERS OF
NON-HUMAN PRIMATES WE HAD AND WE
COLLECTED BLOOD AND CELLS.
WE FIRST LOOKED AT THE MUCOSAL T
CELLS AND IN LOOKING AT THE
MUCOSAL T CELLS WE NOTE THE
VIRUS WE CREATED INDUCED
SIGNIFICANT LEVELS OF CENTRAL
MEMORY CELLS RELATIVE TO THE
PRELEVELS AND SO DID THE DELETED
VIRUS AND THE VIRUS WE CREATED,
THE LEVELS WERE HIGHER THAN THAN
THE WILD TYPE VIRUS.
WE THEN LOOK AT DIFFERENT
ANTIBODIES AND LOOKED AT IG1 AND
THAT'S THE MOST FUNCTIONAL
ANTIBODY IN NON-HUMAN PRIMATES
AND THIS IS THE AREA UNDER THE
CURVE AND THE VIRUS WHERE WE
DELETE AND YOU CAN SEE THE LINES
IN BOTH CASES ARE HIGHER THAN
THIS LINE WHICH IS THE WILD TYPE
AND THE E1B.
THIS JUST SHOWING THE
QUANTIFICATION OVER TIME AND
WHAT YOU'LL NOTE IS THE LEVELS
DROP OFF FOR THE OTHER VIRUSES
RAPIDLY BUT MAINTAINED FOR THE
A4 DELETED VIRUS WE CREATED.
THIS IS SHOWING FROM THE GP120.
AND THE CYTOKINES PRODUCED IN
NON-HUMAN PRIMATES AND THE
DURATION AND THE VIRUS WE'VE
CREATED, WELL, IT CAN PRODUCE
HIGHER LEVELS OF ANTIBODIES FOR
A LONGER DURATION THAN THE
PARENT VIRUS AND THE WILD TYPE
VIRUS.
AND WE DELETE THE GENES AND JUST
DELETING THESE GENES CHANGED THE
RATIO OF ANTIBODIES OR THE
ANTIBODIES MADE.
AND STARTED TO EVALUATE HOW OR
WHERE THE VIRUSES WORK.
>> WE NEED TO SPEED UP A LITTLE
BIT.
WE NEED TO HEAR OUR LAST
SPEAKER.
>> RIGHT, RIGHT.
>> WHAT WE LEARNED FROM THIS THE
IDEA WE START OUT JUST LOOKING
AT THE DNA CONTENT AND RELATIVE
TO OTHERS AND SAYING OH, YOU
HAVE HIGHER LEVELS IT SHOULD BE
MORE IMMUNOGENIC BUT IT'S MORE
THAN THAT AND THEY CONTROL DNA
DAMAGE RESPONSES.
SO WHAT WE END UP THINKING OR
LOOKING AT IS THE IDEA THAT IF
THE VIRUS CANNOT CONTROL DNA
DAMAGE RESPONSE THEN THEY ARE
IMMUNOGENIC.
THESE VIRUSES HERE THEY CAN'T
CONTROL THE RESPONSE THE DNA
DAMAGE RESPONSE.
AND THIS WAS IMPORTANT FOR THE
VIRUSES AND CAN INHIBIT THIS AND
LOSE THIS.
AND WHAT WE DISCOVERED WAS THE
SIGNALLING PATHWAY THAT OCCURS
IN THESE INFECTED CELLS SUCH AS
THE SIGNAL TO THE KINASE THAT
STIMULATES THE MF KAPPA B AND
E43 INHIBITS PML AND ALLOWS ATM
TO AGAIN ACT ON NF KAPPA B.
WORK TOGETHER THEY REPROGRAM NF
KAPPA B AND HAVE YOU MORE CELL
SURVIVAL AND IT'S MORE ON
SURVIVAL AND LESS ON IMMUNE
RESPONSE AND HERE LESS SURVIVAL
AND MORE IMMUNE RESPONSE.
WE TESTED THE IDEA THAT IF WE
DELETE THE SIGNAL HERE THEN WE
SHOULD GET MORE IMMUNE RESPONSES
AND SO WE DELETED E41 SHOWING
THE DELETION AND IT CHANGED TO
SOME LEVEL OF CYTOKINES AND THAT
WAS ENOUGH TO SAY LET'S GO ON TO
THE NON-HUMAN PRIMATES.
WE IMMUNIZED NON-HUMAN PRIMATES
AND WHEN WE LOOKED AT THE
MUCOSAL MEMORY CELLS, CD4, CD8
IT INDUCED HIGHER LEVELS OF
CYTOKI
CYTOKINES.
WE LOOKED AT ANTIBODIES, THIS IS
IG, THERE'S NOT MUCH DIFFERENCE
BUT WHEN WE LOOK AT IG1 THE
RELEVANT IG1 ANTIBODY OR IGA --
>>
[OFF MIC]
>> AND THE LAST SLIDE WAS WE
SHOWED YOU ADENOVIRUS HAS THE
SAME SURFACE PROTEIN WE CAN
CHANGE THE GENES AND WE CAN TEST
THEM IN MICE AND INFORM US FOR
WHAT WE SHOULD SEE IN MICE AND
THAT INFORMS US FOR WHAT WE
SHOULD SEE IN NON-HUMAN PRIMATES
AND DATA SUGGESTS SO FAR IS
ADENOVIRUS EARLY GENES CONTROL
INNATE MUCOSAL AND SYSTEMIC
VACCINE INDUCED IMMUNE RESPONSES
AND THEY DID MOST THE WORK WITH
THE VACCINE AND A LOT OF THIS
STARTED THAT ALLOWED ME AND ITS
WAS FUN FOR ME AND THIS IS WHAT
WE HAVE DONE SINCE THEN.
>> THANK YOUVIC.
A LOT OF WORK AND IF THERE'S NO
PRESSING QUESTIONS I THINK WE
SHOULD GO STRAIGHT TO LAST L LASTLY
ABIMIKU WAS THE FIRST POST-DOC
OF MARJORIE AND GOING TO
BETHESDA TO WORK WITH BOB AND
THEN THERE WAS A BRILLIANT IDEA
TO WORK WITH MARJORIE AND THE
REST IS HISTORY.
THANK YOU FOR COMING.
>> THANK YOU FOR HAVING ME.
THOUGH I'M AN IN UNCOMFORTABLE
POSITION BECAUSE I'M STANDING
BETWEEN YOU AND LUNCH AND EARLY
LUNCH IT WAS SUPPOSED TO BE BUT
I HAVE MARJORIE STILL TO PROTECT
ME AGAINST BOB.
AND TODAY I THOUGHT IT SHOULD BE
IT'S APPROPRIATE TO SHARE WITH
THE AUDIENCE HOW MUCH MARJORIE'S
IMPACTING WHAT WE DO GLOBALLY AS
WELL IN TERMS OF WHAT SHE'S DONE
IN RESEARCH.
AND WE HAVE THE NORTH SIDE
COLLABORATION STARTING WITH THE
POST-DOC IN MARJORIE'S LOB.
AND THERE'S AN GLOBAL
INTERNATIONAL PROGRAM AND
CURRENTLY WE'RE IN EIGHT AFRICAN
COUNTRIES DOING LARGELY WORK IN
HIV AND TB.
OUR PROGRAM INCORPORATES A LOT
OF ACTIVITY AND THE PEP FAR
PROGRAM WHICH HAS PROVIDED
TREATMENT FOR INDIVIDUALS THAT
ARE INFECTED WITH HIV.
WE HAVE A LOT OF WE'VE DONE AND
WE HAVE HEALTH SYSTEMS TO ENSURE
THE DATA BEING PRODUCED IS
APPROPRIATE BY SECURITY AND ONE
AREA THAT WE HAVE ALSO BUILT IN
MORE RECENT YEARS AND AS A
RESULT OF OUR COLLABORATION IS
THE AREA OF RESEARCH.
THIS IS MOST DEVELOPED IN
NIGERIA WHERE I'M FROM.
IN NIGERIA, WHAT WE'VE DONE IS
THAT WE BUILT A PROGRAM THAT IS
BASED ON TREATMENT OF HIV
INFECTED INDIVIDUALS MOST
CO-INFECTED WITH TB BUT ALSO
HAVE A GLOBAL FUND PROGRAM THAT
FOCUSSED ON TB.
DUE TO THE COLLABORATION WITH
THE NORTH, IT WAS IMPORTANT THAT
WE OVERLAID SOME OF THE
ACTIVITIES THAT WE'RE DOING FOR
SERVICE PROGRAM USING THE
INFRASTRUCTURE THAT WE'VE BUILT
TO ALSO PROMOTE RESEARCH.
BUT IN TERMS OF RESEARCH AND
THIS IS PART OF MY FOCUS IN 2016
AND THE INSTITUTE OF HUMAN
VIROLOGY IN NIGERIA WAS SET UP
IN 2004.
OVERLAYING A LOT OF WORK WE'VE
DONE WITH RESEARCH WAS MY FOCUS
IN 2016 THAT WE ESTABLISHED THE
INTERNATIONAL RESEARCH CENTER OF
EXCELLENCE.
AND ONE THING IN TERMS OF THE
RESEARCH AREA IS TO FOCUS ON
CLINICAL TRIAL COHORT STUDIES
AND MOLECULAR IMMUNOLOGY AND
FOCUSSING ON HIV AND TB.
AND THEN WE PARTNERED WITH
INTERNATIONAL RESEARCH
INDIVIDUAL AND MORE IMPORTANTLY
ALSO WITH ACADEMIC FACULTY
WITHIN THE COUNTRY.
THE PURPOSE OF SETTING UP THIS
IS TO ENSURE RESEARCH IS
PROMOTED AND MAINLY FOCUSSING
WITH THE INSTITUTE OF HUMAN
VIROLOGY AND THE UNIVERSITY OF
MARYLAND WITH HIV NIGERIA
PROVIDING ONE-CLASS PLATFORM TO
MAKE SURE IT'S USING
INTERNATIONAL SCIENTIFICAL AND
ETHICAL NORMS AND RESPECTING THE
NORMS WITHIN THE COUNTRIES.
IT WAS MAINLY TO FIRST
COLLABORATE BETWEEN THE NORTH
AND SOUTH BUT ALSO WITHIN THE
NIGERIA AREA RESEARCHERS AND
LIKE I SAID INTERNATIONAL
RESEARCH INSTITUTES.
BUT YOU KNOW, WE DIDN'T START
WITH SUCH ELABORATE RESEARCH
CAPACITY.
OUR INTERACTION AND SETTING UP
THE COLLABORATIONS AND
INFRASTRUCTURE IN NIGERIA
STARTED REALLY IN 1991 WHEN I
CAME TO THE LTCB LABORATORY FOR
MY POST-DOC.
THE USUAL NUMBER WAS THAT IT
WOULD ALLOW TO YOU CIRCLE
THROUGH THE LABORATORIES AND HIS
SENIOR SCIENTISTS AND THEN YOU
HAD A MEETING WITH HIM AT THE
END OF IT TO KIND OF INFORM HIM
OF THE LABORATORY YOU'VE DECIDED
TO WORK IN.
AND I REMEMBER CERTAINLY THROUGH
AND HAVING AN EXTENDED
DISCUSSION WITH MARJORIE, I WAS
CERTAINLY INTERESTED IN
IMMUNOLOGY BECAUSE THAT WAS PART
OF MY TRAINING AS MASTER'S
BEFORE I DID MY Ph.D. BUT THE
OTHER ASPECT I WAS REALLY
INTERESTED IN WAS SETTING UP AN
INFRASTRUCTURE IN MY HOME
COUNTRY TO ALLOW ME TO CONTINUE
TO DO RESEARCH.
AND THAT WASN'T THE KIND OF
THING YOU DISCUSS WITH BOB GALLO
WHEN COUPLE TO HIS LAB BECAUSE
IT'S A VERY RENOWN LABORATORY ON
MAINLY RESEARCH.
CAPACITY BUILDING AT SOME STAGE
BUT YOU KNOW EVENTUALLY BUT THE
FIRST FOCUS HAS TO BE RESEARCH.
IT WAS WONDERFUL TO HAVE
MARJORIE THAT WANTED TO SUPPORT
ME AND WITH ME TREMBLING AT THE
KNEES AS SHE WAS ABLE TO
CONVINCE BOB THAT I COULD DO
BOTH.
AND SO I GOT SUPPORT FROM BOTH.
THAT BUFFER STILL WORK TODAY.
AND JUST TO MENTION THAT
ACTUALLY OUR FIRST ENTRY INTO
NIGERIA WAS CONVINCING THE
GOVERNMENT THEN THAT THERE WAS
HIV.
THIS WAS REALLY AS A RESULT OF
THE WOULD BLOOD TEST THAT WAS
DEVELOPED IN BOB'S SLIDE AND WE
STARTED WITH A ROOM AND TWO
DOORS AND TWO ROOMS I REMEMBER
HAVING THE NATIONAL TELEVISION
COME IN AS I DRAW BLOOD AND TEST
INDIVIDUALS TO CONVINCE THE
COUNTRY THEN THAT THERE WAS HIV.
WE'VE MOVED SINCE THEN.
WE HAVE A TWO-STORY RESEARCH
FACILITY IN 2001 AND IT PROBABLY
WAS THE MAIN RESEARCH FACILITY
THAT WAS DOING HIV TESTING AND
CONFIRMATION FOR MOST OF THE
NORTHERN PART OF THE COUNTRY.
SUBSEQUENTLY NOW WE HAVE TWO
TWIN TOWER BUILDINGS, EIGHT
FLOORS EACH, WHICH IS ON BOARD
AND WE'VE CONTINUED TO BUILD THE
RESEARCH PROGRAM.
BUT HOW DID MY POST-DOC IN
MARJORIE'S LAB TRAIN ME FOR SOME
OF THE THINGS I WAS THEN ABLE TO
DO?
LIKE I MENTIONED EARLIER, ONE OF
MY INTERESTS WAS REALLY BUILDING
CAPACITY AND INFORMATION THE
EPIDEMIC IN MY HOME COUNTRY AS
WELL.
AND THIS PAPER IS ONE THAT WAS
PUBLISHED THE FIRST SUBTYPES IN
NIGERIA SHOWING VERY CLEARLY
THAT IT WAS A SUBTYPE GROUP GENE
AND LATER ON IT IS A STANDARD TO
AG RECOMBINANT BUT THIS G SUB
TYPE HAS CONTINUOUSLY REMAINED
AS THE PREDOMINANT STRAIN AND
ONE OF THE THINGS THAT MARJORIE
DID WAS REALLY TO LINK ME UP
WITH OTHER MEMBERS OF THE TEAM
AND I LEARNED A LOT OF MOLECULAR
VIROLOGY IN THE LAB AS WELL.
WE WENT ON SUBSEQUENTLY TO
ESTABLISH AND DID AGAIN THE
COLLABORATIONS THAT REALLY WAS
BROUGHT TOGETHER AS A RESULT OF
MARJORIE.
BUT ALSO IN WORKING WITH THE
RHESUS MACAQUE GIVES ME AN
OPPORTUNITY TO LEARN A LOT OF
CELL MEDIATED IMMUNITY THAT
LATER ON DESCRIBED FROM THE
FOUNDATION OF WHAT WE'VE
ESTABLISHED IN NIGERIA.
SO WHAT DO WE HAVE NOW?
WE HAVE A GROUP OF FACULTY AND
WE'VE CONTINUED TO COLLABORATE
IN INTERNATIONAL RESEARCH.
WHAT ARE THE AREAS THAT WE'RE
FOCUSSING ON AS I MENTIONED
EARLIER OUR FOCUS IS ON HIV AND
WHAT YOU SEE HERE THAT IS
STARRED AND NIH FUNDED GRANTS
WE'VE BEEN ABLE TO GET AND WHAT
I'M SHOWING HERE IN RED IS SOME
OF THE FUNDING WE'VE GOTTEN
THROUGH NIH TO BUILD THE
CAPACITY FOR TRAINING AND
RAISING UP POST-DOC AT
INDEPENDENT RESEARCHERS.
OUR FUNDING HAS CONTINUED TO
EXPAND.
IN 2016 THE FUNDING WE HAS WAS
AS A RESULT OF COLLABORATION
WITH A NUMBER OF THE FUNDED AS
NIH BUT SINCE IT'S A LOW AND
MIDDLE INCOME COUNTRIES THERE'S
OTHER FUNDING MECHANISMS IN
EUROPE AVAILABLE TO FACULTY IN
NIGERIA NOT NECESSARILY
AVAILABLE TO OUR COLLEAGUES HERE
AT IHV.
WE'VE BEEN ABLE TO EXPAND AND
SOME IS INITIATED FROM OUR
RESEARCH CENTER IN NIGERIA.
SOME OF ANY NEXT SLIDES WILL
POINT TO SOME OF THE THINGS THAT
HAVE REALLY MADE AS OUR ABILITY
TO CONDUCT RESEARCH AT AN
INTERNATIONAL LEVEL WITHIN
RESOURCE LIMITED COUNTRY.
SOME OF THEM HERE PEOPLE TAKE IT
FOR GRANTED BECAUSE HAVE YOU
POWER ALL THE TIME AND SOME HAVE
HAD FOUR POWER BACKUPS TO MAKE
SURE WE HAVE POWER ALL THE TIME
AND ON TOP OF THAT WE HAVE A
LICK ID NITROGEN PLANT JUST IN
CASE AND THE VSL3 FACILITY WAS A
CONTINUIZED LABORATORY PUT
TOGETHER HERE IN FLORIDA AND
SHIPPED AND ASSEMBLED AND WE'RE
ABLE TO DO SEQUENCE IN THE
FUNDED LAB.
THE WORK WE DO INCLUDES -- OKAY.
A SHARED FACILITIES ESPECIALLY
ENSURING THE DATA IS OF HIGH
QUALITY.
WE REALLY HAVE TAKEN ADVANTAGE
OF OUR INTERNATIONAL PARTNERS
AND COVID HAS TAUGHT US HOW TO
DO A LOT OF THINGS VIRTUALLY.
SO THERE'S A LOT OF TRAINING
ESPECIALLY IN DATA THAT HAPPENS
VIRTUALLY AND HERE WHAT I'M
SHOWING IS A LOT OF THE
TRAININGS THAT WE DO ALSO.
SO THE LAST FEW SLIDES I WOULD
LIKE TO JUST UNDER SCORE HOW
MUCH WE HAVE BUILT CAPACITY
ESPECIALLY RESEARCH CAPACITY
THROUGH THE FOGARTY PROGRAM.
THIS REALLY STARTED WITH BILL
BLATNER THE CO-FOUNDER WITH BOB
AND HAD ABOUT 15 YEARS WHERE A
NUMBER OF MASTERS AND Ph.D.
INDIVIDUALS WERE TRAINED.
A LOT OF THE COURSES ARE
RELEVANT TO THE KIND OF RESEARCH
THAT WE DO TODAY.
BUT WHAT IS INTERESTING IS WE'VE
CONTINUED TO BUILD ON BILL'S
FOUNDATION AND WE'RE ABLE TO
ALSO CONTINUE THIS TO TRAIN
ADDITIONAL MASTER'S IN Ph.D. AND
WE HAVE THOSE MASTER'S IN Ph.D.
STATIONED OR ABSORBED INTO OUR
RESEARCH CENTER AT HIV IN
NIGERIA.
THAT WAY WE DO NOT HAVE THEM ALL
SCATTERED AROUND AND WE BEGIN TO
BUILD ON THAT POOL.
AND MORE RECENTLY IN FACT IT WAS
FEBRUARY THIS YEAR, NADIA AND I,
BY THE WAY NADIA -- AND USED TO
CALL ME THE AFRICAN WOMAN WHEN I
JOINED HIS LAB BUT NADIA AND I
GOT ANOTHER FOGARTY PROGRAM
FOCUSSING ON IMPLEMENTING
SCIENCE AND THE PLAN IS TO
CONTINUE DEVELOP THE LONG-TERM
POST-DOCTORAL GETTING TO THE
CORE OF INDIVIDUALS THAT WOULD
THEN NEED TO USE THE CORE
INFRASTRUCTURE THAT WAS SET UP
IN THIS RESEARCH BUILDING TO
CONTINUE TO ALSO COMPETE FOR
THIS VARIOUS RESEARCH
ACTIVITIES.
AND LASTLY WE -- THIS JUST SHOWS
IN THIS RECENT ONE SOME OF THE
MODELS WE'LL BE FOCUSSING ON ONE
CERTAINLY ON IMPLEMENTATION
SCIENCE AND WE ALSO WANT TO
FOCUS ON GRANT WRITING AND
MOVING THE INDIVIDUALS TO
INDEPENDENT RESEARCH.
THAT'S LEARNING FROM SOME OF THE
EXPERIENCES I'VE GONE THROUGH.
LASTLY, WE DID COMPETE FOR THE
DATA SCIENCE IN AFRICA AND WE'RE
VERY PROUD OF THIS.
LIKE I SAID, THESE ARE THE BIG
5.
I DON'T KNOW HOW MANY IN THE
AUDIENCE CAN IDENTIFY THE
ELEPHANT BUFFALO LIONESSE AND
WE'RE PROUD OF THIS AND WE JUST
GOT FUNDED IN OCTOBER.
AND WE'LL ESTABLISH THE
MULTI-LAYER INTERACTIONS BETWEEN
COVID AND SOCIAL INTERACTIONS
AND WE'RE USING TOOLS AND
MOBILITY DATA TO SEE HOW WE CAN
OVERLAY THE TRANSMISSION AND THE
SPREAD AND THE HOT SPOT OF COVID
WITH THE DATA THAT HAS BEEN
ACCUMULATE
ACCUMULATED AND OVERLAY THE HIV
AND SEE THE IMPACT OF AN
EXISTENT PANDEMIC ON ONE COMING
IN.
THIS IS INFORM AFRICA.
WE HAVE FIVE YEARS OF DOING DATA
SCIENCE WHICH IS VERY EXCITING
FOR US.
SO I'LL JUST END BY THANKING
YOU, MARJORIE, FOR EVERYTHING
YOU'VE DONE.
ONE THING I LEARNED DURING THE
POST-DOC IN YOUR LAB IS THE
IMPORTANCE OF EXCELLENCE AND THE
IMPORTANCE OF BUILDING OTHER
PEOPLE UP.
I KNOW YOU TRIED TO TEACH ME
PATIENCE BUT I STILL YELL AT
PEOPLE SO I STILL HAVE SOME WORK
TO DO BUT JUST TO SHOW THAT WHAT
YOU HAVE INVESTED IN ME IS
REALLY HAVING AN IMPACT GLOBALLY
SO WE WANT TO THANK YOU FOR THAT
AGAIN.
THANK YOU.
[APPLAUSE]
>> SO I THINK WE SHOULD TAKE
ADVANTAGE OF YOUR PRESENCE HERE
TO LEARN MORE ABOUT AFRICA AND
THE BEAUTIFUL WORK THAT IS
HAPPENING THERE.
IT'S AN EXTREMELY IMPORTANT
PROJECT LONG TERM AND WE SEE HOW
IMPORTANT IT IS NOW WITH ALSO
ADDRESSING AS YOU JUST MENTIONED
IN PASSING, COVID IN AFRICA.
PERSONALLY, I WOULD LOVE TO KNOW
A LITTLE BIT MORE ABOUT WHAT IS
HAPPENING IN AFRICA IN COVID.
WE HEAR A LOT OF DIFFERENT
THINGS AND IT WOULD BE GREAT IF
YOU CAN TELL US MORE SINCE YOU
WERE SO MUCH BELOW YOUR ASSIGNED
TIME YOU'RE NOT FREE TO GO YET.
>> I WAS TERRIFIED OF DELAYING
LUNCH.
I'M GLAD I DID SAVE US SOME
TIME.
I THINK THE WORLD REALLY KNOWS
WHAT IS HAPPENING IN SOUTH
AFRICA BECAUSE THERE'S GOOD DATA
AND GOOD INFRASTRUCTURE THAT'S
COME --
>> THERE ARE OTHER PLACES.
>> PLACES LIKE NIGERIA OF 200
MILLION THAT WE DO NOT SEE A
SIGNIFICANT AMOUNT OF
SYMPTOMATIC COVID.
I KNOW THERE IS UNDER REPORTING.
I KNOW THERE'S UNDER TESTING BUT
YOU CANNOT ARGUE WITH THE FACT
THAT THE HOSPITALS AT SOME STAGE
WHEN COVID STARTED WERE ALL
PACKED WITH INDIVIDUALS THAT
WERE REALLY SICK AND DYING OF
COVID AND THOSE ISOLATION UNITS
ARE PRETTY MUCH EMPTY NOW.
VACCINATION LEVEL IS VERY LOW.
IT'S 8% TO 10% OF INDIVIDUALS
THAT ARE VACCINATED.
WE KNOW FOR A FACT THAT THE
RURAL AREAS BECAUSE MOST OF THE
TIME EVERYBODY'S OUTDOOR IN THE
PONDS AND EVERYTHING HARDLY USE
ANY MASK.
VERY LITTLE COMING FROM RURAL
AREAS.
MOST THE INFECTIONS ARE WITHIN
THE CITIES AS YOU WOULD EXPECT
BUT REALLY NOT AS MUCH
DEVASTATION THAT WAS PREDICTED.
>> BUT HOW MUCH OF THAT DO YOU
THINK IS CAUSED BY THE YOUNG
POPULATION WHICH IS TOTALLY
DIFFERENT THAN EUROPEAN
ETCETERA, CHINESE SOCIETIES.
>> I THINK AT THE BEGINNING OF
THE PANDEMIC THAT WAS THE
BIGGEST IMPACT UNTIL THE VARIOUS
MUTANTS STARTED INFECTING THE
YOUNGER POPULATION AND I KNOW
THERE'S SOME NERVOUSNESS IN
TERMS OF THE LOW VACCINE
COVERAGE AND THERE'S A STRONG
PUSH TO IMPROVE ON THAT.
BUT I THINK A LOT OF THE OUTDOOR
KINDS OF INTERACTIONS WOULD HAVE
HELPED A LOT.
>> HOLD ON.
NOT EVERYONE IS IN THE TREES IN
NIGERIA.
>> IF YOU GO TO THE RURAL AREAS
HARDLY ANY MASK AT ALL AND YET
IT'S VERY LOW.
MOST THE RURAL AREAS ARE -- I GO
WITH MY MOM AND EVERYBODY'S IN
THE CORRIDOR BECAUSE IT'S SO
WARM.
>> WHAT IS HAPPENING WITH
VACCINATION IN AFRICA?
A YEAR AGO IT WAS THE MOST
URGENT THING IN THE WORLD AND
SOME ARE SAYING HOLD ON TO THE
VACCINES UNTIL WE BUILD
INFRASTRUCTURE.
WHAT DO YOU THINK IS THE ROLE IN
P
PEPFAR OR HOW MUCH HELP IS
NEEDED AND WHAT'S HAPPENING WITH
THE NEW VARIANTS?
AS YOU SAID THE NEW VARIANTS ARE
DOING A TOTALLY DIFFERENT JOB
THAN INITIALLY.
>> ABSOLUTELY.
THESE ARE ALL VERY GOOD POINTS.
ONE THING PEPFAR HAS DONE REALLY
WELL IS TO MOVE AND USE THE
INFRASTRUCTURE FOR ALL THE PCR
TESTING.
FOR INSTANCE, ALL OF OUR
LABORATORIES THAT WERE FOR VIRAL
LOADING AND EARLY DIAGNOSIS WERE
ALL CONVERTED TO TESTING FOR
COVID.
NOW WE HAVE THE ANTIGEN-BASED
TEST THAT IS HAPPENING.
I THINK THE NEGATIVE PART OF THE
HARM THAT WAS DONE IN TERMS OF
VACCINE UPTAKE IS SOCIAL MEDIA.
AND ALL OF THE -- THERE ARE SO
MANY RELIGIOUS COUNTRIES.
NIGERIA IS DEFINITELY RELIGIOUS
AND WHEN YOU HAVE PASTORS AND
INDIVIDUALS SAYING THIS IS NOT
GOOD FOR YOU THEN IT REALLY
IMPACTS.
SO ONE OF THE THINGS THEY'RE
DOING THAT IS REALLY TRYING TO
EDUCATE.
IN TERMS OF CAPACITY IT'S LIKE
YOU MENTIONED THE FREEZERS AND
EVERYTHING THAT HAS BEEN PUMPED
INTO A LOT OF THE COUNTRIES
THROUGH PEPFAR HAVE BEEN
SUPPORTED.
>> DO YOU THINK THAT THERE WILL
BE MORE THAN USUAL RESISTANCE TO
VACCINATION BECAUSE OF SOME OF
THE REASONS THAT YOU MENTIONED?
>> I THINK THERE'S STILL A LOT
OF RESISTANCE.
THERE'S A LOT OF EDUCATION THAT
HAS TO GO IN AND MORE AND MORE
PEOPLE -- IT'S REALLY SAD
BECAUSE YOU WONDER IF WE HAVE
HAD A WORSE IMPACT MAYBE PEOPLE
WOULD HAVE TAKEN VACCINATION
MORE SERIOUSLY.
YOU DON'T WISH THAT FOR ANYBODY
SO IT HAS TO BE A LOT OF
EDUCATION AND YOU'VE GOT TO
BRING ALL THE OPINION LEADERS
AND THEY MUST PENETRATE -- WE
MUST PENETRATE THE CHURCHES AND
THE MOSQUES OTHERWISE WE'RE NOT
GOING GET THE VACCINATION.
>> OKAY.
ANY OTHER QUESTIONS?
BOB IS HUNGRY.
OKAY.
THANK YOU SO MUCH.
>> LET ME INTRODUCE THE SPEAKER,
FROM THE COALITION OF ACADEMIC
PREPAREDNESS FROM NORWAY WHO
WILL TELL US ABOUT NON-UTILIZING
ANTIBODIES FOR EPIDEMIC
PREPAREDNESS.
THANK YOU.
>> GOOD AFTERNOON, EVERYONE.
WHAT A CHALLENGE TO BE THE FIRST
SPEAKER AFTER LUNCH.
I WILL TRY TO KEEP US ALL AWAKE.
FIRST THANK YOU TO THE
ORGANIZERS FOR INVITING ME TO
PARTICIPATE IN THIS SYMPOSIUM.
I HOPE THAT IT'S -- I ADD TO
CELEBRATION OF MARJORIE'S CAREER
AND I WILL TRY TO BE BRIEF IN
THE PRESENTATION OF WHAT
REGARDING ADCC PRIMARILY, ADCC
AND HOPEFULLY SHED REFLECTIONS
ON ADCC AND EPIDEMIC
PREPAREDNESS INNOVATIONS.
THIS IS THE OVERVIEW OF MY TALK,
I WILL TALK PRIMARILY ABOUT
ADCC, HISTORY OF ADCC IN
MARJORIE'S LAB AND BRIEFLY
MENTION OTHER FC MEDIATED
MECHANISMS OF PROTECTION.
