WE'LL BEGIN WITH GENERAL ANNOUNCEMENTS
WE USUALLY MAKE, AND WHEN THE
COUNCIL MEMBERS ARE ABLE TO JOIN
WE'LL GO AHEAD WITH THE
PRESENTATION FROM TIM.
WE'RE VERY FORTUNATE TODAY TO
HAVE AN OPEN SESSION COUNCIL ON
SOME OF THE T CELL EPITOPE WORK
GOING ON AND B CELL WORK AS
WELL.
TIM GANDRAY LEWIS IS GOING TO
INTRODUCE OUR SPEAKERS FOR THAT
PART OF THE OPEN SESSION AGENDA.
WE'RE ON A TIGHT TIME SCHEDULE
BECAUSE ONE OF THE SPEAKERS IN
THE SESSION THIS AFTERNOON HAS A
COMMITMENT AT 1:30 THAT WE WILL
ADJUST OUR SCHEDULE TO
ACCOMMODATE HIM.
THE MAIN THINGS I NEED TO
ANNOUNCE RIGHT NOW ARE THAT
COUNCIL MEMBERS SHOULD REMAIN ON
SCREEN WITH YOUR CAMERA ON, IF
YOU SPEAK, AND ANYBODY WHO USES
THE CHAT SHOULD ONLY BE FOR KIND
OF INCIDENTAL COMMENTS.
WE REALLY DO NEED ALL THE
COMMENTS YOU MAKE TO BE ON
SCREEN AND VERBAL.
AND WE HAVE A FEW SBIR CONTRACT
INITIATIVES THAT WE WILL NOT BE
PRESENTING THIS AFTERNOON, AS
IT'S OUR CUSTOM NOT TO PRESENT
THOSE SBIR TOPICS BUT I NEED TO
MENTION THEM.
ONE IS A PROGRAM FOR REAGENTS OF
NON-MA'AM MALLIAN AND
UNDERREPRESENTED MAMMALIAN
MODELS, AND DISEASES AGAINST
INFECTIOUS AND IMMUNE IMMEDIATE
INDICATED DISEASES, THIRD FOR
ADJUVANT DEVELOPMENT FOR THE
SAME TOPICS.
SO THAT SHOULD COVER IT UNLESS
THERE ARE SPECIFIC QUESTIONS
LATER ON IN THE SESSION WHEN WE
HAVE MORE TIME.
WE CAN PERHAPS COVER THOSE.
WE'LL ALSO BE PUTTING IN THAT
PART OF THE REVIEW FOR COUNCIL
MEMBERS, TWO TOPICS FROM OUR POD
SET OFFICE, OFFICE FOR
BIOINFORMATICS AND TECHNOLOGY
DEVELOPMENT.
ONE OF THOSE IS SOFTWARE FOR WEB
SERVICES, TO AUTOMATE METADATA
ENRICHMENT, AND STANDARDIZATION
FOR DATA ON INFECTIOUS DISEASES
AND IMMUNITY TO DISEASES.
AND THE SECOND TOPIC IS SOFTWARE
OR WEB SERVICES TO REREPRESENT
EXISTING SCIENTIFIC KNOWLEDGE
INTO A KNOWLEDGE CRAFT FORMAT.
WITH THOSE ANNOUNCEMENTS, I'M
GOING TO ASK TIM GANDRAY LEWIS
TO GO AHEAD WITH A BRIEF
AFTERNOON FOR THE AFTERNOON
SESSION AND OUR SPEAKERS.
TIM?
>>THANKS, DAN.
GOOD AFTERNOON, EVERYONE.
I'M GOING TO INTRODUCE THE
SESSION, AGAIN AS DAN SAID, I'LL
TRY TO KEEP IT VERY BRIEF.
MOST OF YOU KNOW THE STRUCTURE
OF OUR EPITOPE DISCOVERY
PROMISE, B CELL EPITOPE DISCOVER
AND MECHANISMS, OF ANTIBODY
PROTECTION PROGRAM, LARGE SCALE
T CELL IMMUNE UP HOPE DISCOVERY
PROGRAM, BOTH FEED IN TO OUR
DATABASE AND ANALYSIS RESOURCE,
DO AN EXPERT JOB OF CURATION OF
EPITOPES FROM THE LITERATURE.
PROGRAM OBJECTIVES ARE TO
CHARACTERIZE IMMUNITY TO
INFECTIOUS AGENTS AND VACCINES,
AND IMMUNE MEDIATED DISEASES,
PRODUCTS FROM DISCOVERS,
PEPTIDES, CAN BE USED TO MAKE
REAGENTS SUCH AS TETRIMERS TO
MONITOR PROTECTIVE IMMUNITY.
WE'VE BEEN VERY FORTUNATE WITH
THIS PROGRAM THAT WE'VE BEEN
ABLE TO ADAPT TO EMERGING
INFECTIOUS DISEASES SUCH AS
SARS-COV-2, AND UTILIZE THIS
PROGRAM TO IDENTIFY PEPTIDES AND
PROTECTIVE IMMUNE MECHANISMS FOR
SARS-COV-2.
AND ALSO DEVELOPMENT OF
THERAPEUTIC MONOCLONALS FOR
THERAPIES.
OVER HISTORY THE PROGRAM HAS
BEEN VERY ADAPTABLE IN
ADDRESSING EMERGING PATHOGENS
SUCH AS MULTIDRUG-RESISTANT
TUBERCULOSIS, WEST NILE, ZIKA
VIRUS, AND MOST RECENTLY
SARS-COV-2.
IT'S AN INCREDIBLY PRODUCTIVE
PROGRAM.
WE'VE HAD, YOU KNOW, OVER 140
PUBLICATIONS FROM THE TWO T CELL
AND B CELL PROGRAMS OVER THE
PAST -- REALLY THIS IS JUST
THREE YEARS THAT IT'S BEEN GOING
ON IN THIS RENEWAL SO THIS IS
JUST FOR THE LAST THREE YEARS,
AND CLOSE TO 3300 EPITOPE
RECORDS HAVE BEEN DEPOSITED.
ONE OF OUR CONTRACTORS YOU'LL
HEAR FROM LATER TODAY DR.
SETTAY, MADE MEGA POOLS OF
PEPTIDES FROM SARS-COV-2
AVAILABLE TO RESEARCH COMMUNITY
AND OVER 200 INDEPENDENT
INVESTIGATIVE TEAMS, 32
COUNTRIES, HAVE ACTUALLY
UTILIZED THOSE.
ANOTHER PROJECT IS A LICENSED
MONOCLONAL TO ASTRAZENECA, UNDER
EXTENDED USE AUTHORIZATION FOR
SARS-COV-2, AND IMMUNE EPITOPE
DATABASE HAS BEEN DOING
TREMENDOUS WORK INCLUDING
META-ANALYSES AND SARS-COV-2 TO
REALLY UNDERSTAND THE ROLE IN
PEDIATRIC MYOCARDITIS.
AND SO WITH THAT, I AM GOING TO
BRING IN OUR SPEAKERS, OUR FIRST
SPEAKER TODAY IS DR. JAMES CLOW
JR., DIRECTOR OF VANDERBILT
VACCINE CENTER.
HE'S PROFESSOR OF PEDIATRICS AND
PATHOLOGY, MICROBIOLOGY AND
IMMUNOLOGY AT VANDERBILT, HOLDS
THE ANN SCOTT HAROLD CHAIR AT
VANDERBILT.
I WILL TURN IT OVER TO DR.
CROWE.
JIM?
>>OKAY.
THANK YOU FOR HAVING ME.
IT'S GREAT TO BE WITH YOU.
I'M FLYING TO D.C. TODAY, I'M
GOING TO BE AT THE ZIKA MEETING
IN THE MORNING.
I'M ALL ABOUT NIH THIS WEEK.
IF I CAN SHARE MY SCREEN.
YOU'VE GOT MY SLIDES AND I'M ON
AUDIO I GUESS.
>>LOOKS GREAT.
>>THANK YOU.
YOU KNOW, AS TIM SAID, I'VE BEEN
INVOLVED IN THIS PROGRAM SINCE I
THINK I WAS ON THE ADVISORY
THING BEFORE IT HAPPENED AND
THROUGH EVERY, ITERATION.
WE WENT INTO IT, EVERYONE
THOUGHT, OKAY, WE'RE GOING TO
GET LIST, DATABASES OF EPITOPES,
THAT COULD BE SLIGHTLY BORING,
BUT I WANT TO CHANGE YOUR MIND
ABOUT WHAT EPITOPE MAPPING IS OR
EPITOPE DISCOVERY IS AND THINK
MORE ABOUT THE DISCOVERY OF
MECHANISMS AND SITES OF
VULNERABILITY, CORRELATES OF
PROTECTION, ALL THAT'S COME OUT
OF THESE CENTERS AND WE'VE HAD A
B CELL MAPPING ON AND OFF FOR
QUITE A WHILE.
I'M GOING TO IMPRESS ON YOU THE
IMPACT OF MAPPING STUDIES.
I THINK WHEN YOU HEAR THAT YOU
MIGHT THINK, OH, THIS MIGHT BE
BORING.
BUT IT'S NOT.
IT'S ACTUALLY BEEN THE CRUX OF
THE FIELD, I BELIEVE.
SO, ONE OF THE THINGS ABOUT B
CELL EPITOPE MAPPING OR ANTIBODY
MAPPING, I THINK, IS THAT WE
WERE AT A TIPPING POINT IN
HISTORY WHERE ANTIBODIES ARE
GOING TO START BEING ONE OF THE
PRINCIPAL TOOLS WE USE IN
EMERGING INFECTION MANAGEMENT.
IN FACT, MAYBE IN ALL INFECTIOUS
DISEASES, AND IT'S ODD THEY
HAVEN'T BEEN BECAUSE YOU'LL
REMEMBER THE FIRST NOBEL PRIZE
WAS GIVEN FOR ANTI-TOXINS, SO
ANTIBODIES WERE DISCOVERED IN
THE CONTEXT OF INFECTIOUS
DISEASES, POLYCLONAL IMMUNE
GLOBULINS USED SINCE THE
1880s, STARTING WITH RABIES
AND TETANUS AND SO ON.
BUT WHY DID CANCER INSTITUTE AND
AUTOIMMUNITY COMMUNITIES, YOU
KNOW, THEY HAVE OVER 100
LICENSED MONOCLONALS, ALL OF A
SUNDAY YOU'LL SEE A -- SUDDEN
YOU'LL SEE THEM COME INTO THE
FACE.
YOU ALREADY HAVE.
THEY ACTUALLY COVER MULTIPLE
DRUG CLASSES, SO ANTIBODIES, NOW
WE KNOW, CAN BE USED FOR
PREVENTION SO THE PRODUCT TIM
MENTIONED CAME OUT OF OUR
LABORATORY OUT OF OUR PROGRAM,
CONVERTED BY ASTRAZENECA TO
LONG-ACTING ANTIBODIES SO THE FC
WAS MODIFIED, OVER A 90 DAY
HALF-LIFE.
WITH PROPER POTENCY IN FACT
THESE ANTIBODIES OUTPERFORMED
mRNA VACCINES IN TERMS OF
EFFICACY AND DURABILITY, WHICH
IS AMAZING.
THEY ARE BETTER VACCINES THAN
VACCINES.
I KNOW YOU DON'T LIKE ME SAYING
THAT, BUT IT'S THE TRUTH, IF YOU
LOOK AT THE DATA.
I THINK WE NEED TO GRAPPLE WITH
THAT BECAUSE IN THE FUTURE
ANTIBODIES FOR SMALL OUTBREAKS
WILL BE QUICKER, BETTER, FASTER,
THEY ALSO ARE -- YOU CAN USE
THEM IN BABIES AND
WASHINGTON-YEAR-OLD PEOPLE, STEM
CELL TRANSPLANTS, MUCH MORE
VERSATILE THAN VACCINES IN
CERTAIN SITUATIONS AND THEY CAN
BE MORE DURABLE AND EFFECTIVE.
WE SAW THAT WITH COVID.
YOU CAN USE THEM IN
POST-EXPOSURE PROPHYLAXIS.
WHICH MOST VACCINES DON'T WORK
WELL.
WE DO USE IT IN PERINATAL HEP B
AND RABIES, PART OF THE REGIMEN,
BUT ANTIBODIES REALLY CLEARLY
CAN BE USED IN POST EXPOSURE
PROPHYLAXIS, THAT'S BEEN SHOWN
CLEARLY IN COVID.
YOU CAN USE IT AS A TREATMENT
LIKE YOU WOULD USE AN ANTI-VIRAL
SMALL MOLECULE DRUG BUT THEY
WILL WORK BETTER THAN DRUGS, AND
YOU CAN TAKE THEM INTO THE
HOSPITAL OR ONTO VENTILATED
PATIENTS.
EVUSHELL LONG ACTING DRUG WAS
LOOKED AS IN ACTIV TRIAL
SPONSORED BY NIH, CAUSED 30%
REDUCTION IN MORTALITY AND
HOSPITALIZED PATIENTS.
SAME DRUG AS VACCINE WORKS AS
ANTI-VIRAL THERAPIES, PRETTY
IMPRESSIVE.
I CAME UP WITH A GOOD PROFILE
FOR VACCINE OR PREVENTION ON TOP
IN BLUE AND TARGET PRODUCT
PROFILE FOR TREATMENT LIKE A
SMALL MOLECULE DRUG OR SOMETHING
FOR COVID.
I DON'T HAVE TIME TO TRICK YOU
HERE.
THESE ARE THE ACTUAL RESULTS OF
THE PHASE 2 PHASE 3 PREVENTION
TRIAL THAT MANY OF YOU KNOW OF
AND WERE PART OF, AND BOTTOM IS
THE TACKLE PHASE 3 RESULTS FROM
SORT OF TREATMENT SCENARIO.
AGAIN, IT'S THE SAME COMBINATION
OF TWO ANTIBODIES.
I'M GOING TO TELL YOU MORE ABOUT
THEM.
AT SIX MONTHS YOU GET AN 83%
REDUCTION IN HIGH RISK
INDIVIDUALS AGAINST ALL
SYMPTOMS, THIS IS NOT JUST
SEVERE DISEASE BUT THIS IS
INCLUDING UPPER RESPIRATORY
INFECTION, WHICH FAR EXCEEDS
WHAT A TWO-DOSE mRNA VACCINE
DOES IN HEALTHY PEOPLE AND COULD
BE USED IN IMMUNE COMPROMISED,
AND TREATMENT SCENARIO WHEN
GIVEN EARLY, NOT LATE BUT EARLY,
YOU GET 90% PROTECTION AGAINST
PROGRESSION TO SEVERE COVID.
I THINK THE VERSATILITY OF
ANTIBODIES MAKES THEM WORTHY OF
OUR ATTENTION.
A LOT OF THINGS THAT HAPPENED IN
THE LAST FIVE YEARS IN THE
ANTIBODY SPACE, ONE IS SPEED.
SO, THROUGH THE EPITOPE MAPPING
PROGRAM WE DID GET SOME
SUPPLEMENTAL FUNDING VERY EARLY,
I THINK THE PROGRAM FLEXED TO
COVID AND WE STARTED TALKING TO
PROGRAM IN FEBRUARY OF 2020, AND
WORK ON THIS SPACE.
IN TERMS OF SPEED, THESE ARE TWO
WORKFLOWS ON THE LEFT, 10X
GENOMICS SINGLE CELL RNAseqO
THE RIGHT IS A BERKELEY LIGHTS
SINGLE CELL DEVICE.
WE RAN TWO WORKFLOWS LIVE FIRE
DURING COVID, TAKE PBMCs FROM
SURVIVORS, ISOLATE B CELLS, USE
FLOW CYTOMETRY TO LABEL CELLS
WITH RECOMBINANT SPIKE PROTEIN,
AND THEN SORT THOSE, WE EARED
EXPANDED THEM IN T CELL-LIKE
MILIEU OR JUST WENT TO SINGLE
CELL RNAseq.
I'M GOING TO CONTINUE THOSE
WORKFLOWS, YOU END UP WITH GENES
YOU SYNTHESIZE, PUT IN PLASMA,
DO MICROSCOPE EXPRESSION, DO
BONDING AND NEUTRALIZATION.
ON THE RIGHT ON THE BEACON COULD
SEE INDIVIDUAL CELLS MAKING
ANTIBODY, AND EVEN WE DID SOME
FUNCTIONAL TESTS, SINGLE CELL
LEVEL, NEUTRALIZATION OR
RECEPTOR BLOCKING AT THE SINGLE
CELL LEVEL, REALLY FOR THE FIRST
TIME THAT I'M EVER AWARE OF.
ONE WAY OR THE OTHER, END UP
WITH RECOMBINANT ANTIBODIES.
AND SO THIS IS SORT OF A
SCHEMATIC.
B CELLS, MEMORY B CELLS, HAVE
THE B CELL RECEPTOR ON THEIR
SURFACE WHICH IS THE ANTIBODY
THEY WILL LATER SECRETE.
THAT'S HOW WE CAN LABEL THEM
WITH A RECOMBINANT PROTEIN LIKE
RBD OR SPIKE PROTEIN, ACTIVATE
ON CD LIGAND AND IL-12 OUT OF
BASIC SCIENCE STUDIES SPONSORED
BY NIH WE USE THOSE TO STIMULATE
A GERMINAL CENTER OR LYMPH NODE
IN VITRO AND PUT THOSE ON THIS
DEVICE, BERKELEY LIGHTS BEACON,
THEY DROP SHIPPED US ONE.
WE'VE NOW BOUGHT IT.
FIRST TIME WE EVER USED IT WE
MADE THE ANTIBODIES, PRETTY
AMAZING.
YOU LABEL BEADS, WITH SPIKE
PROTEIN, AND THOSE ARE PUT OUT
IN A CHANNEL AND YOU PUT A
SINGLE B CELL IN EACH OF THE
LITTLE WELLS, THEY CALL THEM
PENS, AND THE CELLS ARE
STIMULATED SO THEY FILL THIS
VERY SMALL VOLUME, VERY RAPIDLY,
SECRETED ANTIBODY, WHICH GETS
OUT INTO THE CHANNEL.
BEADS LIGHT UP, FLUORESCENTLY,
YOU CAN DO MULTI-COLOR.
WE DID RBD AND SPIKE AT THE SAME
TIME.
THIS IS A VIDEO OF ABOUT 30
MINUTES ON THE INSTRUMENT
LOOKING DOWN.
THE CHIPS ACTUALLY HOLD ABOUT I
THINK 18,000 OF THESE LITTLE
PENS, SO EACH VERTICAL
DISTINCTION IS A SINGLE CELL.
AND I'M JUST GOING TO SHOW YOU A
COUPLE HUNDRED HERE.
OTHERWISE YOU LOSE RESOLUTION.
THIS IS 30 MINUTES STARTING NOW.
YOU SEE A PLUME, THIS IS A BLACK
AND WHITE IMAGE, A PLUME OF
ANTIBODY LABELING THE BEADS THAT
ARE SPIKE OR RBD ON THE CHANNEL
SO UNDERNEATH EACH OF THESE
PLUMES LIKE A VOLCANO PLUME WE
GO TO THE BOTTOM, SEE THE B
CELL, THE INSTRUMENT, YOU
SURROUND THE CELL WITH A SQUARE
OF LIGHT AND THERE'S ENOUGH
ENERGY IN THAT LIGHT BOX TO MOVE
THE LIGHT, THERE'S ENOUGH ENERGY
IN THAT LIGHT BOX TO MOVE THE
CELL OUT.
YOU GET THE ANTIBODY VARIABLE
GENE SEQUENCES OUT.
USING THE TWO WORK FLOWS PBMCs
TO SINGLE CELL OR 10 X GENOMICS
OR BERKELEY LIGHTS, GO TO NEXT
GEN SEQUENCING PROTOCOL WHICH WE
USE ILLUMINA, YOU COULD USE
VARIOUS THINGS.
WE FTP SEQUENCES ON THE SERVERS,
TWIST, SYNTHESIZE, GAVE THEM OUR
EXPRESSION VECTOR, ANTIBODY
PLASMID, GIVES ASSEMBLY CLONE
SYNTHESIZED DNA INTO THE PLASMA
OF THE INSTRUMENT.
WE OPEN THE FED EX BOX, THERE'S
96 WELL PLATES WITH PLASMIDS
READY TO GO, TRANSFECT AND WE'RE
READY TO EXPRESS MONOCLONALS.
POLY PEPTIDES, FUNCTIONAL
MAPPING IS EXTREMELY IMPORTANT
FOR THERAPEUTIC OR PROPHYLACTIC
ANTIBODIES.
FOR COVID THERE WAS NO TEST AT
THE TIME WE DID THIS.
SO ON THE FLY, IN MARCH 2022, WE
DEVELOPED THIS TEST WHICH IS
CALLED REALTIME ANALYSIS, AND
WHAT THIS IS, IT'S A 384 PLATE,
THERE'S ELECTRODES THAT MEASURE
FLOW OF ELECTRICITY ACROSS THE
WELL.
IF YOU PUT CELLS ON THERE, A
TISSUE CULTURE TREATED PLATE,
CELLS BLOCK THE ELECTRICITY, AND
YOU GET A PROPERTY CALLED
IMPEDANCE.
YOU BLOCK.
AND SO FROM ZERO TO 80 HOURS ON
X-AXIS YOU CAN SEE WE PUT CELLS
ON AND SIGNAL GOES OUT BUT IT'S
NORMALIZED CELL INDEX, IT'S
IMPEDANCE OR BLOCKING
ELECTRICITY.
IF YOU ADD VIRUS IMPEDANCE GOES
DOWN BECAUSE YOU GLOW UP THE
CELLS, GET CYTOPATHIC EFFECT,
ELECTRICITY FLOWS AGAIN.
BUT IF YOU ADD VIRUS AND
MONOCLONAL ANTIBODY, IT
NEUTRALIZES THE VIRUS, WHEN THE
INTEGRITY OF THE CELL MONOLAYER
IS MAINTAINED, YOU MAINTAIN
IMPEDANCE.
WE DID THIS ACTUALLY IN ROUGH
DILUTIONS IN QUADRANTS, 1 TO 20,
500, USING CPE ASSAY IN REAL
TIME WITHOUT REAGENTS.
ALL WE HAD WAS VIRUS.
AND CELLS.
AND YOU PUT THOSE TWO THINGS
TOGETHER AND DO NEUTRALIZATION
WITHOUT ANY DETECTING
MONOCLONALS, NO
NEUROFLUORESCENCE, 24-36 HOURS
COULD SEE IT HAPPEN.
IT'S AMAZING.
ANOTHER PROPERTY THAT'S GOING TO
BE REALLY IMPORTANT FOR RNA
VIRUS THE IN THE FUTURE BECAUSE
WE'RE ALL EXPERTS ON VARIANTS
NOW, WE KNEW PROBABLY FOR RNA
VIRUS THERE ARE GOING TO BE
VARIANTS.
POTENTIAL FOR ESCAPE.
SO WE KNEW WE WANTED ANTIBODY
COMBINATIONS TWO OR MORE, SO
THAT WAS A GIVEN THAT WE WOULD
HAVE NON-COMPETING ANTIBODIES IN
THE COMBINATIONS THAT WE COULD
USE FOR THERAPY OR PROPHYLAXIS.
BUT ONE THING OUR GROUP IN
PARTICULAR HAS BEEN
INVESTIGATING IN THE RECENT
MAPPING CONTRACT, FOR FLU,
SARS-COV-2 AND SOME EBOLA STUFF
I'LL SHOW YOU, IS GETTING
SYNERGY OR COOPERATIVITY,
MAPPING AREAS ON THE VIRAL
GLYCOPROTEINS THAT CAN BE
INTERROGATED WITH ANTIBODY, TO
CHANGE THAT ENTIRE GLYCOPROTEIN
BY THE BINDING OF FIRST ANTIBODY
SO VIRUS IN SOME CASE THE
GLYCOPROTEIN BECOMES MORE
ACCESSIBLE OR SUSCEPTIBLE TO
NEUTRALIZATION, BY THE SECOND
ANTIBODY.
SO THE BINDING OF THE FIRST
MAKES THE ACTIVITY OF THE SECOND
BETTER.
THIS WAS CALLED SYNERGY.
IT'S COMMONLY LOOKED FOR IN
SMALL MOLECULE DRUGS AND DRUG
INDUSTRY.
YOU RARELY SEE IT BECAUSE NUMBER
ONE IT'S RARE.
YOU CAN'T BIND A GLYCOPROTEIN
ANYWHERE ON THE SURFACE AND GET
AN ALLOSTERIC EFFECT AT SITE OF
VULNERABILITY.
YOU HAVE TO MAP THAT.
THAT'S WHAT WE DID.
WE WENT THROUGH THOUSANDS OF ABS
THAT -- ANTIBODIES THAT WE GOT
FOR SARS-COV-2 AND LOOKED FOR
COMPETITION FIRST AND HAD
NON-COMPETING PAIRS, LOOKED AT
NEUTRALIZATION WHEN YOU ADD
TOGETHER.
THIS IS A SYNERGY PLOT.
IT'S A DOSE-RESPONSE.
IF YOU GO FROM ZERO TO 250
NANOGRAM ON THE X-AXIS OF 1
ANTIBODY, THE SECOND ON THE
Y-AXIS, CAN YOU SEE THERE'S
CONCENTRATIONS WHERE FOR
INSTANCE THIS ANTIBODY AT 8
NANOGRAMS PER MILL NO
NEUTRALIZATION, AND THIS 6 3, NO
ACTIVITY.
IF YOU COMBINE THEM AT THAT
CONCENTRATION YOU GET NEARLY
100% NEUTRALIZATION.
SO YOU CAN SORT OF SEE IF YOU
REVERSE THAT, YOU IS SEE A
SYNERGY MAP OF A STOICHIOMETRY
AND CONCENTRATION WHERE YOU GET
ENERGY.
THIS TURNED OUT TO BE VERY
IMPORTANT SO HERE'S THE
COMBINATION OF ANTIBODIES CALLED
2196 AND 2130 WE DISCOVERED THAT
ENDED UP IN THE ECUSHELL
PROPERTY, THIS IS SINGLE
RECONSTRUCTION AT VANDERBILT,
BOTH BIND AT THE SAME TIME.
RBD IS NOT VERY LARGE.
YOU HAVE TO BE CAREFUL TO GET
THE ANTIBODIES JUST RIGHT.
THESE ANTIBODIES WERE
SYNERGISTIC.
WE WENT ON ABOUT A MONTH LATER
AND GOT THE CRYSTAL STRUCTURE OF
BOTH ANTIBODIES ON THE RBD,
WE'RE SHOWING IT HERE IN
COMPLEX -- WELL, IN CONTEXT OF
THE TRIMER, WITH OPEN RBD, BUT
WE CAN SEE HOW THESE WORK.
THIS IS -- THIS TYPE OF MAPPING
INCLUDING EM, CryoEM, OR HIGH
RESOLUTION STUDIES WITH
CRYSTALLOGRAPHY OF COMPLEXES HAS
TURNED OUT TO BE A VERY
IMPORTANT PART OF THE MAPPING
ENTERPRISE SO THEN WE CAN GET
CONTACT RESIDUES, POTENTIAL
CONTACT RESIDUES, ANYTHING
WITHIN 4 1/2 ANGSTROMS OF THE
ANTIBODY AND THE VIRUS AND THEN
WE CAN TAKE THOSE VIRAL CONTACT
RESIDUES AND GO OUT TO NEXT
STRAIN OR SOMEPLACE WHERE FIELD
ISOLATES ARE BEING CATALOGED AND
WE CAN PREDICT WHAT'S GOING ON
IN TERMS OF ESCAPE OR FUNCTION.
NOW, ANOTHER REALLY EXCITING
THING WE DID IN THE MAPPING
ENTERPRISE HERE, WE WORK WITH
JESSIE BLOOM'S LAB AT FRED
HUTCH.
JESSIE PUT EVERY AMINO ACID, ALL
20, AT EVERY POSITION OF THE RBD
AND PUT THAT IN A YEAST LIBRARY.
WE WERE ABLE TO DO SORT OF A
THREE-STEP PROCESS.
ONE, WHAT RESIDUES WERE
TOLERATED SO THAT THE PROTEIN
EVEN FOLDED?
IF YOU PUT A RESIDUE IN, PROTEIN
DOESN'T FOLD, THAT'S NOT A
VIRUS.
YOU DON'T HAVE TO WORRY ABOUT
THAT.
SECOND, JESSIE ALREADY MADE THE
LIBRARY AND LOOKED FOR ACE-2
BINDING SO IF THAT RECEPTOR
BINDING DOMAIN DOES NOT MIND
RECEPTOR THAT'S NOT GOING TO BE
A VIRUS, THAT'S GONE.
WE ALREADY CULLED THROUGH THE
SEQUENCES.
DOESN'T FOLD, DOESN'T BIND
RECEPTOR.
WHAT WAS LEFT, THEN WE COULD
INTERROGATE ESCAPING THE TWO
ANTIBODIES WE WERE FOCUSING ON,
YOU CAN SEE ON THE BOTTOM 2196
ON LEFT, 2130 ON RIGHT, WEB LOGO
PLOTS OF AMINO ACIDS THAT WOULD
ALLOW ESCAPE FRACTION WERE
ALMOST NON-EXISTENT SO WE WERE
CATALOGING OUR ANTIBODIES AND
FOCUSING ON THE SYNERGISTIC
PAIRS THAT COULD BIND
SIMULTANEOUSLY AND SYNERGIZE AND
WERE POTENT AND NEUTRALIZING,
SELECTED DIFFICULT TO ESCAPE A
PRIORI.
YOU COULD PREDICT WHAT WAS GOING
TO HAPPEN IN THE FUTURE,
JESSIE'S GROUP COULD.
WE SELECTED THESE ANTIBODIES.
THE REST IS MORE CONVENTIONAL.
MIKE DIAMOND, THERE WERE NO
SMALL ANIMAL MODELS IN MARCH OF
2020, SO MIKE IS A COLLABORATOR
WITH ADENOVIRUS, HUMAN ACE-2
RECEPTOR IN MICE, ISOTYPE
CONTROL, TREATED MAINTAINED
WEIGHT, YOU SEE HISTOPATHOLOGY
IN UNTREATED, CLEAR LUNGS IN
TREATED.
WE WORKED WITH DAN BAROUCH'S LAB
IN THE MONKEYS MODEL, GRAVE
PROPHYLACTICALLY.
LOOKED IN NOSE AND LUNGS
CHALLENGED WITH ORIGINAL VIRUS,
USE BE SUBGENOMIC RNA TEST,
NEWLY MADE RNA, NOT A SINGLE
MOLECULE OF NEWLY MADE RNA
DETECTED IN PROPHYLAX
ANTIBODIES.
THE FINAL GO SIGNAL.
THESE ANTIBODIES LOOK GREAT.
AND NOW BACK TO THE SPEED THING
I MENTIONED, WE GOT THE SAMPLE
FROM THESE CONVALESCENT
INDIVIDUALS INFECTED IN WUHAN
CHINA IN DECEMBER 2019, TRAVELED
TO NORTH AMERICA.
SAMPLE ARRIVES IN NASHVILLE, BY
APRIL 8 HAD GIVEN ASTRAZENECA
THE SEQUENCES.
WE THINK WE CAN TAKE IT DOWN TO
MAYBE 14 DAYS, BUT WHO CARES,
14, 25, THIS STUFF IS FAST.
OTHER GROUPS DID IT AS WELL.
YOU SAW REGENERON AND OTHER
GROUPS THAT ARE IN COMMERCIAL
SPACE WORK VERY QUICKLY TOO.
AND THAT ENDED UP BEING THESE
TWO ANTIBODIES, UNTIL THIS WEEK
THE e-WAY WAS WITHDRAWN,
ANTIBODIES COVERS EVERY VIRUS
THROUGH BA.5.
PEOPLE SAY ALEXANDRIA FAILED. A.
THAT'S NOT TRUE.
THEY LASTED LONGER THAN VACCINES
AND ANTIBODIES.
IF YOU CHECK AND CHECK, WE GOT
SEVERAL YEARS OUT OF THOSE
ANTIBODIES WHICH IS AMAZING.
AND THEN OTHER TOPIC I WANTED TO
TALK ABOUT TODAY IS THE SCALE.
SO THAT'S SHIFTED DURING THE
CURRENT CONTRACT PERIOD, AT
LEAST B CELL EPITOPE MAPPING.
YOU CANNOT IMAGINE THE SCALE WE
CAN WORK AT NOW.
THIS IS EBOLA STUDIES THAT WERE
SUPPORTED IN OUR CONTRACT.
WE DO THE SAME THING, TAKE
BLOOD, IN THIS CASE SURVIVOR
FROM NIGERIA, WHO WAS A
PHYSICIAN TAKING CARE OF A CASE
AND BECAME INFECTED HIMSELF, IN
2014.
PBMCs WERE ISOLATED.
B CELLS, PARAMAGNETICALLY, FLOW
SHORT EBOLA GP, 10X WITH SINGLE
CELL.
WE SAW 100,000 EBOLA GP-SPECIFIC
CELLS AND SORTED THEM, RECOVERED
HEAVY AND LIGHT CHAIN FOR 15,000
FROM ONE SAMPLE.
SO MANY SEQUENCES, YOU CAN'T
JUST MAKE ALL THOSE.
YOU'RE TALKING ABOUT TENS OF
MILLIONS OF DOLLARS JUST TO
SYNTHESIZE DNA.
WE CLUSTERED.
SOME ANTIBODIES WERE RELATED,
FAMILIES, CLONE OH TYPES, WE DID
BIOINFORMATIC WORK.
IF I SHOW YOU THE UNIVERSE OF
15,000 ANTIBODIES COLOR CODED BY
FUNCTION, SO ORANGE IS
PAN-EBOLA, BLUE IS STRAIN
SPECIFIC, THE SIZE OF THE CLONAL
FAMILIES, AN AMAZING UNIVERSE.
I COULD GO ON AND ON ABOUT THE
TESTING THAT WAS PUT INTO THIS.
AND EVEN THE RELATIONSHIPS
GENETICALLY AND FUNCTIONALLY,
BRINGING NEUTRALIZATION WITH
SEQUENCE DIVERSITY, THERE'S A
LOT GOING ON IN TERMS OF
INFORMATICS IN THE SPACE.
IT'S AN EXCITING FIELD,
COMPUTATIONAL BIOLOGY.
ANOTHER TECHNIQUE THAT'S COME
INTO THE FIELD, CELL SIGNAL,
GEORGE GIORGIO, CAN YOU CAPTURE
ANTIBODIES NOT AS GENES FROM B
CELLS OR PLASMA CELLS BUT GRAB
THE ANTIBODIES, FROM PROTEIN OR
SERUM, FROM THE SAME INDIVIDUAL
WE CAPTURED 6,000 MONOCLONAL
ANTIBODIES OUT OF THE PLASMA
ONTO EBOLA GP, ELUTED THEM AS
PROTEINS, SEQUENCED THEM BY MASS
SPEC, YOU CAN COMPARE WITH
PROTEINS.
OPEN QUESTION, IS B CELL MEMORY
POOL THE SAME AS PLASMA CELLS
SECRETING INTO THE SERUM, VERY
IMPORTANT.
WE SEE SUBSTANTIAL OVERLAP.
THEY ARE NOT COMPLETELY
OVERLAPPED BUT HIGHLY
OVERLAPPED.
I THINK MY TAKE ON THIS IS THAT
B CELL MEMORY DOES TO A LARGE
EXTENT REFLECT AT LEAST THE POOL
FROM WHICH LONG LIVED PLASMA
CELLS ARE RECOVERED.
NOW, AM I OUT OF TIME?
DO I NEED TO END?
THESE ARE THREE OF THE CLONES IN
THIS INDIVIDUAL.
IT'S AMAZING ON THE TOP IS HEAVY
CHAINS.
THESE ARE POINT MUTATIONS, SO
THIS IS JUST INTERESTING THAT
INDIVIDUALS MAKE NOT ONLY A
MONOCLONAL, IN THE LINEAGE,
SYME'S THEY ARE MAKING --
SOMETIMES THEY ARE MAKING
HUNDREDS OF VARIANTS, WE WORRY
ABOUT VIRAL VARIANTS BUT YOU SEE
EQUIVALENT OR GREATER EXTENT OF
ANTIBODY VARIANTS.
NOW, SINCE WE HAD SO MANY
ANTIBODY SEQUENCES, 15,000 FROM
THIS INDIVIDUAL, TWO INDIVIDUALS
REPRESENTED HERE IN THE CIRCOS
PLOT.
WE WERE ABLE TO COMPARE DATA
FROM EMORY GROUP, SCOTT BOYD AND
RAFAEL MADDEN AND OTHERS HAVE
LOOKED AT REPERTOIRE OF EBOLA
SURVIVORS AND OTHERS VSV
VACCINEES, CHIMP, THE LINE
CONNECTS.
IT'S GRATIFYING TO SEE NATURAL
IMMUNITY OVERLAPS WITH THAT OF
VACCINE-INDUCED IMMUNITY.
THEY ARE NOT COMPLETELY
DIFFERENT.
THERE'S A LOT OF PUBLIC
CLONOTYPES.
THAT'S SOMETHING THAT'S HAPPENED
IN THE CURRENT CONTRACT LOTS
OF
PEOPLE ARE SEEING, YOU AND I
SHARE AN ANTIBODY SEQUENCE OR
LINEAGE, USED TO BE THOUGHT
RARE, NOW WE SEE IT'S THE NORM.
LOTS OF OVERLAP.
YOU CAN TAKE THOSE PUBLIC CLONE
CLONOTYPES AND DO FUNCTIONAL
GRIDS.