I WILL END ONSO I THINK
AS A BRIEF INTRODUCTION OF WHAT
WE WILL TALK ABOUT HERE I WANT
-- I RAN A QUICK SEARCH JUST
PUTTING IN THE WORDS HIV AND
HDCC ON PUBMED, YOU GET ROUGHRY
150 PUBLICATIONS BETWEEN 1985
AND RECENTLY.
FROM THIS GRAPH THEY PRETTY MUCH
LIE WITHIN THREE PERIODS OF
TIME.
THE FIRST PERIOD OF TIME WAS
SHORTLY AFTER THE VIRUS WAS
DISCOVERED AND FOR THE FIRST 15
YEARS YOU SEE PUBLICATIONS FROM
SWEDEN.
THERE'S A LOT OF OBLIGATIONS
FROM (INAUDIBLE) LAB, SEVERAL
PUBLICATIONS FROM U.S. FOLKS AND
COUPLE OF PUBLICATIONS FROM
CANADIAN FOLKS.
THIS IS THE FIRST 15 YEARS OR
SO.
THEN YOU CAN SEE THAT THERE WAS
A PERIOD OF TIME WHEN
PUBLICATION, I WOULDN'T SAY THEY
SUBSIDE BUD THEY WERE LOWER.
BUT NOT LESS IMPORTANT.
OF COURSE, THIS IS ACTUALLY A
PERIOD OF TIME WHEN MARJORIE
STARTED PUBLISHING ON THIS, YOU
ALSO HAVE SEVERAL OTHER NAMES
FROM THE US. DAVID MONTEFURI,
DENISE BURTON AND YOU CONTINUE
TO HAVE CANADIAN FOLKS, YOU
BEGIN TO HAVE AUSTRALIAN FOLKS,
SOME IN SWITZERLAND BUT MOST
IMPORTANTLY GETTING INITIAL DATA
ON MULTI-CENTER TRIALS WHICH IS
QUITE IMPORTANT.
THEY START EXAMINING THE ROLE OF
ADCC IN CONTEXT OF HIV AIDS AND
HIV VACCINE TRIALS.
THEN YOU HAVE I WOULDN'T CALL
EXPLOSION BUT A LARGER SET OF
PUBLICATIONS HERE AND THIS IS
WHERE YOU ENTER A -- THE BROADER
INTERNATIONAL SCENE I WOULD SAY
BEYOND JUST THE U.S. AND EUROPE
YOU START SEEING A LOT OF
PUBLICATIONS FROM CHINA FOR
EXAMPLE, YOU SEE A LOT OF GROUPS
PUBLISHING IN SYSTEMS IMMUNOLOGY
APPROACHES AND WE WILL HEAR THAT
FROM OTHER SPEAKERS.
BUT AGAIN JUST TO SAY THAT
NON-NEUTRALIZING ANTIBODY
RESPONSES OF COURSE GO BEYOND
ADCC.
THIS IS PERHAPS THE THIRD PERIOD
OF TIME SHED LIGHT ON ADDITIONAL
MECHANISMS OF POTENTIAL
PROTECTION.
SO I WAS VERY HONORED TO BE IN
MARJORIE'S LAB IN -- TRYING TO
SEE THE POINTER HERE.
I WAS IN MARJORIE'S LAB HERE.
IN THIS PERIOD, MORE OR LESS
BETWEEN 2001 AND 2005 BUT
SEVERAL PUBLICATIONS OF THE WORK
THAT WE DID TOGETHER HAPPENED
MUCH LATER THAN THAT, THEY
CONTINUE HAPPENING AND I WAS
VERY HAPPY AND HONORED TO BE
PART OF THE LAB DURING THAT
TIME.
I THINK ONE OF THE ITEMS THAT I
WANTED TO GET ACROSS HERE WAS
WHEN I ARRIVED INTO MARJORIE'S
LAB I DID ASK -- I REMEMBER
ASKING MARJORIE IS ANYONE
LOOKING AT ADCC?
SHE SAID WE HAVE LOOKED AT IT
BUBO PRETTY MESSY ASSAYS THEY
REQUIRE CHROMIUM RELEASE, I
DIDN'T WANT TO WORK WITH
CHROMIUM RELEASE ASSAYS.
I WANTED TO AVOID THEM.
SO SHE SAID WHY DON'T YOU COME
UP WITH WHAT AN ASSAY, VERY
SUPPORTIVE OF THIS PROCESS
COMING UP WITH SAY ASSAY AND
WHAT WE STARTED TO DOING IS
ADAPTING ASSAY AND REBRANDED FOR
ADCC AND WE NAMED IT RAPID FLOUR
METRIC ADCC ASSAY.
IT WAS SIMPLE, LOOKS RUDIMENTARY
COMPARED TO WHAT OTHERS ARE
DOING BUT AT THE TIME WE DID
BASICALLY LABELING DOUBLE
LABELING TARGET CELLS, THESE ARE
TARGET CELLS WE WERE DOUBLE
LABELING WITH MEMBRANE BODY
WHICH IS PKH 26.
PERMEABILITY WHICH IS CFSC.
WHAT YOU WOULD SEE BY LABELING
THE EFFECTOR CELLS YOU SEE IN
THIS QUADRANT AND THEY ARE UNDER
TWO FLOW CYTOMETRY ASSAY.
IN THE PRESENCE OF ADCC ANTIBODY
YOU SEE KILLING AND BY GATING
YOU WOULD BE ABLE TO CALCULATE
THE PERCENTAGE OF KILLING.
AND YOU WOULD HAVE CONTROLS IGG
1 CONTROL WHICH WHICH WE THOUGHT
WAS APPROPRIATE BECAUSE IT
ALLOWED US NOT TO WORK WITH
RADIO ACTIVITY WHICH I DID NOT
WANT TO DO AND IT WAS SIMPLE FOR
THE TIME, ALSO SEEMED RELATIVELY
HIGH THROUGH PUT AND ENABLED US
TO START ANSWERING SOME OF THE
QUESTIONS THAT WE WANTED TO
ADDRESS.
SO THE FIRST PUBLICATION THAT WE
GOT OUT USING THIS ASSAY, THIS
IS THE PUBLICATION FROM JOURNAL
IMMUNOLOGY IN 2006.
WE WENT BACK AND LOOKED AT SERUM
SAMPLES FROM COLLEAGUES PRIMED
AND BOOSTED, PRIME WITH VARIOUS
ADENOVIRUSES, VACCINES AND
BOOSTED WITH PROTEIN AND WE SAW
IF THEY TESTED THE SAMPLES USING
THIS ASSAY YOU CAN SEE VERY HIGH
LEVELS OF ADCC ACTIVITY,
ACTIVITY ACTUALLY TITRATES ALL
THE WAY -- YOU CAN DETECT SOME
ACTIVITY AT ONE TO 100,000
DILUTION BUT THE LINEAR RANGE I
WOULD SAY WOULD BE AROUND AT THE
TIME AROUND 10 -- 1 IN 10,000.
WE SAW THAT THIS ACTIVITY IN --
WAS PRESENT IN THE -- PRIME
BOOSTED WITH VARIOUS VACCINES,
THIS WAS DISTINGUISHABLE FROM
THE CONTROL ANIMALS WHICH WERE
JUST PRIME BOOSTED WITH
NON-ADENOVIRUS VACCINE, EMPTY
VECTOR VACCINE AND NOT BOOSTED
WITH PROTEIN.
YOU CAN SEE SOME LEVEL OF
ACTIVITIES IN COWS PRIME WITH
ADENOVIRUS SIV AND VACCINE BUT
BOOSTED ONLY WITH VECTOR.
SO THIS WAS THE INITIAL
UNDERSECURITIZATION WE SAW SOME
ACTIVITY.
BUT THE IMPETUS FOR LOOKING AT
THIS THAT MACAQUES WERE LOOKED
AT PREVIOUSLY UNDER OTHER
PUBLICATIONS AND THAT YES KNEW
THERE WAS PROTECTION -- WE KNEW
THERE WAS PROTECTION DURING
ACUTE VIEW REAMIA, IT WAS
IMPORTANT ADENOVIRUS VACCINES AT
LEVEL OF PROTECTION, BUT THIS
COULDN'T BE EXPLAINED BY
NEUTRALIZING ANTIBODIES BECAUSE
SIV IS EXTREMELY DIFFICULT TO
NEUTRALIZE, NEUTRALIZING
ACTIVITY WAS MEASURED BUT NOT
PRESENT.
SO THE IMPETUS, I WANT TO
EMPHASIZE THAT HERE, THE IMPETUS
WAS TRYING TO FIND SOME LEVEL OF
ASSOCIATION OR CORRELATE OF
PROTECTION AGAINST SIVMAC 251 IN
THIS MODEL.
WHAT WE DID TOGETHER DAVID'S
GROUP BY STATISTICS WE STARTED
DRAWING CORRELATIONS AND SAW
THAT ADCC ACTIVITY CORRELATED
WITH LOWER ACUTE VIRAL LOADS.
THIS IS -- THIS WAS TESTED
AGAINST S SIVMAC 251 H 9 CELLS AND
CORRELATED WITH GTB ANTIBODY
BINDING -- GP 120 ANTIBODY
BINDING TITRATES.
SO THIS WAS VERY GOOD FINDING
FOR US, WE CONTINUED TO BE
INTERESTED ON THIS, MARJORIE
CONTINUED TO BE SUPPORTIVE OF
LOOKING AT ADDITIONAL
EXPERIMENTS AND I BELIEVE IN THE
LAB THAT THAT KEPT HAPPENING
QUITE SOME TIME.
ADDRESSING TWO OTHER POSITIONS
LATER BE U STILL ABLE TO
FOLLOW-UP ON SOME EXPERIMENTS,
AND ONE OF THE POINTS I WANTED
TO GET ACROSS WITH THIS SLIDE IS
KINETICS YOU CAN ACTUALLY SEE
SOME MEASURABLE ADCC ACTIVITY,
NOT VERY HIGH BUT YOU SEE SOME
MEASURABLE ADCC ACTIVITY RIGHT
AFTER PRIMING WITH THE VECTOR.
OF COURSE YOU SEE THE ACTIVITY
GOING UP BEING HIGHER AFTER YOU
BOOST AND ESPECIALLY RESPONSE TO
CHALLENGE.
THIS WAS REPLICATED IN HIV AND
ADENOVIRUS HIV RECOMBINANT
VACCINE MODEL USING THE
CHIMPANZEE MODEL.
YOU CAN SEE COMPARISON HERE
WHERE YOU SEE AFTER PRIMING WITH
THE ADENOVIRUS HIV RECOMBINANT
VACCINE YOU SEE AGAIN SOME LEVEL
OF ADCC ACTIVITY, NOT HIGH BUT
YOU SEE IT AND THEN YOU SEE THAT
GOING HIGHER AFTER BOOSTING WITH
GP 140 PROTEIN WHICH IN THIS
CASE WAS GIVEN IN I BELIEVE IN
59 ADJUVANT.
YOU CAN SEE CERTAIN DIFFERENCES
BETWEEN REPLICATION COMPETENT
AND REPLICATION DEFECTIVE HIV.
HIV
VACCINES.
SO THAT IS INITIAL PART THAT I
WOULD SAY WOULD SUMMARIZE THE
INITIAL EXPERIMENTS IN TERMS OF
ADCC AND THAT WERE DONE IN
MARJORIE'S LAB.
FAST FORWARD IN TIME AGAIN LIKE
I SAID, THERE ARE SEVERAL OTHER
HC MEDIATED MECHANISMS OF
PROTECTION.
ADCC'S ONLY PART OF THE PICTURE.
WE SEE IT HERE.
SEVERAL OTHER GROUPS HAVE
DESCRIBED MANY, MANY OTHER WAYS
OF POTENTIAL PROTECTION FROM --
WITH FC MEDIATED FUNCTIONS.
WE ARE INVOLVING FC MEDIATED
FUNCTION.
SO HOW DOES THAT FIT IN TO WHAT
IS HAPPENING NOW?
WHEN I WAS INVITED TO GIVE THIS
TALK IT WAS MORE THAN TWO YEARS
AGO.
AND THIS WAS BEFORE COVID-19 SO
MY TALK WOULD HAVE BEEN VERY
DIFFERENT FROM WHAT I'M
PRESENTING TODAY BUT I DID WANT
TO PUT THIS IN THE CONTEXT OF
HOW THE WORLD IS NOW.
AND WHERE WE ARE POTENTIALLY
GOING.
SO COALITION FOR THE COALITION
FOR EPIDEMIC PREPAREDNESS
INNOVATIONS AND WE ARE TRYING TO
INNOVATE IN THAT SENSE.
ONE OF THE QUESTIONS REGARDING
INNOVATION WOULD BE NEUTRALIZING
ANTIBODIES IMPORTANT IN THE
CONTEXT OF EPIDEMIC PREPAREDNESS
PARTICULARLY PATHOGENS WITH ANY
PANDEMIC POTENTIAL.
MY RESPONSE WOULD BE YES AND I
WILL PRESENT SOME EVIDENCE THAT
IS NOT MINE, I REVIEWED IT
BEFORE COMING TO THE MEETING SO
BEAR WITH ME IF I'M NOT AN
EXPERT BUT THOUGHT IT
APPROPRIATE TO RAISE THIS.
SO THIS IS EVIDENCE FOR EBOLA
AND MARBURG AND THIS IS REVIEWD
BY (INAUDIBLE) AND ERICA
SAPPHIRE.
PUT THE REFERENCE THERE.
BUT IN THE EBOLA MODEL BASICALLY
WHAT THEY HAVE SEEN IS THAT IF
YOU -- THERE HAVE BEEN
ANTIBODIES NEUTRALIZING IN
VITRO.
THEY HAVE VERY POTENT
NEUTRALIZING ANTIBODY IN VITRO
AND THOUGHT THESE ANTIBODIES
WOULD HAVE SOME LEVEL OF
PROTECTION AGAINST EXBOW LA.
THEY DIDN'T.
WHEN THEY TESTED THEM IN EBOLA
MODEL THEY DIDN'T.
WHEN THEY STARTED TESTING
COMBINATIONS OF NEUTRALIZING AND
NON-NEUTRALIZING ANTIBODIES OR
ANTIBODIES POORLY NEUTRALIZING
THEY WERE PROTECTED IN THE EBOLA
MODEL SO THIS IS PIECE OF
SHOWING PANDEMIC POTENTIAL YOU
SEE A ROLE FOR NON-NEUTRALIZING
ANTIBODIES TO HAVE PROTECTION
HERE.
THESE OTHER ANTIBODIES SHOWN IN
THE BOTTOM SHOW THREE PRODUCTS
THAT HAVE BEEN APPROVED BY WHO
FOR EPIDEMIC RESPONSE DURING
EBOLA AND ALL THESE HAVE BEEN
VERY WELL CHARACTERIZED IN TERMS
OF NEUTRALIZING AND
NON-NEUTRALIZING CAPACITY.
WHAT THE AUTHORS CONCLUDE HERE
IS THAT IF YOU LOOK AT THE
GLYCOPROTEIN AT EBOLA
GLYCOPROTEIN YOU CAN DIVIDE INTO
THREE TIERS.
AND THEN THE ANTIBODIES THAT
MEDIATE FC MEDIATED PROTECTION,
PRESUMABLY BIND TO TIER 1 OR
VERY TOP GLYCOPROTEINS AND THE
OTHER ANTIBODIES THAT ARE
NEUTRALIZING AND MAY NOT BE AS
PROTECTIVE OR THAT MAYBE
RECOMMENDATION PROTECTIVE AND
NON-PROTECTIVE BINDING MORE TO
THE BOTTOM PART OF THIS TALK OF
THE GLYCOPROTEIN TALK SO I'M NOT
EXPERT ON THIS, NOT MY DATA BUT
PUTTING THAT REFERENCE THERE FOR
THE INTEREST OF THIS AUDIENCE.
THE CONCLUSION WAS THE
NEUTRALIZATION ALONE WAS
INCOMPLETE PREDICTOR OF IN VIVO
SUCCESS WHICH FOR US IN THE HIV
VACCINE FIELD IS NOT NECESSARILY
SURPRISE.
WHAT ABOUT LASA?
THERE IS ONE PIECE OF EVIDENCE
THAT THIS IS AN ARTICLE
PUBLISHED RELATIVELY RECENTLY.
AGAIN, NOT MY DATA, BUT THIS IS
A MODEL OF RABIES VECTOR
VACCINE, IT IS A RABIES VEHICLER
TO OF LASSA VACCINE.
YOU CAN SEE IN THIS SWISS
WEBSTER PUB MODEL THERE'S
VARIOUS LEVELS OF PROTECTION.
WHEN THEY EXAMINE THE
NEUTRALIZING ANTIBODY ACTIVITY
WHICH IS SHOWN HERE, THERE IS
NOT ANY LASSA VIRUS NEUTRALIZING
ANTIBODY ACTIVITY BUT THEN WHEN
THEY TAKE THESE SERA AND THEY
HAVE VARIOUS LEVELS OF NK CELLS
YOU SEE KILLING OF LASSA VIRUS
INFECTED CELLINGS SO THERE IS
ANOTHER PIECE OF EVIDENCE FOR
LASSA, WHICH IS ANOTHER PATHOGEN
OF EPIDEMIC OR PANDEMIC
POTENTIAL NON-NEUTRALIZING
ANTIBODIES COULD BE IMPORTANT.
I DON'T HAVE TO GO OVER THIS,
GIVE YOU -- YOU HAVE PROBABLY
SEEN MANY PIECES OF EVIDENCE ON
HOW NON-NEUTRALIZING ANTIBODIES
HAVE BEEN INVOLVED IN PROTECTION
AGAINST COVID-19.
THIS IS DATA FROM (INAUDIBLE)
LAB AND THERE IS DATA FROM A
GROUP IN CHINA WHICH THEY LOOK
AT LONGITUDINAL ADCC RESPONSES
AND YOU SEE SEVERAL KINETICS
HAPPENING DURING THE FIRST 21
DAYS OF INFECTION YOU SEE
DIFFERENCES BETWEEN THE PATIENTS
WHO DO BETTER THAN OTHERS.
SO THERE IS A -- SOME
ASSOCIATION BETWEEN ADCC
RESPONSES AND PROGNOSIS.
WHETHER INFECTION IS MILD OR NOT
AS MILD, ALSO SAW EVIDENCE FOR
NEPA VIRUS, HEARSA THIS IS ONE OF MY
FAVORITE VIRUS, THEY ARE NOXIOUS
TO BATS BUT TO US THEY ARE
LETHAL.
THESE ARE THE RANGE -- THIS IS A
FLYING RANGE OF BATS, BUT AREAS
SEQUENCES FOUND IN THE WORLD SO
MANY THINK THE NEXT PANDEMIC
COULD BE BY A NIPAH VIRUS OR
NIPAH VIRUS LIKE VIRUS.
SO THERE HAS NOT BEEN ANY
REPORTS OF ADCC AGAINST NIPAH
VIRUS OR HENDRA VIRUS BUT MAYBE
IMPORTANT TO ADDRESS A QUESTION
OF COULD ADCC PLAY A ROLE IN
ACHIEVING A RAPID RESPONSE OR
RAPID IMMUNE RESPONSE TO VIRUS?
AND THE REASON I ASK THIS
QUESTION IS BECAUSE IF YOU LOOK
AT THE WHO TARGET PRODUCT
PROFILE FOR THESE VACCINE, THEY
WANT YOU TO HAVE AN EFFICACY OF
90%, PREVENTING DISEASE AND TO
HAVE A RAPID ONSET OF PROTECTION
PREFERABLY WITHIN A FEW WEEKS
AFTER ADMINISTRATION.
SO THAT IS A QUESTION THAT I'M
ASKING, WOULD IT BE POSSIBLE
NON-NEUTRALIZING ANTIBODY
RESPONSE COULD CONTRIBUTE TO
PANDEMIC PREPAREDNESS OR
EPIDEMIC PREPAREDNESS AGAINST
THIS VIRUS.
ALSO IN TERMS OF SEPPE, PLEASE
REMIND IF I'M CROSSING MY TIME
BUT IN TERMS OF CEPI MISSION AS
YOU MIGHT HAVE HEARD, CEPI IS
CURRENTLY PREPARING TO --
ACTUALLY CHANGED STRATEGIC PLAN.
AND IT HAS IT WILL BE TRYING TO
ACHIEVE A GOAL TO DEVELOP
VACCINE WITHIN 100 DAYS SO THIS
IS SOMETHING THAT IS RATHER
CONTROVERSIAL BUT HAS BEEN
APPROVED BY BOARD AND SECRETARY
AND ON THIS FIGURE HERE, WHAT
CEPI IS TRYING TO SHOW IS FOR
COVID 326 DAYS TRANSPIRED
BETWEEN IDENTIFICATION OF
SEQUENCE AND DEVELOPMENT AND
APPROVAL, REGISTRATION OR
APPROVAL FOR EMERGENCY APPROVAL
FOR VACCINE SO TRIERING TO BRING
THE TIME TO 100 DAYS.
IS THIS ACHIEVABLE, WE DON'T
KNOW BUT WE CAN TRY AND THE
CONCEPT TO DO THIS WOULD BE TO
FOCUS ON 25 VIRUS FAMILIES AND
TO TRY TO HAVE VACCINES OR
VACCINE LIBRARIES AGAINST 25
PROTOTYPIC VIRUS FAMILIES SO YOU
CAN POTENTIALLY HAVE PREPARED SO
TO SPEAK AND PULL THEM UP TO THE
SHELF IN TERMS IN CASE EPIDEMIC
HAPPENED AND CONTINUE WITH
RESEARCH AND PRODUCTION BUT
BASIC CONCEPT IS THERE.
THE QUESTION TO BE ADDRESSED
HERE COULD ADCC AND OTHER
NON-NEUTRALIZING ANTIBODIES
EFFECTOR FUNCTION PLAY A ROLE IN
ACHIEVING THESE 100 DAY MISSION.
I THINK THEY WOULD -- THEY
SHOULD BE LOOKED AT BECAUSE
AGAIN, IT IS NOT ALL ABOUT
NEUTRALIZING ANTIBODIES.
I THINK THE HISTORY OF MANY
DISEASES HAS SHOWN US THAT MAYBE
THE CASE.
I WANT TO END WITH REFLECTIONS
HERE.
I DON'T KNOW HOW MANY PEOPLE IN
THE AUDIENCE READ THIS BOOK BUT
I HIGH LIE RECOMMEND IT.
THIS IS A BOOK BY ALEX
(INAUDIBLE) AND HE'S THE
PRESIDENT OF WORLD PEACE
FOUNDATION ACTIVITIES
UNIVERSITY, NEW PANDEMICS OR
POLITICS.
IT IS RELEVANT IN THE CONTEXT OF
THIS AUDIENCE BECAUSE SEVERAL --
I WON'T SPOIL THE BOOK BUT
SEVERAL KEY POINTS ADDRESSED IN
THIS BOOK AND ONE IS WHENEVER WE
TRY TO FIGHT EPIDEMIC USING A
MARSHALL SCRIPT LIKE WAR DISEASE
SCRIPT THAT GENERALLY FAILS AND
THE AUTHOR SHOWS YOU MANY
EXAMPLES OF THAT.
BUT WHAT I REALLY LIKE ABOUT
THIS BOOK AND THINK IT IS
PARTICULARLY IMPORTANT IN THE
CONTEXT OF THIS GROUP, HE ARGUES
E MANS PATORY APPROACH USED IN
HIV AIDS MITIGATION ALLOWED US
TO LIVE WITH HIV WITH DISEASE
FOR MORE THAN 40 YEARS.
AND MANY OF THESE HAVE ALSO
ALLOWED US TO COME UP WITH
COUNTER MEASURES AGAINST THE
VIRUS.
BUT THIS EMANCIPATION APPROACHES
INVOLVE COMMUNITY ACTIVISM
EMANCIPATION SCIENCE AND
POLITICS, DIVERSITY INCLUSION
AND THIS WILL LEAD TO ACCESS TO
PREVENTION AND TREATMENT.
AND I THINK THAT IS QUITE
RELEVANT BECAUSE IT IS SOMETHING
THAT WE CAN LEARN FROM HIV AIDS
THAT IS SOMETHING PANDEMIC
PREPAREDNESS CAN BENEFIT FROM IN
LOOKING AT EXAMPLES OF HIV AIDS.
JUST TO FINISH MY TALK CLOSING
REMARKS, THERE IS A WEALTH OF
KNOWLEDGE ON THE ROLE OF ADCC
AND NON-NEUTRALIZING ANN BODIES
AGAINST HIV AIDS.
MARJORIE'S LAB WAS KEY.
IN GENERATING SCIENTIFIC
EVIDENCE ABOUT THIS.
THERE IS CLEAR EVIDENCE OF ROLE
OF ADCC NON-NEUTRALIZING
EFFECTOR MECHANISMS AGAINST
DISEASES OF PANDEMIC AND
EPIDEMIC POTENTIAL, IT IS NOT
ALL ABOUT NEUTRALIZING
ANTIBODIES AND THERE IS AN
OPPORTUNITY FOR ADCC TO PLAY A
ROLE IN HOW WE ARE
CONCEPTUALIZING THE UNIVERSE OF
EPIDEMIC PREPAREDNESS AND
RESPONSE.
LAST BUT NOT LEAST LIKE IN HIV
AIDS, COMMUNITY ACTIVISM
EMANCIPATION POLITICS, SHOULD BE
A PART OF PREPAREDNESS AND
RESPONSE AND IN THIS SENSE
DIVERSITY IN THE SCIENCE
ACTIVISM SHOULD PLAY A ROLE IN
GENERATING A -- I PERSONALLY
BELIEVE YOU CANNOT HAVE ACCESS
IF YOU DON'T INVOLVE ALL THE
COMMUNITIES THAT WILL ULTIMATELY
BENEFIT FROM THE DEVELOPMENT OF
THESE COUNTER MEASURES.
IN THIS REGARD I DO WANT TO SAY
THANK YOU, MARJORIE BECAUSE THE
LABORATORY WHERE YOUR LABORATORY
WAS PROBABLY IS NOT THE MOST WAS
PROBABLY ONE OF THE MOST DIVERSE
LABORATORIES WHERE OPINIONS FROM
EVERYONE WHERE VALUABLE I HAVE
FOND MEMORIES OF MANY PEOPLE
FROM MANY DIFFERENT
NATIONALITIES SITTING AT THE
SAME TABLE WITH YOU DISCUSSING
SCIENCE SOMETIMES POLITICS OR
SCIENCE BUT EVERYONE WAS HURT IN
YOUR LAB AND YOU CREATE AD VERY
HIGH BAR FOR ME AS I MOVE ON IN
OTHER CAREERS BECAUSE I WAS
ACTUALLY QUITE SPOILED BY YOU.
IN TERMS OF BEING ABLE TO HAVE
VERY EQUITABLE ACCESS TO GETTING
AN OPINION AND BE RESPECTED FOR
THE OPINIONS SO THEY HAVE -- SO
THANK YOU VERY MUCH AND I WISH
YOU VERY GOOD LUCK IN THE
JOURNEY AHEAD.
[APPLAUSE]
>> COULD YOU GO INTO MORE
DETAIL, REGARDING THE HUNDRED
DAYS?
WHAT IS THE BOARD THINKING THAT
WOULD -- WHAT SORTS OF THINGS
FROM THE STANDARD OR EVEN
ACCELERATED VACCINE DEVELOPMENT
WOULD THEY MODIFY TO ENABLE YOU
TO GET THERE?
IF IT TOOK 60 DAYS FOR mRNA TO
GET INTO CLINIC, THAT'S TINY
PART OF A YEAR AND ALL THE REST
WAS ACCRUAL OF PHASE 1, 2, 3
DATA AND THEN FINALLY APPROVAL.
SO WHAT THINGS WOULD THEY HAVE,
WOULD THEY BE ABLE TO GO -- HAVE
ALREADY PROTOTYPE PLATFORMS FOR
EXAMPLE THAT WOULD GO STRAIGHT
TO PHASE 2?
OR WHAT ARE THEY THINKING ABOUT?
>> IT IS A COMBINATION AND IT IS
PROBABLY WON'T BE A ONE SIZE
FITS ALL TO BE HONEST.
I THINK IT DEPENDS ON THE
PATHOGEN.
FOR EXAMPLE, IF WE PREFERRED TO
BETA CORONA VIRUS, WE KNOW WE
HAVE SUPPORTED VACCINES
DEVELOPMENT OF VACCINES AGAINST
VIRUS, WE HAVE VIRUS COVE 1 AND
SOME VACCINES COULD BE FAST
FORWARD THAT IN THAT REGARD AND
THEN WE HAVE SARS COV-2 VACCINE
AND VARIANTS.
FROM THAT PARTICULAR BETA CORONA
VIRUS FAMILY THE APPROACH COULD
BE SOMEWHAT DIFFERENTLY THAN
FROM NEPA VIRUS VACCINE APPROACH
WHERE YOU ARE NOT ABLE TO RUN
PHASE 3 TRIALS.
FOR EXAMPLE.
FOR NEPA VACCINES YOU NEED TO
RUN A PHASE 1 PHASE 2 AND THEN
HAVE IT READY IN CASE THE RNA
CHANGE TYPE OF 1 TO TEST IN THE
POPULATION BUT OR PURSUE OTHER
MECHANISMS OF REGULATORY
APPROVAL.
SO I THINK IT IS NOT A ONE SIZE
FITS ALL, DEPENDS ON THE
PARTICULAR VIRUS FAMILIES.
IT WOULD BE A COMBINATION OF
SEVERAL PROCESSES.
>> OKAY.
NO MORE QUESTIONS THEN THANK YOU
VERY MUCH.
VERY NICE PRESENTATION.
GEORGE, SO NOW WE ARE GO GEORGE
LEWIS WHO WILL TALK ABOUT
(INAUDIBLE) THAT WILL TELL US
ABOUT OBSERVATIONAL STRUCTURE
AND FUNCTION OF FC MEDIATED
EFFECTER FUNCTION WITH SPECIAL
THANKS TO MARJORIE.
>> OKAY.
THANKS FOR THE OPPORTUNITY TO
SPEAK AT THE SYMPOSIUM TODAY.
WHAT I'M GOING TO DO IS AS BETH
SAID TELL YOU ABOUT SOME SORT OF
NEW OBSERVATION, SOME VERY NEW
WE HAVE COME UP WITH IN THE
INSTITUTE AND WITH COLLABORATORS
ON STRUCTURE AND FUNCTION FC --
WHAT IS GOING ON WITH FC
MEDIATED EFFECTOR FUNCTION.
I WILL FOCUS ON TROGO CYTOSIS OF
ALL THINGS YOU WILL SEE WHY IN A
MINUTE.
I WANT TO BEGIN BY TALKING ABOUT
SPECIAL THANKS TO MARJORIE, I
WAS AROUND BACK IN THE EARLY
DAYS OF HIV, I WASN'T WORKING ON
IT BUT I WAS GETTING READY TO
GET INTO IT AND I WAS TRAINED AS
A QUASI STRUCTURAL CELLULAR
IMMUNOLOGIST CHEMIST AT HEART.
AND ONE OF THE THINGS THAT YOU
FIRST START LOOKING FOR IS
SOMETHING LIKE THIS AT LEAST I
DO, NEUTRALIZING ANTIBODIES.
THINKING ABOUT IT.
AS BOB POINTED OUT TODAY,
EARLIER, THAT THERE ARE PAPE --
HAIR PAPER WAS THE FIRST
DEMONSTRATION, CLEAR
DEMONSTRATION OF NEUTRALIZING
ANTIBODIES IN AIDS AND AIDS
RELATED COMPLEX, VERY IMPORTANT
PAPER.
BUT I THINK ALSO EQUALLY
IMPORTANT, IS THE FOLLOW-ON
PAPER WHERE THEY SHOWED
DEMONSTRATED NEUTRALIZATION
ESCAPE.
THIS IS SOMETHING THAT I KNEW
ABOUT YEARS AGO BECAUSE WE USED
TO USE HAPPEN CONJUGATED
BACTERIA PHAGOS, SENSITIVE ASSAY
FOR MEASURING ANTIBODIES AND I
KNEW ABOU ABOUT PHAGE NEUTRALIZATION
FROM THEN.
THIS WAS IMPORTANT, THERE'S
STILL SOME INFORMATION IN THE
PAPERS, THERE IS A FOLLOW ON
PAPER FROM MARJORIE AND MARV,
THE FIRST AUTHOR ON IT, WHERE
THEY MAP THE POINT MUTATION THAT
CONTROL NEUTRALIZATION
SENSITIVITY GLOBALLY TO THE
VIRUS.
NOW, WHEN I SAW THIS, I THOUGHT
GREAT, THIS IS TELLING YOU ABOUT
THE STRUCTURE BECAUSE APPARENTLY
IT WAS IN THE NEUTRALIZATION
SITE SO THERE'S SOMETHING ABOUT
THE ENVELOPE STRUCTURE THAT WAS
INTERESTING.
YOU WERE SEEING EPISTATIC
EFFECTS WITH NEUTRALIZATION.
SURE ENOUGH NOW THAT WE HAVE
GOTTEN STRUCTURE, YOU CAN SEE
WHERE THE MUTATION MAPS.
THIS IS A CRYOEM STRUCTURE FROM
STEVE HARRISON, MEMBRANE
REMEMBER FROM HIV AND IF YOU
LOOK THERE WHERE THE YELLOW
ARROWS POINT THAT IS POINT
MUTATION IN -- THE PAPER, IT'S
IN -- THE A IN THE AVERY
SEQUENCE IN THE THING, THIS IS
DEEP IN THE INTERIOR OF THE
MOLECULE.
SO SOMEHOW OR ANOTHER THOSE
RESIDUES POINTS MUTATION ARE
GLOBALLY CHANGING THINGS WHEN
AND I WENT AND LOOKED, WORK A
BIT ON NEUTRALIZATION, ALSO
PARTICULARLY (INAUDIBLE) THE
LEAD PROFESSOR THAT WORK US WITH
US AND I COULDN'T FIND ANYTHING
WHERE ANYTHING HAS TRIED TO
FIGURE OUT WHAT THIS DOES NOW
THAT WE HAVE GOOD PANELS OF
BROADLY NEUTRALIZING ANTIBODIES,
ET CETERA SO SOMETHING WORTH
LOOKING INTO AFTER ALL THESE
YEARS.
INFORMATION GENERATED CLOSE TO
40 YEARS AGO NOW, REALLY
VALUABLE.
THE OTHERWISE I WANT TO THANK IS
FOR THE CONTRIBUTIONS FC
MEDIATOR FUNCTION, AND
PROTECTION AGAINST SIV AND SHIF.
WE TALKED ABOUT IT A MINUTE AGO.
SO BY THE TIME THESE PAPERS CAME
OUT, I FIGURED ALREADY STARTED
IN WORKING WITH TONY AND BOB GAL
LOW AND OTHERS TRYING TO COME UP
WITH VACCINE CONCEPT.
WE HAD ONE THAT CAME OUT OF
TONY'S GROUP.
BUT THE PAPER APPEARED ON
VACCINE ELICITED OR MEDIATE ADCC
THAT CORRELATE WITH PROTECTION.
VIREMIA AND RHESUS MACAQUES.
AND IT WAS USING THE ASSESS THAT
WE TALKED ABOUT RFADC ASSAY, I
THINK IT IS A TERRIFIC ASSAY, WE
USE IT EVERY DAY, WE MODIFY IT
AND WE CAN DO PRETTY MUCH
ANYTHING WE WANT TO DO WITH IT
RIGHT NOW.
AND I SEE NO REASON TO CHANGE
WITH IT.
JUST EXTREMELY QUANTITATIVE AND
I WILL COME BACK TO THIS IN A
MINUTE.
SO THIS IS A MAJOR TOOL WE USE
FOR LOOKING EFFECTOR FUNCTION,
THE ONLY THING WE HAVE DONE TO
IT IS TRICKED OUT THE TARGET
CELL LINE A LITTLE BIT SO WE NO
LONGER HAVE TO LABEL WITH
CYTOPLASMIC DYE OR MEMBRANE DYE,
WHAT WE HAVE IS SAME TARGET CELL
EXPRESSING GFP INTERNALLY, CELL
LYSIS RELEASES GFP.
WE PUT A COMMERCIALLY AVAILABLE
CCR 5 SNAP TAG DYE, SNAP TAG IS
A BINDING PROTEIN YOU BELIEVE
LABEL PUT A DIMER IN EASILY.
CCR 5 IS ALSO FUNCTIONAL.
WE DEVELOPED THE ASSAY OUR GROUP
DID, SO THIS OBVIATES STEPS OF
HAVING TO LABEL THE CELLS.
AND IT IS ALSO HIGH THROUGH PUT
AND WE SET THINGS UP VERY FAST
BUT SOMETHING CAME OUT OF THIS
UNSUSPECTED TO US, WE FIRST
STARTED DOING THIS WE WERE
READING JUST LIKE IN THE
ORIGINAL PAPER.
AND WHAT WE DIDN'T KNOW AT TIME
WAS THAT THIS ASSAY REALLY
SIMULTANEOUSLY PICK UP TWO
EFFECTOR MECHANISM, ONE PICKS UP
TRUE KILLING AND THAT'S MEDIATED
BY NK CELLS, IN THE PREPARATION
USING PDMC AS EFFECTERS.
WE FOOLED WITH A LOT OF CELL
LINES NK LINES OR WHAT NOT
TRYING TO GET THOSE TO WORK AS
EFFECTORS AND THEY ARE AT LEAST
IN OUR HANDS TRICKY, WHEREAS
THIS ALWAYS WORKS.
BUT THE OTHER THING THAT HAPPENS
IS THERE ARE MONOCYTES IN THERE,
OFF THE EFFECTOR -- TARGET CELL
SURFACE ON TO THE EFFECTOR
CELLS.
WHY WE DO THE ASSAY TURNS OUT WE
DIDN'T KNOW THIS FOR A LONG TIME
BUT USING A FLOW SIGH TOM TRY WE
FOUND OUT WITH (INAUDIBLE) DOWN
AT DUKE BY AREA SCALING YOU CAN
DISCRIMINATE BETWEEN NK EFFECTS
AND MONOCYTE EFFECTS IN THE SAME
PREPARATION OF PBMCs.
TURNS OUT THAT THE MONOCYTES
MAKE TIGHT CELL JUNCTIONS.
THAT ACTUALLY SURVIVE FIXATION
THROUGH FLOW.
SO LOOK FOR DOUBLEETTES THOSE
MONOCYTE, THOSE CELLS ARETROPE
CYTOING WHEREAS WITH NK KILLING
YOU WILL SEE DEAD CELLS.
THE OTHER WAY IT IS SET UP IS
GFP IN CYTOPLASM.
WHAT WE NOTICED WAS WHEN WE
CLEANED UP ASSAY AND STARTED
DOING WITH PURIFIED MONOCYTES WE
WEREN'T SEEING KILLING.
THE GFP STAYED IN CYTOPLASM.
WE SAW TRANSFER OF MEMBRANE TO
THE EFFECTOR CELL MONOSITE.
MORE DATA ON THAT IN A MINUTE
BUT HOW IMPORTANT THIS HAS BEEN
FOR US. THESE ARE ALL STUDIES
DONE IN RHESUS MACAQUES GOING
BACK TO THE FIRST ONE, THAT WE
REPORTED IN 2007 WHERE WE TOOK
VACCINE I WILL TELL YOU ABOUT IN
A MINUTE, MICHAEL HAS SAID A
LITTLE BIT ABOUT IT WE IMMUNIZED
MACAQUES AND GOT PROTECTION
AGAINST HIGH DOSE CHALLENGE
EXEMPLIFIED BY POST INFECTION
CONTROL, EXACTLY WHAT YOU SAW
WITH MARJORIE, WHAT ALSO THIS IS
WHAT THE SHIV 162P 3 CHALLENGE.
THAT'S HARD TO NEUTRALIZE VIRUS.
WE GET NEUTRALIZING ANTIBODIES
THAT NEUTRALIZE WHAT WE WERE
TALKING ABOUT AT THE TIME BEING
A NOT LAB ADAPTED VIRUS BUT
PRIMARY VIRUS, NO TIERED
ANYTHING AT THE TIME.
THAT'S WHY TONY WE PUSH THE
STUDY, BUT TURNS OUT THERE ARE
NO NEUTRALIZING ANTIBODIES
AGAINST CHALLENGE VIRUS BUT YET
THERE WAS PROTECTION.
THAT PROTECTION CORRELATED WITH
SPECIFICITY AGAINST CD4 INDUCED
EPITOPES.
WE DIDN'T DO ADCC FIRST, WE DID
IT LATER BUT WE WENT BACK AND
LOOKED AT IT AND IT CONFIRMED
EVERYTHING WE HAVE SEEN LATER.
ONCE WE GOT THE APHASIA GOING OR
WHAT NOT.
SO THERE'S A WHOLE SERIES OF
STUDIES ONE HERE AND MANY FOLKS
IN THE ROOM CO-AUTHORS ON,
PAPERS ONE HERE WITH A NUMBER OF
-- I THINK THREE OR FOUR PRIMATE
STUDIES ONE SIV AND THE REST
SHIV.
OVER TIME WE HAVE DONE A LOT OF
THINGS WITH COLLABORATORS ON
THIS.
LOOKING AT THINGS FOR EXAMPLE
LIKE ONE WITH MARGE ACKERMAN AND
CHRIS KELLOGG LOOKING AT SYSTEMS
IMMUNOLOGY AND THEY FOUND
ANTIBODY FC FUNCTIONS IN THIS
VACCINE STUDY SIV CORRELATED
WITH BETTER PROTECTION THAN JUST
ABOUT ANYTHING ELSE AND ALSO
WITH YUNG SU AND JAY AND TONY
AND BOB AND OTHERS WHERE WE
FOUND TRAINED IMMUNITY EFFECT.
WHICH MAKES A LOT OF SENSE.
SO THAT'S VERY IMPORTANT FOR US.
THE RFA -- OUR RDADACC ASSAYS.
NOW DONE STUDIES IN IN HUMANS.
THE FIRST THING WHERE WE TRY TO
USE IT IN FACT IT WAS APPROVED
TO BE USED, IT WAS IN THE
CORRELATE STUDY FOR RV 144,
THERE WAS A INITIAL VETTING OF
METHODS IN THAT THING WHERE THEY
SENT OUT SPECIMENS AND WE WERE
PART OF THE EVALUATION TEAM THAT
HAD THAT HAPPEN, THE RFADCC
ASSAY SCORED BEAUTIFULLY, THE
BEST IN CLASS FOR ADCC ASSAYS.
WE WOUND OUT NOT GOING AHEAD AND
DOING THOSE IN THE RV 144 TRIAL
BECAUSE WE DIDN'T HAVE THE BAND
WITHAND THERE WERE NO OTHER
PEOPLE LOOKING AT CD 4I
ANTIBODIES IN THAT TRIAL.
TONY'S GROUP LOOK AT A 32 LIKE
ANTIBODIES AND WE LOOK AT
CORECEPTOR BINDING ANTIBODIES.
YOU GOT TWO OTHER ADCC ASSAYS
THERE.
EPITHAT WOULD BE THE STORY.
SORRY NOW WE DIDN'T LOOK.
THEN COME ON DOWN, COMPARATIVE
STUDY, THE ONLY ONE OUT THERE
THAT I KNOW OF, STILL MORE
RECENT ONE, THAT WAS DONE WITH
YUNG HUNG AND MARGIE ACKERMAN'S
TEAM WHERE WE LOOK AT DIFFERENT
ASSAYS AND THE ASSAY SHOOK OUT
THERE AND CORRELATED CLOSE WITH
FERRARI TYPE ASSAY BUT NOT
OTHERS.
THE OTHERS THING WE USE THIS
ASSAY FOR, WE GOT PASSKEYS GROUP
WHEN HE WAS HAVE US AT MARYLAND
NOW UNIFORM SERVICES, GOT A
BUNCH OF MONOCLONALS OUT OF RV
144, 306 STUDY AND SAW CRYSTAL
STRUCTURE ANTIBODY LIGAND.
TURNS OUT THAT THESE ANTIBODIES
THAT COME UP IN RV 144 LOOK LIKE
THE ONES WE HAVE WITH VACCINES
IN MACAQUES AND WITH PEOPLE.
AND THOSE STRUCTURE OF THESE
THINGS I WILL TELL YOU IN A
MINUTE.
SO THEN FINALLY, THIS YEAR WE
COMPLETED THE FIRST PHASE 1
TRIAL FOR OUR VACCINE.
SUCCESSFULLY DONE, DONE ENTIRELY
INSIDE THE IHV AT THIS POINT.
GOT RESPONSES LIKE WHAT WE SEE
IN RHESUS MACAQUES.
AND THE VACCINE IS NOW BEING
EVALUATED FOLLOW ON STUDY BY
MHRP IN THAILAND.
PHASE 1B STUDY, COME BACK TO
THAT SO WE DID ONE MOUSE STUDY
THAT'S FORGETTABLE, AND SO BUT
WHAT WE HAVE ALSO DONE, WE HAVE
DONE STRUCTURE FUNCTION STUDIES
THIS IS ALWAYS NEAR ANNEDIDAE TO
MY HEART BECAUSE IT DOSE -- NEAR
AND DEAR TO MY HEART BECAUSE IT
GOES BACK TO BEGINNINGS OF MY
CAREER, WHAT I WAS SORT OF
TRAINED TO DO.
FIRST THING, WE PUBLISHED A
STUDY IN 2013 WHEREBY THIS TIME
WE WERE GOING AFTER CD4 INDUCE
ANTIBODIES ISOLATING MONOCLONALS
WE WORKED OUT PROCEDURE WHEN IT
WAS HARD TO DO THEM.
AND CAME UP WITH A LOT OF
ANTIBODIES OUT OF LEAD CONTROL,
(INAUDIBLE) GROUP HAS.
AND THIS IS WHERE WE IDENTIFIED
A COMPETITION GROUP OF
ANTIBODIES THAT COMPETED WITH
SOME OF JANE'S ROBINSON'S
EARLIER ANN BODIES, WE CALL IT
EPITOPE CLUSTER A IN THIS
PARTICULAR STUDY AND THAT IS
BECOME WIDELY USED NOW.
WE KNOW A LOT ABOUT STRUCTURE OF
THESE ANTIBODIES.
THAT WAS THE FIRST PAPER.
AND THEN ANOTHER PAPER WAS ONE
WE STARTED LOOKING AT THE ASSAY,
THERE WAS SOME INTERESTING
THINGS ABOUT THE DOSE RESPONSE
CURVES THAT WOULD JUST LIKE AN
IGE BASOPHIL DOSE RESPONSE CURVE
IF YOU DOSE THOSE ASSAYS AND
FAMILIAR WITH THOSE FROM THE OLD
DAYS.
AGAIN WE PUBLISHED A NUMBER OF
STRUCTURE ALL AT THESE POINT
WITH MARZANA.
WE HAVE NEW PERSON IN OUR GROUP
WHERE DOING CRYOEM STRUCTURE ON
THE VACCINE ANTIGEN GETTING
CLOSE WITH THAT.
BUT STUFF ALL WITH THEIR GROUP.
SO THESE -- THIS WAS ALL USED
FOR THESE STUDIES.
AND THERE IS ANOTHER FOLLOW-ON
STUDY AGAIN WITH MARJORIE --
MARGIE ACKERMAN'S GROUP LOOKING
AT EFFECTOR ACTIVITIES PREDICTED
BY PHYSICAL PROPERTIES AND THE
ASSAY SHOOK OUT IN THAT GAVE US
INTERESTING INFORMATION.
THERE'S OTHER PAPERS UNDER
SUBMISSION RIGHT NOW, FOLLOWING
ON FOR THESE.
SO WHAT I'M GOING TO POINT OUT
ALSO, GLAD THIS WAS MENTIONED.
WE HAVE A GUY IN OUR GROUP WHO
IS A LONG TERM FRIEND FROM FIRST
DAYS WHEN, A GUY NAMED ALLEN
SMALLJOHN.
IF YOU KNOW ABOUT EBOLA OR
MARKBURG, ALAN WAS IN T MILITARY
USAMRID FOR YEARS BUT HE WAS IN
MARYLAND WHEN I FIRST CAME.
HE PUBLISHED THE FIRST CLEAR CUT
DEFINITIVE STUDY SHOWING THAT
NON-NEUTRALIZING ANTIBODIES
COULD PROTECT.
PUBLISHED IN NATURE IN 1982.
AND IS WHEN PEOPLE WERE FIRST
STARTED MAKING MONOCLONAL
ANTIBODIES. EVERYBODY THOUGHT
NEUTRALIZATION HAD TO BE IT.
HE WAS WITH JERRY COLE AT
HOPKINS.
JERRY CAME TO MARYLAND JUST
BEFORE I DID.
ANDEN ALLEN MAKES MONOCLONALS
FOR THE VIRUS, NEUROTROPIC VIRUS
AND SHOWED CLEARLY THAT HE COULD
GET PROTECTION, BEST PROTECTIVE
ANTIBODY, DIDN'T NEUTRALIZE.
DIDN'T KNOW WHAT IT DID.
HE LOOKED INTO IT LATER AND
TRIED TO FIGURE IT OUT BUT GO
LOOK IN THE LITERATURE HE WAS A
PERSON WHO ISOLATED ONE ANTIBODY
IN COCKTAIL IN THE EBOLA.
C 13 C 6, TO THIS DAY SO WE SET
UP THE SYSTEM BECAUSE ALLEN WENT
INTO RETIREMENT BUT CAME BACK
OUT AND WITH US AT THE INSTITUTE
AND SO WE SET UP MODIFIED RFACD
ASSAY, WE PUT GP ON THE SURFACE
SO THAT LED TO A NUMBER OF
STUDIES, ALL COLLABORATIVE
STUDIES AT THIS POINT.
ONE WAS THE FIRST WITH
HAIRIOTROBINSON WITH HER EBOLA
VACCINE ENCODES VIRUS LIKE
PARTICLE IN MVA AND SHE GETS
PROTECTION AGAINST EBOLA.
IN THAT MODEL IN MONKEYS. TURNS
OUT WE RAN THE ADCC, IT
CORRELATES WITHOUT, AND I
HAVEN'T ANYTHING PUBLISHED BUT
(INAUDIBLE) WAS PART OF THE
STUFF FUNDED BY MILITARY FOR
DOING THINGS WITH THE VACCINES
FROM THE VSV VACCINE THAT MERCK
HAS.
AND FALLS OUT FOR THAT.
THEN WITH JIM CROW, AT
VANDERBILT PUBLISHED SEVERAL
STUDIES WHERE WE HAVE DONE THE
ADCC ASSAY ON THE INTERESTING
ANTIBODIES THAT JIM AND GROUP
HAS ISOLATED.
AND IT TURNS OUT THAT IT
CORRELATES PRETTY WELL IN MANY
WAYS WITH THESE THINGS.
NOW WE HAVE A REALLY NICE
COLLABORATION WITH DANIEL
(INAUDIBLE) AT SCRIPS FORMER
STUDENT OF ERICA SAPPHIRE AND HE
IS THERE POST DOC WITH ANDREW
WARD.
SO THAT'S HOW WE ARE CONTINUING
THE EBOLA WORKS SO WHAT I WILL
SHOW YOU NOW FOR HIV WE HAVE
ALSO SEEN FOR EBOLA.
THAT IS OUR SITE SO TO SPEAK.
HERE IS VACCINE, TALKING ABOUT
BEFORE, SINGLE CHAIN WE CALL IT
CONFIRMATIONALLY CONSTRAINED GP
120.
FIRST DESCRIPTION OF THAT WAS IN
2000 WHEN TONY DESIGNED IT BUNCH
OF US ALL WORKING TOGETHER
LOOKING AT DIFFERENT VACCINE
CONCEPTS WITH THE INSTITUTE WAS
FIRST STARTED.
CAME UP WITH THIS CONCEPT, AND
WHEN WE WENT THROUGH VETTING
PROCESS, ALL THE THINGS WE LOOK
AT FELL BY THE WAYSIDE.
THIS WAS CHOSEN AS THE PRIMARY
CANDIDATE.
SO THE FIRST STUDY I MENTION WAS
PUBLISHED IN 2007.
WHERE ANTIBODIES AGAINST CD4
INDUCE SITES AND GP 120 CONTROL
SHIV CHALLENGE.
I HAVE NO RELIGION ABOUT
NEUTRALIZATION,
NON-NEUTRALIZATION.
THIS IS SIMPLE OBSERVATION.
WE SET UP THIS STUDY, WHEN WE
GOT IT GOING, SOLE GOAL IS WE
GOT PROTECTION WE WORK WITH --
WE DO SOME MORE WITH IT WE
DIDN'T GET PROTECTION IN MONKEY
MODEL WE WOULDN'T.
SO THIS IS A SHIV 16 P 3, SO
PASSES AT THE TIME, POST
INFECTION CONTROL OF VIREMIA, WE
WONDERED HOW THESE THINGS
WORKED.
THERE WAS A LITTLE REVIEW HERE
IN CURRENT HIV RESEARCH WHERE
TONY POINTED THIS OUT.
THAT THE NEUTRALIZATION PROBABLY
WAS AND SOMETHING LIKE ADCC WAS
PROBABLY GOING ON.
AND THAT IS WHY WHEN THE ASSAY
CAME OUT WE JUMPED ON IT.
AND STARTED USING IT.
AND I AM COMFORTABLE WORKING
WITH CHROMIUM, THAT STUFF BUT
HAVEN'T DONE CTL ASSAYS FOR
SEVERAL YEARS, I SAID OH HELL
NO, THEY ARE A MESS.
I WAS CO-AUTHOR ON ONE OF THE
FIRST ADCC PAPERS THAT WAS
PUBLISHED IN 1982 THAT WAS DONE
WITH CHROMIUM RELEASE, US
THOUGHT IT WAS THE MOST BORING
THING THAT I WAS PART OF AT THE
TIME, DIDN'T DO ANYTHING WITH IT
FOR YEARS.
YOU GET WISER AS YOU GET OLDER.
SO ANYHOW, FULL ON SINGLE
CHAINS, THIS EXPOSES THE MOST
CONSERVED PARTS OF GP 120.
AND THAT INCLUDES CORECEPTOR
BINDING SITE BUT ALSO INCLUDES
THIS REGION DOWN THERE WE DEFINE
AS EPITOPE CLUSTER A, EPITOPE
CLUSTER A IN NATIVE TRIMER IS
THE INTERNAL DOCKING SITE FOR GP
41.
SO WHEN GETS TRIGGEREDDED YOU
WIND UP TWO EPITOPE REGIONS COME
TOGETHER.
FIRST THE 32 SIDE OF IT, AGAIN
ONE OF JAMES ROBINSON'S ANTIBODY
AND SECOND IS C 11.
WE KNEW IT STRUCTURALLY BUT AT
THE TIME OF PUBLISH WE KNEW IT
CORRELATE WITH PROTECTION.
THAT'S WHAT WE KEPT ZYSEING.
I MENTION THIS, WE JUST
PUBLISHED THE FIRST VACCINE
TRIAL WHERE WE GOT THESE
RESPONSES, CORECEPTOR BINDING
SITE ANTIBODIES AS WELL AS
CLUSTER A ANN BODIES AND WE
HAVEN'T DONE CRYSTAL STRUCTURES
OR CRYOEMs ON THE ANTIBODIES
OUTS OF THESE PEOPLE BUT BY
COMPETITION ANALYSES WHICH TURN
OUT TO BE GOOD.
THESE LOOK LIKE THE ONES THAT
CAME UP IN RV 144 THAT CAME UP
IN MONKEY STUDIES.
SO THIS IS PART OF THE TRIAL 46
I MENTION WHERE IT IS A LATE
BOOST OF RV 306, BOOST OF RV 144
VOLUNTEERS, COLLABORATION WITH
MMRP AND DUKE WITH BART HAYNES
TEAM AT DUKE AND (INAUDIBLE) ET
AL.
SO THAT IS ONGOING, JUST STARTED
ENROLLING AND IMMUNIZING ABOUT
THREE MONTHS ACHE, IN IS GOING
ON IN THAILAND.
SO THAT'S GOING -- WE ARE
MANAGING THE DRUG PRODUCT, ET
CETERA, AND SPENT TIME DOING
THAT.
BUT HERE IS WHERE I AM HEADED.
WE SAW THESE KINDS OF DATA FOR
EXAMPLE, THIS PAPER PUBLISHED
WITH TIM PHOTS LEAD AUTHOR A LOT
OF FOLKS CO-AUTHORS.
WE FOUND IN OUR PROTECTION
STUDIES THAT WE SEE A SWEET
SPOT, THAT THERE WAS -- IF YOU
HAD ENOUGH ANIMALS, ENOUGH
INDEPENDENT TYPES OF STUDIES YOU
CAN SEE A SITUATION WHERE IF YOU
PLOT NUMBER OF CHALLENGES, LOW
DOSE CHALLENGES OF SHIV 1623P OR
SIV THERE IS A SWEET SPOT FOR
PROTECTION.
WHERE THE ADCC RESPONSE TO THE
CD4I EPITOPES IS HIGH ENOUGH,
BUT YOU DON'T HAVE TOO HIGH OF
T-CELL RESPONSE.
WE DON'T KNOW ABOUT T-CELL
PHENOTYPE THERE BUT WE HAVE SOME
INDICATION WHAT IT IS NOW AND IT
IS WHAT YOU WOULD EXPECT IT TO
BE.
SORT OF LIKE WHAT YOU SAW WITH
THE QUOTE UNQUOTE RV 144 REPEAT
THAT DIDN'T REPEAT.
ANYHOW SO WE HAD THAT, SO WE HAD
THE ABILITY AND INTEREST TO DO
ALL THE CRYSTALLOGRAPHY, ET
CETERA, MONOCLONALS PULLED OUT
OF PEOPLE AND NOW MACAQUES TO GO
PICK APART WHAT THESE EPITOPE
REGIONS LOOK LIKE AND NOTHING
WAS KNOWN ABOUT THEM UNTIL WE
STARTED THIS EXCEPT POINT
MUTATIONS. THE POINT MUTATIONS
ARE SHOWING REGIONS WE KNOW ARE
OUTSIDE THE EPITOPE FOOTPRINT.
WHAT THIS IS, IS THIS IS GP 120,
ACTUAL STRUCTURE, FOR CORECEPTOR
BINDING SITE ANTIBODIES IN 12I 2
THAT WORKS ON PASSIVE
IMMUNIZATION STUDY FOR POST
INFECTION CONTROL, DOESN'T
NEUTRALIZE THE VIRUS.
EPITOPE CLUSTER A THERE WAS
COMPETITION DATA THAT SAID THAT
THE ANTIBODIES A 32 AND C 11
WOULD CROSS COMPETE BUT
NON-RECIPROCAL FASHION.
SO THERE WAS EPITOPE OVERLAP OR
THESE TWO EPITOPES WERE
PHYSICALLY CLOSE ENOUGH TO ONE
ANOTHER WITH FULL LENGTH
ANTIBODY, IF ONE BOUND THE OTHER
TRY TO COME IN, THERE'S
CANDIDATES REDUCE, BY CRYSTAL
STRUCTURE WE SHOW THESE THINGS
RECOGNIZE ADJACENT EPITOPES, IN
THIS REGION THAT FORMS ON GP 120
AFTER LETS GO GP 41 DURING VIRAL
ENTRY OR DURING EXIT IF COMPELS
HAVE CD4 ON IT.
WHAT I AM NOT SHOWING YOU IS WE
HAVE MONKEY MONOCLONAL, ONE OF
JAMES,NA RECOGNIZES THE HYBRID
REGION IN HERE, A LOT OF THESE
ANTIBODIES COMING OUT OF ONE
FROM RV 144 RECOGNIZE FOOTPRINT
THAT OVERLAPS THESE.
SO WE HAVE THESE STRUCTURE, WE
HAVE BINDING ANGLES WE HAVE
PHYSICAL CHEMICAL INFORMATION
PLUS WE HAVE A ROBUST ASSAY THAT
IS QUANTITATIVE SO ALLOWED US TO
HAVE FUN WITH THERE'S THINGS.
THIS IS WHERE I GO OFF OF HIV.
WHICH TAKE PRIME MARE HUMAN
MONOCYTES, -- PRIMARY HUMAN
MONOCYTES AND WE WILL PUT THEM
TOGETHER AND DO THE TROGO
CYTOSIS BUSINESS AND LOOK IT IN.
SO WE DO WE ARE ABLE LOOK IN
TISSUE NOW AND THICK TISSUE
SECTION, ABLE TO DO THIS BY LIVE
IMAGING LIVE CELL IMAGING, DO IT
BY STANDARD CONFOCAL AND SUPER
RESOLUTION CONFOCAL MICROSCOPY
SO I WILL SHOW YOU DATA FROM
THIS.
AND WHAT IS GOING ON WITH THIS,
IS TROGO CYTOSIS PROCESS, YOU
GET MEMBRANE FRAGMENTS FROM
TARGET CELL TRANSFERRED OVER ON
TO THE EFFECTOR CELL BUT YOU
ALSO GET ANTIGEN ANTIBODY
COMPLEXES GOING OVER AS WELL.
NO ANTIBODY IN THERE OR ANTIGEN
IN THERE, YOU DON'T GET TROGO
CYTOSIS.
WHAT IT MEANS, I DON'T KNOW.
IT CORRELATES IN OUR STUDIES.