THE AMOUNT OF INFORMATION WE'RE
GETTING IS ENORMOUS.
WE CAN TAKE IT BACK TO MORE
CONVENTIONAL MAPPING THINGS, AND
WE TOOK THE TOP 15 CLONOTYPES
SHARED BETWEEN INDIVIDUALS BEING
VACCINATED OR INFECTED, DID E.M.
STUDIES.
YOU CAN SEE THESE INDIVIDUALS
ARE DECORATING THE ENTIRE
GLYCOPROTEIN, TOP, SIDE, BOTTOM,
STEM.
THESE INDIVIDUALS MAKE
ANTIBODIES TO EVERYWHERE ON THE
GP WHICH IS VERY COOL.
BECAUSE THE MONOCLONALS ARE
SNAPSHOTS.
WHEN YOU MAKE THEM AT THIS SCALE
YOU SEE HOW BIG IT IS.
WE TOOK A LOT OF CLONOTYPES INTO
MOUSE STUDIES, SO THIS IS EBOLA
ZAIRE, LETHAL INFECTION,
CONTROLS ALL DIED.
BUT MANY OF THESE
MONOTHERAPY-TREATED GROUPS WITH
PUBLIC CLONOTYPES ARE COMPLETELY
PROTECTED.
I THINK THIS IS REASSURING, LOTS
OF PEOPLE MAKE THE SAME
ANTIBODIES AND ARE PROTECTED
AGAINST EBOLA.
THE HUMAN SYSTEM IS COMPLEX AND
CAPABLE.
I WANT TO REEMPHASIZE THE
REVOLUTION IN CryoEM, A LOT OF
MAPPINGS GOING ON AT VERY HIGH
RESOLUTION OF ANTIBODIES AND
COMPLEX PARTICLES, BUT ALSO HGX,
YOU KNOW, PROTEOMIC STUDIES,
SINGLE PARTICLE RECONSTRUCTION,
CO-CRYSTAL STRUCTURES, SO MUCH
STRUCTURAL GOING ON.
WE CAN FIND ANTIBODIES EXTREMELY
FAST, USED AS VACCINES TO
PREVENT.
EVUSHELL WAS GIVEN IM, WE
LEARNED IV IS A LOSING STRATEGY,
WE DON'T HAVE ENOUGH INFUSION
SYSTEMS BUT IM IS COMPLETELY
POSSIBLE.
WE CAN GET INTO CLINIC RAPIDLY
WITH ANTIBODIES.
I HAVE A PROGRAM, WE'RE TRYING
TO MAKE ANTIBODIES FOR 100 AHEAD
OF TIME, DON'T HAVE TO WAIT.
NOW YOU MAY KNOW THAT WE DID A
STUDY WITH MODERNA, THE FIRST
ANTIBODY BY mRNA INTO HUMANS,
IT'S BEEN DONE IN HUMANS, WE
WORKING TOWARD NEBULIZED, $10 A
DOSE, ANTIBODIES ARE THE FUTURE.
YOU'RE GOING TO SAY DO I WANT A
VACCINE, ANTIBODY, BOTH?
IT'S NOT EITHER/OR.
IT'S BOTH/AND.
ANTIBODIES ARE GOING TO RISE IN
PROMINENCE AND USE, I THINK
YOU'LL SEE THAT.
THIS IS A PICTURE OF MY GROUP AT
VANDERBILT.
I MENTIONED MIKE DIAMOND, JESSIE
BAROUCH, A LOT OF COMPANIES
HELPED US.
NIH HAS BEEN OUR PRINCIPAL
FUNDING SOURCE FOR THE MAPPING
STUDIES, BUT DOLLEY PARTON GOT
US GOING, OUR LOCAL CELEBRITY,
WHO HELPED US GET GOING ON
COVID.
I'LL STOP THERE AND MAY HAVE
USED MY TIME BUT I'M HAPPY TO
ANSWER QUESTIONS IF THERE IS
TIME.
>>JIM, THANK YOU.
THAT WAS TERRIFIC.
WE'RE PAST THE TIME.
YOU SAID YOU HAD ANOTHER
COMMITMENT.
IF THAT'S CHANGED, WE HAVE TIME.
>>I THINK I'VE GOT MINUTES.
LET'S DO IT.
>>QUESTIONS FROM COUNCIL
MEMBERS?
>>JIM?
>>YEAH, JIM, TERRIFIC.
I WAS INTERESTED IN YOUR
PRESENTATION OF SYNERGY, AND HOW
FAR DOES THAT GO?
DO YOU SEE SYNERGISTIC
INTERACTIONS WHERE AN ANTIBODY
CAN ACTUALLY OPEN UP AN EPITOPE
THAT'S OTHERWISE INACCESSIBLE?
>>YES, YOU CAN SEE THAT.
IN THE PREVIOUS ITERATION OF
THIS CONTRACT WE WERE MAPPING
FLU ANTIBODIES AND DISCOVERED
TRIMER INTERFACE ANTIBODIES.
SO IN OLIGOMERS LIKE FLU,
TRIMERS, SURFACES ARE OBSCURED,
YOU CAN'T SEE THEM IN CRYSTAL
STRUCTURE BUT FOUND ANTIBODIES
THAT WOULD RECOGNIZE THOSE SITES
AND WORK IN VIVO AND ARE
PROTECTIVE.
WE'VE BEEN LOOKING.
THERE'S SOME DYNAMICS THEREFORE
TO HA, THEY OPEN DURING FUSION
BUT TURNS OUT EVEN BEFORE FUSION
THEY ARE MOVING AROUND, CRYSTAL
STRUCTURES WE THINK OF THEM AS
MARBLE STATUES BUT THEY ARE NOT
LIKE THAT.
THESE THINGS ARE WIGGLING AND
OPENING.
SINCE THAT TIME WE FOUND SIMILAR
ANTIBODIES FOR NEAPA, DIMERS
BIND DOWN, NOT SHOWN ON THE
SURFACE OF THE CRYSTAL STRUCTURE
OF DIMER BUT THOSE ARE EVEN
NEUTRALIZING AND WE SEE
ANTIBODIES TO OTHER SURFACES
PROLONG OPENNESS THAT'S ALREADY
THERE, IT'S NOT JUST PROVOKING
IT, LIKE IT'S ALREADY THERE,
SORT OF LOCKS IT MORE OPEN OR
TENDS TO BE MORE OPENNEN.
DIMER, TRIMER, TETRA VIRUS,
PROBABLY EVERY VIRUS HAS
INTERFACE EPITOPES NOT
UNDERSTOOD YET, AND THESE
COMBINATIONS ARE PLAYING WITH
THAT.
THEY ARE CHANGING DYNAMICS OF
THE OLIGOMERS, THAT'S AN
IMPORTANT FUTURE DIRECTION, HOW
TO HARNESS THAT.
>>GREAT.
THANK YOU.
>>JIM, THERE'S A LOT BEEN
PUBLISHED RECENTLY ABOUT
VACCINAL EFFECTS WITH HIV,
BROADLY NEUTRALIZING ANTIBODIES.
WONDERING IF YOU'VE HAD THE
OPPORTUNITY TO LOOK AT THAT WITH
SOME OF THE SARS-COV-2
ANTIBODIES AND ANY OTHER
THOUGHTS ON ENGINEERING YOUR
ANTIBODIES TO ENHANCE A VACCINAL
EFFECT IF YOU'VE DONE THAT OR
HAVE PLANS TO DO THAT?
>>YEAH, I'M HESITANT TO ANSWER
BECAUSE I'M AN OUTLIER IN THIS
FIELD.
WHEN WE PUT EVERYTHING TOGETHER
WITH AZ, WE ELECTED TO KNOCK OUT
ALL FC GAMMA RECEPTOR
INTERACTIONS.
OF COURSE THERE WAS CONCERN
ABOUT ADE, HIGHER REPLICATION AT
THAT TIME BUT IT TURNED OUT I
THINK TO BE THE RIGHT THING.
IF YOU LOOK AT HOSPITALIZED
PATIENTS THE ONLY ANTIBODY DID
NOT FAIL FOR FUTILITY AS YOU
WELL KNOW WAS EVU SHELL AND
CAUSED 30%.
YOU DON'T NEED FC MEDIATED, YOU
23450ED NEUTRALIZATION.
IF YOU HAVE EARLY OUTPATIENT
MILD DISEASE AND LOW NUMBER OF
INFECTED CELLS, PROBABLY IN YOUR
UPPER AIRWAY, SURE, HAVE AT IT,
KILL ALL THE CELLS, MAYBE TUNING
UP IS A GOOD IDEA BECAUSE YOU
WANT TO ELIMINATE THE FACTORIES.
WHEN YOUR LUNG IS HIGHLY
INFECTED, YOU'RE IN THE HOSPITAL
BECAUSE YOU HAVE LUNG
INFLAMMATION OR YOU'RE ON THE
VENTILATOR, DO YOU WANT TO POUR
OIL ON THE INFLAMMATORY MILIEU
BY BRINGING IN EFFECTOR CELLS
AND TURNING THEM UP?
I THINK THE ANSWER IS NO.
YOU WANT TO TURN THEM OFF,
THAT'S WERE PEOPLE WERE USING
GLUCOCORTICOIDS, YOU ALREADY
HAVE INFLAMMATION, MY OWN
OPINION WHICH NOT MANY PEOPLE
SHARE, I DON'T THINK, IS YOU
WANT TO TURN IT OFF.
YOU WANT A SILENT ANTIBODY.
BUT TAKE IT FOR WHAT IT'S WORTH
>>JIM -- I'M SORRY.
>>GO AHEAD.
>>I WANT TO FOLLOW UP.
THANK YOU VERY MUCH, A GREAT
PRESENTATION.
TO FOLLOW UP ON DAN'S QUESTION,
COULDN'T YOU HAVE DIFFERENT
ANTIBODIES FOR DIFFERENT STAGES?
SO MAYBE FOR PREVENTIVE
MONOCLONAL MAYBE YOU DO WANT
SOMETHING THAT HAS A VACCINAL
EFFECT BECAUSE THEN YOU'RE
NEUTRALIZING BUT THEN YOU'RE
ALSO MAYBE -- WHEN PEOPLE GET
EXPOSED YOU COULD BE EXPANDING
THE RESPONSE IF YOU WERE GETTING
SOME KIND OF VACCINAL EFFECT?
YOU WOULDN'T WANT TO USE THAT
WHEN SOMEBODY IS IN THE
HOSPITAL, RIGHT?
>>I AGREE WITH THAT.
THIS TOUCHES ON -- THERE'S A
LARGE LITERATURE ON
PASSIVE/ACTIVE IMMUNIZATION,
VERY POPULAR IN THE '60s.
ACTUALLY DID SOME WORK IN THE
INTRAMURAL PROGRAM IN
CHIMPANZEES WITH A POLYCLONAL
ADDING LIVE ATTENUATED VACCINES.
IF ANTIBODY NEUTRALIZATION TITER
IS HIGH IN SETTING OF
VACCINATION YOU SUPPRESS PRIMARY
RESPONSE.
IT'S NOT GOOD FOR THE PRIME.
WELL, THE ANTIBODY RESPONSE TO
THE PRIME.
BUT IF YOU GET THE STOICHIOMETRY
JUST RIGHT YOU ENHANCE THE
PRIMING AND ENHANCE THE MEMORY
RESPONSE TO A SECOND DOSE OF
VACCINE.
THAT WAS WORKED OUT WITH
MEASLES, WHEN THE VACCINE WAS
TOO HOT, PEOPLE WERE GIVEN
IMMUNOGLOBULIN AND VACCINE, CHOP
DID STUDIES.
IF ANTIBODIES ARE HIGH,
SUPPRESS.
IS IT STOICHIOMETRY IS PERFECT
YOU GET ENHANCEMENT.
WHEN ANTIBODIES ARE LOW YOU
DON'T SEE ANY EFFECT.
PART IS NOT JUST TIMING BUT
UNDERSTANDING THE FUNCTIONAL
STOICHIOMETRY.
AND MAYBE -- SOME PEOPLE TRIED
TO COUPLE THE ANTIBODY IN
VACCINE TO TARGET DEN DENDRITIC
CELLS.
YES, FOR VACCINATION THERE'S A
SWEET SPOT OF ANTIBODIES THAT'S
BETTER THAN ANTI-GEN ALONE,
EXACTLY WHY I'M NOT SURE.
>>IS THERE ANYTHING IN THE
OFFING THAT WOULD, YOU KNOW,
LEAD TO A NEW TECHNOLOGY OR
PLATFORM FOR ENHANCING ANTIBODY
DURABILITY THAT, YOU KNOW,
THERE'S SOME NICE ADVANCES THERE
ALREADY BUT IS THERE ANYTHING
YOU'RE AWARE OF THAT'S COMING IN
THE FUTURE?
>>THE CURRENT THINGS BEING USED
LIKE YTELS, THOSE GET YOU TO,
WELL, EACH ANTIBODY IS DIFFERENT
BECAUSE THE FV INTERACTS WITH
FCRN SO DEPENDING ON THE
SEQUENCE BUT YOU MIGHT ENVISION
110 PHYSICAL -- 110 SOMEDAY
PHYSICAL HALF-LIFE, WHERE YOU
TOP OUT, THE BENEFITS HERE, I
DIDN'T HAVE TIME TO TALK ABOUT
OUR PROGRAM, I WANTED TO, WE'VE
DONE DISCOVERY WHERE YOU CAN
FIND ANTIBODIES, SOME FOR RIP
WHERE POTENCY IS .01 NANOGRAM
PER mL.
THE PREDICTED, WE DOSE DOWN IN
ANIMALS, PREDICTED EFFECTIVE
DOSE IN HUMAN IS LESS THAN 1
MILLIGRAM, IT'S 700 MICROGRAMS.
SO IF YOU GAVE A HUMAN BEING 300
MILLIGRAMS STANDARD DOSE OF THAT
ANTIBODY, YOU ALREADY HAVE A
HALF-LIFE THAT'S ENORMOUS, IT'S
YEARS.
IF YOU ADDED YTE TO THAT NOW
YOU'RE OUT TO 5 TO 10 YEARS.
I THINK THE POTENTIAL GAINS ARE
NOT NECESSARILY ENGINEERING THE
FCRN FOR PHYSICAL HALF-LIFE,
IT'S ACHIEVING A LONGER
FUNCTIONAL HALF-LIFE BY DROPPING
THE IC 50, INCREASE POTENCY, YOU
HAVE A LONGER FUNCTIONAL
HALF-LIFE RATHER THAN PHYSICAL
HALF-LIFE.
THAT'S WHERE THE GAINS ARE TO BE
MADE.
>>GREAT.
THANK YOU.
>>EXCELLENT.
>>TIM, SEEMS LIKE WE SHOULD
MOVE ON.
>>THANK YOU FOR HAVING ME.
>>I'LL SEE SOME OF YOU TOMORROW
AT THE ZIKA MEETING.
>>SEE YOU THEN.
>>THANKS, JIM.
>>ALL RIGHT.
SO OUR NEXT PRESENTATION IS FROM
DR. SETEE, PROFESSOR IN CENTER
FOR AUTOIMMUNITY AND
INFLAMMATION AND CENTER FOR
INFECTIOUS DISEASE AND VACCINE
RESEARCH AT LA JOLLA INSTITUTE
FOR IMMUNOLOGY.
AND ALEX, ARE YOU READY?
>>YES.
LET ME SHARE MY SCREEN.
AND HOPEFULLY YOU CAN SEE IT.
AND LET ME GO ON PRESENTATION
MODE.
OOPS.
YOU SEE THE SCREEN?
>>LOOKS GREAT, ALEX.
>>OKAY, GOOD.
WHAT I THOUGHT I WOULD DO IS
TALK ABOUT TWO DIFFERENT
EXAMPLES OF WHAT HAS BEEN
ACCOMPLISHED IN THE CONTEXT OF
THE EPITOPE IDENTIFICATION AND
DISCOVERY, ONE RELATED TO
DENGUE, THE OTHER SARS-COV-2
MORE RECENTLY.
AND SPECIFICALLY, FIRST HAVE A
SHORT OVERVIEW OF APPROACHES FOR
T CELL EPITOPE IDENTIFICATION
AND VALIDATION AND GO INTO TWO
SPECIFIC EXAMPLES HERE.
SO, YOU ALL KNOW DIFFERENT
ASSAYS ARE UTILIZED TO MEASURE
T-CELL RESPONSES, SO PROBABLY
THE MOST SIMPLE BUT CONVERSELY
LESS INFORMATIVE ASSAY IS THE
WHOLE BLOG INTERFERON GAMMA
RELEASE ASSAY, USED IN TB AS
QUANTUM THEOREM ASSAY, IT WORKS
BUT DOESN'T TELL WHAT YOU KIND
OF CELLS ARE PRODUCING
INTERFERON GAMMA.
NEXT STEP UP IS THE ELISPOT
ASSAYS, ROBUST, RELATIVELY
HIGH-THROUGHPUT, BUT FAR LESS
SENSITIVE OF SOME OF THE MORE
SECOND GENERATION ASSAYS THAT
I'M GOING TO DISCUSS IN A
SECOND.
AND ALSO THEY DON'T GIVE ANY
INFORMATION ABOUT THE PHENOTYPE
OF A RESPONDING T CELL WHERE
THEY ARE CD4 OR CD8 AND SO
FORTH.
EXTRACELLULAR CYTOKINE STAINING
INSTEAD DETECT FUNCTIONAL
RESPONSES, AND THEY ARE -- THEY
GENERATE PHENOTYPIC INFORMATION.
NOT ONLY YOU DETECT INTERFERON
GAMMA OR IL-5 OR IL-10 BUT ALSO
KNOW WHAT IS THE CELL THAT MAKES
CD4, CD8, YOU CAN MEASURE ALSO
OTHER PHENOACTIVIC MARKERS ON
THE CELL SURFACE.
POTENTIAL PROBLEM IS ICS DETECTS
PRODUCTION OF CYTOKINE, A
PROBLEM, IN THE CASE FOR EXAMPLE
OF FOLLICULAR T HELPER CELLS
BECAUSE NOTORIOUSLY BOTH CELLS
DON'T MAKE A LOT OF CYTOKINES.
YOU WOULDN'T DETECT, IN FACT WE
DEVELOP THIS AIM ASSAY IN
COLLABORATION WITH SHANE CROWLEY
THE MAIN DRIVING FORCE TO
OVERCOME THE ISSUES.
AIM ASSAY DETECTS UPREGULATION
OF MARKERS WHEN THE T CELL IS
ENCOUNTERED THE COGNATE ANTIGEN,
IT'S NOT DEPEND ON DETECTING
CYTOKINES, THE OTHER PROBLEM IF
YOU RUN AN ICS FOR INTERFERON
GAMMA, THE CELL IS NOT MAKING
INTERFERON GAMMA BUT MAKING A
DIFFERENT CYTOKINE, YOU WOULD
MISS.