IT'S CLEARLY THE PHENOMENON
ITSELF CLEARLY IMPORTANT FOR
BOTH POSITIVE AND NEGATIVE
OUTCOMES FROM CANCER ANTIBODY
TRIALS.
ALSO THERE IS A PROBLEM IN NK --
CAR T-CELL MEDIATED KILLING,
ITSELETCETERA.
THERE'S LIKE 10,000 GRANTS THAT
HAVE PHAGOCYTOSIS OR ADCC IN IT,
EIGHT HAVE TROGO CYTOSIS.
AND THEY ARE ALL ON MAINTENANCE
WHICH IS WHERE THE PHENOMENON
WAS DISCOVER SOD IT HAS THAT
ATTRACTION.
SO THINKING NOW, BACK FROM A
FUNCTIONAL BASIS WHICH
STRUCTURAL BASIS, I ALWAYS TRY
TO DO THIS BUT PLOTTED HERE, WE
SET THE SYSTEM SO THAT WE CAN
VET IN A SINGLE ASSAY ANTIBODIES
AGAINST ALL PARTS OF ENVELOPE,
ON THE TARGET.
WE DID THIS BY SPINNING
(INAUDIBLE) ON TOP OF TARGET
CELLS SO WE CAN PICK UP CD4 1,
ET CETERA, AND MADE THESE
ANTIBODIES AND EXPRESSED IN SAME
SYSTEM, PURIFIED THE SAME WAY ET
CETERA.
AND WHEN WE WENT TO LOOK TO SEE
WHAT ROLE SPECIFICITY PLAYED IN
EFFECTOR FUNCTION IN THIS ASSAY,
BELIEVE OR NOT THIS WAS THE
FIRST STUDY I COULD FIND WHERE
ANYBODY COMPREHENSIVELY LOOKED
AT ANY ANTIBODIES IN TERMS OF
SPECIFICITY OF STANDARD SYSTEM.
TO DO THAT.
SO HERE IS WHAT WE CAME UP WITH.
FIRST, THINKING ABOUT IT, UP
HERE, AT THE TOP IS INTRINSIC
AFFINITY RANGE OF FC GAMMA
RECEPTORS BINDING TO PURIFIED
HUMAN IGG 1 WE ARE TALKING ABOUT
HERE.
AND SO IT IS ALL UP IN THE HIGH
-- LOW MICROMOLAR RANGE, ALL THE
WAY UP TO ESSENTIALLY THE
MILLIMOLAR RANGE DEPENDING WHICH
PAPERS YOU LOOK AT AND SO THAT
IS UNLIGANDED FREE ANTIBODY, IT
BINDS. IF YOU LOOK HERE, THERE
WAS A BIG DIFFERENCE BIG SPREAD
CYTOTOXICITY VERSUS EC 50, HALF
MAX VALUE FOR DIFFERENT ANTIBODY
CLASS SPECIFICITY CLASSES.
TURNED OUT THEST ONES IN THIS
ASSAY FORMAT, TURNED OUT TO BE
CONSISTENTLY CLUSTER A
ANTIBODIES.
THESE WERE ONES WE ISOLATED PLUS
JAMES ANTIBODIES THERE WAS A
BUNCH WE PULLED OUT.
THE NEXT BEST WERE CO-RECEPTOR
BINDING SITE ANTIBODIES IN THIS
PARTICULAR ASSAY.
WE CAN GET THINGS, THIS WOULD
PICK UP SOME ANTIBODIES AGAINST
CD4 BINDING SITES WHICH ARE
INTERMEDIATE IN GP 481 ANTIBODY
ARE NOT GOOD FOR THIS AND MANY
WAYS NOT PARTICULARLY GOOD IN A
LOT OF DIFFERENT TYPES OF
EFFECTOR ACTIVITIES SO THIS IS
WHAT INTERESTED ME.
WHAT IS GOING ON HERE?
THESE ARE MUTANT COMPLEXES,
GIVING YOU A BIOLOGICAL GAME FAR
AND ABOVE INTRINSIC BINDING
CONSTANT IS FOR FREE IGG.
THAT MAKES SENSE, YOU ARE
SWIMMING IN FREE IGG.
SO THERE IS A TONIC BINDING BUT
IT'S LOW AFFINITY SO WILL IS A
AFFINITY COMPONENT TO THIS BUT
OTHER STUFF GOING ON AS WELL, WE
WANT TO KNOW WHAT THIS IS.
FIRST WE GOT STRUCTURALLY
DEFINED MOLECULES THAT ARE DOING
THIS.
WHAT ARE THE IMMUNE COMPLEXES
LOOK LIKE?
THAT DO THIS?
THIS IS WHAT WE HAVE BEEN
WORKING, THIS SHOWS YOU IF YOU
LOOK AT EC 50 OF THE CLUSTER A
ANTIBODIES, THERE IN BLUE, IT IS
10,000 FOLD DIFFERENCE FROM WHAT
YOU SEE BY INTRINSIC --
DIFFERENT FC RECEPTORS FROM
HUMAN IGG 1.
WE USE THIS ASSAY, WHAT WE
DECIDED TO DO WE STARTED IMAGING
WHAT'S GOING ON, REALLY GET A
PICTURE OF THE KINETICS OF THE
ASSAY, WHO IS DOING WHAT.
BEAUTY OF IT IS, WE GET
INTERESTING BIOLOGY WITH ONE
ANTIBODY, ONE ANTIGEN, TWO
CELLS.
AND YOU ONLY SEE PART IN FLOW
CYTOMETRY, LOOK IN CULTURE A LOT
OF STUFF GOING ON FLO ON IN THE FLOW
SIGH TOM TEAR IF YOU DON'T DO IT
RIGHT.
A FEW MONTHS IN TOM'S LAB AS
PARTS OF GATES FOUNDATION THING,
AND SO YOU CAN SEE THE CLASSIC
SORT OF SYNAPSE FORMATION, OVER
TIME AND THEN YOU CAN SEE
TRANSFER OF THE MEMBRANE
FRAGMENT WHICH IS CCR 5, IN THIS
CASE JUST LOOKING AT GP 120 HERE
AS IT COMES ACROSS.
SO YOU CAN DEFINE THESE
DIFFERENT STEPS, YOU ALSO SEE
THIS HAPPENING IN LIVE CELL
CULTURE.
AND YOU CAN ALSO DO GREATER
IMAGING ON THIS, AND FOLLOW
THESE EVENTS DURING FLOW
CYTOMETRY.
SO WE ARE CALLING THIS ANTIBODY
MEDIATED TROG CYTOSIS.
THIS IS THE CLUSTER A.
AND ONE OF THE THINGS WE NOTICE
IN THESE ASSAYS IS THEY SHOW PRO
ZONES YOU DON'T ALWAYS SEE A PRO
ZONE ASSAY IN ADCC ASSAY BUT
OFTEN YOU DO, WE ARE STARTING TO
UNDERSTAND WHY YOU SEE IT NOT
OTHERS BUT THESE ANTIBODIES DO
IT AND PRO ZONES IN THE DOSE
RESPONSE CURVE TO ME THAT LOOKED
LIKE A BASE FILL ACTIVATION BY
IGE AND CROSS LINKING IGE
ANTIBODY, THERE'S LITERATURE ON
THAT.
SO THIS PAPER POSITION PIECE PUB
ESTABLISHED ON IT WHAT IS GOING
ON HERE, WE THINK THIS LOOKED
LIKE JAY AND I HAD A PRECIPITOUS
REACTION AND SOMETHING AROUND
FOR OVER A HUNDRED YEARS,
SOMETHING SIMILAR TO THIS GOING
ON YOU WILL SEE IN A SECOND.
SO WHAT HAPPENS IN THE SWEET
SPOT WHERE YOU GET BIOLOGY IS
ASCENDING PART OF THE CURVE, EC
50 UP TO PLATEAU CYTOTOXICITY,
WHATEVER THAT IS.
SO HERE THIS IS EQUIVALENT
PREDICTION WISE EQUIVALENT TO
ZONE OF EPITOPE ACCESS OR
ANTIGEN EXCESS PRECIPITOUS
REACTION SO THE ANTIBODIES
COMING IN ARE (INAUDIBLE) BUT
THEY HAVE TO DO SOMETHING WITH
IT TO MAKE THE RIGHT COMPLEX.
THEN THINGS GET -- YOU HAVE SOME
OPTIMAL PRESENTATION OF THE
IMMUNE COMPLEX.
AS YOU ADD MORE ANTIBODY IN, THE
COMPLEXES BECOME INCREASINGLY
VALENT AND LESS EFFECTIVE
BIOLOGICALLY SO YOU SEE PRO ZONE
GOING DOWN.
WITH HE IS DIFFERENT ANTI-DO
BODIES YOU SEE DIFFERENT PRO
ZONES WITH IT.
LOOK AT THE EC 50s OR DOWN
INTO THE EXTREMELY POTENT WITH
THESE ANTIBODIES ADCC WE ARE
DOWN IN THE -- ALMOST INTO THE
LOW PICA MOLAR RANGE WHICH IS
WAY BELOW WHAT YOU SEE WITH ANY
NEUTRALIZATION ASSAY.
SO WHAT WE HAVE DONE THEN, SHOW
YOU THE NEXT SLIDE HERE.
SO WHAT WE HAVE DONE WE HAVE
LABELED ANTIGEN, LABELED
ANTIBODY AND LABELED MEMBRANE.
SO WE HAVE -- LOOK AT TROGO
CYTOSIS EFFECT AND SO THAT'S
SHOWN IN RED THERE IN THE CURVE
AND YOU SEE IT IS A BELL SHAPED
CURVE JUST WHAT WE SEE, IF WE
ARE READING FLOW CYTOMETRY,TROGO
CYTOSIS ASSAY.
THAT'S WHAT IT LOOKS LIKE FOR C
11 ANTIBODY, MOST POTENT.
AND THAT IS INTERESTING, IF YOU
LOOK AT ANTIGEN ON THE EFFECTOR
CELL THIS IS ALL EFFECTOR CELL
NOT TARGET CELL, LOOK AT AMOUNT
OF ANTIGEN ON EFFECTOR CELL IT
ROUGHLY CORRELATES.
YOU SEE THE SAME BELL SHAPED
CURVE WHICH MAKES SENSE, WITH
THIS.
BUT WHEN WE WENT AND LOOKED AT
TRACK ANTIBODY IN THE SAME CELLS
SAME TIME, WE SAW MORE ANTIBODY
WE PUT IN, THE MORE ANTIBODY
AFTER POINT WE FOUND ON THE
EFFECTOR CELL BUT AT SOME POINT
LESS ANTIGEN.
SO THAT SAID LET'S LOOK AT RATIO
OF ANTIGEN AND ANTIBODY SO I
NORMALIZED EVERYTHING, WHEN WE
GET TO STUDY WE DIDN'T CALIBRATE
INSTRUMENT TO GET NUMBERS OF
MOLECULES PER CELL BUT IN THE
PROCESS OF DOING THAT.
SO THIS EMERGED TO BE VERY
INTERESTING TO US.
SO THOSE ARE THE CURVES TOGETHER
SO IF YOU LOOK UP IN PURPLE
THAT'S THE RATIO CURVE OF
ANTIGEN TO ANTIBODY OR ANTIBODY
TO ANTIGEN ON THE EFFECTOR CELL.
THAT'S WHAT'S COME OVER TROGO
CYTOSIS.
SO DOWN HERE IN THE LINEAR
ASCENDING OR EXPONENTIAL
ASCENDING RANGE OF DOSE RESPONSE
CURVE WHAT YOU SEE IS COMPLEXES
COMING ON TO EFFECTOR CELL ARE
ROUGHLY ONE TO ONE FOR ANTIBODY
ANTIGEN.
THAT DOESN'T MEAN IT'S MOLAR 1
TO 1 JUST SORRY .1, THAT DOESN'T
MEAN WE DON'T KNOW WHAT THE TRUE
STOICHIOMETRY IS BUT POINT IS
THAT REGION INCREASING
BIOLOGICAL ACTIVITY PUTTING MORE
ANTIBODY, THAT RATIO OF ANTIGEN
TO ANTIBODY IS COMINGEN EFFECTOR
CELL IS CONSTANT.
THEN YOU GET OUT TO MAXIMUM
TROGO CYTOSIS EFFECT AND WHAT
YOU GET PAST MAXIMUM THAT RATIO
STARTS TO GO DOWN.
SO YOU START GETTING MORE
ANTIBODY AND LESS ANTIGEN ON THE
TARGET CELL.
SO THIS IS VERY INTERESTING
BECAUSE THIS IS EXACTLY WHAT YOU
PREDICT FROM A LITERATURE
EXACTLY WHAT THOUGHT IN THE
PAPER.
MADE US START WONDERING WHAT IS
GOING ON NOW ON THE TARGET CELL
SIDE AND ON THE EFFECTOR CELL
SIDE WE HAVE THE TOOLS TO LOOK
AT THIS.
THIS IS AN EARLIER PAPER ALL OUR
COMPLEXES ARE MADE WITH GP 120
BINDING TO CD4 IN THIS
PARTICULAR MODEL.
SO WHERE IS -- HOW CD4 ARRANGED
ON CD4 POSITIVE T-CELL SURFACE?
TURNS OUT THAT THE -- IN
UNINFECTED CELLS FOR EXAMPLE,
THEY RANGE IN NANOCLUSTERS THAT
ARE ROUGHLY 50 NANOMETERS ACROSS
AND USUALLY HAVE TYPICALLY FOUR
CD4 MOLECULES OF NANOCLUSTER.
PUT HIV ON THEM, OVER THERE THE
GREEN PUT HIV ON THEM, THEN THE
NANOCLUSTERS GET TO BE 200
MICRONS AND YOU WIND UP WITH 16
COMPLEXES IN THE CLUSTER.
SO BECAUSE WE DO STRUCTURAL
STUFF WE BUILT MODELS, BUILT --
THERE IS NO CREE OWE EM OF
ANYTHING IN CD4 BUT USE STITCHED
TOGETHER THE BEST -- CRYOEM OF
ANYTHING IN CD4.
THAT CD4 IN RED IN THE CIRCLE
THERE IS THE CELL MEMBRANE,
MICRODOMAIN, IF YOU WILL, THE
GREEN IS GP 120 BINDING.
WHAT WE WILL DO IS DOCK ON OUR
ANTIBODIES AGAINST CLUSTER A WE
HAVE THE STRUCTURE FOR.
SO THERE IT IS.
THAT IS THE 7SN ANTIBODY TAKEN
FROM STRUCTURE THAT IS ONE OF
THE STRUCTURE THAT WAS FROM
SMALL ANGLE X-RAY SCATTERING
STUDY DONE BY PERKINS THAT'S FC
RECEPTOR COMPETENT FOR BINDING.
SO YOU IT BINDING THERE AND THEN
WHAT WE DID HERE, THE PURPLE WE
HAVE DOCKED IN AN FC RECEPTOR.
AND THE C TERMINUS OF THE FC
RECEPTOR TOWARD EFFECTOR CELL IS
ORIENTED DISTAL TO THE TARGET
CELL MEMBRANE.
WE HAVE OTHER ANTIBODIES WITH
DIFFERENT BINDING ANGLING.
WE PUBLISHED A COUPLE OF PAPERS
ON THESE AND WE FIND THE FORMER
A-- FARTHER AWAY FROM TARGET
CELL THAT FC DOCKING SITE IS,
THAT CORRELATES WITH ACTIVITY.
RIGHT NOW WE ARE BUILDING A
VECTOR SPACE MODEL SO THAT WE
CAN ACTUALLY QUANTIFY THAT WITH
SOME PRECISION.
SO MADE US WONDER WHAT IS GOING
ON IN MICRODOMAINS SO BASED ON
THE INFORMATION FROM OUR
MODELING STUDIES WE KNOW ROUGHLY
THE SIZE OF -- THIS IS A TWO
DIMENSIONAL ARRAY, OF THE
CLUSTERS THERE ABOUT 20 MICRONS,
20 NANOMETERS IN SIZE.
SO WHAT I'M SHOWING HERE IS WHAT
A MICRODOMAIN LOOKS LIKE, IF YOU
GOT 16 MOLECULES OR 15 MOLECULES
CD4 AND YOU HAVE THESE THINGS
DOCKING ON THERE.
SO THIS IS A RELATIVELY SPARSE
BASE.
BUT THESE THINGS ACTIVATE FC
RECEPTOR THERE HAS TO BE CROSS
LINKING.
BUT WHAT WE FOUND FROM ALL THIS
IS THAT WE UNCOVERED AN
ALLOSTERIC EFFECT THAT FAVORS
CROSS LINKING UNDER SPARSE
CONDITIONS.
WE FOUND THIS FROM THIS KIND OF
STUDY, THIS IS DONE BY
(INAUDIBLE) RAY A BIOPHYSICIST
IN OUR GROUP AND TIARA, WE TOOK
MONOMERIC ANTIBODY WITH AND
WITHOUT ANTIGEN AND ASKED TO
BIND TO FC RECEPTORS ON A
JERKETTE CELL YOU PUT FC
RECEPTORS ON SO BINDING WE KNOW
ANTIBODY IS MONOMERIC BECAUSE OF
SEVERAL THINGS.
WE DO CORRELATION SPECTROSCOPY
THAT GIVES YOU DIFFUSION
COEFFICIENT SO MONOMERIC
ANTIBODY, WHEN IT BINDS ANTIGEN,
IT IS NOT MAKING HIGHER ORDER
POLYMERS, MOST BINDING TO
INDEPENDENT ANTIGEN MOLECULES
AND ONE ANTIBODY BUT MOSTLY ONE
TO ONE.
WE HAVE NOW GOTTEN EQUIVALENT TO
DO SIZE EXCLUSION CHROMING TOTY
LIGHT ANGLE LIGHT SCATTERING
ANALYSES.
IT SAYS THE SAME THING.
MAKING THESE COMPLEXES WE ARE
MAKING ACTUALLY SOME OF THE
CLINICAL MATERIAL THAT WENT INTO
THE PRACTICE RUN THAT WENT INTO
CLINICAL TRIALS.
WHAT WE ARE SHOWING HERE IS
SIMPLY C 11 FLUORESCENT C 11
BIENSTOCK JERKETTE CELLS ON THE
LEFT WITHOUT ANTIGEN AND THEN
WHERE WE SIMPLY GIVEN ONE
MOLECULE OR TWO MOLECULES OF
FULL LENGTH SINGLE CHAIN WHERE
IT IS NOT CROSSING, YOU SEE A
BIG DIFFERENCE BY FLUORESCENCE
INTENSITY.
THAT COMPLEX EVEN UNDER THESE
CONDITIONS YOU GET SOME BINDING
OF FREE ANTIBODY, YOU GET
BINDING INCREASE BINDING THERE.
YOU CAN ALSO SEE BY ELISA,
MARGIE ACKERMAN'S GROUP ALSO GOT
THIS OBSERVATION WITH SPR DATA.
ANYHOW, SO THAT TELLS ME
SOMETHING INTERESTING RIGHT
THERE BUT THE OTHER THING YOU
CAN DO IS YOU CAN DO LIFETIME
FLUORESCENCE.
WHICH MEASURES THE LIFETIME OF
THE DECAY SIGNAL.
AND TURNS OUT THAT INTENSITY
FLUORESCENCE IS INDEPENDENT OF
THE ENVIRONMENT.
WHEREAS LIFETIME IS AFFECTED BY
LOCAL ENVIRONMENT.
SO WHAT IT IS REALLY GOOD AT, IF
YOU SEE LIFETIME SIGNAL SAYING
CONFIRMATIONAL CHANGE.
IN THE MOLECULES THAT -- WHERE
YOU GOT YOUR DYE.
SO YOU LOOK HERE, WE HAVE GOT A
REALLY FLOOD FLUORESCENT
LIFETIME SIGNAL THERE. IS A
CONFIRMATIONAL CHANGE AND THAT
COMPLEX OF SOME SORT THAT'S
ASSOCIATED WITH THE BINDING TO
THE FC RECEPTOR.
AGAIN, NO ROSALYNING HERE AT
THIS POINT.
NO CROSS LINKING.
SO BY USING SUPER RESOLUTION
MICROSCOPY, (INAUDIBLE) WHO DID
THIS, WE FED THESE SAME CELLS
NOW TWO DIFFERENT CLUSTER A
ANTIBODIES N 5 I 5 A 32 LIKE AND
WE LOOK FOR THE CO-LOCALIZATION
OF THESE TWO OVER SEPARATE
DISPOSITION OF THESE TWO ON THE
CELLS AND IF YOU JUST SEE RED OR
GREEN BY ITSELF, THAT MEANS
COMPLEXES BIND INDEPENDENT LET
AND NOT COLOCALLIZING.
THAT DOESN'T MEAN THE SAME
COMPLEX JUST CLOSE TO ONE
ANOTHER.
WE DON'T KNOW YET WHAT'S GOING
ON THERE THEY COULD BE IN THE
SAME COMPLEX BECAUSE THESE CAN
BIND SIMULTANEOUSLY TO THE FULL
ON SINGLE CHAIN BUT THIS SHOWS
YOU BINDING TO MICRODOMAIN
ROUGHLY THE SIZE WHAT YOU WOULD
BE TO SEE FOR CD4 ON TARGET CELL
AND SAME THING GOES ON IN THE
EFFECTOR CELL.
SO WITH THIS, WHEN WE GOT THIS
OBSERVATION, BECAUSE I REMEMBER
ALL THE OLD TIMES HIKING ABOUT
THIS STUFF, MAYBE THERE IS AN AL
LORALLOSTERIC CHAIN.
LET ME QUICKLY SAY WHAT WE DID
IS WE PUBLISHED, WE FOUND AN
ALLOSTERIC CHAIN BY HYDROGEN
EXCHANGE AND OTHER METHODS,
SHOWING THAT THE BINDK ANTIBODY
TO ANTIGEN, THOUGH MONOMERIC
MAKES FC RECEPTOR ASSUME
CONFIRMATION MORE FAVORABLE TO
ANTIBODY, SO IT'S SIMULATING THE
LOCAL MICRODOMAIN SO ANYHOW,
JUST SKIP PAST THIS AND TELL YOU
THE IMPORTANT PART.
THE END OF IT YEAH.
THANK YOU, MARJORIE.
[APPLAUSE]
SORRY FOR RUNNING OVER.
>> THE LAST SPEAKER BEFORE THE
BREAK IS MARGIE ACKERMAN, AND
SHE IS (INAUDIBLE) GOING TO TELL
US COMPLIMENTS TO MARJORIE AND
(INAUDIBLE).
>> THANK YOU.
SO I'M GRATEFUL FOR THE
OPPORTUNITY TO TO PARTICIPATE IN
THIS CELEBRATION OF MARJORIE'S
CAREER.
WHICH I THINK I HAVE SUB
CONSCIOUSLY EMULATED A LITTLE
BIT, OR AT LEAST ASPIRE TO NOW
THAT I'M AWARE OF IT.
HAVING DONE MY Ph.D. WORK IN
THE SETTING OF CANCER AND
TRANSITIONED INTO ANTIBODIES AND
FROM THERE TO EXTRA NEUTRALIZING
ACTIVITIES OF ANTIBODIES.
I DIDN'T TRAIN WITH MARJORIE,
BUT I CAME INTO THE FIELD OF HIV
RESEARCH AT MAYBE THE SECOND
PEAK, THE BEGINNING OF SECOND
UPSWING OF INTEREST IN
NON-NEUTRALIZING ANTIBODY
FUNCTIONS THAT WE TALKED ABOUT
AND LOOKING BACK AT WHO WAS IN
THAT SPACE BEFORE THAT BOOM.
IT WAS MARJORIE.
SO THE -- FOR BETTER OR WORSE
THE FIRST GRANT EVER GOT WAS A
BIG ONE AND IT HAD AN SAB.
SO MARJORIE WAS THE PERSON WE
ASKED TO ADVISE US IN THAT
EFFORT.
I QUICKLY CAME TO RECOGNIZE HER
AS A PERSON WHO DIDN'T HAVE THE
LOUDEST VOICE IN THE ROOM BUT
HAD THINGS THAT I WANTED TO HEAR
AND WANTED TO THINK ABOUT.
SO DESPITE BEING AT DISTANCE, I
REPRESENT MANY, MANY, MANY
DOZENS IF NOT HUNDREDS OF JUNIOR
SCIENTISTS WHO HAVE FELT
POSITIVE EFFECTS OF YOUR CAREER
IN LESS DIRECT WAYS THAN
TRAINEES DESCRIBED TODAY.
SO WE HAVE ALREADY HEARD ABOUT
TODAY A NUMBER OF NON-HUMAN
PRIMATE AND HUMAN IMMUNIZATION
STUDIES IN WHICH QUALITATIVE
ASPECTS OF THE IMMUNE RESPONSE
RATHER THAN NEUTRALIZATION HAVE
RELATED TO PROTECTION FROM
INFECTION.
SO MY GROUP HAS BEEN USING BIG
DATA AND MACHINE LEARNING
APPROACHES TO TRY TO INTEGRATE
UNDERSTANDING ABOUT FEATURES OF
THE ANTIBODY RESPONSE AND
FUNCTIONS OF ANTIBODIES, ACROSS
DIVERSE INDIVIDUALS TO REALLY
UNDERSTAND WHAT MAKES FOR AN
EFFECTIVE HUMORAL IMMUNE
RESPONSE.
BY DOING SO, WE REALLY HOPE TO
USE DATA DRIVEN DISCOVERY TO
FIND ROBUST CORRELATES OF
PROTECTION TO ADVANCE VACCINES
AS WELL AS PROVIDE INSIGHT HOW
MONOCLONAL ANTIBODY BASED
INTERVENTIONS MIGHT BE IMPROVED.
SO THE WORK I WILL TALK ABOUT
TODAY STARTS WITH A STORY YOU
HAVE ALREADY HEARD ABOUT DNA
PRIME VACCINE BASED STUDY
CONDUCTED BY FOLKS AT IHV AND
PROSPECTUS BIOSCIENCES DNA
ENCODED ADJUVANTS WERE COMPARED.
ONE ARM OF THE STUDY THE ONE
WITH ADJUVANTED WITH IL 12 WAS
ASSOCIATED WITH REDUCED RISK OF
INFECTION AFTER REPEATED LOW
DOSE INTRARECTAL CHALLENGE.
WHAT WAS REPORTED IN THE
ORIGINAL STUDY WAS ANIMALS WITH
HIGH ADCC BUT LOW INTERFERON
GAMMA EXPRESSION THOSE SHOWN IN
RED WERE BETTER PROTECTED THAN
THOSE WITH ADCC BUT HIGH
INTERFERON GAMMA EXPRESS SHOWN
IN GREEN OR NO ADCC ACTIVITY AT
ALL IN BLUE SO THE CONCLUSION
WAS YOU NEEDED BALANCE BETWEEN
ARMS ADAPTIVE IMMUNE SYSTEM.
SO THIS FELT LIKE MAYBE AN IDEAL
SETTING IN WHICH TO USE THESE
MULTI-VARIANT ANALYTICAL
TECHNIQUES TO TRY TO UNDERSTAND
HOW ASPECTS OF THE ANTIBODY
RESPONSE RELATE TO EACH OTHER
AND RELATE TO OUTCOMES.
I THINK THAT OBSERVATION
RESULTED FROM SOMEWHAT AD HOC
ANALYSIS.
WHEN YOU READ THROUGH YOUR PILE
OF DATA AND TRY TO DISCOVER WHAT
ON EARTH RELATES TO OUTCOME
OBSERVED SO WE WANTED TO DEPLOY
ASSAYS AND ANALYSIS MORE
SYSTEMATICALLY AND BROADLY.
BUT AT THE SAME TIME DO IT IN
STATISTICALLY PRINCIPLED WAY
WHICH GETS TRICKIER THE MORE
MEASUREMENTS YOU HAVE SO FOR
THAT WE TURN TO SYSTEMS SEROLOGY
TOOLS THAT AIM TO ENABLE US TO
CAST THAT BROAD NET ACROSS
ANTIBODY PHENOTYPES AND ANTIBODY
FUNCTION AND THEN USE RIGOROUSLY
CONTROLLED MACHINE LEARNING
TECHNIQUES TO UNDERSTAND
RELATIONSHIPS.
SO WE USED CROSS VALIDATED
MACHINE LEARNING APPROACHES TO
PREDICT RISK OF INFECTION.
HOW THIS WORKS IS THAT WE TAKE A
SUBSET OF THE ANIMALS IN THE
STUDY AND WE IDENTIFY FEATURES
AND COEFFICIENT WEIGHTS FOR
THOSE FEATURES THAT PREDICT
INFECTION RISK AND WE TEST THEIR
ACCURACY ON THE ANIMALS THAT
WERE HELD OUT FROM THE MODELING.
SO IT IS NOT A DATA FITTING
EXERCISE, TRULY A PREDICTIVE
EXERCISE, SACRIFICING SOME OF
THE DATA LEARNING RULES AND
PREDICTIVE MODEL AND APPLYING TO
EXAMPLE ANIMALS THAT WERE HELD
OUT.
SO YOU CAN DO THIS AGAIN AND
AGAIN AND AGAIN WITH DIFFERENT
COMBINATIONS OF ANIMALS,
DIFFERENT COMBINATIONS OF
FEATURES AND SEE WHAT HAPPENS.
SO WHEN WE DID THAT WITH
RESPONSE MAGNITUDE OR TITER
ALONE, THERE WAS REALLY NO
ABILITY TO PREDICT RISK OF
INFECTION.
O SHOWN HERE PREDICTIVE RISK
VERSUS CHALLENGE OUTCOME ACROSS
THE DIFFERENT IMMUNIZATION
GROUPS IN THIS STUDY AND THERE'S
NO WISHFUL THINKING, THERE'S A
RELATIONSHIP HERE OR NOT.
AND THERE WAS ALSO NO ABILITY TO
RECAPITULATE WHAT WAS OBSERVED
THIS TRIAL, REDUCED RISK OF
INFECTION OBSERVED IN IL 12
ADJUVANTED GROUP SO USING
ANTIBODY TITTEDDER MEASUREMENTS
THAT AMOUNT OF IGG SPECIFIC TO
HIV ANTIGENS, IN THIS STUDY
DIDN'T GIVE ANY ABILITY TO
PREDICT THE OUTCOME.
IN CONTRAST IF WE INCLUDE THE
QUALITATIVE FEATURES OF
ANTIBODIES THAT WERE RAISED AND
FUNCTIONS OF THOSE ANTIBODIES
MEASURED IN CELL BASED ASSAYS,
WE NOT ONLY SAW GOOD CORRELATION
BETWEEN PREDICTIVE RISK AND
NUMBER OF CHALLENGES REQUIRED TO
ESTABLISH INFECTION AT LEVEL OF
INDIVIDUAL ANIMALS WHICH IS
SHOWN AT THE LEFT, PREDICTED
RISK THOSE ANIMALS EACH DOT IS
AN INDIVIDUAL ANIMAL.