THERE ARE OTHER ASSAYS SUCH AS
TETRIMERS, TCR SEQUENCING WHICH
I'M NOT GOING TO DISCUSS HERE
TODAY, I TRUST DAVID WHO FOLLOWS
ME, NEXT SPEAKER, WILL DISCUSS
SOME OF THIS, ALSO VERY GOOD.
ONE POINT I WANT TO TOUCH ON IS
BEYOND WHAT PARTICULAR ASSAY YOU
MAY USE, YOU HAVE TO CONSIDER
DIFFERENT FORMATS TO BE
CONSIDERED.
SO, ONE FORMAT IS EX VIVO
ASSAYS.
SO THAT IS A FORMAT WHERE YOU
JUST ASSAY FRESH CELLS OR FROZEN
PBMCs, WITHOUT ANY IN VITRO
EXPANSION.
THIS IS OBVIOUSLY MOST
PHYSIOLOGICALLY BECAUSE DOESN'T
ALTER PHENOTYPE.
THESE ARE LIVE CELLS, CAN BE
SORTED AND ISOLATED, SO IT'S
VERY AMENABLE TO DOWNSTREAM
ANALYSIS WITH TRANSCRIPTOMICS,
SINGLE CELL AND SO FORTH.
IT DOES REQUIRE A LITTLE BIT
MORE CELLSN IN COMPARISON TO IN
VITRO.
WHAT'S TYPICALLY DONE, ONE- OR
TWO- WEEK IN VITRO CULTURE, AND
THIS IS VERY EFFECTIVE IN
EXPANDING ANTIGENS FOR T CELLS.
THAT'S ALLOWING YOU TO DETECT
WEAK RESPONSES, YOU CAN STILL DO
TCR SEQUENCING, YOU CAN DO IT EX
VIVO.
IT'S MORE LABORIOUS, CERTAIN
CELL TYPES LIKE TREG DO NOT
EXPAND REEL IN VITRO.
PHENOTYPE IS ALTERED.
THE LAST THING TO CONSIDER IS
WHAT ANTIGEN ARE YOU GOING TO
USE TO CONFRONT THE T CELLS IN
THE ASSAY FORMAT AND IN SPECIFIC
ASSAY.
AND I'M GOING TO DISCUSS THIS A
LITTLE BIT FURTHER BECAUSE WE
NEED TO ANSWER THE QUESTION OF
WHAT ANTIGENS ARE WE GOING TO
USE AND WHAT IS A RELEVANT
ANTIGEN TO USE IN MOST
SITUATIONS, WHAT PROMPTED US TO
DEVELOP THIS MEGAPOOL APPROACH.
THIS STARTS FROM CONSIDERATION
THAT IN REALITY HUMAN T CELLS
RECOGNIZE A LOT OF DIFFERENT
ANTIGENS, SO YOU GO FROM 25
DIFFERENT POTENTIAL ANTIGENS, IN
THE CASE OF SARS-COV-2, TO
HUNDREDS OF DIFFERENT EXPRESSED
IN POXVIRUSES OR HERPES VIRUSES,
TO THOUSANDS OF DIFFERENT
ANTIGENS EXPRESSED IN THE CASE
OF BACTERIA SUCH TUBERCULOSIS,
BORE DA TELL A, SO THIS IS
FURTHER COMPLICATED BY THE FACT
HUMANS EXPRESS HLA TYPES, SIX
DIFFERENT IN MOST HUMANS, SIX
DIFFERENT CLASS 1, EIGHT CLASS 2
TYPES.
SO IT'S A VERY LARGE DEGREE OF
HETEROGENEITY WHICH PROMPTED US
TO REALLY PUSH THE TECHNOLOGY TO
SEE HOW MANY DIFFERENT PEPTIDES
AND CANDIDATE EPITOPES WE COULD
COMBINE A SINGLE POOL, HAD TO
USE SEQUENTIAL STRATEGY TO AVOID
TOXICITY AND NOW CAN ROUTINELY
USE POOLS OF HUNDREDS OF
DIFFERENT PEPTIDES.
DEPENDING ON THE PARTICULAR
APPLICATION, YOU CAN USE
OVERLAPPING PEPTIDES, SPANNING
THE ENTIRE SEQUENCE OF ANTIGEN
OF INTEREST, OR IF THE NUMBER OF
ANTIGENS
ANTIGENS TO BE CONSIDERED IS TOO
LARGE OR CONVERSELY AVAILABLE TO
STUDIES TOO SMALL YOU CAN USE
PREDICTIONS MAKING SURE YOU DO A
GOOD JOB REPRESENTING IT, IN THE
DIFFERENT LOCI OR IF THE TARGET
IS STUDIED EXTENSIVELY AND THE
EPITOPES RECOGNIZED BY HUMAN T
CELLS ARE ALREADY KNOWN TO A
LARGE EXTENT, YOU CAN USE
UNDATED EVIDENCE.
THIS MEGAPOOL APPROACH HAS BEEN
DESCRIBED AND VALIDATED FOR MANY
DIFFERENT VIRUSES BUT MICRO
BACTERIA, ALLERGENS,
AUTOIMMUNITY.
AS WAS MENTIONED EARLIER, WE
HAVE DEVELOPED REAGENTS, AND
SHARED THESE REAGENTS WORLDWIDE
WITH STARTING COLLABORATIONS,
SOMETIMES SENDING TO PEPTIDES,
IN THE CASE OF SARS-COV-2 THIS
WAS IMPORTANT TO CATALYZE BASIC
AND APPLIED RESEARCH, WE SHARED
THIS WITH HUNDREDS OF LABS IN
163 DIFFERENT LABS, SIX
CONTINENTS, IF YOU KNOW OF
ANYBODY THAT HAS A LAB IN
ANTARCTICA WE'LL HAVE HAVE EVERY
CONTINENT WHERE WE SENT THE
MEGAPOOLS.
LET ME GIVE AN EXAMPLES OF HOW
WE USE THESE APPROACHES TO
CHARACTERIZE T CELLS IN THE CASE
OF DENGUE IS, A MAJOR HEALTH
PROBLEM, A MEMBER OF THE
FLAVIVIRUS GENUS, WHICH HAS
SEVERAL VIRUSES OF PUBLIC HEALTH
CONCERN SUCH AS ZIKA, YELLOW
FEVER, WEST NILE, SO FORTH.
AND DENGUE IS -- THERE ARE FOUR
SEROTYPES, ONE THROUGH FOUR,
HUNDREDS OF MILLIONS OF
INFECTIONS PER YEAR, AND LIKELY
THE LIFE-THREATENING SEVERE
DISEASE IS LIMITED TO SMALL
FRACTION OF THOSE CASES, AND
CURIOUSLY THIS SEVERE DISEASE IS
ASSOCIATED WITH AN HETEROLOGOUS
SETTING.
SOMEONE GETS INFECTED WITH
DENGUE 2 AT FIRST TIME,
REINFECTED A SECOND TIME WITH
SAY DENGUE 3, THE RISK FOR
SEVERE DISEASE IS HIGHEST.
WE STUDIED QUITE A BIT DENGUE IN
TWO ENDEMIC AREAS, ONE
NICARAGUA, THE OTHER SRI LANKA,
BY DESIGN BECAUSE WE WANTED TO
STUDY TWO COMPLETELY DIFFERENT
POPULATIONS ASSOCIATED WITH
DIFFERENT HEALTH CARE, DIFFERENT
ETHNICITIES, HLA DISTRIBUTIONS,
TO FOLLOWING BUT REALLY FINDINGS
THAT WOULD BE TRUE AND VERIFIED
IN SUCH DIVERSE SETTINGS WOULD
BE REALLY UNIVERSAL.
SO, WE APPLIED ETHICAL
IDENTIFICATION STRATEGIES AND WE
EVENTUALLY DEFINED ABOUT 300
EPITOPES IN NICARAGUA ENDEMIC
AREA POPULATION AREA, ABOUT 400
IN SRI LANKA, AND THEN WE LOOKED
AT THE OVERLAP AND WE ZOOM IN ON
ABOUT 250 DIFFERENT EPITOPES
WHICH ACCOUNTED FOR 90% OF
ACTIVITY, IN BOTH POPULATIONS
ABLE TO GENERATE A CD8 MEGAPOOL,
WE THEN SHOWED THIS MEGAPOOL
COULD BE USE TO DETECT T-CELL
RESPONSES IN ANY POPULATION, IF
YOU WENT IN BRAZIL, INDIA OR
WHATEVER, STILL WERE ABLE TO USE
THIS MEGAPOOL TO DETECT
RESPONSES AND SIMILAR EPITOPE
IDENTIFICATION STUDIES WHICH I'M
NOT GOING TO DISCUSS FOR
INTEREST OF TIME LED TO
DEFINITION OF MEGA POOL AT CD4
EPITOPES RECOGNIZED IN HUMAN
POPULATIONS AND ENCODED BY
DENGUE.
SO, ONE IMPORTANT THING THAT
CAME OUT OF THIS STUDY, NOW WE
COULD ACTUALLY MEASURE RELIABLY
HUMAN T-CELL RESPONSES IN
PATIENT POPULATIONS WITHOUT
HAVING JUST ONE EPITOPE OR
PARTIAL PRESENTATIONS, AND THIS
WAS RELEVANT BECAUSE I'VE BEEN
IN THE FIELD FOR A LONG TIME,
THIS CONCEPT, BUT WAS ACTUALLY
NOT REALLY SUPPORTED
EXPERIMENTALLY BY SOLID DATA, IN
MY OPINION, THAT T CELLS WERE
RESPONSIBLE FOR THE SEVERE
DISEASE IN ACUTE PHASE AND THAT
THE T CELLS WERE THE BAD ACTORS.
IN FACT IN MY OPINION ALSO
SOMEWHAT REALLY UNFORTUNATE
CONSEQUENCES BECAUSE IN THE
VACCINE SETTING PEOPLE SENDED TO
SHY AWAY FROM INDUCING T-CELL
RESPONSES.
BUT THEN WE WERE ABLE TO
ACTUALLY ISOLATE THE
ANTIGEN-SPECIFIC CD4 AND CD8 T
CELLS AND ASK THE QUESTION, IS
THERE A DIFFERENCE IN IMMUNE
SIGNATURE?
ARE THEY CAUSING HAVOC IN THE
ACUTE PHASE AND SEVERE DISEASE?
AND THAT WAS NOT TRUE.
BOTH FOR CD4 AND CD8.
SO THIS WAS A VERY IMPORTANT
ADVANCE, IN MY OPINIONS,
UNDERSTANDING IMMUNOPATHOLOGY
ASSOCIATED WITH SEVERE DENGUE
INFECTION.
FURTHERMORE, WHEN YOU LOOK AT
THE TARGETS OF IMMUNE RESPONSES,
TURNS OUT THAT IMMUNE RESPONSES
ON TOP OF CD8 AND ON THE BOTTOM
IS CD4, FOR SAKE OF
ILLUSTRATION, IS THAT THE
ENVELOPE PROTEIN WHICH IS THE
TARGET OF ANTIBODY RESPONSES,
WHICH ARE OF INTEREST, TO
PREVENT INFECTION, IS ACTUALLY
NOT A GOOD TARGET FOR EITHER CD4
AND T CELLS.
IN THE CD4 T CELLS ACTUALLY
RECOGNIZE CAPSID AND NS 3 MOST
DOMINANTLY, CD8 SIMILARLY NS3 IS
DOMINANT TARGET FOLLOWED BY NS5.
THIS IS RELEVANT IN THE CONTEXT
OF VACCINE DEVELOPMENT.
AND SO THERE ARE THREE DIFFERENT
MAIN VACCINE CONSTRUCTS AND
DESIGNS THAT HAVE BEEN DEVELOPED
FOR DENGUE AND ONE IS DENGVAXIA
DEVELOPED BY SANOFI PASTEUR, ONE
BY TAKEDA, AND TV003 FOLLOWING
THE WORK OF STEVE WHITEHEAD AT
NIAID, AND THEN MERCK.
ALL RELY ON FOUR DIFFERENT
ENVELOPES FROM FOUR DENGUE
SEROTYPES.
IN THE CASE OF THE DENGVAXIA THE
BACKBONE OF YELLOW FEVER WHILE
TAKEDA AND NIAID USED THEM
(INDISCERNIBLE).
I WANT TO REMIND YOU WHAT I
SHOWED IN THE PREVIOUS SLIDE ACY
HUMAN T CELLS NOT RECOGNIZED
ENVELOPE VERY WELL, THERE WAS
ALSO ANOTHER STUDY TO GET TO THE
BOTTOM WHERE WE LOOKED AT
WHETHER THE T CELLS INDUCED BY
YELLOW FEVER WOULD RECOGNIZE
DENGUE SEQUENCING.
THIS SAYS IF YOU WANT T CELLS,
IF YOU WANT ALSO CD8 TO KILL
INFECTED VIRUS, CD4 TO SUPPORT
THE ANTIBODY RESPONSE, ENVELOPE
BY ITSELF AND/OR ENVELOPE BY
ITSELF OR A YELLOW FEVER
CONSTRUCT IS NOTE -- NOT GOINGE
ENOUGH.
WE EVALUATED T-CELL RESPONSES
INDUCED BY 003 TETRAVALENT
VACCINE, THAT INDUCES STRONG
T-CELL RESPONSES, ACTUALLY EVEN
HIGHER THAN WHAT WE'RE SEEING IN
CONVALESCENT PEOPLE FROM
NICARAGUA AND OTHER AREAS.
ALSO MOST T-CELL RESPONSE
OBSERVED IN THE CONTEXT OF
TETRAVALENT VACCINE WAS AGAINST
SEQUENCES SHARED BY FOUR
SEROTYPES.
SO REALLY INTERESTING FINDING,
SUCH AS COMBINATION OF DIFFERENT
DENGUE BACKBONES REALLY FOCUSES
BY FAR
THE BEST.
ALSO EARLY CD8 RESPONSE
CORRELATED, IF YOU LOOK AT EARLY
RESPONSE, ANTIBODIES DO NOT
CORRELATE WITH A PARTICULARLY
MILDER OUTCOME.
THIS WAS CONFIRMED BY THE GROUP
IN SINGAPORE WHICH ADDED THE
FACT THAT EARLY T-CELL RESPONSES
WERE ASSOCIATED WITH LOWER VIRAL
LOAD, USING SAMPLES FROM
OUTBREAK, EARLY WUHAN EARLY IN
ITALY IN 2020, WE SAMPLED,
CONFIRMED WITH LARGER COHORT AND
SAW THIS IMPORTANT PHENOMENON
THAT ANOTHER IMPORTANT CORRELATE
OF MILD DISEASE WAS TO HAVE A
POLYFUNCTIONAL RESPONSE, SO
PEOPLE WITH T-CELL RESPONSE NOT
ONLY AGAINST SPIKE BUT ALSO
AGAINST OTHER ANTIGENS DID MUCH
BETTER THAN THE RESPONSE FOCUSED
ON SPIKE, A RECENT DATA BUT FOR
EMPHASIZE IN CERTAIN CONDITIONS
YOU CAN EVEN HAVE A PROTECTION
ASSOCIATED WITH T CELLS FROM
INFECTION.
SO, AT THIS POINT, WE MOVED TO
MEASURE RESPONSES AGAINST
VACCINATION, USING THE MEGAPOOLS
DEVELOPED IN THE PREVIOUS
STUDIES, COMPARED TWO mRNA
PLATFORMS, PROTEIN ADJUVANT
VACCINE, THIS IS A NICE STUDY IN
MY OPINION, SUBJECTS THAT WERE
ALL RECRUITED AND DONATED BLOOD
IN OUR CLINICAL CORE AND ALL
ASSAYS RUN ON THE SAME
INSTRUMENT, SAME DAYS, BY THE
SAME PEOPLE.
IT'S TIGHTLY COMPARABLE, AND
WHAT YOU SEE HERE THE THE
ANTIBODY RESPONSE GOES UP AND
DOWN, RISK WANES OVER SIX MONTHS
FOR BOTH mRNA PLATFORM, THE
VIRAL VECTOR IS NEVER AS GOOD IN
TERMS OF NEUTRALIZING ANTIBODY
BUT HAS A DIFFERENT KINETIC AND
GOOD NEUTRALIZING SIMILAR TO
mRNA.
I WANT TO POINT OUT THE
DIFFERENCE IN KINETICS OF T-CELL
RESPONSES, FOR DIFFERENT
PLATFORMS ARE MUCH MORE DURABLE.
THEY WANE MUCH LESS RAPIDLY.
NOW, THIS FITS WITH WHAT WAS
OBSERVED BEFORE THE VARIANTS
BECAME PREVALENT, WERE ALREADY
IN A NUMBER OF STUDIES FOR
EXAMPLE KAISER PERMANENTE WAS
OBSERVED THAT THE RESPONSE
INDUCED BY VACCINATION OR
NATURAL INFECTION TENDED TO PAIN
IN TERMS OF PROTECTION AGAINST
INFECTION, BUT THE VACCINES
STILL RETAINED SUBSTANTIAL
PROTECTION AGAINST SEVERE
DISEASE AND DEATH.
WHAT ABOUT VARIANTS?
WE DID STUDIES TO IDENTIFY CD4
AND CD8 EPITOPES, AND WE ENDED
UP DEFINING CLOSE TO A THOUSAND
EPITOPES, WE'RE NOT THE ONLY ONE
DOING THE STUDIES.
IF YOU MAKE A MEGA ANALYSIS OF
ALL THE DATA THAT WAS CONTAINED
IN THE IDP IN EARLY 2021,
ALREADY 2,000 DIFFERENT CD4, CD8
EPITOPE EXPERIMENTALLY DETECTED
IN HUMANS.
BY NOW WE'RE ABOUT 3,000
EPITOPES.
SO WHY AM I HARPING ON THIS
LARGE NUMBER OF EPITOPES BEING
RECOGNIZED?
BECAUSE THIS IS DIFFERENT FROM
WHAT YOU HEARD JIM TELLING YOU
WITH THE RBD REGION IS SMALL, SO
RELATIVELY FEW MUTATIONS IN THE
REGIONS THAT ARE RECOGNIZED BY
NEUTRALIZING ANTIBODIES CAN LEAD
TO A SUBSTANTIAL LOSS OF
ANTIBODY OVERACTIVITY OF
NEUTRALIZING ANTIBODIES.
AND CONVERSELY FOR T CELLS THIS
IS NOT REALLY FEASIBLE BECAUSE
SARS-COV-2 WOULD HAVE TO MUTATE
CONSTANTLY FROM EPITOPES TO
COMPLETELY ESCAPE HUMAN T CELL
RECOGNITION.
SO, IN FACT, THAT'S EXACTLY WHAT
YOU SEE.
WE SAW IN THIS STUDY WHERE WE
LOOKED AT ALL DIFFERENT VARIANTS
FROM ALPHA TO OMICRON, BUT
ESSENTIALLY 85% OR MORE OF T
CELL ACTIVITY WAS PRESERVED WITH
DIFFERENT VARIANTS.
THE LARGE MAJORITY OF T CELL
EPITOPES WERE 100% CONSERVED.
AND IN FACT IF WE DID REPERTOIRE
STUDIES WE ALSO SHOWED IN
INDIVIDUAL SUBJECTS THE MAJORITY
OF T CELL EPITOPES RECOGNIZED IN
THAT PARTICULAR SUBJECT WERE NOT
IMPACTED BY MUTATIONS IN THE
VARIANTS.
THIS WAS IN CONTRAST TO WHAT WAS
SEEN IN THE CASE OF NEUTRALIZING
ANTIBODIES, DRAMATICALLY
IMPACTED.
THIS WAS DONE, WAS CONFIRMED IN
MANY DIFFERENT STUDIES IN
COLLABORATION THAT INVOLVED LABS
ALL OVER THE WORLD, AND AT THIS
POINT IT'S I THINK WELL
ESTABLISHED THIS DICHOTOMY,
IMPACT IS MUCH GREATER AT THE
LEVEL OF NEUTRALIZING ANTIBODIES
WHILE T CELL ACTIVITY IS
ESSENTIALLY PRESERVED AT THE
POPULATION LEVEL.
THIS FITS WITH WHAT WE'RE
SEEING.
WE ALL SEE EVERY DAY WHERE NOW
THE VARIANTS HAVE PROTECTION
INDUCED BY VACCINE OR EFFECT OF
DIFFERENT VARIANTS IS MUCH
DECREASED IF YOU LOOK AT
PROTECTION FROM INFECTION, BUT
PROTECTION FROM SEVERE DISEASE
AND FROM DEATH IS MUCH -- IT'S
STILL LARGELY PRESERVED.
WHERE DO WE GO?
THIS IS ILLUSTRATING SOME OF THE
MEGAPOOLS WE CONTINUE TO
OPTIMIZE.
THIS IS A COMBINATION OF POOLS
THAT WE HAVE USED TO DETECT
IMMUNE RESPONSES, SO IT'S
DAYS BASICALLY A MEGA POOL THAT
DETECTS SPIKE, ANOTHER ONE THE
REST OF THE GENOME.
THIS COMBINATION PLOTS TWO
DIMENSIONAL -- IN TWO DIMENSIONS
CAN ACTUALLY TELL YOU A HISTORY
OF INFECTION.
SO PEOPLE ARE VACCINATED ONLY
REACT ONLY AGAINST SPIKE.
PEOPLE THAT HAVE BEEN INFECTED
BUT NOT VACCINATED OF COURSE SEE
BOTH SPIKE AND THE REST OF THE
GENOME.
PEOPLE THAT HAVE BREAKTHROUGH
INFECTION, FOR EXAMPLE, OR
HYBRID IMMUNITY, WE'VE SEEN BOTH
SPIKE TWICE, ONE BECAUSE OF
VACCINATION, ONE BECAUSE OF
INFECTION, THEY HAVE SO TO SPEAK
A HIGHER CONTENT OF SPIKE.
WE CAN ACTUALLY DO THIS AND
WE'VE SHOWN THIS IS MUCH MORE
SENSITIVE AT ANY SEROLOGICAL
ASSAY.
I WANT TO CLOSE HERE WITH THE
NOTION THAT ALL THIS DATA TELLS
US THAT A T CELL INDUCING
VACCINE COMPONENT COULD HAVE
REAL VALUE, TO ENHANCE
PREPAREDNESS AGAINST FUTURE
POSSIBLE PANDEMICS, AGAIN IN THE
CONTEXT OF PREVENTING SEVERE
DISEASE, NOT INFECTION,
SYNERGISTIC, THIS COULD BE
APPLICABLE TO MANY DIFFERENT
VIRUS FAMILIES THAT ARE PANDEMIC
CONCERN LISTED HERE WHERE YOU
COULD FOCUS ON WHAT ARE THE
CONSERVEED T CELL EPITOPES TO
DEVELOP BROADLY APPLICABLE
PANDEMIC PREPAREDNESS REAGENT.
SO, WHERE WE REMAINING
CHALLENGES AND TARGETS EMERGING
AND REEMERGING TARGETS I DID NOT
HAVE TIME TO GO INTO Mpox.
WE PUBLISHED A STUDY AT THE END
OF A YEAR, CHARACTERIZING AND
DESCRIBING THE FIRST ROUND OF
POOLS THAT CAN BE USED TO
DEVELOP -- TO DETECT RESPONSES
AGAINST Mpox IN THE
VACCINATION INFECTION SETTING,
PANDEMIC PREPAREDNESS, ALSO
BACTERIAL TARGETS LESS WELL
CHARACTERIZED, ALSO MANY OF THE
VACCINE TARGETS THAT ARE
REGISTERED OR IN DEVELOPMENT ARE
LESS WELL CHARACTERIZED.
I'D LIKE TO THANK THE PEOPLE
THAT HAVE BEEN INVOLVED IN THESE
STUDIES, STRONG COLLABORATION
WITH SHANE CROWLEY, AND MANY
OTHERS, THE IDP, AND THANK YOU
ALL AND I'LL TAKE QUESTIONS.
>>THANK YOU VERY MUCH, ALEX.
I'D LIKE TO OPEN THIS UP FOR
QUESTIONS.
ARE THERE ANY QUESTIONS FROM THE
COUNCIL MEMBERS OR FROM THE
OTHER ATTENDEES?
>>THIS IS JIM.
YOU MADE THE POINT PUTTING BROAD
RANGE OF T CELL EPITOPES COULD
BE HELPFUL FOR DURABILITY OF
PROTECTION AGAINST SEVERE
DISEASE.
WHAT ABOUT HOST CHARACTERISTICS?
AND ARE THERE SPECIAL
CONSIDERATIONS, YOU KNOW, FOR
DIFFERENT CATEGORIES OF HIGH
RISK INDIVIDUALS THAT YOU NEED
TO FACTOR INTO THEIR APPROACH.
>>YEAH, THAT'S A VERY GOOD
QUESTION.
CERTAINLY ONE THING THAT IS
IMPORTANT TO CONSIDER, I MEAN,
PRAGMATICALLY IF YOU SEE NOW THE
PEOPLE THAT THE MAJORITY OF
PEOPLE THAT HAVE SEVERE DISEASE
TODAY ARE OLDER PEOPLE, PEOPLE
THAT DIDN'T ALL GET BOOSTED, IF
YOU WANT TO BE OVERGENERALIZING.
THAT ALREADY SUGGESTS WHAT ARE
SOME OF THE TARGETS FOR
INTERVENTION.
WHICH LEAD TO YOUR QUESTION --
WHICH LINK TO YOUR QUESTION, FOR
EXAMPLE, IS THERE A DIFFERENCE
IN T-CELL RESPONSE AND MEMORY
T-CELL RESPONSE IN OLDER PEOPLE?
IT'S A COMPLICATED THING.
OLDER PEOPLE MAY RESPOND LESS,
RIGHT?
JUST BECAUSE THEY ARE OLDER.
CONVERSELY OLDER PEOPLE BECAUSE
WE TEND TO HAVE MORE SEVERE
DISEASE MAY GET MORE ANTIGEN
LOAD SO WHEN YOU LOOK AT SOME OF
THE CONVALESCENT THERE'S
INDICATION THEY HAVE BETTER
T-CELL RESPONSE BECAUSE
(INDISCERNIBLE) BY THE VIRUS,
THAT OPENS THE QUESTION HOW LONG
DOES THIS IMMUNE RESPONSE -- DO
THEY -- IS THERE A DIFFERENCE IN
HOW QUICKLY OR STABLE THEIR
T-CELL RESPONSE EVOLVES OVER
TIME?
SO THOSE ARE VERY GOOD
QUESTIONS.
STILL I THINK OPEN TO A LARGE
DEGREE.
>>THANK YOU.
ARE THERE ANY OTHER QUESTIONS?
>>YEAH, TIM, I HAD A QUESTION.
ALEX, YOU ALLUDED TO PANDEMIC
PREPAREDNESS.
I'M WONDERING WHAT YOUR THOUGHTS
ARE ON WHERE ARE THE
BOTTLENECKS, AND SHOULD WE BE,
YOU KNOW, ASSEMBLING PEPTIDE
MEGAPOOLS FOR FUTURE
POSSIBILITIES OR IS IT
SUFFICIENT TO GET THINGS GOING
VERY QUICKLY WHEN THE NEXT
PATHOGEN EMERGES?
>>RIGHT.
I BELIEVE CERTAINLY A LOT OF
GROUND WORK COULD BE DONE TO
DEFINE THE EPITOPES POTENTIALLY
SHARED BETWEEN DIFFERENT
FAMILIES OF CONCERN, AND HAVE
BEEN VALIDATE AND READY TO GO.
OBVIOUSLY YOU CAN DO THAT AFTER
THE FACT, BUT YOU COULD DO THE
WORK AND HAVE INFORMATION READY
AND AVAILABLE AND READY TO GO,
OBVIOUSLY IN THE CONTEXT OF
PANDEMIC PREPAREDNESS, WE --
DEVIL'S ARGUMENT, WE DON'T KNOW
WHERE THE PANDEMIC CAN COME
FROM.
WELL, YES, BUT IF WE CAN COVER
SEVEN OR EIGHT FAMILIES THAT ARE
OF REASONABLE CONCERN THAT'S
ALREADY OF VALUE.
I MEAN, CERTAINLY IF ONE -- ONE
COULD EVEN ENVISION THAT THESE
ANTIGENS OR FRAGMENTS OF DOMAIN
IDENTIFIED OF BEING OF
PARTICULAR INTEREST, ARE
IDENTIFIED, ONE CAN DO LIMITED
PHASE 1 TRIALS TO SEE IF
DIFFERENT VACCINES ENCODING THEM
WORK OR SOMETHING LIKE THAT.
BUT CERTAINLY THE GROUND WORK
COULD BE DONE AND I THINK BE A
LOT OF VALUE IN DOING IT.
>>THANK YOU.
>>THANK YOU VERY MUCH.
ARE THERE ANY OTHER QUESTIONS
BEFORE ALEX GOES?
IF NOT, I WANT TO THANK YOU,
ALEX, FOR A GREAT PRESENTATION.
WE APPRECIATE IT.
NEXT SPEAKER TODAY IS DR. DAVID
KOELLE, THE PROFESSOR AT -- A
PROFESSOR AT THE UNIVERSITY OF
WASHINGTON, DEPARTMENT OF
MEDICINE, DIVISION OF ALLERGY
AND INFECTIOUS DISEASES.
HE'S AN ADJUNCT PROFESSOR OF
GLOBAL HEALTH, AND A JOINT
PROFESSOR IN LABORATORY OF
MEDICINE IN DEPARTMENT OF
MEDICINE.
HE'S ALSO AFFILIATE INVESTIGATOR
AT THE FRED HUTCHINSON CANCER
RESEARCH CENTER.
AND DAVID'S SLIDES LOOK GREAT.
DAVID, IF YOU'RE READY?
>>THANK YOU.
CAN YOU HEAR ME?
>>GREAT.
>>THANKS FOR THE INVITATION.
IT'S A TALL ORDER TO FOLLOW JIM
AND ALEX BUT I'LL TRY MY BEST.
I'M GOING TO SHIFT EMPHASIS A
LITTLE BIT TOWARDS T-CELL
RECEPTOR SIDE OF THINGS, AS WELL
AS EPITOPE DISCOVERY, FORTUNATE
TO HAVE A CYCLE, AND TURNING TO
HOST SITE AND ACKNOWLEDGMENT
SLIDE TO SHOW A COUPLE FOLKS
WHOSE WORK I'M GOING TO
EMPHASIZE TODAY.
MY TITLE INCLUDED THE CONCEPT OF
TISSUE T CELLS, AND BECAUSE
NATURE DOES A NICE THING FOR US,
WHICH IS ANTIGEN SPECIFIC T
CELLS AT THE SITE OF THE
DISORDER, SO RATHER THAN
NECESSARILY FISHING RARE T CELLS
OUT OF THE BLOOD OR SENSITIVE
TECHNIQUES CAN SOMETIMES BIOPSY
THE SITE AND USE THIS
PRESUMPTIVE NATURAL ENRICHMENT,
AND IF I HAVE TIME I'LL SHOW YOU
WORK FROM SYPHILIS, CARCINOMA,
SHINGLES BUT STARTING WITH
COVID, ANTIGEN SPECIFIC T CELLS
IN THE NOSE, I BELIEVE.
ON THE RIGHT OF COURSE THIS IS
WHAT WE'RE TALKING ABOUT.
WITH ADVANCEMENTS IN SEQUENCING
CAN USE THE SEQUENCE OF CDR 3
REGION AS A BARCODE TO HELP US
TRACK T CELLS FROM THE BLOOD TO
TISSUES AND BACK AND FORTH.
SO, OUR STRATEGY OVER THE YEARS
HAS STARTED WITH BLOOD AND
TISSUE, WHEN WE START WITH BLOOD
WE TEND TO ENRICH RARE
PATHOGEN-SPECIFIC T CELLS OUT OF
THE BLOOD, GUARANTEEING THE T
CELLS WE'RE STUDYING REACT TO
THE WHOLE PATHOGEN.
SO IT'S NOT JUST A CIRCULAR
ARGUMENT WHERE YOU ENRICH WITH
PEPTIDE, TEST WITH PEPTIDE, WE
HAVE THE PATHOGEN IN THE CYCLE,
EXPAND OR EXPRESS REPORTER CELLS
AND GO TO ANTIGEN AND EPITOPE
DISCOVERY.
WITH TISSUE-BASED SYSTEM WE CAN
USE THIS NATURAL IN SITU ENRICH.
-MENT.
WE CAN EXPAND CELLS OR SEQUENCE
AND GO TO DISCOVERY.
THIS IS BLOOD EMPHASIS.
WE USE AIM.
ALEX INTRODUCED AIM, IN THE
EARLY DAYS WERE USING CD137 AS
OUR MARKER.
HERE IS A BLOOD SAMPLE, SOME
CELLS ARE EXPRESSING IN RESPONSE
TO ANTIGEN I MENTIONED.
AND WHEN WE EXPAND WE'RE 50%
ANTIGEN REACTIVE, FROM .1 OUT OF
THE BODY TO NOW 50%, NOW
DISCOVERING EPITOPES, IT'S LIKE
SHOOTING FISH IN A BARREL.
WE CAN GO ON TO FULLY DEFINE
WHAT ANTIGENS AND EPITOPES ARE
IDENTIFIED.
THIS IS A SCREEN, GENOME WIDE
SCREEN FOR EVERY IN THE ZOSTER,
HERE IS THE SCREEN BEFORE THE
VACCINE, 28 DAYS AFTER THE
VACCINE.
YOU CAN SEE THERE DOES SEEM TO
BE A VACCINE RESPONSE, THIS IS A
WHOLE VIRUS VACCINE, THE
ATTENUATED OLD ZOSTRAVAX.
HERE IS DAY ZERO ON THE LEFT,
YOU CAN SEE WHEN WE GO TO DAY
28, A ONE-DOSE VACCINE, WE HAVE
A LOT BROADER RECOGNITION AFTER
DAY 28 AND THIS LASTS OUT TO DAY
182.
MECHANISM OF ACTION FOR THIS
VACCINE AND NEWER SHINGLES
VACCINE IS THOUGHT TO BE A T
CELL MECHANISM OF ACTION, IN
CONTRAST TO ALMOST ALL VACCINES
THAT WE TALKED ABOUT PREVIOUSLY,
PROBABLY IT'S THE T CELLS THAT
MATTER HERE.
WHEN WE HAVE HITS AT THE OPEN
LEVEL, OR GENE LEVEL, EXAMPLE
FROM GENE 68, THE GLYCOPROTEIN,
FAIRLY ROUTINE, FIGURING OUT
WHAT THE PEPTIDES ARE.
YOU CAN FIGURE OUT
HLA-RESTRICTING LOCUS, USEEL
SINGLE ANTIGEN PRESENTING T
CELLS, ANTIGEN PRESENTING,
PARDON ME.
ONLY ANTIGEN PRESENTING CELLS
THAT EXPRESS WERE ABLE TO
PRESENT THE EPITOPE.
ONE OF THE PAIN POINTS IS WHAT
ARE YOU GOING TO USE TO ENRICH
YOUR T CELLS OUT OF THE BLOOD.
WE WOULD LIKE TO START WITH
WHOLE PATHOGEN.
WE WANT OUR T CELLS TO SEE THE
WHOLE PATHOGEN.
PREVIOUS EXAMPLE USING CD137,
ANOTHER FROM ANOTHER PAPER WITH
ZIKA, CD137, STARTED GETTING
MORE PICKY, COVID WORK, REQUIRED
COORDINATE EXPRESSION OF 69 AND
137, SORTING CELLS, HERE IS
WHOLE KILL VIRUS.
WITH HERPES VIRUS CAN CAUSE
SEVERE DISEASE IN IMMUNE
COMPROMISED PEOPLE REQUIRED
COORDINATED EXPRESSION OF THREE
MARKERS IN ORDER TO SELECT THE
FINAL CELLS FOR DOWNSTREAM
STUDY.
IN THE LAST THREE YEARS WITH
SARS-COV-2 AND MANY PEOPLE
WORKING IN THE FIELD THERE'S
BEEN SORT OF A LITERATURE
EXPLOSION OF THESE
ACTIVATION-INDUCED MARKERS THAT
PEOPLE USED TO ENRICH NOW CELLS
OUT OF THE BLOOD.
I WON'T SHOW DOWNSTREAM WORK BUT
I'LL SUMMARIZE THAT IT MATTERS
WHICH MARKERS YOU USE IN TERMS
OF THE PHENOTYPE OF THE
DOWNSTREAM CELLS AND
SPECIFICITY, THE GROUP USING
COMBINATION, ICS WITH SURFACE
MARKER, COMBINATION, AIM ICS AND
MATTERS HOW LONG YOU STIMULATE
BEFORE YOU SORT AND DO
DOWNSTREAM STUDIES, AN
INTERESTING PAIN POINT OR
CONTROVERSY POINT GOING FORWARD.
SWITCHING GEARS TO COVID,
GETTING INTO TRMs, TISSUE T
CELLS, GOOD LUCK TO FOLLOW
COHORT OF PEOPLE FOR TWO-YEAR
PERIOD, WITH HYBRID IMMUNITY.
INFECTED EARLY IN THE PANDEMIC,
GOT ROUTINE TWO-DOSE mRNA
VACCINATION, A YEAR LATER A
BOOSER, AT THE END WE DID A
NASAL SWAB LOOKING FOR
SARS-COV-2-SPECIFIC T CELLS.
THIS IS USING T-CELL RECEPTOR
BARCODES FOR TRACKING RATHER
THAN AS FUNCTIONAL ASSAY.
A PLOT OF EVENTS, THESE
INDIVIDUALS, 53 INDIVIDUALS.
THIS IS THE VACCINATIONS, THE
CIRCLE HERE.