THOSE WITH RELATIVELY HIGH RISK
WERE INFECTED EARLIER IN SERIES,
THOSE WITH PREDICTED LOWER RISK
WERE INFECTED LATER WE CAN
RECAPITULATE OBSERVATION THE IL
12 ADJUVANTED GROUP HAD A LOWER
RISK OF INFECTION.
SO THIS WAS PRETTY EXCITING.
THEN WE LOOKED UNDER THE HOOD OF
THE MODEL AND FOUND FEATURES
ASSOCIATED WITH REDUCE RISK OF
INFECTION THOSE HIGH AND SHOWN
IN RED, AMONG INFECTION
RESISTANT ANIMALS SHOWN IN
BRIGHT GREEN, THOSE WERE
UNINFECTED OR DARK GRAY, WERE
ASSOCIATED WITH THE C 1Q BINDING
ABILITY OF HIV SPECIFIC
ANTIBODIES.
SO THIS ANALYSIS REALLY
IMPLICATED A ROLE OF THE
COMPLIMENT SYSTEM YOU ALREADY
HEARD ABOUT TODAY IN PROTECTION
MEDIATED BY THIS VACCINE.
A LOT THESE NON-HUMAN PRIMATE
VACCINATION STUDIES HAVE RAISED
QUESTIONS ABOUT WHETHER THESE
RELATIONSHIPS ARE INTERESTING
ASSOCIATIONS OR MECHANISTICALLY
RELEVANT.
EVEN AS THESE OBSERVATIONS PILE
UP FROM DIFFERENT STUDIES, THEY
STILL DON'T ENABLE YOU TO
DIRECTLY CONCLUDE WHAT ACTIVITY
WAS RESPONSIBLE FOR PROTECTION.
ASSOCIATIVE EVIDENCE.
SINCE THIS WAS THE FIRST BUT NOT
THE ONLY TIME WE HAVE SEEN
COMPLIMENT RELATED SIGNATURE WE
WANTED TO ASK WHAT DO WE KNOW
ABOUT COMPLIMENT IN VACCINE OR
ANTIBODY MEDIATED PROTECTION
FROM HIV INFECTION OR ANY OTHER
INFECTION.
THE ANSWER IS, IT IS
COMPLICATED, THIS IS THE
SIMPLEST DIAGRAM, CLEANEST I
HAVE SEEN IN THE COMPLIMENT
CASCADE.
IT IS TRULY A HUMONGOUS MESS.
AND UNPREDICTABLE.
IT IS AFTER LUNCH SOME OF US ARE
SHIELDING OUR EYES FOR
SLEEPINESS REASONS BUT I HAVE
GIVEN YOU ANOTHER ONE.
LOTS OF WAYS TO ACTIVATE
COMPLIMENT, SOME MEDIATED BY
ANTIBODIES, MANY IMMEDIATE
YATESED BY OTHER FACTORS, MANY
WAYS TO INTERRUPT AND
PREMATURELY TERMINATE COMPLIMENT
CASCADE BEFORE IT RESULTS IN
LYSIS OR CLEARANCE OF A TARGET
PARTICLE.
AND ALSO WAYS COMPLIMENT
PROTEINS INFLUENCE EACH OTHER
AND INFLUENCE OTHER EFFECTOR
CELLS PHENOTYPES.
NORBERT AND FOR WORSE.
FOR BETTER AND FOR WORSE.
THIS IS A COMPLICATED SYSTEM
THAT'S BEEN TRICKY FOR US TO TRY
TO PARSE OUT IN VITRO.
I WILL SPEND A LITTLE BIT OF
TIME TALKING ABOUT THAT.
BUT SINCE IT IS COMPLICATED, IT
IS ALSO INTERESTING AND SO
MARJORIE HAS WORKED ON IT AND SO
HERE IS MAYBE ONE STUDY THAT
HASN'T BEEN MENTIONED YET TODAY,
MILLION WHICH A MODEST
CORRELATION OF VIRAL LYSIS WAS
ASSOCIATED WITH REDUCED
INFECTION RATE IN A SEX
DEPENDENT FASHION.
SO THAT'S THE GRAPH SHOWN AT THE
LEFT YOU SEE VIRION LYSIS
MEDIATED BY ANTIBODIES IN
COMPLIMENT, ON THE Y AXIS AND
THE NUMBER OF CHALLENGES TO
ESTABLISH INFECTION ON THE X.
ALSO SOME EVIDENCE OF
RELATIONSHIP BETWEEN VIRAL LYSIS
AND PEAK VIRAL LOAD FOLLOWING
INFECTION SHOWN ON THE RIGHT.
SO THIS WAS PART OF WHAT
INSPIRED US TO TRY TO DIG INTO
WHETHER THESE OBSERVATION -- WE
COULD MECHANISTICALLY BOLSTER
THESE ASSOCIATIVE OBSERVATIONS.
PART OF THE REASON WHY THIS
PATHWAY IS SO COMPLICATED TO
UNDERSTAND OR PREDICT ITS
ACTIVITIES LIKELY RELATES TO THE
STRUCTURAL BIOLOGY INVOLVED SO
WE KNOW FROM SOME ELEGANT
STRUCTURAL BIOLOGY STUDIES THAT
IGG CAN HEXAMERRIZE THROUGH FC
DOMAIN AND THIS CAN BETTER
ENGAGE WITH THE 6 GLOBULAR HEADS
OF Y 1Q, IT VARIES WHETHER BOTH
FAB ARMS ARE BOUND TO ANTIGEN OR
NOT BUT IT IS INCREDIBLY
COMPLICATED AND NOT PERFECTLY
UNDERSTOOD.
THIS IS A -- INSTEAD OF SPATIAL
INFLUENCES THAT ARE DIFFICULT TO
HAVE A SENSE FOR AFRE A PRIORI
CERTAINLY IN CONTEXT OF
POLYCLONAL ANTI-BOUGH BODIES
WHERE STUDIES ARE PRESENT.
BUT STUDIES OF ANTI-CANCER MAPS
IT VARIES ALONG MAPS THAT TARGET
THE SAME ANTIGEN.
AND OF COURSE THE SPARSITY OF
ENVELOPE AND ITS MULTIPLICITY OF
CONFIRMATIONAL STATES ONLY ADD
COMPLICATION IN THE CONTEXT OF
HIV INFECTION.
SOMETHING ELSE WE KNEW FROM THE
CONTEXT OF HIV WAS THAT THERE
WAS A LOT OF SOMEWHAT
CONFLICTING CONFUSING EVIDENCE
RELATED TO NATURAL INFECTION.
BUT THERE WAS ONE DEFINITIVE
PASSIVE TRANSFER STUDY, SEMINOLE
ONE IN THE FIELD WHICH AT THE
SAME TIME IS INDICATED
ACTIVITIES OTHER THAN
NEUTRALIZATION DID CONTRIBUTE TO
PROTECTION FROM INFECTION
EXCLUDED COMPLIMENT FROM BEING
IMPORTANT MECHANISM OF ACTION SO
THE SO ANTIBODY IN RED IS KA
MUTANT AND ABLATES INTERACTION
BETWEEN ANTIBODIES AND C 1Q THAT
DIDN'T DECREASE B 12 TO PREVENT
INFECTION AT ALL.
KNOWING ALL THE ASPECTS, WE
WANTED TO HEAVY SIT THIS GIVEN
HOW NUANCED COMPLIMENT
ACTIVATION APPEARED FROM OTHER
FIELDS.
SO BEASKED THE QUESTION, UNDER
WHAT CIRCUMSTANCESES DOES B 12
EXHIBIT COMPLIMENT ACTIVITY SO
WE SET PANEL OF B 12 FC VARIANTS
USED IN ORIGINAL PASSIVE
TRANSFER EXPERIMENTS, LALA AN FC
GAMMA REACCEPT TO AND C 1Q KNOCK
OUT, KA IS A C 1Q BINDING KNOCK
OUT AND ENHANCED VARIANTS, EG
AND EFATE WHICH ARE SHOWN IN
BLUE, REPORTED TO ENHANCE
COMPLIMENT ACTIVATION BY DIVERSE
MECHANISMS.
SO THE ANSWER TO THE QUESTION
WHETHER B 12 WAS COMPLIMENT
ACTIVE WAS SOMETIMES SOME WAYS
SO SHOWN AT LEFT IS ANTIGEN
BINDING TO TWO RECOMBINANT
ENVELOPE PROTEINS FOUR WHICH
VARIANTS SHOWED EQUAL BINDING.
AND SHOWN IN THE CENTER, IS
BINDING OF C 1Q TO IGG BOUND TO
TARGETS, AS WELL AS TO ANTIBODY
DIRECTLY CODING IN ELISA PLATE
IS WHAT WAS SHOWN IN THE
ORIGINAL NHP PASSIVE TRANSFER
EXPERIMENT SO WILD TYPE B 12
SHOWS SOME ACTIVITY WHEN BOUND
TO ELISA PLATE AND THE EFTAE AND
EG VARIANTS SHOWED MORE BUT WE
REALLY DIDN'T SEE ANY ACTIVITY
AGAINST ANTIBODY WHEN BOUND TO
ANTIGEN.
WHETHER THAT'S BECAUSE OF WHAT
GEORGE INTENDED TO TELL US ABOUT
WHAT HE CUT OUT OF HIS TALK OR
SOMETHING ELSE.
I DON'T KNOW.
BUT THAT WAS THE OBSERVATION.
SO WILD TYPE B 12 COULDN'T DO
THIS IN THIS RECOMBINANT ANTIGEN
ASSAY.
WE ALSO LOOK AT C 3D DEPOSITION
IN THIS CONTEXT.
AND IT WAS DIFFERENT THAN THE C
1Q BINDING.
SO THAT SUGGESTED THAT WHATEVER
PART OF THE QUOTHMENT CASCADE
YOU LOOKED AT YOU MIGHT GET
DIFFERENT PATTERNS.
SO SEEMS DIFFICULT TO
EXTRAPOLATE.
OF COURSE WHAT'S SHOWN IN THE
LOWER RIGHT CORNER IS THE IMPACT
THAT ANTIGEN CAN HAVE.
SO HIGHLY GLYCOSYLATED ANTIGEN
LIKE HIV ENVELOPE DIRECTLY
INTERACT WITH COMPLIMENT CASCADE
PROTEINS AND TRIGGER C 3 D
DEPOSITION.
BUT IF TREATED THOSE ANTIGENS
THEN WE HAD BETTER RESOLUTION TO
SEE ANTIBODY DEPENDENT ACTIVITY.
SOMETIMES A SMIDGE OF COMPLIMENT
WAS ACTIVATED BY WILD TYPE B 12.
WHAT IF WE TARGETED FREE
VIRIONS?
ALL THE ANTIBODIES BOUND TO
VIRUS BUT C 1Q BINDING COULD BE
DETECTED FOR EFTAE ENHANCED
VARIANT AND NOT FOR WILD TYPE B
12 AND CERTAINLY NOT FOR KNOCK
OUT.
GIVEN WILD TYPE DIDN'T HAVE THAT
ACTIVITY.
SIMILAR FOR C 3 DEPOSITION
THOUGH A SMIDGE OF ACTIVITY OF
WILD TYPE B 12 BUT NONE OF THESE
VARIANTS OF B 12 LYSED VIRUS IN
OUR HANDS AND POSITIVE CONTROL
HERE WAS 10 A 8V 4 WITH
ENHANCING MUTATIONS.
WHAT ABOUT ON EXPRESSING CELLS>>
HERE IS HEC 293 CELLS EXPRESSING
ON SURFACE AND WE SAW AGAIN
REASONABLY EQUIVALENT ANTIGEN
BINDING.
C 1Q BINDING OF THESE CELLS ONLY
FOR ENHANCED VARIANTS, NOT FOR
WILD TYPE B 12.
C 3 DEPOSITION AND C 5B 9
FORMATION OF ENHANCED VARIANTS
AND NOT WILD TYPE AND SIMILAR
OBSERVATION MADE IN TERMS OF
CELL LYSIS.
COMPLIMENT DEPENDENT
CYTOTOXICITY.
ALSO MADE FOR B 12, WE COULDN'T
SEE ANY VIR ROLLSIS OR SIGH
TOLLSIS INDUCED BY B 12.
FIRST SET OF CONCLUSIONS ARE B
12 DIDN'T EXHIBIT COMPLIMENT
ACTIVATING ACTIVITY CERTAINLY
NOT ROBUSTLY.
EXPECTED TRENDS WERE GENERALLY
OBSERVED ACROSS THE VARIANTS BUT
THE MAGNITUDE AND EXTEND OF
DIFFERENCES BETWEEN ENHANCED
WILD TYPE AND KNOCK OUT WERE
VARIED, DETAILS WERE DIFFERENT.
SO COMPLIMENT DEPENDENT
ACTIVITIES KIND OF VARIED BY
EVERYTHING -- EVERY PARAMETER WE
HAD EXPLORED.
WHICH WAS A LITTLE BIT
FRUSTRATING.
SOMEONE SITTING IN ENGINEERING
SCHOOL WHO TENDS TO THINK OF
THINGS AND MAYBE A LITTLE BIT
REDUCTIONIST FASHION.
SO THE CONCLUSION WE CAME TO WAS
THAT FURTHER IN VIVO EXPERIMENTS
ARE PROBABLY GOING TO BE
NECESSARY TOSS TRY TO UNDERSTAND
WHETHER THIS IS IMPORTANT ASPECT
OF ANTIBODY BIOLOGY NORTH AND
WE'LL CONTINUE TO SEE
CONFLICTING OBSERVATION AN SPIN
WHEELS IF WE KEPT ALL OF OUR
INVESTIGATION IN VITRO.
IT ALSO SUGGESTED THAT THAT
ORIGINAL PAPER USING B 12 WHICH
SHOWED COMPLIMENT PLAYED NO
ROLE, B 12 DIDN'T HAVE A GOOD
ABILITY TO INDUCE COMPLIMENT
ANYWAY SO YOU ARE KNOCKING OUT
SOMETHING THAT WAS ALREADY
FUNCTIONALLY DEAD, WE THOUGHT
MAYBE THIS IS GENERALLY TRUE
GENERALLY TRUE BNAB BUT MAYBE
NOT AND IT IS CERTAINLY
SOMETHING WORTH TESTING.
IF B 12 WAS A DUD WHAT ELSE
COULD WE USE?
FOR US TIMOTHY AND OLIVIA
SCHWARZ WERE WORKING ON
ANSWERING THAT QUESTION FOR US.
SO WE PICKED TO INVESTIGATE 10E
8 WHICH SEEMS TO WORK BETTER IN
VIVO THAN IN VITRO IN TERMS OF
ITS WHAT ITS NEUTRALIZATION
POTENCY PREDICTS, IT IS A LITTLE
MORE PROTECTIVE IN VIVO THAN
EXPECTATION AND SHOWED A LITTLE
COMPLIMENT DEPENDENT
CYTOTOXICITY AND 10101074 THE
ANTIBODY THEY TESTED THAT HAD
THE BEST LITEDDIC ACTIVITY AMONG
BNABS SCREENED.
I WON'T SAY MUCH ABOUT
BIOLOGICAL CONCLUSIONS ABOUT THE
PASSIVE EPIFUSION STUDY.
BUT -- INFUSION STUDY THAT ANN
AND I CONDUCTED BECAUSE IT WAS
PUBLISHED EARLIER THIS YEAR, BUT
WE COMPARED WILD TYPE FUNCTIONAL
KNOCK OUT AND ENHANCED VARIANT
INFUSE TO ANIMALS AND CHALLENGE
WITH SINGLE HIGH DOSE VIRUS AND
THIS EXPERIMENT SHOWED THAT
EFFECTOR FUNCTION LIKELY
PHAGOCYTOSIS CONTRIBUTED TO
REDUCED SET POINT VIRAL LOADS.
BUT INSTEAD I WOULD LIKE TO
FOCUS ON SOME OF THE OTHER
LESSONS THAT WE LEARNED.
AND THAT TRANSFER TO OTHER
STUDIES WHICH WERE THE ANTIBODY
VARIANTS SHOWED PRETTY DIVERGENT
PK PROFILES SO CHANGING FC IS
NOT NEUTRAL TO PLASMA HALF LIFE
AND BIODISTRIBUTION SO THE KNOCK
OUT SHOWED GREATER SERUM
PERSISTENCE WHICH IS SOMETHING
OBSERVED BEFORE AND THE FC
RECEPTOR AND COMPLIMENT BINDING
ENHANCED VARIANT SHOWED
ACCELERATED CLEARANCE.
BUT BECAUSE THE ANIMALS IN THIS
STUDY WERE INFECTED COMPARISON
AMONG ANTIBODY TREATMENT GROUPS
WAS DIFFICULT.
SO THIS MADE US HEAVY SIT OTHER
VARIANTS HOVARIANTS WE RETESTED, DRIVEN BY
WILD TYPE 101-0374 AT LEFT AND
THREE ENHANCED MUTANTS, Y 300D
AND E 345R ON THE RIGHT.
WE RECAPITULATED OLIVER'S
COMPLIMENT AT COMPLIMENT AND CAN
DO BETTER IN CONTAINING
COMPLIMENT AND ENHANCING
MUTATION LIKE FTAE AND OTHERS.
BUT EACH OF THESE MUTATIONS WE
KNEW OR EXPECTED TO HAVE PK
PROBLEMS SO WE EXCLUDED FROM
CONSIDERATION.
SO THAT LEFT US WITH THE E 435G
MUTATION.
WHICH WAS ORIGINALLY DESCRIBED
BY DEJONG.
THAT DOESN'T CLANG THE INHERENT
ATIN -- CHANGE THE INHERENT
AFFINITY FOR THE C 1Q HEAD BUT
ALTERS IGG HEXAMERRIZATION
PROPENSITY BUT NOT IN SOLUTION.
THIS IS THE TIERRY BEHIND
ANTIBODIES, IT TAKES BINDING TO
ANTIGEN AND A CERTAIN EFFECTIVE
LOCAL CONCENTRATION THEN IT WILL
FAVOR HEXAMERRIZATION OF THE FC
DOMAIN.
AND POTENTIATED INTERACTION WITH
C 1Q AND COMPLIMENT DEPOSITION.
SO DEFINITELY OLDER AND LIKE
DEBATEICALLY WISER BECAUSE WE
WERE WORKING WITH COMPLIMENT
FROM OUR PRIOR STUDIES IN B 12
AND 10 BEFORE, WE DESIGNED AN
EXPERIMENT WHICH ANIMALS WERE
TREATED THREE DAYS PRIOR TO
CHALLENGE WITH FOUR MGS PER KG
OF 101074 WILE TYPE KNOCK OUT
AND COMPLIMENT ENHANCED VARIANTS
SUB Q.
PRIOR TO REPEATED LOW DOSE
CHALLENGE COMPRISED OF WEEKLY
INTERRECTAL EXPOSURE TO 30 CIB
50s OF SHIV SF 162 P 3.
THIS WAS THE RESULT.
SO OUR ENHANCED VARIANT DID
BETTER THAN THE ISOTYPE CONTROL,
IT WAS A NEUTRALIZING ANTIBODY
AFTER ALL.
RECEIVE BUT LESS WELL THAN
EITHER WILD TYPE OR COMPLIMENT
KNOCK OUT KA FORM.
WHICH HAD LARGELY OVERLAPPING
KAPLAN MIRE CURVES SO ON ITS
FACE THIS RESULT SUGGESTS THAT
NOT DID ENHANCING COMPLIMENT
ACTIVITY OFFER NO BENEFIT,
ELIMINATING IT HAD NO EFFECT.
, IT WASN'T CONTRIBUTING.
DESPITE THIS BEING BEST MUTATION
ANTI-BODY ENGINEERING HAD TO
OFFER US IN IN TERMS OF
ENHANCING ACTIVITY, WE HAD THE
STICKY PROBLEM OF VARIABLE PK.
AND THE PEAK CONCENTRATION OF
OUR COMPLIMENT ENHANCED EG
VARIANT WAS LOWER AND CLEARANCE
ELIMINATION OF EG WAS FASTER
THAN EITHER WILD TYPE OR KA
KNOCK OUT MUTANT.
YOU CAN SEE THAT THE TIME POINT
WHICH EACH ANTI-BITTY
CONCENTRATION DROPPED BELOW THE
PER ML VARIED CONSIDERABLY
ACROSS THE PANEL.
SO THIS WAS DISAPPOINTING
OUTCOME.
THIS IS LIKE A REAL GOLDBURG
DESIGN EXPERIMENT TO COME UP
WITH A CONCLUSION THAT ANTIBODY
DOSE AT TIME OF CHALLENGE
MATTERS.
WE WONDERS WHAT CAN WE LEARN?
FROM THIS EXPERIMENT?
WE WANTED TO CONSIDER IF THERE
WAS ALTERNATIVE WAY OF ANALYZING
THIS EXPERIMENTAL DATA THAT
WOULD BETTER ACCOUNT FOR THE
EFFECT OF VARIABLE PK FOR THESE
FC MODIFIED ANTI-BODIES.
SO FOR THAT WE LOOK TO WORK
COMING OUT OF THE VRC AND SOME
ARE PAGU'S WORK WHICH CAP PLAN
MIRE CURVES MODIFY TO REPORT THE
LEVEL OF ANTIBODY NEUTRALIZATION
ACTIVITY AND ITS RELATIONSHIP TO
LEVEL PROTECTION.
SO BY DOING THIS ANALYSIS YOU
CAN ARRIVE AT IN VIVO EC 50
NEUTRALIZATION ACTIVITY.
SO FOR THIS PARTICULAR VIRUS, IT
IS MORE SENSITIVE TO THE
SPECIFIC MAB THAN CD4 BINDING
SITE OR V 1 V 2 MAB, YOU CAN SEE
THE LEFT SHIFT OF THE CURVE, IT
TAKES LOWER IN VITRO NEW TITER
TO PROTECT HALF THE ANIMALS WITH
THAT HOE WITH SPECIFIC MABS THAN
CD4 OR V 1 V 2.
WE THOUGHT IF YOU COMPARE
ANTIBODIES WITH DIFFERENT
SPECIFICITIES IN THIS WAY
COMPARE THE SAME SPECIFICITY BUT
WITH DIFFERENT FC MODIFICATIONS.
SO RELATING THE LEVEL OF
PROTECTION, OBSERVED, DIRECTLY
TO THE LEVEL OF ANTIBODY
REQUIRED.
SO IF WE DO THAT, FOR THE 101074
DATA SET AND PLOT CONCENTRATION
OF ANTIBODY PRESENT AT TIME OF
BREAK THROUGH, IN THE ANIMALS
TREATED WITH EACH ANTIBODY, WE
WOULD COME -- WE MIGHT COME TO A
VERY DIFFERENT CONCLUSION.
SO AS MEDIAN CONCENTRATION OF
BNAB PRESENT AT TIME OF
INFECTION WAS .6 MICROGRAMS PER
ML WILD TYPE IT WAS THREE TIMES
LOWER THAN THAT FOR THE EG
COMPLIMENT ENHANCED VARIANT.
IT WAS TWO TIMES HIGHER FOR THE
KNOCK OUT VARIANT.
SO WHY WOULD NOT SURE WE CAN
EXCLUDE THE EFFECTIVE SYSTEMATIC
BIASES IN ALTERED PK FROM
IMPACTING THIS ANALYSIS, I THINK
IT DOES SUGGEST FIRST COMPLIMENT
MAY PLAY A ROLE IN 101074
MECHANISM OF ACTION, AND THAT
THIS ACTIVITY MIGHT BE ABLE TO
BE ENHANCED IN VIVO THROUGH
ANTIBODY ENGINEERING.
OF COURSE ALTERED PK IS A REAL
PROBLEM IN CLINICAL TRANSLATION
OF ANY OF THESE INSIGHTS.
SO IN CONCLUSION I STILL THINK
COMPLIMENT IS TRICKY AND IF
MARJORIE HAD ANOTHER CAREER TO
INVEST IN THIS SPACE MAYBE CHECK
FIGURE IT OUT.
PK DIFFERENCES ARE CONFOUNDER
TRYING TO UNDERSTAND MECHANISM
OF ACTION USING ANTIBODY
MUTATIONS.
WHETHER NAY ARE SINGLE HIGH DOSE
OR REPEAT LOW DOSE CHALLENGE
MODELS, BUT I THINK IT IS STILL
IMPORTANT TO USE THE NHP MODEL
TO DO THIS.
AND WE THINK GIVEN THE
COLLECTION OF STUDIES THAT HAVE
BEEN REPORTED TO DATE, THE
MECHANISMS OF ACTION OF
DIFFERENT BNABs ARE GOING TO
DIFFER.
SO WE WOULD WANT TO LOOK AT
THESE KINDS OF ANALYSES IN THE
CONTEXT OF ALL CLINICALLY
RELEVANT BNABs.
WITH THAT I WANT TO THANK BEN
THE Ph.D. STUDENT IN THE
GROUP, DEFENDING I THINK TWO
WEEKS FROM TODAY IN THIS
HALLOWEEN PHOTO OF THE LAB HE IS
HIDING IN THIS RED SUIT.
WHEN I WENT TO LOOK FOR A BETTER
PHOTO I FOUND THIS ONE OF HIM
HIDING FROM MONEY.
BEN HAS BEEN RESPONSIBLE REALLY
FOR ALL OF OUR WORK MANY THE
CONTEXT OF MABS AND COMPLIMENT,
HE DIDN'T DO THE WORK ALONE, IT
IS THE RESULT OF A LONG STANDING
COLLABORATION WITH ANN AT HSU
AND FORMER POST DOC DAVID
SPENCER AND EARLIER WORK
MOTIVATING THIS FOLLOW-UP IN
TERMS OF CORRELATES OBSERVED IN
VACCINE STUDIES, REALLY CAME
FROM THE FLSC VACCINE GEORGE
TALKED ABOUT AND AS WELL AS SOME
OF MARJORIE'S OBSERVATIONS.
AND SO I WANTED TO THANK
MARJORIE FOR THAT OPPORTUNITY TO
COLLABORATE.
ON THAT STUDY AND ALSO FOR
SENDING YOUR FANTASTIC TRAINEES
OUT INTO THE WORLD, IT'S BEEN
GREAT TO HEAR FROM SOME OF THEM
TODAY.
THANK YOU.
[APPLAUSE]
>> SO VERY INTERESTING OBVIOUSLY
COMPLEX AS YOU SAID STUDY.
MAYBE I MISSED IT BUT DID YOU
LOOK AT DIFFERENCES BETWEEN FC
GAMMA R 1 R 2 AND R 3 AS IN THE
WORK OF JEFF RAVAGE WHERE
OBVIOUSLY THOSE FUNCTIONS ARE
DIFFERENT AND THAT COULD PLAY A
ROLE?
>> SO FOR EACH OF THE PASSIVE
TRANSFER EXPERIMENTS WE HAVE
DONE WE HAVE ALSO LOOKED AT
FINDING AFFINITY OF THE VARIANTS
TO FC GAMMA Rs AND FCRN.
SOME WE TRANSFERRED ANTIBODIES
THAT HAD ENHANCEMENTS TO BOTH FC
GAMNA RES AND C 1Q IN THE
PUBLISHED STUDY, I DIDN'T TALK
ABOUT AS MUCH TODAY.
WITH EG MUTANT, THE BINDING TO
FC GAMMAR IS NOT SUBSTANTIALLY
IMPACTED.
BUT I THINK THIS IS AN IMPORTANT
-- AN IMPORTANT TO THINK ABOUT
THIS.
WHERE PEOPLE HAVE SEEN ANTIBODY
ENGINEERING ENHANCE OR FC
ENGINEERING ENHANCE PROTECTION
IN ANIMAL MODELS IT HAS BEEN IN
MICE.
PEOPLE HAVE NOT SEEN THAT HAPPEN
IN THE NHP MODEL THOUGH THE SAME
-- SO PGG 121 IN PUBLICATIONS
YOU CAN ENHANCE ITS PROTECTIVE
CAPACITY IN VIVO IN MICE WHEN
PEOPLE HAVE LOOKED AT THIS IN
THE NON-HUMAN PRIMATE MODEL
MULTIPLE LABS THEY DON'T -- THEY
HAVEN'T SEEN A ROLE.
I THINK THE EXPERIMENTS DONE
LOOKING AT OTHER FC RECEPTOR
BINDING ENHANCED ANTIBODIES THEY
HAVE ALSO NOT PANNED OUT IN VIVO
IN THE NON-HUMAN PRIMATE MODEL.
THAT MAY BE BECAUSE OF
CHALLENGES RELATED TO LIKE
FUNDAMENTALLY TO CHALLENGES
RELATED IN PC.
SO I THINK IT WOULD BE REALLY
INTERESTING TO HEAVY SIT THOSE
EXPERIMENTS -- RE-VISIT THOSE
EXPERIMENTS FOR UNDERSTANDING
THE CONCENTRATION ON BOARD BNAB
INFECTION TO SEE IF THERE IS
MORE EVIDENCE THIS IS POSSIBLE
IN NON-HUMAN PRY MATES.
-- PRIMATES.
>>WE ARE IN THE LAST SESSION
HERE ON SEX BIAS AND MICROBE
SIDES, I WOULD LIKE THE FIRST
INTRODUCE ISKRA TUERO, FORMER
POST DOC WITH MARJORIE, NOW
ASSOCIATE PROFESSOR IN LIMA,
PERU.
>> GOOD AFTERNOON, EVERYONE.
SO FIRST I WANT TO THANK THE
ORGANIZER FOR INVITING ME.
I'M REALLY HONORED TO BE HERE
FOR THIS VERY SPECIAL OCCASION.
THE TITLE OF MY TALK IS SEX
DIFFERENCE TO BE CONSIDERED IN
THE DESIGN OF HIV VACCINES.
MY NAME IS ISKRA TUERO.
I WAS POST DOC AT DR.
(INAUDIBLE) LAB FROM 2011 TO
2016 AND I'M PROFESSOR AT
UNIVERSIDAD PERUANA CAYETANO
HEREDIA IN PERU.
IT IS WELL KNOWN THE SEX HORMONE
IMMUNE RESPONSE THROUGH SPECIFIC
RECEPTOR EXPRESS IN IMMUNOCELLS,
THE MECHANISM IS MODULATED BY
THE SEX HORMONE BY IMPACT THE
VACCINE OUTCOME AND THIS --
MIGHT DEFINE THE SEX DIFFERENCE.
BUT THE SORRY I WILL TELL YOU
NOW WASN'T REALLY INTENDED TO
ELEVATE SEX DIFFERENCE.
SO THIS IS -- WITH THIS PAPER,
WHICH WE DEMONSTRATE FOR THE
FIRST TIME THAT EFFICACY SHARED
IN VACCINATED FEMALES IN MALES
VERSUS MACAQUE, THIS PAPER WAS
PUBLISHED IN 2016.