TIME OF FIRST VACCINE, ABOUT A
YEAR AFTER THEIR INFECTION, SOME
PEOPLE HOSPITALIZED IN PURPLE
BAR, MOST PEOPLE WEREN'T
HOSPITALIZED.
EACH ONE OF THE MARKERS
BASICALLY IS A TIME POINT WE
HAVE FOR BLOOD SAMPLE ON THESE
PEOPLE.
WE DID CAREFUL SEROLOGY TRACKING
ON THESE INDIVIDUALS TO MAKE
SURE THEY REALLY DID HAVE HYBRID
IMMUNITY.
ONE PERSON SNUCK IN WHO PROBLEM
WASN'T INFECTED.
IN LATE 2020, TITERS WENT UP.
EVERYBODY ELSE ALSO HAD, WENT UP
WITH VACCINE.
WE CAN ALSO SEE THREE PEOPLE WHO
HAD BREAKTHROUGH INFECTIONS EVEN
AFTER VACCINATION, THEY HAD
COVID, GOT THEIR VACCINES,
ANTI-TITER WENT UP, EXCLUDED
FROM DOWNSTREAM ANALYSIS WHO
WERE OUTLIERS IN THE COHORT.
HOW ARE WE GOING TO NOMINATE
T-CELL RECEPTORS OR T CELLS AS
CANDIDATE SPIKE-T CELLS?
WE DID SEQUENCING WITH
IN-PERSON, ANALYZE DIFFERENCE
BETWEEN BEFORE VACCINE DOSE 1
AND THIS WAS ABOUT THREE WEEKS
AFTER VACCINE DOSE 2.
SO SHOWN HERE ON THESE RED
ARROWS ON THE LEFT.
WE GOT BLOOD ON THE X-AXIS FROM
BEFORE THE VACCINE WAS GIVEN,
THEY HAD MEMORY.
AND THEN AFTER TWO DOSES OF
VACCINATION, EACH DOT IS A T
CELL PROTOTYPE THAT'S AT THE
BETA CHAIN THAT'S DETECTED BY
NEXT GENERATION SEQUENCING USING
ADAPTIVE BIOTECHNOLOGIES
PLATFORM.
YOU SEE LOG SCALE HERE, ONE OUT
OF 100 T CELLS, ONE OUT OF
100,000 T CELLS, 10 TO THE MINUS
FIFTH.
ALL RED DOTS HAVE EXPANDED BY
FACTORS OF SOMETIMES 100, 1,000,
EVEN 10,000, TEMPORALLY IN
RESPONSE TO VACCINATION.
THESE ARE CANDIDATE VACCINE
REACTIVE T-CELL RECEPTORS.
IF WE LOOK BRIEFLY AT TWO
INDIVIDUALS, NOW LOOKING AT THE
KINETICS, THREE OR FOUR WEEKS
APART, THIS PERSON HAD A NICE
BUMP BEFORE AND AFTER FIRST
DOSE, FURTHER BUMP AFTER THE
SECOND DOSE, ALL OF THESE HERE.
IF YOU SUMMARIZE ACROSS BOTH
DOSES NICE RESPONSE ACROSS BOTH
DOSES, SO THE INDIVIDUAL HERE
DID NOT HAVE MUCH RESPONSE AFTER
FIRST DOSE, SOME CLONOTYPES DID
GO UP, NICE RESPONSE AFTER THE
SECOND DOSE, OVERALL IF YOU LOOK
ACROSS THE WHOLE RESPONSE MOST
OF THE RESPONSES SEEM TO COME
OUT OF A NAIVE POOL BECAUSE VERY
FEW WERE PRESENT IN THE MEMORY
POOL AND TOOK TWO SHOTS TO BRING
THESE OUT.
SOME PEOPLE ARE NICE MEMORY
AFTER ONE, OTHER TOOK TWO DOSES
AND DIDN'T RECRUIT TOO WELL OUT
OF PREVIOUS MEMORY.
THERE'S ONE METHOD TO NOMINATE
TCRs.
LET'S LOOK AT ORTHOGONAL
DIFFERENT METHOD, GOING BACK TO
ACTIVATION INDUCED MARKER
SERVING, HOT TIME POINT, THREE
OR FOUR WEEKS AFTER THE SECOND
DOSE OF VACCINE.
AND IF I THROW IN SPIKE PEPTIDE
POOL, I DON'T GET MUCH WITH
ACTIVATION MARKERS.
IF I THROW IN -- PARDON ME.
IF I THROW IN SPIKE PEPTIDE POOL
I'VE GOT A HUGE RESPONSE AND WE
COULD PUT THESE CELLS INTO 10 X
PLATFORM FOR SEQUENCING, GOT
30,000 PAIRED T CELLS CANDIDATE,
THERE ARE PAIN POINTS OR
PROBLEMS.
WE DON'T KNOW WHETHER CHOOSING
69 AND 137 MISSED A BUNCH OF
TCRs, WE DON'T KNOW HOW
SPECIFIC THIS PLOT HERE IS
BECAUSE OF THE PHENOMENON OF
ACTIVATION.
THIS IS NICE.
WE NEED TO DO FURTHER
CONFIRMATION.
ONE OF THE THINGS WE COULD DO TO
DO THIS WAS TO LOOK AT THE
OVERLAY.
I'M SHOWING YOU A VERY BIG
OVERLAY.
HERE IS THE BEFORE DOSE.
THERE'S THE AFTER DOSE HERE.
ALL OF THE DOTS ON THE UPPER
LEFT ARE POTENTIALLY VACCINE
RESPONSE FROM THE FIRST METHOD I
SHOWED YOU.
WE OVERLAID WHETHER WE FOUND
THEM USING SINGLE CELL TCR Seq
AND THE SIZE OF THE MARKER IS
NUMBER OF SISTER CELLS WE WERE
ABLE TO RECOVER, QUITE A NICE
CORRESPONDENCE HAVING
CONFIDENCE.
COULD LOOK AT TRAJECTORY, NOW WE
HAVE THIS BARCODE FROM THE
SINGLE CELL METHODOLOGY, YOU CAN
LOOK BACK OVER THIS TWO-YEAR
TIME FRAME OF THESE INDIVIDUALS,
ASK DO THESE CLONOTYPES GO UP?
FOCUS ON THE RIGHT, AVERAGE
INCREASE OVER THE COURSE OF THE
VACCINATION, AND CORE FINDING
HERE IS WHEN WE GIVE THE mRNA
VACCINE, WE SEE NICE INCREASE IN
ABUNDANCE IN THE BLOOD BUT NOT
NEARLY AS IMPRESSIVE.
THIS IS FROM U PENN, TRYING TO
RESOLVE WHAT THE PLUSES AND
MINUSES MIGHT BE AND HOW TO SORT
THIS OUT.
WHEN WE GATHER SEQUENCES WE CAN
CLUSTER THEM IN BIOINFORMATIC
SEQUENCE SPACE, A
REPRESENTATION.
THIS IS FROM 17 PEOPLE, EACH
SHOWN IN DIFFERENT COLOR.
IF THE SEQUENCES ARE CLOSE TO
EACH OTHER, THEY WILL BE JOINED
BY A LINE, JOINING INTO
CLUSTERS, AND WE CAN CALL THIS
LIKE A SANITY CHECK THAT THESE T
CELLS FROM DIFFERENT INDIVIDUALS
ARE IDENTICAL OR SIMILAR TO EACH
OTHER.
SOME OF THE SANITY CHECKS WE CAN
DO ARE THESE CLUSTERS CO-SORT
WITH H ALLELES OR PHENOTYPES AND
SOME RECAPITULATE WILL IT --
LITERATURE.
THIS CLUSTERS, SEEING THIS HOT
A2 EPITOPE.
WE WANTED TO TEST OTHER
CLUSTERS.
WE CHOSE SOME OF THESE SHOWN
WITH THE ARROWS THAT WERE A-3
RESTRICTED, SHOWN ON THE RIGHT
THESE ARE INDIVIDUAL PATIENTS,
THE FREQUENCY OF CELLS IN
CLUSTERS DID GO UP OVER TIME
WITH VACCINATION.
SO HERE WE'RE GETTING TO ONE OF
THE NICE TECHNOLOGIES IN THIS
AREA BUT ALSO A PAIN POINT WHICH
IS TCR RECONSTRUCTION.
WE PICK SIX AND RECONSTRUCTED
THEM AND PUT THEM IN REPORTER
CELLS, BOTTOM LINE IN RED IS WE
PREDICTED THIS TCR WOULD BE
RESTRICTED BASED ON ASSOCIATION
WITH PEOPLE WHO WERE HLA 3, WHEN
WE TRANSFECT, BOTH HLA 3 AND 4
SPIKE, REPORTER CELL HAS A NEON
GREEN SIGNAL THAT GOES UP WHEN
THE CELL IS ACTIVATED THROUGH
T-CELL RECEPTOR.
WE USE T-CELL RECEPTORS, PUT IN
LENTIVIRUSES, FORCED INTO THESE
REPORTER CELLS.
AND WE WERE ABLE TO DO THIS WITH
SIX RECEPTORS, ONE THROUGH SIX,
EPITOPE IS HERE, THIS K THROUGH
L AMINO ACID EPITOPE, LOOKING AT
VARIANTS IN THE INTEREST OF
TIME, DIFFERENT TRUNCATIONS, YOU
CAN SEE SUBTLE DIFFERENCES HERE
IN REACTIVITY WITH 100% TOP
REACTIVITY WITH SUBTLE
DIFFERENCES IN THE T-CELL
RECEPTOR LENGTH AND IDENTITY.
WE CAN VALIDATE T-CELL RECEPTORS
OUT OF THE TENS OF THOUSANDS
WE'VE NOMINATED BY
RECONSTRUCTION BUT DIFFICULT TO
DO THEM ALL.
LET'S GET BACK TO TO THIS.
WE SWABBED NOSES, MADE DNA,
SEQUENCED, LOOK FOR OVERLAP BY
COMPARING THE NOSE SEQUENCES TO
LOOKUP TABLES I'LL MENTION NEXT
THAT TO -- TO COUNT SARS-COV-2
SPECIFIC T CELLS.
ONE INDIVIDUAL ON THE LEFT, HERE
IS T-CELL RECEPTOR SEQUENCE FROM
THE NOSE, THE MOST ABUNDANT, ONE
OUT OF 50, AND YOU CAN SEE
TCRs WE RECONSTRUCTED WERE
ALSO FOUND IN THE NOSE, EXACT
MATCHES.
WE CAN SEE HYBRID IMMUNITY, YES
LOCALIZING TO THE SITE YOU NEED
HOST DEFENSE AGAINST INCOMING
SARS-COV-2 INFECTION.
ON THE RIGHT THESE ARE EVERY
COLORED CIRCLE HERE, THE SAME
ABUNDANCE HISTOGRAM, EACH ONE OF
THE COLORED CIRCLES IS A
SARS-COV-2, MOSTLY CD8s, A FEW
CD4s FOUND IN THE NOSE.
IN A LOOKUP TABLE LOOK AT TCRs
MORE ABUNDANT.
THIS IS SEQUENCING ONLY.
HERE IS EXAMPLE WHERE THE D CELL
RECEPTORS IN RED WERE FOUND IN
THIS AFTER GREATER THAN BEFORE,
COMPARATIVE BIOINFORMATIC
ANALYSIS.
FINALLY IF ALL HAVE YOU IS A
SWAB, YOU DON'T HAVE BLOOD, WANT
TO DO LESS WORK, YOU CAN CONSULT
A LOOKUP TABLE, PUBLISHED BY ONE
OF THE FIVE TECHNOLOGY THE
COMPANIES.
THIS COMPANY PUBLISHED A LONG
LIST.
T-CELL RECEPTORS THEY THINK ARE
SARS-COV-2 SPECIFIC.
NASAL SWABS FOUR PEOPLE.
IF WE LOOK WE CAN SEE THERE ARE
HITS FROM NASAL SWAB, SEQUENCING
THE NASAL SWAB THAT APPEAR TO BE
SARS-COV-2-SPECIFIC.
THE PROBLEM IS THE DATA IN THE
LOOKUP AVAILABLE IS NOT SUPER
WELL VETTED AND CAN SOMETIMES
HAVE DIFFICULTIES OR QUESTIONS
AND NEEDS TO BE IMPROVED.
FROM THE COVID WORK PRE-VERSUS
POST VACCINATION, COULD NOMINATE
SOME TCRs, THE OVERLAP MAKES
SENSE.
WE CAN DO LIMITED FUNCTIONAL
CONFIRMATION BUT THIS IS A PAIN
POINT THAT COSTS MONEY, TAKES
TIME.
CAN USE THIS DATA TO GO BACK TO
NASAL SWABS TO CONFIRM TISSUE
RESIDENT MEMORY T CELLS, VIA
USING TCRs AS BARCODE CAN BE
FOUND IN THE NOSE.
THIS IS A VIRUS THAT CAUSES A
RARE CANCER, EXPRESSED IN CELLS,
CAN BE STAINED FOR HERE, POOR
PROGNOSIS CANCER, YOU CAN SEE
DEPENDING ON PRESENTING STAGE.
THERE'S 400 AMINO ACID PROTEIN
SPACE EXPRESSED, REQUIRED FOR
ONCOGENESIS IN THIS CANCER.
WE'VE BEEN ABLE TO DO T CELL
EPITOPE MAPPING IN THIS, IT'S
TWO ISOFORMS OF THE PROTEIN.
WE CAN COVER ABOUT 95% OF THE
POPULATION WITH KNOWN CD8 T CELL
EPITOPES IN THIS ONCOPROTEIN.
SO, USING THIS APPROACH WE CAN
SHOW THAT INDIVIDUALS WHO HAVE
TETRAMER POSITIVE CELLS IN THE
BLOOD HAVE BETTER OUTCOME WITH
CHECKPOINT THERAPY AND FORM
CLUSTERS IF WE CLUSTER BY
SEQUENCE SPACE.
AS OUTCOME OF THIS WORK, WE HAVE
A CLINICAL TRIAL UNDERGOING
USING A VACCINE AND WE'RE
PLANNING ON USING TCR SEQUENCING
AS BIOMARKER FOR THIS
INDICATION.
WAY AT THE OTHER END OF THE
SPECTRUM OF A SMALL PROTEIN
SPACE.
WE'RE RUNNING LATE, TIM.
I WANT TO SWITCH GEARS TO
CHICKEN POX.
IT CAN CAUSE CHICKEN POX AND
LATER ON SHINGLES.
WE'RE INTERESTED IN STUDY LOCAL
T-CELL RESPONSE IN THIS CASE,
AND SEE WHETHER THERE'S A
DURABLE IMMUNE RESPONSE AFTER
INFECTION.
WE HAVE A PROTOCOL THAT LASTED
FOR A YEAR WHERE WE DID BIOPSIES
OF SHINGLES SKIN, HEALED SKIN,
FOR UP TO A YEAR AFTER SHINGLES.
AND WE WOULD TEST THE T CELLS
WITH THE WHOLE PROTEOME AND
EPITOPES TETRAMERS, SEQUENCE,
AND GOING BACK TO STAIN THE SKIN
FOR ANTIGEN SPECIFIC T CELLS.
SO, IN THIS PARTICULAR CASE WE
WERE ABLE TO PERFORM WHOLE
PROTEOME SCREENS AND EXPRESSED
ALL OF THE FRAMES AND STUDIED
THE T CELLS IN SKIN BIOPSIES OF
HEALED SHINGLES SKIN, TESTING
FOR REACTIVITY WITH EVERY
INDIVIDUAL OR AS YOU CAN SEE THE
SUBJECT ON DAY 45 AFTER RECOVERY
FROM SHINGLES HAD A LOT OF T
CELL REACTIVITY AGAINST MANY
INDIVIDUAL PROTEINS.
TO SUMMARIZE AN INDIVIDUAL HERE
AT DAY 45, DAY 90 AND 60, THREE
HLA ALLELES, SOME REACTIVITY
AGAINST PROTEINS ON HEALED
SHINGLES SIDE, VERY LITTLE ON
CONTRALATERAL.
ON TOP ARE SHINGLES SKIN DAY 45,
DAY 90, DAY 360, ONE TO 500.
WE HAD DIFFERENT HITS, LOOKING
AT CONTROL SKIN WE HAVE VERY
MUCH LESS REACTIVITY OVER IN THE
CONTROL SKIN.
SO, WHAT WE ARE -- THIS IS A
SUMMARY OF OVERALL DATA, CLEAR
WINNER, ORF 9.
MY MAIN POINT HERE IS TO SAY
THAT WE NEED TO LOOK ACROSS THE
PROTEOME, THIS LARGE PATHOGEN,
HARDER THAN SARS-COV-2 WITH
ABOUT 24 PROTEINS, THIS VIRUS
HAS ABOUT 70 PROTEINS, BUT WE
TOOK THE PAINS TO DO THIS AND
FOUND A CLEAR WINNER HERE, IN
RED ON THE RASH SITE ONLY A LOT
MORE REACTIVITY OVER ON THE RASH
SIDE THAN ON THE CONTRALATERAL
SITE.
SO, THE CONCLUSION OF THIS IS
THAT THERE IS A SEATING OF SKIN,
IMMUNE MEMORY, LOCAL MEMORY AT
THE SITE OF HEALED INFECTION.
WE FOUND MANY, MANY INDIVIDUAL
EPITOPES AND CONTRIBUTED THIS TO
IMMUNE EPITOPE DATABASE, SOME
EPITOPES LISTED HERE WITH
VARIOUS HLA RESTRICTIONS.
AND WE WERE ABLE TO MAKE AT THE
TETRAMERS, YOU CAN SEE VERY HIGH
REACTIVITY WITH SOME INDIVIDUAL
TETRAMERS AND USING THEM WERE
ABLE TO GO BACK AND STAIN
BIOPSIES OF THE SKIN.
WE'RE PROVING, THIS WAS IN A DAY
45 AND DAY 90, THESE ANTIGEN
SPECIFIC T CELLS ARE RETAINED
LONG TERM IN THE SKIN AFTER
RECOVERY FROM THE VIRAL
INFECTION.
THIS IS AN ASPIRATIONAL GOAL OF
VACCINOLOGY, WOULD LOVE TO GIVE
A SHOT AT I.M. AND HAVE T CELLS
GO TO THE NOSE IN TERMS OF
SARS-COV-2, AND BE
PRE-POSITIONED TO FIGHT AGAINST
INITIAL INOCULATION, WE'RE NOT
THERE YET BUT USING NOTHING BUT
BUT ESPECIALLY WITH IN SITU,
DEMONSTRATE THEY ARE RETAINED AT
THE INITIAL SITE.
I'M GOING TO SKIP THIS A LITTLE
BIT.
I WOULD MENTION THAT WE -- A
CURRENT HORIZON TO TO MOVE TO
CULTURE INDEPENDENT EPITOPE
DISCOVERY, THIS IS USING POWER
OF TCR RECONSTRUCTION, IN THIS
CASE WE DID A CERVICAL BIOPSY,
AND SORTED THESE ACTIVATED CELLS
USING CD137 AS ACTIVATION MARKER
AND DID SINGLE CELL TCR, FOUND A
LENTIVIRUS, A REPORTER CELL ABLE
TO RECOGNIZE WHOLE VIRUS AND
ALWAYS LIKE TO DO THIS TO SHOW
RECOGNITION OF WHOLE VIRUS.
ONCE WE HAVE THIS RECOMBINANT
TCR WE'RE ABLE TO SCREEN IT
AGAINST ALL FOUR OF THE PERSON'S
HLA ALLELES AND HERPES PROTEINS
AND FOUND WHICH PROTEIN THIS
REPRESENTED.
WHILE THE TECHNOLOGIES I SHOWED
PREVIOUSLY ARE KIND OF, YOU
KNOW, DEPENDENT ON HAVING OR
GROWING LIVING T CELLS WITH
ENOUGH PAIN CAN DO THIS PURELY
FROM TCR SEQUENCE.
A FEW SLIDES FROM SYPHILIS, A
NEW INITIATIVE TO STUDY THIS
BACTERIAL DISEASE FOR WHICH
THERE'S NO VACCINE.
IN THE KALES OF SECONDARY
SYPHILIS, OCCURS ABOUT A MONTH
AFTER PRIMARY SYPHILIS, AND SKIN
IS TEAMING WITH BACTERIUM, AND T
CELLS, AND QUESTION IS ARE THERE
ANTIGEN SPECIFIC T CELLS IN
THERE?
THIS IS A BIOPSY OF SECONDARY
SKIN LESION, RED SQUIGGLES ARE
BACTERIA.
YOU CAN SEE THIS SAMPLE IS
RELATIVELY LOADED WITH BACTERIA
AND INFILTRATE CONTRIBUTING TO
INFLAMMATION.
IF WE STAIN THIS BIOPSY FOR
IMMUNE CELLS, WE CAN SEE A LOT
OF CD4 AND CD8 CELLS, INTERESTED
IN STUDYING IF THEY ARE ANTIGEN
SPECIFIC.
THESE ARE BACTERIA.
WHAT ARE THEY DOING HERE?
COULD BE ANTIGEN SPECIFIC OR
SEMI INNATE CELLS, INTERESTED IN
THAT.
THERE'S LOCAL UPREGULATION AT
HLA CLASS 2, MIGHT BE EXPECTED
FROM THE REDNESS OF THE RASH,
EFFECTOR CELLS IN SKIN THOUGHT
TO BE MACROPHAGES, AND
PHAGOCYTOSIS OF ANTIBODY-COATED
THOUGHT TO BE HOW THE HOST
CLEARS THE BACTERIA BECAUSE THE
RASH WILL GO AWAY, MAKING
NEUTRON GAMMA WE HYPOTHESIZE TO
LEAD TO CLEARANCE.
WE TAKE TRADITIONAL APPROACH OF
CLONING IN THIS CASE A THOUSAND
OPEN FRAMES, WE CAN'T CLONE THEM
ALL, WE CLONED 50 HERE IN THIS
EXAMPLE.
WE USED BLOOD TO ENRICH CELLS
OUT OF THE BLOOD REACTIVE WITH
WHOLE TP AND USING AIM, MARKERS,
SORTING AND EXPANDING CELLS WE
DO HAVE SOME HITS HERE,
DUPLICATE HAVE DECODED FOR THE
FIRST TIME IDENTITY OF CD4 T
CELL ANTIGENS, INCLUDING
ACTIVITY OF WHOLE ORGANISMS OVER
HERE, WHOLE MICROORGANISMS, WELL
BEHAVED, I WON'T GO INTO DETAILS
BUT DR RESTRICTED, PREDICTED TO
BE PREDICTED BY DR BETA 7, SO WE
CAN DRILL DOWN INTO INDIVIDUAL
EPITOPES AND SHOW TRADITIONAL
HLA RESTRICTION.
WHAT'S INTERESTING HERE WAS
TISSUE RESIDENT MEMORY T CELLS
ARE LEFT BEHIND AFTER PENICILLIN
TREATMENT FOR SECONDARY
SYPHILIS, SOME INDIVIDUAL CAME
IN, STUDY WITH BOTH BLOOD AND
SKIN BIOPSY, DAY ZERO, ALSO ON
DAY 30 AND DAY 180 AFTER
PENICILLIN THERAPY.
WITH SKIN WE DIDN'T NEED
ENRICHMENT, GREW CELLS OUT OF
THE SKIN, NO CHECK FOR
ENRICHMENT.
IN MICE CONSISTENCY OVER THE
TIME, BLOOD AND SKIN, FOR THE
SAME OPEN READING FRAMES TEND TO
BE DETECTED AT BOTH TISSUES.
IN THE BLOOD AND SKIN BIOPSY BUT
INTERESTINGLY HOW TODAY 180
AFTER RESOLUTION AND PENICILLIN
TREATMENT WE SEE FUNCTIONAL
EVIDENCE FOR THESE STILL BEING
HERE.
AND THIS IS THE WORK OF TARA
BRINKLEY, MD/PHD IN THE LAB,
NOMINATING UNNAMED -- A BLANK
SPACE FOR THE ANTIGENIC TARGETS,
DOUBLE MEMORY, INNER AND OUTER
MEMBRANE POSITIVE SPACE.
WE HAVE SEVERAL PROTEINS HERE,
SOME ARE VACCINE CANDIDATES, WE
HAVE SUPPORT FROM THE NIAID AND
U19 PROGRAM TO SUPPORT FURTHER
DEVELOPMENT OF SOME OF THESE AS
VACCINES.
SO JUST TO SUMMARIZE HERE, THIS
IS THE TRIAD, PAIN POINTS, WISH
LIST.
SINGLE CELL IS GREAT BUT
EXPENSIVE, TAKES A LOT OF
HANDS-ON TIME.
VALIDATION IS EXTENSIVE AND
SLOW.
PEOPLE ARE USING CRISPR/CAS9,
LENTIVIRUS, WE NEED BETTER
TECHNOLOGY.
DATABASES OF ANTIGEN SPECIFIC
TCRS ARE A MESS.
THERE'S REDUNDANCY, FALSE
POSITIVES, METADATA IN THERE IS
VARIABLE AND SOMETIMES POOR.
THERE ARE MANY POORLY DOCUMENTED
OR DISTRACTING TCRs IN THESE
DATABASES.
FINALLY ANOTHER PAIN POINT, I
THINK, THAT THERE'S A LOT OF
KNOWLEDGE WITH INDUSTRY, YOU
HAVE A TCR, WHAT DOES IT SEE,
THAT IS NOT MAYBE PUBLICLY
AVAILABLE.
FOR SUMMARY, I'M SORRY FOR GOING
SO LONG, WHAT WE'VE TRIED TO
INCORPORATE IS BIOLOGICALLY
INFORMATIVE STUDY DESIGNS,
LOOKING WITHIN SUBJECT,
LONGITUDINALLY PRE AND POST
VACCINE, AS IN COVID, AND THEN
CROSS-SECTION, BLOOD VERSUS
SKIN, LOOK FOR PUBLIC AND
SIMILAR TCRs, I SHOWED THE
CLUSTERING, USING TCR SEQUENCE,
WE'D LIKE TO MATCH TO PEPTIDE
HLA LIGANDS ON THE RIGHT, THE
HOLY GRAIL TO MATCH UP TRIADS,
TCR SEQUENCING AT SINGLE CELL OR
BULK LEVEL, FUZZY MATCHING DOES
SEEM TO HELP.
AND IT'S NICE TO HAVE METADATA
THAT GOES WITH EACH ONE OF THOSE
CELLS, AND LASTLY THE WET LAB
TOOLS FOR FIGURING OUT IF YOU
HAVE T CELLS, FIGURING OUT
PEPTIDE AND HLA, THAT'S HEREN MY
OPINION.
THANKS TO ALEX AND OTHERS,
SOFTWARE TOOLS, YOU CAN KNOW HLA
AND OPEN READING FRAME, FOR
PICKING EPITOPES, THESE ARE
LARGELY HERE.
BUT LOOKUP TABLES, IF YOU WANT
TO KNOW HAS ANYBODY SEEN IT
BEFORE OR WHAT DOES IT SEE,
THOSE ARE A MESS AND BETTER
LOOKUP TABLES ARE NEEDED.
FINALLY ASPIRATIONALLY WE'VE GOT
HUNDREDS OF THOUSANDS OF TCR
SEQUENCES, CAN WE FOLD AND
FIGURE OUT, LET'S SAY WE KNOW
PATHOGEN, WHAT PEPTIDE DOES IT
SEE?
THAT'S REALLY A FUTURE HORIZON
THAT SOME COMPUTATIONAL BIOLOGY
COLLEAGUES ARE WORKING ON, BUT
MAYBE WE'LL GET THERE AND WE'LL
BE ABLE TO FIGURE OUT A TCR SEES
ARE VARIABLE LOOPS, TO PREDICT
GREEN AND BLUE LIGAND DOWN HERE.
THAT IS ALL I WANTED TO SAY.
THANK YOU, TIM.
>>THANKS, DAVID.
THAT WAS GREAT.
I'D LIKE TO OPEN THIS UP TO
EVERYONE FOR QUESTIONS PLEASE.
SO, DAVID, ONE OF THE THINGS
THAT WAS INTERESTED IN T
PALLIDUM STUDIES, YOU WERE ABLE
TO SEE THE RESPONSES IN SKIN, I
GUESS EVEN UP TO WHAT WAS IT
120?
>>DAYS, YEAH.
>>WHICH IS FASCINATING.
IS THERE RESIDUAL ANTIGEN?
WHAT IS DRIVING THAT.
>>THE TISSUE RESIDENT
PHENOMENON HAS COME OUT OVER THE
LAST 10 OR 15 YEARS, IN OUR
SHINGLES STUDIES DID SURFACE
SWABS UP TO A YEAR.
BY DAY 360 WE COULDN'T DETECT
DNA ANYMORE BUT PRESENCE OF
ANTIGEN OR PEPTIDE CAN'T BE
RULED OUT.
WHERE THEY ARE LACKING MOBILITY
RECEPTORS, MOBILITY RECEPTORS
SEEM TO BE TAKEN UP BY CD69,
BINDING, ALSO LACKING A LOT
OF -- ALSO EXPRESSING CD103
WHICH TENDS TO LOCK THEM INTO
THE SKIN BY BINDING REALLY MAKES
IT SUCH THAT PROBABLY CONTINUING
ANTIGEN ISN'T REQUIRED ANYMORE.
IT'S KNOWN THOSE TISSUE RESIDENT
MEMORY T CELLS PATROL LOCALLY BY
LIVE-CELL IMAGING BUT TEND NOT
TO EXIT VERY WELL TO THE BLOOD.
>>EXCELLENT.
>>DO YOU HAVE ANY EVIDENCE OF
ENRICHMENT OF SPECIFIC TISSUE
SEQUENCES FOR SKIN OR FOR
SPECIFIC TISSUES?
>>WE OBTAINED SPECIFIC TCRs
FROM THE BLOOD AND LOOKED AT
COMPARATIVE ABUNDANCE IN THE
SKIN, USING THE SKIN AS SITE OF
ENRICHMENT, COMPARED TO
ABUNDANCE IN THE SKIN TO BLOOD,
MUCH, MUCH MORE ABUNDANT IN THE
SKIN THAN BLOOD, BY A FACTOR OF
100 OR DID 1,000 FOLD, PUBLISHN
NATURE COMMUNICATION.
WE'VE SHOWN FOR ACUTE GENITAL
HERPES CELLS ARE MORE ABUNDANT
IF YOU BELIEVE TCR Seq COUNTS.
WITH NATION ALCOVED SITUATION,
HYBRID IMMUNITY VERSUS PEOPLE
INFECTED AND HAD THE SHOT, THE
DOUBLE BOOST, IF YOU LOOK AT
NASAL SWABS VERSUS ABUNDANCE IN
BLOOD THEY ARE MORE ABUNDANT IN
NASAL SWABS THAN IN THE BLOOD.
>>BUT IS IT DRIVEN BECAUSE OF
VIRUSES THERE OR WHAT I'M SAYING
IS THAT IS THERE EVIDENCE OF --
WITHOUT ANTIGEN LOAD TO BE
DRIVEN TO SPECIFIC TISSUE?
>>THAT'S A TOUGH QUESTION.
>>THERE'S SOME EVIDENCE IN THE
GUT, MORE IN THE GUT THAN
SOMEWHERE ELSE.
>>IN WHICH TISSUE?
>>IN THE GUT.
>>OH, IN THE GUT.
DR. NO CHEMOKINE RECEPTORS
ASSOCIATED WITH LAMINA, BUT YOU
CAN GUIDE RESPONSE, WHETHER THEY
KNOW THEY ARE SUPPOSED TO GO TO
GUT OR SKIN, THAT'S ASPIRATIONAL
AND I HAVEN'T SEEN EVIDENCE FOR
THAT, YOU KNOW, IN THE ABSENCE
OF CHEMOKINE SIGNAL OR ABSENCE
OF LOCAL INFLAMMATION OR CERTAIN
TCRs PRONE TO CERTAIN TISSUES.
>>THANK YOU.
>>DR. AUGUSTINE?
>>DO WE HAVE TIME?
THANK YOU FOR THAT PRESENTATION.
I THOUGHT I HEARD YOU SAY THAT
YOU DON'T THINK VACCINATION IS
GOING TO INDUCE -- RIGHT NOW WE
CAN'T USE VACCINE TO TRIGGER
TISSUE SPECIFIC T-CELL RESPONSES
BUT WITH YOUR SARS-COV-2 STUDIES
THOSE PEOPLE WERE INFECT AND
THEN YOU WERE ABLE TO GET T
CELLS FROM THE NOSE BUT THEY
WERE ALSO VACCINATED.
DO YOU REALLY THINK THAT THE
VIRUS INFECTION WAS TRIGGERING
THAT MIGRATION TO THE NOSE OR
VACCINES HAD A ROLE TO PLAY AND
I DIDN'T CATCH HOW LONG T CELLS
RESPONSES REMAINED IN THE NOSE.
>>UNFORTUNATELY WE ONLY ABLE TO
STUDY NASAL RESPONSES AT ONE
TIME POINT.
WE STUDIED THREE OR FOUR WEEKS
AFTER THE BOOST, THE CANONICAL
THIRD DOSE.
I CAN'T TELL YOU BECAUSE WE
DON'T HAVE A PARALLEL STUDY BUT
THERE ARE STUDIES IN SINGAPORE,
I THINK VACCINE-ONLY, IF I
RECALL CORRECTLY, THEY COULD
RECOVER USING ELISPOT OR ICS
COULD RECOVER T CELLS FROM THE
NOSE.
THEY DIDN'T HAVE A QUANTITATIVE
TCR Seq.
THE LYLE McKINNON GROUP FROM
CANADA DID FUNCTIONAL STUDIES
THAT MIGHT HAVE BEEN ON
INFECTION ONLY BUT I THINK
YOU'RE ASKING A BIGGER QUESTION,
FASCINATING, ONCE T CELLS ARE
PRIMED BY TISSUE SPECIFIC
INFECTION LET'S SAY RESPIRATORY
TRACT ARE LEFTOVER MEMORY CELLS
IN THE BLOOD OR CENTRAL MEMORY
CELLS SOMEHOW ADDRESSED TO GO
BACK TO THAT TISSUE OF
EDUCATION?
AND WE KNOW THAT'S TRUE WITH
GENITAL HERPES.
A SUGAR MODIFICATION IS ON SOME
T CELLS.
IF YOU SAMPLE MY BLOOD, 50 TO
70% OF THOSE TETRAMER POSITIVE
CELLS HAVE CLA ON THEM READY TO
GO BACK TO SKIN.
THE IDEA A TISSUE-SPECIFIC
INFECTION CAN FIRST PRIME CELLS
FOR HOMING, COULD THEN BE TAKEN
ADVANTAGE OF WITH VACCINE,
THERE'S SOME SORT OF STABLE
IMPRINTING THAT'S GOING ON AFTER
A GEOGRAPHICAL OR ANATOMIC
LIMITED INFECTION IN G.I. TRACT
OR RESPIRATORY TRACT, TRUE FOR
SKIN IN HERPES, ALSO MELANOMA.
>>THANK YOU VERY MUCH.
>>THAT WAS GREAT.
I WANT TO THANK DR. KOELLE FOR A
GREAT TALK AND THE OTHER
SPEAKERS, DOCTORS CROWE AND
SETTE FOR A WONDERFUL AFTERNOON
SESSION AND I'LL TURN IT OVER TO
DAN.
DAN, ARE YOU --
>>YES, I'M HERE.
>>THANK YOU.
MY THANKS ALSO TO ALL THREE
SPEAKERS FOR COVERING REALLY
INTERESTING DATA AND ALSO SOME
VERY PROVOCATIVE FORWARD-LOOKING
THOUGHTS.
WE HAVE FOUR CONCEPTS FOR
CLEARANCE DISCUSSION BY COUNCIL.
WE'RE OVER ON TIME, WE'LL GO
OVER 3:30 ADJOURNMENT IF WE NEED
TO.
I HOPE STAFF CAN BE BRIEF AND WE
WON'T PUSH PEOPLE BEYOND THAT
TIME.
FIRST OF THESE IS CALLED
INNOVATIONS IN FUNCTIONAL B CELL
EPITOPE DISCOVERY.
AS WE'VE DONE IN THE PAST, I'M
GOING TO ASK FOR AN EN BLOC
RECOMMENDATION AFTER ALL OF
THESE ARE DISCUSSED, UNLESS THAT
CONVERSATION FOR ANY ONE OF THEM
SUGGESTS THERE MIGHT BE TOO MANY
CONCERNS TO GO WITH THAT.
JOE, PLEASE TAKE IT OVER FROM HERE
SO, I'M JOE BREEN.
I'M GOING TO TALK ABOUT THE
RENEWAL OF WHAT'S CURRENTLY
CALLED B CELL EPITOPE DISCOVERY
MECHANISMS OF ANTIBODY
PROTECTION PROGRAM, AND I CAN
KEEP THIS BRIEF BECAUSE YOU
HEARD A VERY INTERESTING
INTRODUCTION BY DR. CROWE
EARLIER.
WANT TO MAKE SURE ALL ASPECTS
ARE CLEAR AS WE REQUEST RENEWAL
OF THIS FY 24 INITIATIVE.
CAN YOU GO TO THE NEXT SLIDE
PLEASE.
SO, THE PURPOSE OF THIS PROGRAM
IS TO IDENTIFY AND VALIDATE
HUMAN B CELL EPITOPES THAT
INDUCE PROTECTIVE OR
PATHOGENNICS RESPONSES IN SOME
CASES THROUGH INFECTIOUS
DISEASES BUT ALSO AUTOIMMUNITY,
ALLERGY MODELS, TRANSPLANT
REJECTION AS WELL.