FOR THIS STUDY WE FOLLOW
CHALLENGE THE INITIAL AIM FOR
THIS STUDY IS THE GP 140
MONOMERIC -- SO THE RATIONALE
FOR THIS STUDY THAT GP 1 FOR COB
SERVE CONFIRMATIONAL EPITOPES
THAT MAYBE MORE PROMISING TO
INDUCE BROAD NEUTRALIZING
ANTIBODY BETTER THAN THE
MONOMERIC GP 120.
SO WE PRIME OR ANIMAL WITH THE
ADENOVIRUS EXPRESS IN DIFFERENT
RECOMBINANT PROTEIN FOLLOWED BY
CORRESPONDING GP 120 OR GP 140
TO FINALLY BE CHALLENGE
REPETITIVE LOW DOSE OF SIV
MAPPED 251.
SO IN THE FIRST ANALYSIS WE
FOUND SOME DIFFERENCE BETWEEN
TWO GROUPS GP 120 AND GP 140.
OTHERS SUGGEST MAYBE SHE IS --
SHE DID SOMEONE LIKE THIS
BEFORE.
BY SEX COMPARE FEMALES WITH
MALE, AND THAT'S THE RESULT I
WOULD PRESENT NOW.
SO ALL ANIMAL -- NOT ALL, ONLY
ONE FEMALE DIDN'T GET INFECTED,
SO HERE YOU CAN SEE THE VIRAL
LOAD IN THE DIFFERENT
IMMUNIZATION GROUPS.
YOU CAN SEE THAT VACCINATED
FEMALES EXHIBITED SIMILAR FOR
THE DIFFERENT IMMUNIZATION
GROUP.
HOWEVER, WHEN WE PERFORM
ANALYSES TO SEE RATE OF A AND B
INFECTION AMONG FEMALES OR MALES
MALES, HERE IS THE RESULT.
WHEN WE ANALYZE ALL
IMMUNORECOGNIZE FEMALE COMPARED
TO CONTROL FEMALE YOU CAN SEE
STATISTICALLY SIGNIFICANT
DIFFERENCE IN THE SIV
(INAUDIBLE) SO FEMALES REQUIRE
MORE CHALLENGE TO BECOME
INFECTED COMPARED TO TH TO IMMUNIZE
MALE.
SO WE ANALYZE DIFFERENCE IN
IMMUNE RESPONSE FOCUS ANTIBODY
RESPONSE.
SO HERE AS A RESULT REGARDING
THE IMMUNIZATION GROUP, YOU CAN
SEE ALSO DIFFERENCE IN THE SIV
APPLICATION MANY THE GROUP GP
120 VERY SIGNIFICANT FOR THE
FEMALES BOTH OF THE CONTROLS BUT
IMAGINE THAT IN GROUP GP 140
IMMUNIZE FEMALE.
BIG DIFFERENCE WAS NOT OBSERVED
IN BOTH IMMUNIZATION GROUP FOR
THE MALE, IMMUNIZED MALES THEN
WE ANALYZED ANTIBODY RESPONSE
ANTIBODY RESPONSE BETWEEN
FEMALES AND MALES, AND WE FOUND
SOME SIMILARITIES.
THESE INCLUDE SIMILAR IN THE
NEUTRALIZING ANTIBODY TITERS, IN
ORDER ACTIVITIES WE MEASURE
CYTOTOXICITY PHAGOCYTOSIS, WE
FOUND SIMILARITIES IN THE LEVELS
OF PLASMA BLAST PLASMA CELLS,
MEMORY B CELLS ENVELOPE SPECIFIC
IGA IN BONE MARROW, ALSO SIMILAR
ENVELOPE SPECIFICALLY MEMORY B
CELLS IN BLOOD AND WHEN WE
EVALUATE ENVELOPE SPECIFIC IGG
AND IGA MUCOSAL SIDE WE DIDN'T
FIND DIFFERENT.
NOW I WILL SHOW YOU THE
DIFFERENCE WE FOUND FOR THE
DIFFERENT -- COMPARISON BETWEEN
FEMALE AND MALES SO I SHOW YOU
FINDING ANTIBODY TITERS THROUGH
GP 120 AND GP 140 AND YOU CAN
SEE IN THE GP 120 GROUP THE MEN
HAVE IODINED ANTIBODY TITERS TO
GP 120 COMPARED TO FEMALES.
THESE ARE THE DIFFERENT IN WEEK
14, 53 AND TWO WEEKS POST
INFECTION M. THE GP FOR THE
TITLER GP 120 WE HAVE OBSERVE
DIFFERENCE BETWEEN FEMALES AND
MALES.
WE DIDN'T OBSERVE DIFFERENCE IN
GP 140 GROUP.
GP 120 AND GP 140.
ALSO WE TESTED HUMAN IMMUNE
RESPONSE IN MUCOSAL SIDE.
SO IN THIS TABLE I SHOW YOU THE
GP 120 SPECIFIC MEMORY B CELLS,
WE TEST THESE -- THIS LEVEL OF
THESE CELLS AT WEEK 53, TWO
WEEKS POST INFECTION AND EIGHT
WEEK POST INFECTION.
AT TWO WEEKS POST INFECTION THE
FEMALES SHOW HIGHER NUMBER OF
PORTION OF ENVELOPE SPECIFIC
MEMORY B CELL COMPARED TO THE
MALES.
THEN AT THIS TIME POINT TWO
WEEKS POST INFECTION, SO WE
TRIED TO ADD -- FIND IF THIS
PORTION OF MEMORY B CELL WERE
ASSOCIATE SO THE SIV -- FOR ALL
IMMUNIZED FEMALES FOR ALL
IMMUNIZE FEMALES AND FOR THE GP
120 GP 140 IMMUNIZE FEMALE YOU
CAN SEE STATISTICAL DIFFERENCE,
POSITIVE CORRELATION BETWEEN THE
ENVELOPE AND SPECIFIC MEMORY B
CELLS VERSUS THE NUMBER OF --
FEMALES REQUIRE MORE CHALLENGE
TO BE COMPARED TO (INAUDIBLE)
SAME FOR PLASMA CELL, THIS IS
MUCOSAL SIDE SO THE SAME FOR
PLASMA CELL ALSO YOU CAN SEE ALL
IMMUNIZE FEMALE HAVE POSITIVE
CORRELATION BETWEEN NUMBER OF
CHALLENGE REQUIRED TO BECOME
INFECTED.
WE COULDN'T FIND ANY
STATISTICALLY DIFFERENT IN THE
ALL IMMUNIZED.
SO WHAT WE TRIED TO FIND THERE
IS A RELATION BETWEEN ENVELOPE
SPECIFIC IGG THAT MEASURE IN
ERECTILE SECRETION WE DID NUMBER
OF CHALLENGE AND IN THE
UNIMMUNIZED FEMALE YOU CAN SEE
-- YOU CAN SEE THAT WE HAVE
STATISTICALLY SIGNIFICANT
DIFFERENCE IN ALL IMMUNIZE
FEMALES AND ONLY IN THE GROUP GP
120 IMMUNIZE FEMALES.
SO OUR RESULT, AT THIS POINT OUR
RESULT HIGHLIGHT THE IMPORTANCE
OF MUCOSAL IMMUNITY ON MEMORY B
CELL OF THE (INAUDIBLE) FOR
PROTECTION AND THIS SHOULD BE
CONSIDERED FOR DEVELOPMENT OF
HIV VACCINE.
THESE WERE THE FIRST REPORT IN
IN SEX BIAS IN RESPONSE TO SIV
VACCINE IN RHESUS MACAQUE.
THIS WAS THE FIRST PAPER SHOWING
THE RESULT BUT THEN MY
COLLEAGUES IN THE LAB FOUND
ADDITIONAL PARAMETERS THAT COULD
HELP UNDERSTAND THE SEX
DIFFERENCE.
HE EVOLUTE THE DIFFERENT IGG OF
CLASS AND FOUND THAT IGG WAS
REALLY HIGH IN THE FEMALES
COMPARED TO THE MALES.
SO THEN HE TRIED TO UNDERSTAND
WHAT IS THE ROLE OF IGG 3 IN THE
SIV APPLICATION AND WHAT HE DID
IS -- ACQUISITION AND HE
CORRELATED LEVELS OF GP AND IGG
3 VERSUS THE DIFFERENT ANTIBODY
FUNCTIONS LIKE CYTOTOXICITY,
PHAGOCYTOSIS AND HE FOUND
POSITIVE CORRELATION IN ONLY IN
THE UNIMMUNIZED FEMALES AND
CORRELATED LEVELS WITH PEAK
VIRAL LOAD, HE FOUND NEGATIVE
CORRELATION IN THAT MORE IGG
MAYBE FACILITATED REPLICATION OF
VIRUS.
FINALLY ADDITIONAL PARAMETER --
SORRY.
ADDITIONAL PARAMETER THAT
EVALUATED WAS REGULATORY B
CELLS.
HE FOUND SECOND DOSE -- POST
INFECTION THE MEN HAVE HIGH
LEVELS OF THE -- COMPARED TO
FEMALES AND THESE LEVELS HIGH
LEVELS OF REGULATORY B CELL
CORRELATE WITH HIGH LEVELS WITH
HIGH VIRAL LOAD IN THE PEAK OF
THE INFECTION.
SO I WANT TO SUMMARIZE THESE
FINDINGS.
THIS FIRST DEMONSTRATION OF 6
DIFFERENT SIV VACCINE PROTECTION
EMPHASIZE THE NEED FOR SIX
BALANCE IN VACCINE TRIALS SO
ALSO A FINDING OF COURSE
INCREASE COMPLEXITY.
IN TO THESE AREAS FOR FUTURE
INVESTIGATION LIKE IGG SUBTYPE,
GLYCOSYLATION, FC POLL MORPHINE
AND THE MICROBIOME MIGHT
INFLUENCE THE VACCINE OUTCOME.
IN ADDITION, WE NEED TO TAKE
ACCOUNT INTO ACCOUNT POTENTIAL
HORMONAL EFFECT BUT PARTICULARLY
EFFECT IN MUCOSAL IMMUNITY WITH
THE SITE OF THE CHALLENGE, ALSO
THINK ABOUT THE VACCINATION THAT
MIGHT BE IMPORTANT FOR THE
OUTCOME.
I THINK IT WOULD BE IMPORTANT TO
IDENTIFY IMMUNOCORRELATE
ASSOCIATED WITH PROTECTION IN
MALE PRIOR TO VACCINE REGIMEN.
ALSO IT IS IMPORTANT TO IDENTIFY
POTENTIAL MECHANISM THAT MIGHT
FACILITATE SIV ACQUISITION AS I
SHOW YOU IN MALES.
FINALLY I WANT TO SAY THANKS TO
MARJORIE, THANKS FOR HER
KNOWLEDGE, MY DEEPEST
CONGRATULATIONS FOR YOUR
INSPIRING CAREER.
THANK YOU FOR MY GRATITUDE
FOREVER.
THANK YOU.
[APPLAUSE]
>> THANK YOU FOR KEEPING YOUR
TALK SHORT.
ANY QUESTIONS?
>> THANK YOU FOR THAT, VERY
INTERESTING.
DID YOU COLLECT ANY INFORMATION
ON CYCLING HOW THE FEMALES WERE
CYCLING?
THIS IS A CHALLENGE, IF HAS BEEN
INTRAVAGINAL YOU WOULD HAVE DONE
--
>> BECAUSE THE STUDY WAS NOT
DESIGN TO CHASE SEX DIFFERENCE
WE PERFORM THAT ANALYSIS FOR
SOME OF THE MACAQUES, BUT WE
DIDN'T HAVE COMPLETE DATA.
SO WITH THIS WE COULDN'T FIND
ANY OR SOMETHING INTERESTING FOR
THE ANALYSIS.
WE JUST HORMONES YES BUT NOT ALL
ANIMALS.
WE DON'T HAVE THIS CYCLING.
IT WAS NOT THOUGHT FOR THAT ONE.
BUT COULD BE INTERESTING TOOL TO
FLOW FUTURE MORE IN DETAIL THIS
PARAMETER.
IN FOLLOW-UP HAVE YOU ABLE TO
APPLY, GIVING HORMONES OR
BLOCKING HORMONES TO SEE TO TEST
WHETHER THIS IS FAMILIAL EFFECT
IN OTHER WORDS YOU CAN BLOCK
TEST TOAST ROANE PRODUCTION OR
BLOCK TESTOSTERONE RECEPTORS OR
GIVE ESTROGEN, ALL THESE THINGS
THAT YOU CAN DO IN ANIMAL MODELS
THAT MIGHT HELP ADDRESS THOSE
QUESTIONS.
>> MAYBE FUTURE STUDY WE NEED TO
CONSIDER ALL THAT, THIS STUDY
WAS NOT THINKING WAS PLANTED
ACROSS HORMONES OR ANY OTHER
PARAMETERS.
>> AS OF LATE THERE HAVE BEEN A
LOT OF THESE SEX DIFFERENCES,
HOWEVER I HAVEN'T SEEN ANY
INDICATOR OF WHAT -- WHY THEY
WOULD BE.
THAT DIFFERENCE X AND Y BUT
BESIDES THAT, WHY IS THERE THAT
DIFFERENCE IN THE OUTCOME, ANY
IDEA?
>> THE SEX HORMONES AFFECT ALL
TISSUES, NOT ONLY IN FEMALE BUT
DRUG HORMONES ELIMINATE IMMUNE
RESPONSE IN MANY IMMUNOCELL
RECEPTOR SO WE COULD PICK THAT
SIX HORMONE MODEL IMMUNORESPONSE
IN ALL TISSUES.
(OFF MIC)
>> I HAVE NO ANSWER FOR YOUR
QUESTION, MAYBE WE CAN DISCUSS
LATER.
>> THERE ARE NO FURTHER
QUESTIONS, THANK YOU.
[APPLAUSE]
>> I FEEL LIKE A BIT OF AN
ENTERLOPER AT THIS MEETING.
I'M FILLING IN FOR TOM MUSIC WHO
WAS UNABLE TO JOIN US, HE WAS
POST DOC WITH MARJORIE BUT
CURRENTLY HE'S DEPLOYED IN SYRIA
SO HE CAN COULDN'T COME BACK FOR
THE MEETING.
TODAY I WILL BE TALKING ABOUT
LACTIN V, A LACTOBACILLUS FOR
PREVENTION OF RECURRENT BV AN
HIV.
THE VAGINAL MICROBIOME IN WOMEN
CAN EXIST IN H TWO STATES.
IT CAN BE LACTOBACILLUS
DOMINATED WHETHER YOU YOU LOOK
AT GRAM STAIN OR BACTERIAL
COMMUNITIES PLOT YOU CAN SEE LOW
DIVERSITY.
PH IS LOW.
AND THIS ANTAGONIZES PATHOGENS
AND THERE'S HIGH LEVELS OF
LACTIC ACID WHICH OF COURSE
DRIVE PH DOWN.
THIS IS A NON-INFLAMMATORY
ENVIRONMENT.
IF YOU CONTRAST THAT TO
BACTERIAL VAGINOSIS, A CONDITION
OF WOMEN DYSBIOSIS OF
MICROBIOME, LOOK AT GRAM STAIN
OR BACTERIAL COMMUNITIES PLOT,
YOU CAN SEE THAT THERE'S HIGH
DIVERSITY, THERE IS A LOSS OF
HYDROGEN PEROXIDE PRODUCING
LACTOBACILLI AND THERE'S HIGH PH
ABOVE 5, THAT IS HIGH FOR
VAGINAL VAULT.
THERE CAN BE ODOR DISCHARGE AND
INFLAMMATION INCLUDING CYTOKINES
AND SOME POLYNOR FIN LEUKOCYTES
AND THIS INCREASE WOMEN OF STI
AND HIVE.
THIS IS TREATED WITH ANTIBIOTICS
BUT RECURRENCE IS COMMON.
THERE'S A NUMBER OF STUDIES
LINKING THE VAGINAL MICROBIOME
TO HIV RISK.
ONE OF THE FIRST WAS FROM CRAIG
COHEN AT UCSF IN 1995, THAT
SHOWED ASSOCIATION OF BV WITH
HIV ACQUISITION IN WOMEN.
COHEN WENT ON TO FIND THAT WOMEN
WITH BV MORE LIKELY TO TRANSMIT
HIV TO THEIR INTIMATE MALE
PARTNERS.
FINALLY A STUDY BIGOSESMAN IN
IMMUNITY FOUND WOMEN WITH EV
WERE FOUR FOLD HIGHER RISK OF
HIV ACQUISITION.
THIS IS JUST A DIAGRAM FOR THE
GOSSMAN STUDY.
WOMEN WITH LACTOBACILLUS
DOMINANT COMMUNITIES ESPECIALLY
LACTOBACILLUS KRISPOTUS HAS
LOWEST LEVELS OF PRO INPROGRAM
FOUR CYTOKINES AND LOWER RISK OF
HIV INFECTION LOWER NUMBERS OF
TARGET CELLS IN LIEU MINUTE OF
VAGINA.
WOMEN WITH DIVERSE BACTERIAL
COMMUNITIES CONVERSELY HAD
HIGHER LEVELS OF
PRO-INFLAMMATORY CYTOKININGS AND
UP TO 17 FOLD INCREASE IN
ACTIVATED CD4 TARGET CELLS FOR
HIV.
AND AGAIN THEY WERE FOUR FOLD
MORE LIKELY HIV INFECTED IN
STUDY.
THIS WAS DONE IN YOUNG SOUTH
AFRICAN WOMEN.
SO OUR HYPOTHESIS WAS THAT WE
COULD ADD A LIVE BIOTHERAPEUTIC
PRODUCT A LACTOBACILLUS KRESATUS
TO SHIFT THE VAGINAL MICROBIOME
TOWARD OPTIMAL STATE TO PREVENT
AND REDUCE HIV RISK.
WHAT IS A LIVE BIOTHERAPEUTIC
PRODUCT AND WHY IS IT DIFFERENT
THAN PROBIOTIC?
THIS WAS A TERM COIND BY THE FDA
LBP IS A BIOLOGICAL PRODUCT THAT
CONTAINS LIVE ORGANISMS SUCH AS
BACTERIA BUT IT IS APPLICABLE TO
PREVENT TREAT AND CURE DISEASES
OF HUMANS.
THUS LBP IS BASICALLY A DRUG, IT
IS MANUFACTURED UNDER GNP
CONDITIONS AN MEETS POTENCY AND
PURITY STANDARDS WHEREAS
PROBIOTICS MAY CONTAIN LIVE
ORGANISMS BUT THEY ARE REGULATE
AS DIETARY SUPPLEMENTS OR FOOD
AND NOT INTENDED TO TREAT
DIAGNOSE OR CURE OR PREVENT ANY
DISEASE IN HUMAN IF YOU READ ON
THE LABEL.
SO LACK TIN V OUR PRODUCT IS A
LACTOBACILLUS CRESTPOTIS.
THAT IS THE STRAIN DESIGNATION.
IT IS DELIVER AS A POWDER IN
VAGINAL APPLICATOR.
IT HAS BEEN IN A PHASE 1 TRIAL
WHICH IS SHOWED SAFETY AND WELL
TOLERATED.
IT'S BEEN IN A PHASE 2A TRIAL
WHICH WAS SPONSORED BY NIAID,
WHERE WE SAW 78% COLONIZATION.
WE RECENTLY COMPLETED A PHASE 2B
TRIAL WHICH I WILL TALK ABOUT
TODAY, THIS WAS 228 WOMEN AND IT
WAS DIRECTED BY CRAIG COHEN
UCSF, THE PAPER WAS PUBLISHED IN
THE NEW ENGLAND JOURNAL OF
MEDICINE.
IN A SUCCESSFUL LACTIN
COLONIZATION OF PHASE 2 WE CAN
SEE THIS WOMAN HAS HIGH LEVELS
OF BACTERIAL VAGINOSIS
ASSOCIATED BACTERIA INCLUDING
MEGASFERA EBAV, COMBINATION OF
BACTERIA, AND LOW LEVELS OF
LACTOBACILLUS INERS.
YOU CAN SEE A DROP IN BV
BACTERIA BUT YOU DONE SEE A
CONCOMITANT INCREASE IN
LACTOBACILLUS UNTIL YOU ADD
EXOGENOUS LACTOBACILLUS AND THEN
YOU SEE LEVELS INCREASE.
AND THIS HELPS DRIVE DOWN AND
KEEP THESE AN AEROBIC BACTERIA
LOW LEVELS SO LACTIN B
ADMINISTERED FOLLOWING
ANTIBIOTICS REPLENISHES THE
MICROBIOME TOWARDS OPTIMAL
STATE.
THIS IS FROM PHASE 2A STUDY, YOU
CAN AGAIN SEE THESE ARE
BACTERIAL COMMUNITIES.
THIS WAS A WOMAN BEFORE
TREATMENT, 28 DAY NOW ALMOST ALL
LAKOTAS BACILLUS C, CRESPOTIUT
SHOWN IN PINK SO SUCCESSFUL
TREATMENT.
SO IN OUR PHASE 2B STUDY, THE
OBJECTIVES WERE TO TREAT -- TEST
TOTAL OF 228 WOMEN RANDOMIZE 2
TO 1 TO RECEIVE LACTIN V OR
PLACEBO.
THEY RECEIVED TOPICAL M&A
RATANIZOL FIVE DAYS OR PLACEBO
FIVE DAYS AND TEN WEEKS OF
TREATMENT TWICE A WEEK.
THEY WERE TREATED TWICE A WEEK
TO BE SURE THAT DURING MENSES WE
WOULD CONTINUE TO ADD
LACTOBACILLUS AN IT WOULDN'T BE
RINSED OUT.
PRIMARY OBJECTIVES OF THE STUDY
WERE EFFICACY OF EB RECURRENCE
AND SAFETY AT 24 WEEKS.
SECONDARY OBJECTIVES WERE TO
DETERMINE COLONIZATION LEVELS,
USER ACCEPTABILITY AND
TOLERABILITY, LONG TERM
COLONIZATION OF LACTIN B 24
WEEKS WHICH IS 12 WEEKS POST
DOSING AFTER THEY RECEIVED LAST
DOSE AND LONG TERM EFFICACY.
BV IS A CLINICAL CONDITION.
THERE ARE SEVERAL DIAGNOSTIC
CRITERIA, THE CLINICAL CRITERIA
IS CALLED AN AMSEL SCORE, A
COMBINATION OF WHETHER THE WOMAN
HAS A HOMOGENOUS DISCHARGE
WHETHER SHE HAS CLUE CELLS WHICH
ARE BACTERIAL VAGINAL CELLS, IS
A LOT OF BACTERIA STICKING TO
THEM.
PH ABOVE 4.5 AND OFTEN A FISHY
ODOR CALLED THE WENT TEST.
THE OTHER IS A GRAM STAIN WHICH
IS CALLED A NUGEN CRY TIERIA.
YOU LOOK FOR GRAM POSITIVE RODS
LACTOBACILLI AND OTHER CURVED
RODS WHICH ARE GARDENERELLA AND
OTHER PLEA OWE MORPHIC RODS.
7 TO 10 MEANS BV SO IN OUR
STUDY, WHEN WE EXAMINE WOMEN
ENROLLED, 51% HAD HAD MORE THAN
5 BV EPISODES IN A YEAR AND
RECURRENT BV IS CONSIDERED THREE
OR MORE EPISODES IN A YEAR.
SO MAJORITY OF WOMEN IN OUR
STUDY REALLY HAD RECALCITRANT
BV.
SO THESE ARE DEMOGRAPHICS.
60% OF THE WOMEN IN STUDY WERE
WOMEN OF COLOR.
BV IS MORE COMMON IN HISPANIC
AND WOMEN OF AFRICAN DESCENT.
SO WE HAD THOSE NUMBERS WELL
REPRESENTED IN THE STUDY.
THOSE KIND OF WOMEN WELL
REPRESENTED.
SO IN OUR PHASE 2 BV CURE WAS
57% IN THE LACTOBACILLUS
CRESPONDIS ARM AND 39% PLACEBO
ARM 12 WEEKS.
THIS CORRELATED WHEN YOU LOOK AT
ALL THE DATA TOGETHER OF ABOUT A
46% REDUCTION IN RECURRENT
BACTERIAL VAGINOSIS.
AND THE SAME WAS TRUE AT 24
WEEKS, THREE MONTHS POST DOSING.
AT THE 12 WEEK VISIT
CRESPONDUCTIONUS WAS DECKED IN
79% OF WOMEN IN LACTOBACILLUS
CRESPONDUS ARM AND ONLY 3% IN
PLACEBO HAD LACK OF
LACTOBACILLUS STRAIN.
WHEN YOU EXOGENOUSLY GIVE IT YOU
CAN DRIVE WOMEN TOWARD THAT AND
TREATING ARE ANTIBIOTICS IS NOT
ENOUGH TO BRING BACK
LACTOBACILLUS.
SO HERE WE ARE LOOKING AT PCR
QPCR FOR TOTAL BACTERIA IN
VAGINA BACK LOW BACILLUS
SPECIES, CRISPATUS OR CTV 05
STRAIN AND PH.
IN IS AFTER WOMEN HAVE BEEN
TREAT WITH ANTIBIOTIC SO WE HAVE
DRIVEN DOWN THE TOTAL NUMBER OF
BACTERIA IN VAGINAL MICROBIOME
UX SEE THE PH STARTED HIGH OVER
HERE ABOUT 6 IN THIS WOMAN.
AFTER TREATED WITH ANTIBIOTICS
THE LEVELS COME DOWN AND SHE IS
TREATED WITH LACTIN V.
ALL THESE QPCR CURVES PARALLEL
SHOWING THAT MOST OF THE
BACTERIA NOW IN THE VAGINA
LACTOBACILLUS CRISPATUS AND R
STRAIN.
IN WOMEN SUCCESSFULLY COLONIZED,
WE COULD SEE USUALLY THEY HAD
ABOVE 10 TO 6 LACTOBACILLI.
YOU CAN SEE HERE THESE ARE WOMEN
THAT WERE BV POSITIVE, THIS IS
12 WEEKS AND 24 WEEKS.
SO AGAIN, BY 24 WEEKS THE WOMEN
THAT DID NOT HAVE LACTOBACILLUS
WERE BV POSITIVE.
SO 10 TO 6 SEEMED TO BE THE
NUMBER SUFFICIENT TO PREVENT
RECURRENCE.
SO IN SOME OF THE NEXT STEPS WE
ARE STARTING TO ANALYZE META
GENOMIC TIME COURSE FOR LACTIN
V, I WILL SHOW YOU THAT DATA.
CHARACTERIZE VAGINAL MICROBIOME
ECOSYSTEMS.
PERFORM SOME INTEGRATIVE
ANALYSIS TO IDENTIFY
ASSOCIATIONS WITH BV, THIS IS
STILL ONGOING.
AND DEVELOP PREDICT SIGNATURES.
PILOT STUDY OF 29 WOMEN WERE
SAMPLES UNDERWAY TO LOOK AT
MICROBIOME BEING DONE AT THE
ZUCKERBURG BIOHUB AND LOOKING AT
CYTOKINES AND CHEM COINS DONE AT
THE CALL LAB IN TORONTO.
CHEMOKINES.
SO IN THAT PRELIMINARY DATA OF
META GENOMIC TIME COURSE YOU CAN
SEE WOMEN TREAT WITH LACTIN V
START WITH A BV PROFILE, HERE
THEY HAVE GOT HIGH LEVELS BV
ASSOCIATED BACTERIA, GARNERELLA
BACKROID DEES AND TREAT WITH
ANTIBIOTICS AT TIME POINT 1, THE
FIRST LACK DOE BACILLUS BECOME
IN WOMEN IS LACTOBACILLUS ENERS.
IT IS NOT CONSIDERED ONE OF THE
MOST BENEFICIAL LACTOBACILLI BUT
IT IS ABLE TO LIVE VERY HAPPILY
WITH BACTERIAL VAGINOSIS
ASSOCIATED BACTERIA.
SO BUT ONCE WE ADD LACTOBACILLUS
CRISPATIS THESE WOMEN CONVERTED
TO LACTOBACILLUS CRISPATUS
MICROBIOME, THIS TIME POINT 7 IS
24 WEEK TIME POINT SO THAT IS
AGAIN THREE MONTHS POST DOSING.
THESE WOMEN CONVERTED TO
LACTOBACILLUS CRISPATUS
PHENOTYPE, THIS HAS
LACTOBACILLUS AT THE SECOND TIME
POINT, DIDN'T CONVERT AT THIS
TIME POINT THEN PICKED UP
CRISPATUS THEN WENT BACK TO THAT
PHENOTYPE STILL CONSIDERED MORE
LACTOBACILLUS THAN -- THIS IS
PRELIMINARY DATA.
WE ALSO FOUND TREATMENT WITH
LACTIN V AND WE COULD LOOK FOR
SOLUBLE IMMUNE FACTORS LACTIN B
WAS ASSOCIATED WITH SUSTAINED
REDUCTION IN VAGINAL LEVELS OF
PRO INFLAMMATORY CYTOKINES
INCLUDING IL 1 AND E CAD HEREIN.
SO LACTOBACILLUS TREATMENT TENDS
TO DECREASE PROINFLAMMATORY
CYTOKINES AND IMPROVE EPITHELIAL
BARRIER FUNCTION.
SO THAT WAS AN INTERESTING
FINDING THAT WAS SORT OF
UNEXPECTED.
IN POST HOCK ANALYSIS, WE FOUND
A CLINICAL CURE WAS KEY TO
SUCCESS.
SO A CLINICAL CURE MEANT WOMEN
WHO WERE TREATED WITH
MATRONIDAZOLE THAT DECREASED
THEIR BUGS TO THE POINT WE COULD
ADD LACK LOW BACILLUS AND IT WAS
ABLE TO -- LACTOBACILLUS AND
ABLE TO DOMINATE OR FULFILL THAT
NICHE.
WHEN WE LOOKED AT THIS DATA WAS
ANALYZED BY MIKE AT EMIS SO
WOMEN WHO WERE CURED WITH
METRODANOZOLE BETTER ABLE TO
STABILIZE AND STAY CURE POST
HOCK ANALYSIS WE HAD 88% CURE
RATE IF WE COULD DRIVE DOWN
THOSE BV BACTERIA, WHEREAS WOMEN
WHO DIDN'T GET A CLINICAL CURE,
THERE IS NO ROOM FOR THEM TO
COME IN AND COLONIZE AND KEEP
WOMEN FROM GETTING BV AGAIN, YOU
CAN SEE PLACEBO AND LACTOB
COMPLETELY OVERLAP HERE.
SO THERE'S A LOT OF REMAINING
CHALLENGES IN BV RESEARCH.
THERE IS INCOMPLETE
UNDERSTANDING OF THE ETIOLOGY OF
BV.
WE KNOW SEX IS A BIG FACTOR THAT
DRIVES BV.
THERE IS A STUDY BY SUPRA META
WHO SHOWED THAT YOUNG UGANDAN
WOMEN HAD HIGH LEVELS BAC LEVELS LEVELS
LACTOBACILLUS CRISPOTES BUT WHEN
HAY STARTED HAVING SEX IT HAD A
EVERYONE THE PHENOTYPE, AFRICAN
WOMEN AND AFTER CANMEN WOMEN HAD
HIGH LEVELS BV AND THEY WENT
WELL THERE'S SOMETHING
INTIMATELY DIFFERENT, WHY THAT'S
WHY THEY HAVE BV BUT IT IS NOT
REALLY THE CASE.
BECAUSE YOUNG WOMEN WHO HAVEN'T
HAD SEX YET DON'T HAVE BV BUT
ONCE THEY HAVE SEX THEN THEY GET
BV SO ROLE OF THE BIOFILM AND
BACTERIA PHAGOAND WHAT EFFECTS
CANDIDA HAS AND WHAT EFFECTS
THIS OTHER LACTOBACILLUS HAVE
ARE STILL UNKNOWNS THAT WE AND
MANY OTHERS ARE LOOKING INTO.
PHYSIOLOGICAL VARIATIONS.
IN THE VAGINAL MICROBIOME CAUSED
BY MENSTRUATION SEMEN WHICH IS
VERY ALKALINE, PREGNANCY,
HORMONE CONTRACEPTION HORMONES
IN GENERAL, DIET AND THE RECTAL
MICROBIOME ALSO PLAY A ROLE.
THE FUNCTION OF THE METABOLOME
FOR THE VAGINAL MICROBIOME IS
SOMETHING WE ARE LOOKING INTO.
FINALLY HOST MICROBE
INTERACTIONS MAY FACILITATE OR
INHIBIT SIV TRANSMISSION OR DRUG
METABOLISM.
VARYING APPROACHES TO BV
DIAGNOSIS THE TOOLS TO DIAGNOSE
BV ARE O AMSEL AND NUGEMT ARE
SUBJECTIVE SO THAT MAKES
EVERYONE'S JOB MORE DIFFICULT.
SO WE ARE PROCEEDING NOW WITH
THE PHASE 3, TWO PHASE 3 PIVOTAL
TRIALS IN U.S., WE WILL ENROLL
WOMEN AGE 16 TO 45, FDA LIKES
YOU TO HAVE A PEDIATRIC GROUP SO
WE TRY TO ENROLL WILL TRY TO
ENROLL AT LEAST A FRACTION OF
YOUNGER WOMEN 16 TO 18 WHICH IS
CONSIDERED A PEDIATRIC GROUP, WE
WILL ENROLL WOMEN WITH
RECURRENTS BV THREE OR MORE
EPISODES IN ONE YEAR PERIOD.
WE WILL CONTINUE TO INCLUDE
WOMEN OF COLOR.
AGAIN MONITOR SAFETY EFFICACY
AND ACCEPTABILITY AND EVENTUALLY
APPLY FOR A BIOLOGICS LICENSE.
I THINK THAT IS MY LAST SLIDE
ACKNOWLEDGE MY COLLABORATORS, I
LEFT OFF RUPERT WHO DID THE
IMMUNE WORK.
I CUT MY TALK SHORT SO THERE'S
TIME FOR LAST SPEAKER.
[APPLAUSE]
>> VERY RECOGNIZE PROGRESS.
DO YOU HAVE ENOUGH DATA, TREATED
PEOPLE TO SEE WHETHER LACK TIN V
RESTORATION OF THE PREDOMINANTLY
L CRISPATUS FLORA ALSO REDUCED
FREQUENCY OF EITHER CANDID A
INFECTIONS OR URINARY TRACT
INFECTIONS?
>> WE DO KNOW LACTOBACILLUS
CRISPATUS DECREASES URINARY
TRACT INFECTIONS, A SEPARATE
PHASE 2A STUDY DONE WITH ANN
STAPLETON AT THE UNIVERSITY OF
WASHINGTON.
AND THAT SHOWED THAT
LACTOBACILLUS CRISPATUS ADDITION
HAD 50% REDUCTION IN WOMEN WITH
RECURRENT URINARY TRACT
INFECTIONS.
AND THAT IS LIKELY BECAUSE E.
COLI WHICH IS THE PRIMARY CAUSE
OF UTI IN WOMEN IS COMES RETRO
GRADE FROM RECTUM, IF YOU HAVE
LOTS OF LACTOBACILLUS PRESENT
YOU WILL PREVENT THAT MOVEMENT
OF THE BACTERIA.
AS FAR AS CANDIDA, WE DON'T HAVE
A LOT OF DATA ON THAT.
WE DID COLLECT THAT IN THE
CLINICAL TRIAL BUT THERE REALLY
WASN'T ENOUGH FOR US TO SHOW
ANYTHING STATISTICALLY.
THE LAST SPEAKER IS DAN APPELLA
FROM NIDDK.
THANK YOU, VERY MUCH.
LET ME JUST SAY IT IS AN HONOR
TO BE SPEAKING TO YOU TODAY.
BEING PART OF THIS SYMPOSIUM TO
HONOR MARJORIE WAS REAL KEY
COLLABORATORS IN THE PROJECT I
WILL TELL YOU ABOUT TODAY.
THIS IS THE LAST TALK SO WILL
TRY TO BE INTERNETTIC AND
ENTERTAIN YOU AT THE SAME TIME
IF I CAN.
SO I WILL TELL YOU ABOUT THIS
MOLECULE.
I WILL CALL IT 247 THROUGHOUT
THIS TALK.
THIS MOLECULE HAS FASCINATED ME
A NUMBER OF YEARS AND THE MAIN
GOAL OF THIS STUDY IS TO REALLY
LOOK AT THE MECHANISM OF ACTION
AND REALLY UNDERSTAND ITS
ANTI-HIV PROPERTIES IN
PARTICULAR, AND WE ALSO ARE
INTERESTED IN PERFORMING AS MANY
PRE-CLINICAL STUDIES AS POSSIBLE
TO TRY TO GET TO FIRST IN HUMAN
CLINICAL TRIAL WITH MOLECULE OR
SOMETHING THAT IS CLOSELY
RELATED TO THE MOLECULE.
COUPLE OF THINGS TO TELL YOU.
I'M AN ORGANIC CHEMIST SO YOU
WILL SEE QUITE A FEW CHEMICAL
STRUCTURE, THAT IS HOW I THINK
AND COMMUNICATE MY MAIN
CONTRIBUTION IS TO REALLY MAKE
THIS MOLECULE WHICH IS WHY I
HAVE $15 PER GRAM.
THIS IS RESULT OF SYNTHESIS
PRODUCTION FOR THIS PARTICULAR
MOLECULE SO NOT TOO HARD TO
MAKE.
I ALSO WANT TO MENTION THAT I
DID NOT INVENT THIS MOLECULE,
NUMBER OF OTHER WITHIN THE
INTRAMURAL PROGRAM OF NIH CAME
UP WITH THIS MOLECULE.
I SEEM TO BE I GUESS THE LATEST
SPOKESPERSON TO ADVOCATE MOVING
THIS MOLECULE FORWARD IN VARIETY
OF WAYS I WILL TRY TO TELL YOU
TODAY.
SO THIS MOLECULE DOES HAVE SOME
ANTI-HIV PROPERTIES, IF YOU LOOK
IN STANDARD CELL BASED ANTIVIRAL
ASSAYS FOR HIV, THE IC 50 VALUES
IN LOW MICROMOLAR RANGE NOT VERY
IMPRESSIVE COMPARED TO POTENT
DRUGS ON THE MARKET.
TOXICITY DOESN'T SEEM TO BE MUCH
ISSUE IN CELL BASED ASSAYS
TOXICITY VALUE ARE WELL OVER 100
MICROMOLAR, WHILE THE ACTIVITIES
MAYBE NOT VERY IMPRESSIVE IT IS
VERY CONSISTENT.
WE HAVE TESTED THIS WITH A WIDE
VARIETY OF DIFFERENT FORMS OF
HIV, DIFFERENT CLADE SUB GROUPS,
EVEN HIV VARIANTS THAT HAVE
KNOWN DRUG RESISTANCE TO
EXISTING DRUGS ON THE MARKET AND
IT IS ALL PRETTY MUCH THE SAME.
WE HAVE DONE SOME ANIMAL STUDIES
WITH THIS MOLECULE, ALWAYS
SHOWING VERY LOW TOXICITY IN
ANIMALS WELL TOLERATED.
ONE FASCINATING THING TO ME FOR
THIS MOLECULE IS AFTER SEVERAL
ATTEMPTS TO TRY TO GENERAL RATE
HIV RESISTANT VIRUS TO THIS
MOLECULE, IT HASN'T BEEN
SUCCESSFUL SO WE CONCLUDE
BASICALLY A HIGH BARRIER TO
GENERATE RESISTANT VIRUS TO THIS
MOLECULE, IT SPEAKS TO A UNIQUE
MECHANISM OF ACTION.
THIS IS A COLLABORATIVE PROJECT
AND TRYING TO ADVANCE AS TOPICAL
MICROBE SIDE TO PREVENT HIV
INFECTION.
ONE COLLABORATOR WAS MARJORIE ON
THIS PARTICULAR WORK AND I WILL
SUMMARIZE HER DATA ON THIS
SLIDE.
THIS IS A MICROBE SIDE CHALLENGE
IN WHICH OUR MOLECULE WAS
FORMULATED IN A JELL, ABOUT .8%
MOLECULE IN HYDROXYHEC GEL.
ANIMALS ARE CHALLENGED WEEKLY.
AND THIS GEL IS APPLIED
VAGINALLY THEN THREE HOURS AFTER
APPLICATION OF MICROBE SIDE THEY
ARE CHALLENGED WITH A VIRUS.
PANELS FOLLOWED A LONG TIME.
AND THIS WAS GREAT DATA TO SEE
WITH MICROBE SIDE YOU SEE QUITE
NICE PROTECTION IN THE TREATED
ANIMALS IN BLUE LINE WHEREAS
UNPLACEBO ANIMALS ARE INFECTED.
WE STARTED THIS COLLABORATION
WITH MARJORIE HOPING WE COULD
COMBINE MICROBE SIDE WITH
ADENOVIRUS VACCINE.
THOSE DATA DIDN'T LOOK SO GOOD.
NOT ENTIRELY ALWAYS SURE WHY
THOSE DATA DESCRIBED IN
THISBLYCATION.
-- PUBLICATION.
IT WAS MARJORIE'S SUGGESTION NOT
TO STOP THE WORK AND SHE
INTRODUCED ME TO (INAUDIBLE)
WITH HER VACCINE.
WE DECIDED TO REDO EXPERIMENTS
WITH BETH'S VACCINE AGAINST
HIV-1 OFFERING AROUND 60 TO 70%
PROTECTION.
USED SAME FORMULATION OF OUR
MICROBE SIDE AND GEL AFTER
MICROBE SIDE ANIMALS ARE
CHALLENGED.
THIS DATA WORKED NICELY.
I WILL GO THROUGH THIS BRIEFLY.
IN THE VACCINE ALONE TREATED
ANIMALS YOU CAN SEE THE EFFECT
OF VACCINE IN BLUE OVER THE
CONTROL GROUP.
MICROBE SIDE ALONE DIDN'T FAIR
SO WELL WITH MINIMAL PROTECTION
BUT WHEN YOU COMBINE VACCINE AND
MICROBE SIDE YOU SAW ENHANCEMENT
FOR PROTECTION, COMPARISON TWICE
COMBINATION TREATMENT AND THE
VACCINE ALONE.
SO WE WOULD LOVE TO MOVE THIS
FORWARD TO SOMETHING THAT WOULD
INVOLVE HUMAN CLINICAL TRIALS.
WE ARE AWARE THOUGH FROM PRIVATE
PREVIOUS CLINICAL WORK WITH
MICROBE SIDES IN PARTICULAR THE
-- CLINICAL TRIAL THAT GEL
FORMULATION IS NOT IDEAL FOR
PATIENT COMPLIANCE.
WE ARE ALSO AWARE FROM THE --
ANOTHER CLINICAL TRIAL THE
ASPIRE TRIAL THAT A FORMULATION
OF MOLECULE IN A MONTHLY VAGINAL
RING WHICH IS SLOW RELEASES
MOLECULE IS MORE TOLERABLE.
THIS IS ONE THAT USED A
DIFFERENT MOLECULE CALLED
DUPIVERINE SHOWING IMPROVEMENT
IN REDUCING HIV INFECTION.
THE GOAL IS TO FORMULATE
MOLECULE INTO SLOW RELEASE FORM,
COLLABORATION WITH (INAUDIBLE)
AT INSTITUTE PROCESS OF
FORMULATING MOLECULE IN HIS RING
WE HOPE TO CLINT TO GENOVEFFA
WOULD TOO MUCH DELAY.
PART OF THE MAIN QUESTIONS IS
WHY DOES THIS WORK?
WELL?
AND WHAT COULD BE GOING ON?
A LOT OF GOALS OF MY WORK IS TO
FIGURE HOW DOES THIS MOLECULE
INTERFERE WITH HIV?
WE LIKE TO USE THIS KNOWLEDGE TO
SUPPORT FUTURE CLINICAL WORK.
AND MAYBE MAKE DIFFERENT
DERIVATIVES THAT PERHAPS WORK
BETTER.
THERE IS A CHALLENGING PROJECT
BECAUSE IT DOESN'T BEHAVE LIKE
ANY DRUG I KNOW OF AGAINST HIV,
I WOULD DOESN'T INTERFERE WITH
ESTABLISHED TARGETS BY THE
EXISTING PANEL OF DRUGS.
WE HAVE TESTED AGAINST ALL THE
HIV ENZYMES, REVERSE TRANSCRIPT
ACE, PROTEASE, VIRAL ENTRY,
DOESN'T DO ANYTHING.
WE ARE ALSO AWARE IN IS A
REACTIVE MOLECULE.
IN TERMS OF PROTEINS.
(OFF MIC)
>> YES WE HAVE TESTED AGAINST
OTHER -- DO YOU WANT ME TO COME
BACK LATER?
I CAN COME BACK LATER AT THE END
OF THE TALK.
WE HAVE TESTED THAT, THE ANSWER
WILL BE A LITTLE MORE CLEAR AT
THE END.
THIS IS A ANOTHER MOLECULE THAT
BINDS TO PROTEINS, BASICALLY AS
FAR AS WE CAN TELL.
MOST DRUGS HAVE A PROTEIN
BINDING INTERACTION THAT'S
NON-COVALENT.
THIS IS A REACTIVE MOLECULE,
MEANING IT DOES A CHEMICAL
REACTION IN A CERTAIN SPECIFIC
MANNER I WILL GO THROUGH IN
DETAIL WITH VARIETY OF CHEMICAL
STRUCTURE.
O THE REACTIVE END IS HERE THE
THYRO ESTER PORTION OF MOLECULE
ATTACHED TO SULFUR AND THE
ACETYLGROUP WINDS UP REACTING IN
PARTICULAR WITH SULFUR.
IN PARTICULAR IN THIS EXAMPLE
MOLECULE WITH THE SULFUR AND
SIDE CHAIN OF CYSTEINE.
SMEW COMBINING OUR MOLECULE WITH
THE MODEL COMPOUND WHAT ORGANIC
CHEM U.S.ES LIKE TO DO WE CAN
OBSERVE AN ACETYLTRANSFER FROM
THE 247 MOLECULE TO SULFUR OF
THE CYSTEINE SIDE CHAIN.
THE ORIGINAL 247 FORMS THIS AS A
BY-PRODUCT WHICH IS JUST THE
PHTHALATE, THIS IS S MINUS
BECAUSE WE MEASURED PKA OF
PROTONATED FORM, 5.8 SO UNDER PH
OF 4 DEPROTONATED AS S MINUS
SHOWN HERE.
AFTER THIS INITIAL TRANSFER, WE
WATCH BY NMR WE CAN SEE IT IS
FAST IT DOESN'T STAY IS A I CAN
THE BUT THERE THERE IS AN AMINE
SHOWN HERE THE ACETYLGROUP
TRANSFERS FROM THE SULFUR TO THE
NITROGEN TO FORM A MORE STABLE
-- WE CAN ALSO OBSERVE BY NMR
THIS IS A SLOWER PROCESS.
SO THERE IS A BASIC MODE OF
REACTIVITY HERE, AND WE KNOW
THAT THIS MOLECULE LIKES TO
RAPIDLY TRANSFER ACETYLGROUP
TOOTHILY GROUP SHORTIAS CYSTEINE
FOLLOWED BY RERANGEMENT TO MEAN
GROUP.
THIS IS THE BAY SING MODEL
REACTIVITY I WILL EXTEND UP TO
SHOW YOU HOW THIS IMPACTS HIV.
IN AN INTERESTING WAY.
NEXT I WILL SHOW YOU HOW AT A
VERY BROAD LEVEL HOW 247
INTERFERES WITH HIV AND WE HAVE
DATA FROM ELECTROMICROSCOPY
TREATED HIV INFECTED CELLS WITH
MOLECULE 247 BY ELECTRON
MICROSCOPY RESULTS IN VIRUS
PARTICLES LOCKED IN IMMATURE
NON-MATURE STATE.
YOU HAVE VARIETY OF GAG AND
PROCESS PROTEINS THAT ARE RING,
THIS IS THE TEXTBOOK IMAGE OF
IMMATURE HIV PARTICLE AND AFTER
MATURATION WITH HIV PROTEASE,
THE VIRUS TAKES ON THIS
MORPHOLOGY AND WITH CONDENSED
CAPSID CORE.
Z SO IN TREATED CELLS YOU CAN
SEE VIRUS BUTTING AND FREE VIRAL
PARTICLES IN SOLUTION THAT
REMAIN LOCKED IN IMMATURE STATE,
VERSUS CONTROL YOU CAN SEE VIRAL
PARTICLES NICELY MATURED ALL THE
WAY.
AND BECOME INFECTIOUS.
IN THE IMMATURE STATE PARTICLES
HIV PARTICLES ARE CONSIDERED TO
BE NON-INFECTIOUS.
SO DRILL DOWN MORE DETAIL TO GAG
POLYPROTEIN WE SEEM TO THINK IS
BEING EFFECTED BY OUR PARTICULAR
MOLECULE.
SO SHOWN HERE IS CARTOON OF GAG,
IT IS A POLYPROTEIN HIV PRODUCES
A LOT OF GAG POLYPROTEIN.
IT HAS THREE DOMAINS MATRIX
CAPSID AND NUCLEAR CAPSID
DOMAIN, NUCLEAR CAPSID BINDS
NUCLEIC ACID, PARTICULAR HIV
RNA.
HIV GAG IS SET UP BEAUTIFULLY TO
SELF-ASSEMBLE INTO A VIRAL
PARTICLE WHICH CONTAINS 2,400
GAG MOLECULES WITH LOCALIZED
CONCENTRATION OF GAG AS THEY ARE
CRUNCHED TOGETHER.
IMMATURE VIRAL PARTICLE IS THEN
DRIVEN TO MATURATION AFTER HIV
PROTEASE IS ACTIVATED HIV
PROTEASE CLEAVE IT IS BONDS
BETWEEN DOMAINS OF GAG WHICH
THEN RESULTS IN THE CORE
CONDENSATION AND THEN INFECTIOUS
VIRAL PARTICLE.
SO THIS IS A BASIC REACTIVITY I
JUST SHOWED.
AND WE THINK THAT THIS BASIC
REACTIVITY TRANSLATES UP TO
INTERACTING WITH GAG.
THIS IS WHAT WE WANTED TO
ACTUALLY SHOW.
BASICALLY THAT OUR MOLECULE HAS
PATTERN OF REACTIVITY IN THE
COMPLICATED HIV PROTEIN.
I WILL GO THROUGH THIS ONE MORE
TIME BRIEFLY.
WE KNOW THE ACETYLGROUP HERE IN
OUR MODEL SYSTEM TRANSFERS TO
CYSTEINE SIDE CHAINS AND
EVENTUALLY TO AMINE SIDE CHAIN,
SO WE WANTED TO STUDY WHETHER
THE ACETYLGROUP HERE DOES
SOMETHING SIMILAR IN THIS
COMPLICATED PROTEIN PUTTING
ACETYLGROUPS ON HIV GAG.
THEY WE STUDY THIS IS MASS
SPECTROMETRY, I WILL MENTION ALL
DONE BY WORK BY LISA JENKINS AT
NCI.
SO I WANT TO GO THROUGH THIS ON
THE STRUCTURE MODEL OF GAG.
I WANT TO ORIENT YOU A LITTLE
BIT AND PROTEIN STRUCTURE SHOWN
UP HERE IS THE MATRIX DOMAIN OF
GAG.
THERE ARE TWO DOMAINS FOR THE
CAPSID, THERE'S THE END TERMINAL
DOMAIN OF CAPSID AND C TERMINAL
AND NUCLEAR CAPSID IS HERE.
HIGHLIGHTED AMINO ACID SIDE
CHANGES ARE RESIDUES THAT
RECEIVE ACETYLGROUP FROM 247
MOLECULE.
I WANT TO ZOOM ON THIS
INDIVIDUAL REGIONS.
GREEN SIDE CHAINS ARE LIE LYSINE
THAT IS ACETYLATED BY THE
MOLECULE, CYSTEINE 57 RECEIVES
ACETYLGROUP.
THIS IS AN EXAMPLE BASIC
REACTIVITY I SHOWED YOU IS
RECAPITULATED IN PROTEIN FORM.
YOU CAN SEE CYSTEINE FOLLOWED BY
LYSINE INDICATING FIRST REACTION
CYSTEINE FOLLOWED BY TYPE OF
INTRAMOLECULAR TRANSFER TO
SURROUNDING LYSINES IN THE
PROTEIN.
IF YOU LOOK AT CAPSID, THERE IS
A SIMILAR SITUATION WITH TWO
CYSTEINES NEARBY FOLLOWED BY
SOME LYSINES IN GREEN, PURPLE
HERE, AND NUCLEAR CAPSID HIGHLY
AS TELLATED.
MORE DETAIL IN NUCLEAR CAPSID IN
THE FOLLOWING SLOT.
BEFORE I MOVE ON I WANT TO
MENTION THERE ARE MORE LYSINES
IN THIS PROTEIN, THEN ARE
ACETYLATED MEANING NOT EVERY
LYSINE RECEIVES ACETYLGROUP,
RATHER THERE SEEMS TO BE A
SPECIFIC REACTIVITY PATTERN WE
THINK WHICH IS GOVERNED BY FIRST
REACTION CYSTEINES FOLLOWED BY
TRANSFER TO LYSINE.
NUCLEAR CAPSID IS WHERE WE THINK
THE ACTUAL BUSINESS END OF THE
ACTIVITY IS TAKING PLACE.
WANT TO GO DOWN TO MORE DETAIL
NUCLEAR CAPSID REPRESENTED BY
BLUE DOT.
NUCLEAR CAPSID IN HIV GAG IS 55
AMINO ACIDS LONG, THE ONLY
STRUCTURE WITHIN THIS SMALL
PROTEIN IS THE COORDINATION OF
TWO ZINCS BY AMINO ACID SIDE
CHAIN SHOWN HERE.
THESE ARE REALLY SMALL ZINC
FINGERS REFERRED TO AS ZINC
KNUCKLES AND WE KNOW THAT THE
MOLECULE 247 REACTS SPECIFICALLY
INITIALLY WITH THE C TERMINAL
ZINC KNUCKLE IN THIS HIV NUCLEAR
CAPSID PROTEIN.
WE THINK WE HAVE AN
UNDERSTANDING AS TO WHY.
WHEN WE LOOK FOR CAREFULLY AT
THIS PARTICULAR ZINC KNUCKLE,
ONE AT C TERMINAL END WE CAN SEE
IT IS IN AN INTERESTING E QUICK
LIB YUM.
WHEN THE MOST OF THE FORM OF
THIS MOLECULE HAS ZINC
COORDINATED BY HISTINE HISTIDINE
SIDE CHAIN THAT IS UNREACTIVE
WITH MOLECULE BUT BY NMR
DETECTED BY 2% SOLUTION HAS ONE
OF THE SULFUR ATOMS OFF ZINC,
FREE THIOL IN SOLUTION.
THIS IS A RAPID EQUILIBRIUM
FAVOR IT CAN COORDINATED FORM
BUT A LITTLE FREE THYOL, IF YOU
RECALL FROM ACTIVITY THE
MOLECULE LIKES TO REACT WITH
THYOLS, WITH THE ACETYLGROUP
INTERFERING WITH FREE FORM THE
TRANSFER GROUP ON TO SULFUR
ATOM.
AFTER THIS REACTION, THIS
PREVENTS THE SULFUR FROM
RECOORDINATING TO ZINC, LOOSENS
COORDINATION.
WE KNOWNA THE ZINC FALLS OUT
RESULTING IN A DENATUREATION,
THE DENATURE PASTE MAYBE
FACILITATED BY/WHICH COORDINATES
TO ZINNC.
WE KNOW ACETYLGROUP DOESN'T STAY
ON SULFUR IT REARRANGES TO
NITROGEN ON SIDE CHAINS NEARBY.
THIS EXPOSES FREE THIOLS IN
SOLUTION AND LEADS TO CROSS
LINKING WITH THE GAG DISULFIDE
FORMATION WHICH IS KEY FOR THAT
PHENOTYPE WE SAW WITH HIV LOCKED
IN IMMATURE STATE.
SO I WANT TO HIGHLIGHT MECHANISM
ONE MORE CARTOON FASHION.
REPRESSURE MEMORY WHEN MOLECULES
-- CELLS INFECT WITH HIV ARE
TREATED ARE MOLECULE IT RESULTS
IN NON-INFECTIOUS IMMATURE
PARTICLES WE THINK THAT WHAT
HAPPENS IS OUR MOLECULE REACTS
WITH GAG, TRANSFERS THE
ACETYLGROUPS NUCLEAR CAPSID
DENATURED EXPOSES FREE THIOLS
AND CROSSES DISULFIDE BONDS THAT
LINK THE POLYPROTEIN TOGETHER.
SO THAT WAS IN VIVO.
THEN WE WANTED TO SEE IS THERE
EVIDENCE THAT THIS
ACETYLTRANSFER MECHANISM HAPPENS
IN AT LEAST IN CELLS.
TO DO THESE EXPERIMENTS WE
TREATED HIV INFECTED CELLS WITH
MOLECULE, THAT PRODUCES VIRIONS
WE ISOLATED AND LOOK AT GAG FROM
THESE VIRIONS THAT WE NOTICE
THAT ALL WAS ACETYLATED IN A
SIMILAR PATTERN TO SOLUTION.
SO THAT WAS EXCITING TO SEE
UNTIL WE WENT BACK AND THOUGHT A
LITTLE MORE ABOUT REACTIVITY
PATTERN WE HAD ESTABLISHED.
IN WHICH WE KNOW THAT OUR
MOLECULE LIKES TO TRANSFER
ACETYLGROUPS TO ANY SULFUR IN
THE CELL.
CYSTEINE SIDE CHAINS GLUE
TATHIONE COA ANYTHING.
SO DIDN'T MAKE SENSE IN A CELL
THIS 247 WITH ACETYLGROUP
SURVIVE LONG ENOUGH TO BOUND TO
GAG AND DO THIS REACTION.
SO THE ANSWER BECAME A LITTLE
BIT MORE CLEAR WHEN WE THEN WENT
TO LOOK MORE AT THE GAG WE
LOOKED AT.
I DON'T KNOW WHAT I DID.
SORRY.
THIS IS WHEN I START APOLOGIZING
FOR THE COMPUTER.
I WILL UNPLUG AND PLUG IT BACK
IN.
I WAS.
PUNCH LINE IS RUINED.
I'M SORRY.
CABLE MIC WENT BAD.
WE ARE BACK.
IT IS TELLING ME TO SPEED UP.
úANYWAY, SO IF YOU REMEMBERWHAT
WHERE I WAS LOOKING AT
ACETYLGROUPS ON THE GAG AFTER
TREATING WITH CELL AND LOOK AT
ACETYLGROUPS ON THE GAG NONE
CAME FROM ORIGINAL MOLECULE, ALL
FROM ACETYLCOA.
WHERE DID THE ACETYLGROUPS GO,
SO IN THIS BASIC REACTION, THIS
PHTHALATE PRODUCED IS NOT
INNOCENT.
RATHER THAT PHTHALATE
REGENERATES THE ORIGINAL
MOLECULE BY INTERACTION WITH
ACETYLCOA.
SO THE WAY THIS WORKS IS THAT
CHAIN CELLS WITH 247 LOSES
ACETYLGROUP TO GLUE TATHIONE AND
PERFORMS A PHTHALATE, THAT
REACTS TO ACETYLCOA TO
REGENERATE ITSELF TO GET
ACETYLGROUP TO REFORM AND CARRY
OUT REACTIONS IN CELLS.
SO THE REALLY INTERESTING
MECHANISM GOING ON IN CELLS,
WHICH I AM DIAGRAMMING HERE, IN
CELLS YELLOW BLOB IS CELL,
INFECTED WITH HIV, WE CAN TREAT
THIS CELL WITH 247 BUT IT
RAPIDLY LOSES ACETYLGROUP TO ANY
AVAILABLE THIOL, GLUE TATHIONE
POLYIN PARTICULAR AND IN CELLS
THIS INTERMEDIATE, PHTHALATE IS
MAJOR SPECIES THAT HANGS OUT A
LOT AND DOES PROBABLY NOT MUCH.
THE MOLECULE CAN INTERACT WITH
ACETYLCOA TO REGENERATE 247 AND
PROBABLY CYCLES BACK AND FORTH
ACETYLATING AND LOSING
ACETYLGROUP MANY TIMES.
WE THINK IT IS ONLY REALLY IN
THE CONTEXT OF WHEN SOMETHING
LIKE HIV GAG CRUNCHES TOGETHER
TO FORM A HIGH EFFECTIVE
CONCENTRATION THAT THERE IS
ENOUGH 247 IN THE CONTEXT OF
ASSEMBLY IT IS ABLE TO GO IN
ACETYLATE THE SMALL AMOUNT OF
NUCLEAR CAPSID AND THE OTHER
THIOLS IN GAG START
DENATUREATION AND LEAD TO CROSS
NIHING WE SEE COMING OUT.
SO THAT WAS AN INTERESTING STORY
BUT I WANT TO GO BACK NOW TO
THESE DATA AND ALSO MY
DISCUSSIONS WITH BETHA HOW DO WE
UNDERSTAND WHY COMBINATION OF
MOLECULES OF VACCINE WORK SO
WELL.
AND IT WAS BETHA WHO POINTED OUT
WHILE ANTIVIRAL ACTIVITY IN
MECHANISMS ARE ALL NICE IT
DOESN'T EXPLAIN THIS IMPROVEMENT
THAT WE SEE.
SO THAT RAISES THE QUESTION IS
THERE ANOTHER MECHANISM FOR THIS
MOLECULE I ADMIT IS ENTIRELY
POSSIBLE.
SO BETHA STARTED LISTING THE
FACTORS THAT SHE CAREFULLY
CHARACTERIZED THAT SORT OF LEAD
TO A REDUCTION IN EFFICACY OF
HER VACCINE.
SHE JUST HAPPENED TO MENTION IN
HER LIST THIS PARTICULAR PROTEIN
ZZ 3H 7A NEGATIVE REGULARLY TORR
OF VACCINE, THE Z PROTEIN FOR
SHORT.
THIS HAPPENS TO BE RNA BINDING
ZINC FINGER DOMAIN, THEY ALL
HAVE ZINC COORDINATED BASISSTEEN
AND HISTIDINE.
THERE'S NOT A TON KNOWN ABOUT
THIS PARTICULAR PROTEIN IT IS
KNOWN mRNA LEVELS OF PROTEIN
INCREASE AND LIPO POLYSACCHARIDE
INDUCE MACROPHAGES BUT MOLECULE
LIKES TO REACTS WITH THESE TYPES
OF ZINC FINGERS.
SO WE THOUGHT THIS COULD BE
ANOTHER TARGET AND GOING TO
BRIEFLY MENTION WE HAVE
COMPLETED A EXPRESSION OF
RECOMBINANT PORTION OF PROTEIN
THAT HAS ZINC FINGERS AND WE DO
KNOW WE OBSERVED THAT OUR
MOLECULE DOES REACT WITH ONE OF
THE FOUR ZINC FINGER DOMAINS IN
RECOMBINANT PROTEIN.
THAT IS YET TO BE DETERMINED
WHETHER IT HAPPENS IN CELLS BUT
I THINK IT MAY BE ENTIRELY
POSSIBLE THAT MOLECULE DOES
KNOCK OUT ONE OF THE NEGATIVE
REGULATORS OF THAT COULD EXPLAIN
ENHANCEMENT WE ARE SEEING.
I STILL HOPE ANTIVIRAL MECHANISM
IS ALSO CONTRIBUTING.
BECAUSE WE DID A LOT OF WORK ON
IT.
SO THAT IS MY TALK.
BRIEFLY TO CONCLUDES SHOW YOU
INTERESTING THINGS, CAN COMBINE
MICROBE SIDE WITH VACCINE TO
IMPROVE PROTECTION IN SIV
INFECTION IN RHESUS MACAQUES.
IT IS A COMPLICATED MECHANISM
FOR THE MOLECULE IN ACETYLATES
AND CYSTEINES IN LYSINES OF HIV
GAG, BOTH IN VITRO AND IN CELLS.
E DENATURES NUCLEOCAPSID IN HIV
GAG, IT INVOLVES ACETYLCOA,
MULTIPLE ACETYLTRANSFERS AND
WERE -- YOU CAN FORMULATE THIS
AS A GEL, TRYING TO FORMULATE
THIS INTO OTHER DELIVERY DEVICES
CURRENTLY.
SO FINALLY, LET ME ACKNOWLEDGE A
NUMBER OF PEOPLE, POST-DOCS IN
MY LAB, MARK HAS BEEN THE PLANS
BILL POE DOC INVOLVED.
THIS IS A HIGHLY COLLABORATIVE
PROJECT, I MENTIONED BETHA AND
MARJORIE KEY NOVEMBERING
FORWARD.
LISA DOING MASS SPECTROMETRY.
MANY COLLABORATORS OVER THE
YEARS CONTRIBUTE TO THIS.
AND FINALLY LET ME THANK YOU FOR
YOUR ATTENTION.
[APPLAUSE]
>> I'M GOING TO TAKE THE FIRST
QUESTION.
>> OKAY.
>> SO IS THERE A WAY YOU CAN
TARGET THIS MOLECULE TO HIV
INFECTED LATENT CELLS IN PEOPLE?
>> SORT OF.
WE PROBABLY COULD IN THEORY.
IF THERE WAS A GOOD TARGETING
AGENT.
THAT YOU WANTED ME TO LINK IT
WITH I PROBABLY COULD.
WHEN PEOPLE ASKED ABOUT LATENCY
I GET A RILLET DEFENSIVE BECAUSE
I DON'T KNOW AND I DON'T THINK A
LOT OF PEOPLE KNOW.
I ALMOST SUGGESTED TO PEOPLE
JUST BLAST SOMEONE WITH A
MOLECULE, IT IS PRETTY WELL
TOLERATED.
AMONG ANIMALS, YOU CAN GO UP TO
HIGH CONCENTRATIONS, THE OTHER
THING I LIKE IS IT IS REALLY
SMALL.
COMPARED TO OTHER DRUGS, WHICH I
ALWAYS FEEL HAVE A BIT OF
PERMEATION PROBLEM THIS IS TINY
SO IT SHOULD OOZE ITS WAY AROUND
REALLY WELL.
I WOULD LOVE TO TEST IT.
SO LET ME SAY I'M A KISS PLAYER
OF THE MOLECULE, IF IT IS
AVAILABLE COMMERCIALLY DON'T BUY
IT COMMERCIALLY TALK TO ME.
I CAN DISTRIBUTE IT OUT.
ALTHOUGH -- YEAH.
I SORT OF PICK AND CHOOSE
BECAUSE I LIKE TO GET DATA BACK
AND I'M ALWAYS INTERESTED TO SEE
ANY MORE FEEDBACK HOW WE CAN
PUSH FORWARD.
>> VERY INTERESTING CHEMISTRY
AND --
>> THERE IS A LONG HISTORY,
THERE'S MANY MOLECULES AS YOU
KNOW THAT INTERACT WITH
CYSTEINES SH GROUPS, PROTEINS
AND WITH GROUPS OF LYSINES AND
SO FORTH.
THAT MIGHT MIMIC SOME OF THESE
EFFECTS AND IN PARTICULAR WITH
ONE OF THE THINGS THAT COMES TO
MIND IS THAT LYNN HURSTBURG FOR
MANY YEARS PUSHED THE IDEA OF
USING N ACETYLCYSTEINE TO TREAT
HIV AND THERE WERE PEOPLE WHO
WERE TAKING HIGH DOSES OF NS
CYSTEINE AS A PROPHYLACTIC
AGAINST HIV INFECTION.
THERE WAS SOME EVIDENCE FOR
POSSIBLE EFFICACY, I HAVEN'T
FOLLOWED IT IN GREAT DETAIL.
BUT I'M WONDERING WHETHER
THERE'S SOME CONNECTION HERE.
>> POSSIBLY.
I KNOW SOMEWHAT FAMILIAR WITH
THE N ACETYLSTORY, PART OF THE
PROBLEM IS BIOAVAILABILITY AND
METABOLISM BODIES AN CELLS KNOW
HOW TO METABOLIZE IT.
AND THAT MIGHT BE PART OF IT.
>> PRIMARILY REDUCING AGENT BUT
PUT TUMOR ETHANOL IN OUR
CULTURES.
>> YEAH.
YEAH.
YEAH VERY HIGH DOSES WERE USED
AND THEY ARE TOLERATED.
I DON'T THINK THE EFFECTS WERE
VERY PROMINENT IF I REMEMBER
CORRECTLY.
THAT IS WHAT I REMEMBER.
BUT IT IS -- YEAH.
>> STILL TARGETING CYSTEINES.
>> YOU ARE TARGETING CYSTEINES.
I HESITATE TO GO IN THIS
DIRECTION BECAUSE IT GETS FUZZY,
IT IS A REDUCING AGENTS AND SURE
OURS IS A A REDUCING AGENT AS
WELL.
THERE IS BY-PRODUCT ACETYLATION
PATHWAYNA MIGHT BE UNIQUE TO
OURS.
I FEEL LIKE IN ADDITION TO SORT
OF CHANGING REDUCING ENVIRONMENT
TO SOME DEGREE, I THINK WE ARE
SHUFFLING THIS ACETYLGROUPS IN
ODD WAY PERHAPS, THERE IS AN
ACETYLOME IN THE CELLS THAT IS
IMPORTANT, MAYBE TWEAKING THAT
ON TOP OF EVERYTHING ELSE.
>> THERE'S ESTERS PEOPLE USE TO
TARGET LYSINES OR TO TARGET
CYSTEINES.
>> SURE.
YEAH.
ABSOLUTELY BUT THOSE ARE -- I
THINK NOT SORT OF -- THOSE ARE
ONE AND DONE REACTIONS IF I
REMEMBER CORRECTLY.
I MEAN, THERE ARE LOTS OF DRUGS
THAT HAVE COVALENT WAR HEAD
WHETHER THEY KIND OF COME IN AND
SLAP ON A GROUP AND THEY ARE
DONE AND USUALLY THEY HAVE A
TARGETING EFFECT WHERE THERE'S
SOMETHING BINDING.
THIS DOESN'T BIND TO ANYTHING.
AS FAR AS I CAN TELL.
IT JUST HAS A REACTION THAT
TAKES PLACE, ALMOST LIKE
METABOLITE.
' IN SOME WAY SO SEEMS TO EFFECT
THINGS AT MUCH SORT OF METABOLIC
WAY.
>> I DON'T KNOW IF YOU MENTION
DON'T YOU EXPECT TO MOLECULAR TO
BE ANGIOGENIC IN LONG TERM
BECAUSE (INAUDIBLE) HOW LONG
HAVE YOU GIVEN IT TO MACAQUES
AND CAN YOU TELL THAT -- BECAUSE
YOU WANTED PROLONG LET'S SAY
EXPOSURE, IS IT REACT GENIC?
>> WE HAVE NOT THIS PARTICULAR
FORMULATION BUT I HAVE ORALLY
BIOAVAILABLE FORMULATION OF THIS
MOLECULE.
WE HAVE GIVEN ORALLY TO MACAQUES
FOR OVER SIX MONTHS AND THEY
SEEM FINE.
THIS COMES UP ALL THE TIME AND
PROBABLY ONE OF THE MAIN REASONS
WHY I HAVE TROUBLE ADVANCING
THIS MOLECULE BECAUSE EVERYBODY
SAYS THIS IS NON-SPECIFIC AND
GOING TO BE TOXIC AND JUST
HAVEN'T FOUND TOXICITY YET.
I'M STILL LOOKING FOR THE
TOXICITY, I HAVEN'T FOUND IT
YET.
NOW, I WILL SAY A COUPLE OF
THINGS.
WE ARE PERTURBING THINGS BUT I
DO THINK THAT THERE IS A
SELECTIVE ENVIRONMENT WHICH THIS
MOLECULE EVEN BECOMES ACTIVE.
I THINK IT IS UNREACTIVE IN MOST
CASES.
THOUGH I TALK ABOUT REACTIVITY I
THINK THE ONLY TIME IT REALLY
BECOMES ACTIVE IS WHEN YOU HAVE
A REALLY HIGH CONCENTRATION OF
PROTEINS SUCH AS WHEN GAG IS
ASSEMBLING TO.
EFFECTIVE CONCENTRATION IS OVER
FOUR MILLIMOLAR IN THOSE
PARTICULAR CASES.
I THINK YOU ALSO NEED TO HAVE
SOMETHING ALONG LINES OF THAT
EQUILIBRIUM IN ZINC FINGERS I
SHOWED YOU.
ONLY 2% OF 1 ZINC FINGER OF HIV
NUCLEAR CAPSID IS EVEN
AVAILABLE.
IF IT IS ALL FULLY COORDINATED
IT IS NOT DOING ANYTHING SO YOU
NEED TO HAVE SOME VERY SPECIFIC
ENVIRONMENTS FOR HIGH PROTEIN
CONCENTRATION, A LITTLE BIT OF
AN AVAILABLE UNLIGATED ZINC
FINGER IN ORDER FOR THE MOLECULE
TO DO ANYTHING.
OTHER THAN THAT, EVERYTHING ELSE
IS BACKGROUND NOISE.
I LIKE TO TALK ABOUT ACETYLATION
PATTERN, I SHOWED YOU'RE PLACES
OF ACETYLATION CYSTEINES AND
LYSINES.
I DON'T THINK CELLS CARE VERY
MUCH ABOUT THAT.
CELLS HAVE DEACETYLASES THEY CAN
DEAL WITH THAT AND I DON'T THINK
IT HAPPENS A WHOLE LOT ACTUALLY.
I THINK IT IS SOME BACKGROUNDS
THAT HAPPEN BUT IT IS ONLY THESE
VERY SPECIALIZED CIRCUMSTANCES
WHERE WE REALLY SEE THIS
HAPPENING.
THAT IS SORT OF MY ANSWER TO
THAT AREA.
>> FASCINATING, FASCINATING
FININGS.
I WAS WONDERING HOW YOUR
APPROACH COMPARES TO THAT OF
LARRY FROM NCI Z WHO USED A T 2
TO TARGET ZINC FINGERS, ACTIVATE
VIRUS PARTICLES.
AT 2 STANDS FOR (INAUDIBLE)
THIOL.
>> THAT IS ADMINISTERED AS A
DISULFIDE, NEEDS TO GET REDUCED.
PROBABLY NOT A LOT DIFFERENT.
I THINK THAT MOLECULE DID HAVE
TOXICITY IN THE END WHICH I
THINK SORT OF BLOCKED THE
ADVANCEMENT.
I THAT MOLECULE WAS ALSO BEING
USED TO STERILIZE CELL
EXTERNALLY FOLLOWED BY
INTRODUCING TO PEOPLE.
I CAN'T REMEMBER THE DETAILS
ABOUT APPROACH OF AT 2.
(OFF MIC)
>> THANK YOU.
>> YEAH.
YEAH.
>> THIS IS A COMMENT BECAUSE IT
IS RELEVANT TO THIS PARTICULAR
MEETING.
THAT THE DATA WE HAVE NOW WHICH
ARE COMING -- NOT FINISHED BUT
SHOW CLEARLY THIS DRUG IS DOES
HAVE EFFECT OF NK ACTIVITY IS
INCREASING GRANZYME AND
PATHOGEN, INCREASING ALSO THE
CYTOSIS THE MONOCYTES TO EAT
NECROTIC CELLS AND THOSE
DECREASING T-CELL ACT
ACTIVATION.
DO YOU THINK A SPECIAL
(INAUDIBLE) AS WELL AS
(INAUDIBLE) SO BECAUSE OF ALL
THIS EFFECT THAT IS MY QUESTION
TO YOU.
WHAT WE THOUGHT ABOUT IS PERHAPS
ZINC BEING FREED RESPONSIBLE FOR
BOTH EFFECT BECAUSE ZINC IS THE
SECOND MESSENGER RNA AFTER
(INAUDIBLE) WE HAVE IN THE BODY
AND THERE IS INCREDIBLE EFFECT
ON IMMUNE SYSTEM SO IT COULD BE
THAT I WAS QUESTION ASKING YOU
DO YOU THINK 2% OF NUCLEAR
CAPSID CAN BE AVAILABLE COULD
EXPLAIN SMALL ANTIVIRAL EFFECT
BUT THEN ZINC FREEING THE ZINC
FROM TRANSCRIPTION FACTOR FROM
OTHER FACTOR IN THE CELLS COULD
REALLY BE IMPORTANT FOR SEEN IN
COMBINATION WITH VACCINE?
>> AT THIS POINT NOTHING
SURPRISES ME ABOUT THIS
MOLECULE.
IT'S SMALL BUT VERY COMPLICATED
IN A LOT OF WAYS.
I COULD ENVISION A SITUATION
WHERE SOME DEGREE WE ARE
SELECTIVELY PERTURBING CERTAIN
POOL OF ZINC SOMEWHERE WHICH
MAYBE HARD TO PIN DOWN
SPECIFICALLY.
BUT SURE I THINK IT IS ENTIRELY
POSSIBLE.
IT IS NOT PERTURBING THINGS TOO
MUCH BECAUSE ZINC CAN BE TOXIC
TOO, IF IT IS TOO STRONG AND TOO
MANY TRANSCRIPTION FACTORS YOU
STOBBINGICS EFFECTS WHICH WE
DON'T SEE.
SO I FEEL THERE COULD BE A ZINC
INCREASE BUT PROBABLY SOMEWHAT
TARGETED AT THE SAME TIME IN A
UNIQUE COMPLICATED WAY.
>> I KNOW YOU MENTIONED THE ZINC
MOLECULE THAT BETHA IDENTIFIED
WHY WITH ALL THESE FACTORS THIS
MOLECULE DOES WHY DIDN'T IT WORK
WITH MARJORIE'S VENOSUM I DON'T
THINK WE KNOW.
>> IN THAT EXPERIMENT WE USED AN
ADJUVANT CONTROL.
GAVE SOME PROTECTION.
>> YEAH.
IN COMBINATION.
>> (OVERLAPPING SPEAKERS)
>> SO WE SHOULD HAVE USED NO
ADJUVANT.
AND WE MIGHT HAVE HAD A MUCH
BETTER RESULT.
BUT IT'S GREAT THAT YOU ARE
USING ANOTHER VACCINE.
>> YEAH.
GETTING BACK TO YOUR ORIGINAL
QUESTION ABOUT OTHER VIRUSES WE
HAVE SCREENED A PANEL OF NUMBER
OF OTHER VIRUSES AND DIDN'T SEE
MUCH ACTIVITY AT ALL.
WE DO SEE SOME EFFECT WITH SARS
COV-2 BUT WHAT IS INTERESTING IS
ALSO SARS COV-2 HAS ZINC
DEPENDENCE THERE IS A LOT OF
ZINC THAT IS VERY IMPORTANT FOR
THAT PARTICULAR VIRUS AS WELL.
(OFF MIC)
>> THAT IS MY GUESS.
ALSO TRYING TO CHASE A PROTEIN
TARGET WITHIN SARS COV-2 AND TRY
TO RECAPITULATE SIMILAR WORK
MASS SPECTROMETRY THAT SHOWS
DISSOCIATES ZINC.
[APPLAUSE]
>> WHILE DOING THAT I WOULD LIKE
TO THANK THE SPEAKERS IN THIS
SESSION AND THE SPEAKERS IN ALL
THE SESSIONS AND WE LIKE TO CALL
MARJORIE UP TO GIVE HER FINAL
THOUGHTS.
>> WELL, THANK YOU ALL.
THANK YOU ALL VERY MUCH FOR
COMING.
I WOULD LIKE TO FIRST THANK ALL
THE SPEAKERS, I THINK ESPECIALLY
THOSE WHO CAME PARTICULARLY LONG
DISTANCE.
THESE DAYS IT STILL IS NOT EASY
TO TRAVEL SO THANK YOU FOR
COMING.
I WOULD LIKE TO MENTION EACH OF
YOU IN TURN JUST WITH A FEW
SENTENCES.
I ALREADY COMMENTED FOLLOWING
BOB'S TALK SOMEHOW I KNEW
SOMEHOW I KNEW SOMETHING WOULD
CALL HIM AWAY.
BUT HE WAS GRACIOUS AND PLANNING
TO STAY THE WHOLE TIME AND
CALLED AWAY BY A POLITICAL
MEETING I THINK.
IN ANY CASE, I WILL STARTS WITH
RUTH RUPRECHT.
WE DID MANY FACT HAVE A NUMBER
OF GREAT COLLABORATIONS BUT ONE
THING THAT I VALUE VERY MUCH
OVER THE YEARS FROM YOU WAS YOUR
ADVICE.
BOTH FORMAL ADVICE AS A
SCIENTIFIC ADVISOR TO THE
VACCINE BRANCH AND ALSO JUST
INFORMAL DISCUSSIONS YOU
COMMENTED ON WHAT WAS GOING ON
WHAT I SHOULD DO, IT WAS VERY
VALUABLE.
THANK YOU VERY MUCH.
GRAB OUR COLLABORATIONS LED TO
INTERESTING EX-- GLAD OUR
COLLABORATIONS LED TO
INTERESTING EXPERIMENTS FOR YOU.
MARK, DELIGHTED YOU ARE GOING
FORWARD WITH REPLICATING
ADENOVIRUS.
I COULDN'T HAVE DONE IT, I'M NOT
A CLINICAL PERSON AND YOU ARE
MOVING US FORWARD, IT IS NOT
EASY, AND THE STORY YOU
PRESENTED WITH THE COVID, THAT
IS GREAT.
I WISH YOU GREAT SUCCESS, I HOPE
YOU HAVE SUCCESS WITH HIV AS
WELL AND LOOK FORWARD TO SEEING
THAT IN THE LITERATURE ONE DAY.
THANK YOU.
THANK YOU FOR YOUR SHARING YOUR
DATA.
MICHAEL TO MASS.
GREAT TO SEE YOU PRESENT SOME
INTERESTING AND IMPORTANT DATA
NOW THAT YOU HAVE LEFT THE NIH
YOU HAVE YOUR OWN GROUP AND I
THINK YOU HAVE ALWAYS BEEN
INTERESTED IN ADENOVIRUSES, YOU
ARE MOVING THEM FORWARD AS
AGENTS FOR VACCINES FOR ALL
KINDS OF INFECTIOUS AGENTS AS
WELL AS CANCER, LOOK FORWARD TO
YOUR FUTURE STUDIES AND FUTURE
DIRECTIONS.
ASHLEY, WE MENTIONED YOU WERE
ONE OF MY FIRST POST-DOCS YOU OF
COURSE AS WE SET UP HERE
INFORMALLY CAME TO WORK WITH BOB
BUT HE HAPPILY SUGGESTED YOU
JOIN MY GROUP AND I THINK WE HAD
A VERY PRODUCTIVE FUN TIME WE
WORKED ON NIGERIAN ISOLATES ON
HIV 2 AND IT IS REWARDING TO SEE
YOUR SUCCESS AT THE IHV AND ALSO
INTERNATIONALLY ESPECIALLY.
BEST OF LUCK.
RRAUL, YOU MADE A GREAT
CONTRIBUTION TO MY GROUP BY
SUGGESTING WE CHANGE THE ASSAY
FOR ADCC, YOU WENT TO DEVELOP
THAT ASSAY AND YOU MADE A VERY
NICE FINDING SHOWING THAT ADCC
ACTIVITY WAS ASSOCIATED WITH
REDUCED ACUTE VIREMIA.
THIS I THINK REALLY DID
STIMULATE THE FIELD AND LET YOU
WARN INVESTIGATORS LOOKING AT
THIS ACTIVITY AND ITS POTENTIAL
SO THANK YOU FOR THAT AND THANK
YOU FOR COMING ALL THE WAY FROM
OSLO TO TALK TO US.
GEORGE LEWIS, I ALSO VALUE VERY
MUCH ALL THE ADVICE YOU HAVE
GIVEN US OVER THE YEARS.
ALSO AS A SCIENTIFIC ADVISOR AND
INDEPENDENTLY OR INFORMALLY.
YOU HAVE BEEN A GREAT
COLLABORATOR AND YOU REALLY DID,
I MEAN, YOUR IMPROVEMENTS ON THE
ADCC ASSAY WERE REALLY VERY
HELPFUL AND YOU HAVE BEEN VERY
GENEROUS IN DISCUSSING THAT
APPROACH, SHARING YOUR REAGENTS
AND IN FACT YOU ARE ALWAYS
TELLING US WHAT YOU ARE DOING
AND SHARING YOUR THOUGHTS AND
THAT'S REALLY A GREAT
COLLABORATOR.
SO THANK YOU SO MUCH.
MARGIE ACKERMAN, RATHER THAN
FOCUSING JUST ON ANTIBODY
ACTIVITIES, I THINK YOU TOOK A
SOMEWHAT DIFFERENT APPROACH LOOK
AT FC RECEPTOR WHICH WAS -- THAT
ALSO STILL LATED THE FIELD
GREATLY.
LEADING TO INSIGHTS TO WHAT WAS
GOING ON, IN THE SYSTEMS BIOLOGY
IT'S BEEN GREAT.
WE DID ACTUALLY HAVE THAT ONE
NICE COLLABORATION ON
COMPLIMENT.
SO THANK YOU FOR THAT.
I WILL ALSO LOOK FORWARD TO YOUR
FUTURE STUDIES.
TWO YEARS AGO ABOUT THE TIME
THAT I WAS THINKING I MIGHT
ACTUALLY RETIRE, THE TOPICS OF
INTEREST TO ME OR WHAT I WOULD
HAVE CONTINUED WOULD HAVE BEEN
TO STUDY THE SEX BIAS
MICROBIOME, AND ALSO
COMBINATIONS OF VACCINES AND
MICROBE SIDE.
SO THAT LAST SESSION WAS MADE UP
OF THOSE THINGS THAT NEVER --
THAT I WOULD STILL BE DOING
BECAUSE I THOUGHT THEY WERE OF
GREAT IMPORTANCE.
SO ISKRA AS SHE DESCRIBED WAS
THE FIRST TO SHOW A SEX BIAS IN
ONE OF OUR MONKEY STUDIES.
THIS REALLY OPENED MY EYES I
THINK TO A WHOLE BODY OF
LITERATURE.
THERE IS A SEX BIAS IN MANY
IMMUNE RESPONSES, ACTUALLY AS I
RECALL, WOMEN RESPOND BETTER TO
VACCINES BUT IN TERMS OF HIV
THEY BECOME INFECTED THEN OR
THEY SUCCUMB TO DISEASE MORE
QUICKLY, THERE IS A LOT WE DON'T
KNOW SO ENCOURAGE YOU TO
CONTINUE FOLLOWING THE EFFECTS
OF THE SEX BIAS ON VACCINE
OUTCOME.
LAUREL, THANK YOU SO MUCH.
WHERE ARE YOU?
YOU ARE AN EXPERT IN MICROBIOME,
THANK YOU FOR COMING.
TOM MUSIC WAS GOING TO SPEAK HE
WAS A POST DOC ANY GROUP, NOW
BACK IN THE ARMY AS YOU
MENTIONED, HE IS MAJOR MUSIC.
HE HAD SOME NICE FINDINGS ON
DIFFERENCES IN THE MICROBIOME
ASSOCIATED WITH VACCINATION.
YOU HAVE ALWAYS BEEN THOUGH WE
HAVE NEVER COLLABORATED DIRECTLY
YOU HAVE ALWAYS BEEN VERY
GRACIOUS AND SHARING YOUR
KNOWLEDGE ANSWERING QUESTIONS
ABOUT STUDIES ONGOING IN MY
GROUP SO THANK YOU VERY MUCH.
FINALLY DANIEL APPELLA, WE DID
START A COLLABORATION ON VACCINE
MICROBE SIDE AS YOU POINTED OUT
AND I MIGHT MENTION THAT
ACTUALLY THAT WAS INITIALLY I
WAS STIMULATED IN THAT BY STUDY
OF CECILIA JANE MAYOR AND ETTA
RAY APPELLA WHO REPORTED A
VACCINE MICROBE SIDE COMBINATION
THAT WAS PROMISING.
CECILIA RETIRED BEFORE WE COULD
DO A COLLABORATION TOGETHER BUT
I WAS HAPPY TO GO FORWARD WITH
YOUR TWO FOUR SEVEN, DIDN'T HAVE
QUITE THE EFFECT THAT I HAD
HOPED THOUGH 247 WORKED GREAT
AND DELIGHTED THAT YOU AND BETHA
HAVE GOTTEN TOGETHER AND ARE
WORKING ON THIS AND REALLY
EXPANDED THE APPROACH, I CAN'T
WAIT TO SEE THE FULL STUDY, IT
WILL BE HOPEFULLY SOON.
I THINK WE WILL BE VERY EXCITING
AND IMPORTANT FOR THE FIELD.
IN MANY WAYS.
FINALLY, I WOULD LIKE TO THANK
THE PIs OF THE VACCINE BRANCH,
BETHA, GEORGE, BARBARA, I WAS
ALSO GOING TO MENTION -- SHE HAD
TO LEAVE OUR CLINICAL PERSON.
YOU HAVE GIVEN A LOT OF SUPPORT
OVER THE YEARS, COLLABORATIVE
HELP, YOU SHARED REAGENTS, YOU
HAVE GIVEN CRITICAL ADVICE.
AND YOU HAVE GIVEN FRIENDSHIP.
THOSE THINGS HAVE BEEN REALLY --
NOW IS THE TIME THAT I GET
EMOTIONAL, THAT IS WHY I HAVE
NOTES.
ALL OF THAT IS REALLY IMPORTANT
TO ME.
I THANK THE ADMINISTRATIVE
STAFF, JACKIE MCVAIL SITTING
OVER THERE MY SECRETARY FOR A
LONG TIME WHO WORKED VERY HARD
IN ORGANIZING THIS MEETING.
I SEE TANYA IS SITTING THERE,
TECHNICAL STAFF.
NOW WORKING WITH JAY WHO HAS
DONE A GREAT JOB.
I THANK ALL POST DOCS WHO HAVE
COME, RUTH, ARIF.
ANYWAY, I CAN'T MENTION
EVERYBODY. BUT I HAVE SO
ENJOYED WORKING WITH Y'ALL.
I JUST AS A FINAL PERSONAL NOTE,
RETIRING IS EASY, YOU JUST
LEAVE, YOU WALK AWAY.
IT IS MAKING THE DECISION TO
RETIRE THAT'S SO DIFFICULT.
BECAUSE BEING A SCIENTIST IT IS
A WONDERFUL CAREER.
YOU COME TO WORK EVERY DAY, YOU
DECIDE WHAT YOU WANT TO DO, YOU
DO IT THE WAY YOU WANT TO DO IT,
SCIENCE IS ALWAYS CHALLENGING,
IT'S NEVER BORING.
IT IS ALWAYS LEADING IN A NEW
DIRECTION, NEW AVENUES TO
EXPLORE.
AND IT BRINGS YOU YOUNG PEOPLE
WHO ARE ENTHUSIASTIC AND
TALENTED AND CONTRIBUTE TO YOUR
STUDIES.
IT BRINGS YOU SENIOR SCIENTISTS
WITH WHOM YOU CAN COLLABORATE
AND ADVANCE THE FIELD HOPEFULLY.
FINALLY, SCIENCE MAKES YOU
INTERACT WITH A WHOLE WORLD AT
LEAST IT HAS FOR ME.
SO THAT'S BEEN GREAT.
SO ALTHOUGH NEW EXPERIENCES
AWAIT ME, LEAVING IS BITTERSWEET
BUT I THANK YOU ALL FOR THIS FOR
TODAY, IT'S MADE IT EASIER TO
LEAVE.
AND THAT IS REALLY ALL I CAN
SAY.
THANK YOU VERY MUCH.
[APPLAUSE]
>> I ALSO WANT TO THANK EVERYONE
FOR PARTICIPATING, ALL THE
SPEAKERS AND EVERYONE ELSE WHO
CAME AND STAYED THROUGH THE
WHOLE THING, IT'S BEEN GREAT.