THE PROGRAM HAS REALLY
EMPHASIZED IDENTIFYING
MECHANISMS OF ANTIBODY MEDIATED
PROTECTION AS WELL AS DISCOVERY
OF EPITOPES, AND THAT FOCUS ON
FUNCTION HAS REALLY INCREASED AS
THIS RATHER LONGSTANDING PROGRAM
HAS MATURED.
AND AS YOU HEARD FROM DR. CROWE
AND OTHER EXAMPLES AS WELL
PROGRAM'S BEEN VERY HELPFUL FOR
MONOCLONAL ANTIBODY DISCOVERY
AND DESIGN, AND WE HOPE WILL
CONTINUE TO INFORM VACCINE
DESIGN OVERALL AS WELL.
NEXT SLIDE PLEASE.
THE CURRENT AWARDEES, THESE ARE
CONTRACTORS, ARE DR. CROWE WHOSE
WORK YOU HEARD, WAS ORIGINALLY
SUPPORTED FOR INFLUENZA WORK BUT
AS TIM MENTIONED WE EXPANDED
WITH SARS-COV-2.
ALSO, THERE'S BEN DORANZ.
THE FIRST THREE, DAVID FREMONT
AT WASHINGTON UNIVERSITY IS ALSO
IN THIS PROGRAM.
THOSE THREE ACTUALLY COLLABORATE
CLOSELY TO UTILIZE STRENGTHS AND
STRUCTURAL AREAS, BIOLOGY, SOME
HIGH-THROUGHPUT ANALYSIS THAT
DR. CROWE TALKED ABOUT.
THE FOURTH AWARDEE IS OUR
BACTERIAL PROJECT, DR. NICK
MANTIS TRYING TO IDENTIFY
BORELLIA BURGDOFERI, LYME
DISEASE, DIAGNOSE DIAGNOSTICS A
CHALLENGE, NO HUMAN DISEASE
CURRENTLY AVAILABLE.
NEXT SLIDE PLEASE.
SO THE PROGRAM'S BEEN VERY
PRODUCTIVE.
TIM MENTIONED THIS IN BOTH THE
T- AND B-CELL PROGRAM.
B CELL PROGRAM WE'VE HAD JUST IN
THE LAST 3 1/2 YEARS ABOUT 70
PUBLICATIONS, YOU HEARD ABOUT
EVUSHELD, NO LONGER AVAILABLE
UNDER EUA.
IN TICK BIOLOGY TRYING TO
DEVELOP AND UNDERSTAND BORELLIA
BURGDOFERI IMMUNITY.
STILL WORKING ON INFLUENZA
PROJECTS AND OTHER PREVIOUS
SLIDES.
NEXT SLIDE PLEASE.
THIS IS A PROPOSED FY 24
INITIATIVE, CONCEPT FOR COUNCIL
APPROVAL, THE SCOPE WIFF -- OF
WHICH WAS A SMALL ENHANCEMENT.
WE REFER TO DISCOVERY AND
VALIDATION IN HUMANS, STILL WANT
THE MECHANISMS OF ANTIBODY
FUNCTION BECAUSE IT'S SO
IMPORTANT, AND CONTINUE TO
PUBLICLY SHARE THE EPITOPE DATA
ANTIBODY AND STRUCTURAL DATA
SUBMISSION THROUGH THE IEDB AND
OTHER PUBLICLY RELEVANT
DATABASES.
AND PROPOSE THE CONTRACTORS CAN
ALSO DEVELOP PREDICTION TOOLS
ALTHOUGH MANY ARE ALREADY
AVAILABLE ON THE IDB SO IT'S NOT
A REQUIREMENT, IT'S AN OPTION.
IN THIS FY 24 WE'VE ADDED
POTENTIAL AND HIGHLIGHTED
PATHOGENS OF PANDEMIC POTENTIAL
PUBLISHED BY NIAID RECENTLY, IN
EIGHT FAMILIES ALEX MENTIONED
EARLIER, THAT COULD BE
CONSIDERED AS WELL AS THE
PATHOGENS IN THE OTHER AREAS
THAT ARE IN NIAID'S INTEREST
THAT WERE ON PREVIOUS SLIDES.
WE DON'T SUPPORT CLINICAL TRIALS
FOR THIS.
WE OF COURSE UTILIZE CURRENTLY
AVAILABLE CLINICAL TRIALS OR
HUMAN SAMPLES ALREADY AVAILABLE
ARE PREFERABLE.
IN PARTICULAR, WE'D LIKE TO
EXPAND THE USE OF TECHNOLOGIES
INCLUDING HIGH-THROUGHPUT
APPROACHES TO MAKE IT MORE
SCALABLE EPITOPE DISCOVERY AND
CHARACTERIZATION AND LOWER COST
AND EFFICIENT, VERY MUCH ALONG
THE LINES OF WHAT DR. CROWE
DESCRIBED, BUT THERE ARE OTHER
APPROACHES AS WELL.
HE DESCRIBED HIS BUT THERE ARE
OTHERS IN PLACE, CURRENT
CONTRACT POOL, WE WOULD ASSUME
THAT OTHERS WOULD APPLY AS WELL.
THE MECHANISM IS BROAD AGENCY
ANNOUNCEMENT FOR FIVE-YEAR TERM,
WORKED WELL, PRODUCTIVE AND
FLEXIBLE FOR US OVER THE PAST --
SINCE THE EVOLUTION OF THIS
PROGRAM AND WE PREFER TO KEEP
THAT THIS WAY.
I'LL STOP AND BE HAPPY TO TAKE
QUESTIONS.
THANK YOU.
AGAIN, SORRY FOR THE AV
PROBLEMS.
OVER.
>>IF THERE ARE NO QUESTIONS
WE'LL MOVE TO TIM, YOU HAVE THE
NEXT CONCEPT CLEARANCE.
>>EXCELLENT.
THANK YOU VERY MUCH, DAN.
SO, THE CONCEPT I'M GOING TO
PRESENT IS T CELL IMMUNE EPITOPE
DISCOVERY AND MECHANISMS OF T
CELL PROTECTION PROGRAM.
THIS IS A RENEWAL OF THE
EXISTING PROGRAM, BUT EXPANDS IT
AND I'LL EXPLAIN HOW THAT WORKS
AS WE GO ON.
BUT PRIMARY OBJECTIVES OF THE T
CELL PROGRAM ARE TO IDENTIFY AND
VALIDATE HUMAN IMMUNE EPITOPES.
THIS IS, YOU KNOW, OF COURSE
INCREDIBLY IMPORTANT IN THE
PROTECTION FROM INFECTIOUS
DISEASE, AUTOIMMUNE DISEASE,
UNDERSTANDING VACCINE RESPONSE
AND UNDERSTANDING REJECTION OF
TRANSPLANTED CELLS AND TISSUES.
WE HELP TO BE ABLE TO CONTRIBUTE
TO A BETTER UNDERSTANDING OF
IMMUNE MEDIATED PATHOGENESIS,
TRIGGERED BY INFECTION,
AUTOIMMUNE PROCESS, MICROBIOME
OR TRANSPLANTATION.
WE FOUND THAT WE'VE BEEN ABLE TO
REALLY UNDERSTAND VIRAL
EVOLUTION AND STABILITY OF
T-CELL RESPONSES WITH THIS
PROGRAM, EVEN WITH RAPID VIRAL
EVOLUTION, AND GET A GOOD SCOPE
OF THE BREADTH OF HUMAN T-CELL
RESPONSES TO A VACCINE OR
PATHOGEN.
WE'VE BEEN ABLE TO LOOK AT
FAMILIES OF RELATED PATHOGENS
AND SEE HOW THERE MIGHT BE
CROSS-REACTIVITY BETWEEN VARIOUS
EPITOPES FROM THESE PATHOGENS
WITH THE IDEA THAT IF YOU SEE
CROSS-REACTIVITY ACROSS FAMILIES
OF PATHOGENS, YOU CAN ESTABLISH
A RATIONAL UNIVERSAL VACCINE FOR
THAT PATHOGEN.
WE'VE LOOKED AT THIS IN TARGETS
OF THERAPEUTIC INTERVENTIONS.
THIS IS A LONGSTANDING PROGRAM
INITIATED IN 2004 AND IN 2004
WERE ABLE TO AWARD 11 CONTRACTS,
ACROSS BOTH T- AND B-CELL
EPITOPES AND THEY ALSO INCLUDED
TWO CONTRACTS FOR EPITOPE
PREDICTION TOOLS.
IN 2009 THE CELLS SEPARATED TO
GET MORE CONTRACTS THAT ARE
INTERESTED IN THE B CELL
MECHANISMS OF PROTECTION, SO
THIS ONE REQUIRED ONLY T CELL
EPITOPE DISCOVERY PLUS EPITOPE
VALIDATION.
IN 2014 A REQUIREMENT THIS BE
VALIDATED IN HUMANS.
IN 2019 INCREASED SCOPE FOR
INFECTIOUS DISEASE AND VACCINE
RESPONSE, AUTOIMMUNE DISEASE,
INCLUDE
INCLUDE IDENTIFICATION OF
PATHOGEN EPITOPES, AND INCREASE
SOLICITATION SCOPE.
AWARDED SIX CONTRACTS, BOTH
VIRAL AND BACTERIA, BECAUSE OF
THE FLEXIBILITY OF THE PROGRAM
WERE ABLE TO ADD SARS-COV-2 WHEN
THAT BECAME EMERGING PATHOGENS
AND FUNGAL PATHOGENS,
UNDERSTUDIED, AND ABLE TO AWARD
A CONTRACT TO STUDY AUTOIMMUNE
DISEASES, AND IN THIS CASE
RHEUMATOID ARTHRITIS.
IT'S BEEN A SUCCESSFUL PROGRAM.
WE'VE IDENTIFIED THOUSANDS OF
EPITOPES OVER THE YEARS AND JUST
RECENTLY GREAT SUCCESS WITH
SARS-COV-2, PERTUSSIS, HSV-1 AND
2, EVEN WITH OUR AUTOIMMUNE
PROGRAM WITH ARTHRITIS, ALL
BEING DEPOSITED INTO THE IEDB
AND BECOME PUBLICLY ACCESSIBLE,
ACROSS CD 4 AND CD8 T CELLS
WE'RE DISCOVERING THEM.
WE MENTIONED PUBLICATIONS
EARLIER AND MENTIONED MEGAPOOL
DISTRIBUTION EARLIER BUT, AGAIN,
THIS IS SOMETHING THAT WAS
ENABLED BY THIS BEING IN THE
CONTRACT MECHANISM.
AND FINALLY, THE FY 24 PROPOSED
INITIATIVE, IT MUST ADDRESS ONE
OF THE THREE RESEARCH AREAS,
EITHER INFECTIOUS DISEASE OR
VACCINE RESPONSES, AUTOIMMUNE
DISEASE OR TRANSPLANTATION
ANTIGENS, ALOE OR AUTOANTIGENS
COULD BE INCLUDED.
SCOPE EXPANSION FOR FISCAL 24
REQUIRES FUNCTIONAL
CHARACTERIZATION OF THE T CELL
SUBSETS AND MECHANISMS OF T CELL
PROTECTION AND PATHOGENS.
THIS IS SOMETHING THAT
INVESTIGATORS HAD PROPOSED
BEFORE BUT NOW WILL BE REQUIRED
COMPONENT OF THIS SOLICITATION.
AS OPTIONAL COMPONENTS, WE ARE
ALLOWING INVESTIGATORS AND
ENCOURAGING IN SOME WAYS TCR
REPERTOIRE ANALYSIS TO BE
INCLUDED, NOVEL EPITOPE
PREDICTION TOOLS HAVE ALWAYS
BEEN AN OPTION FOR THIS, IT'S
STILL INCLUDED, AND WE'RE ALSO
INCLUDING AS AN OPTION LOOKING
AT PANDEMIC PROTOTYPE PATHOGENS
AS WAS MENTIONED BEFORE.
MUCH LIKE THE B CELL WE'RE NOT
ALLOWING CLINICAL TRIALS FOR
THIS.
BUT WE ENCOURAGE INDIVIDUALS
TO -- INDIVIDUAL INVESTIGATORS
TO FORM COLLABORATIONS WITH
OTHER CLINICAL TRIAL GROUPS SO
THAT THEY CAN UTILIZE THOSE
SAMPLES.
WE'RE GOING TO USE THE CONTRACT
BROAD AGENCY ANNOUNCEMENT
MECHANISM, A FIVE YEAR DURATION.
WITH THAT, I'M HAPPY TO TAKE ANY
QUESTIONS.
>>TIM, THIS IS JIM.
ONE CLARIFICATION.
SO THE CORRELATES OF PROTECTION,
IS THAT LARGELY ACCOMPLISHED
THEN THROUGH COLLABORATIONS
SINCE THERE AREN'T CLINICAL
TRIALS THAT ARE INVOLVED OR DOES
THIS ALLOW SOME OBSERVATIONAL
STUDIES TO BE ABLE TO DO THAT?
>>SO, REALLY IT'S
UNDERSTANDING -- SO
UNDERSTANDING WHICH T CELLS ARE
INVOLVED, YOU CAN GET CLINICAL
SAMPLES, SO OF COURSE THERE'S
GOING TO BE LOADS OF CLINICAL
SAMPLES AVAILABLE, MOST OF THE
INSTITUTIONS HAVE EITHER
CLINICAL CENTERS ASSOCIATED WITH
THEM.
... (TECHNICAL DIFFICULTIES,
PLEASE STAND BY)
>>MACAQUE BREEDING COLONIES.
>>WE CAN SEE YOUR SLIDES BUT
CAN'T HEAR YOU.
>>THAT LOOKS ALL RIGHT?
>>YES.
NOW WE CAN HEAR YOU, GREAT.
>>I WAS GOING TO ASK YOU TO PUT
IT IN PRESENTER VIEW.
SORRY ABOUT THAT.
>>IT'S NOT?
>>NOT FROM OUR END.
IF YOU WANT TO GO FORWARD WITH
THIS VIEW, YOU CAN.
IT'S NO NON-PRESENTER VIEW.
>>I SHOULD CHANGE THE DISPLAY?
>>NO.
ON THE ACTUAL POWERPOINT ITSELF,
LOWER RIGHT-HAND CORNER THERE'S
A PRESENTER VIEW BUTTON.
THERE YOU GO, YEAH.
>>I CLICKED BUT IT SHOWS UP ON
A DIFFERENT SCREEN.
>>WE CAN PROCEED WITH THIS
VIEW.
WE'RE PRESSED FOR TIME.
I JUST WANTED TO CLARIFY.
>>SORRY ABOUT THAT.
I'M NOT SURE HOW TO FIX THIS.
GOOD AFTERNOON, I'LL JULIA SHAW,
TRANSPLANTATION BRANCH
PRESENTING NIAID SPF MACAQUE
BREEDING COLONIES FOR CONCEPT
CLEARANCE.
OKAY.
>>STAND BY.
I'LL SHARE THE SLIDES FOR YOU.
>>SORRY.
OKAY.
NEXT SLIDE.
ALL RIGHT.
THE TRANSPLANTATION BRANCH
SUPPORTS ROBUST AND INTERACTIVE
NON-HUMAN PRIMATE PROGRAM THAT
INCLUDES TWO RESEARCH CONSORTIA,
ONE ON PIGS AND NON-HUMAN
PRIMATE TRANSPLANTATION, OTHER
ALLOGENICS, AND SUPPORT ALLELE
DISCOVERY AND TYPING
TECHNOLOGIES CONTRACT AND NIAID
MACAQUE BREEDING COLONY, FOR
COOPERATIVE STUDY GROUP
PROJECTS.
IT WAS ESTABLISHED IN 1999, TO
EVALUATE SAFETY AND EFFICACY OF
FDA-APPROVED AGENTS AND NOVEL
TREATMENTS IN CLINICALLY
RELEVANT TRANSPLANT MODEL.
THIS SUPPORTS THE MAJORITY OF
TRANSPLANT TOLERANCE RESEARCH
CONDUCTED IN NON-HUMAN PRIMATE
MODELS AND INCLUDES WORK IN
KIDNEY, HEART, ISLET, AND LUNG
TRANSPLANTATION.
FINDINGS HELPED TRANSITION NINE
REAGENTS TO CLINICAL TRIALS AND
TRANSPLANTATION AND RENEWAL
AWARDS FOR THIS INITIATIVE ARE
EXPECTED EARLY THIS YEAR.
HOWEVER, AT INCEPTION IT FACED
CONSIDER CHALLENGES OBTAINING
NON-HUMAN PRIMATE DUE TO LOW
AVAILABILITY AND HIGH COST,
DRIVEN BY HIGH DEMAND FOR
MACAQUES FOR HIV/AIDS RESEARCH.
NEXT SLIDE.
STARTING IN 2000 NIAID
ESTABLISHED BREEDING COLONIES
FOR PATHOGENS RHESUS MACAQUES
AND CYNMOLGUS MACAQUES, SEVERAL
YEARS LATER THE AWARD WAS
CONVERTED TO CONTRACT MECHANISM
TO ALLOW FOR ENHANCED NIAID
OVERSIGHT AND DIRECTION.
MORE RECENTLY SUPPORT FOR
COLONIES ONGOING CARE AND
BREEDING HAS BEEN SUPPORTED
THROUGH SEVEN-YEAR COMPETITIVE
CONTRACTS SOLICITATION WITH
AWARDS ISSUED IN 2010 AND AGAIN
IN 2017.
NEXT SLIDE.
THE OVERARCHING GOAL REMAINS THE
PROVISION OF HIGH QUALITY
PEDIGREED MACAQUES.
REQUIREMENT FOR SPF ANIMALS
REDUCES RISK OF OPPORTUNISTIC
INFECTION TO PREVENT ALLOGRAFT
REJECTION.
PEDIGREE ENABLES KNOWLEDGE OF
DEGREE OF DONOR-RECIPIENT
RELATEDNESS, PROMOTING RIGOROUS
DESIGN AND DATA INTERPRETATION,
BETTER APPROXIMATES CLINICAL
PRACTICE AND COLONY MANAGEMENT
PERSPECTIVE INFORMS BREEDING
STRATEGIES.
THE GOAL IS TO ENHANCE RIGOR AND
REPRODUCIBILITY FOR TRANSPLANT
EXPERIMENTS WHILE REDUCING
NON-HUMAN PRIMATE LOSS THROUGH
OPPORTUNISTIC INFECTIONS AND USE
RELATED TO SUBOPTIMAL STUDY
DESIGN.
NEXT SLIDE.
IN ADDITION TO ACHIEVING AND
MAINTAINING SPF STATUS,
CONSISTENTLY STRONG WITH CASE
RESOLUTION OVER 90%.
SHIPMENTS FALL SHARPLY IN 2019
CONTRACT YEAR DUE TO COVID-19
RELATED RESEARCH RESTRICTIONS
SHIPMENTS HAVE RECOVERED,
MATURED AND ABLE TO MEET
RESEARCH NEEDS.
NEXT SLIDE.
YOU MAY APPRECIATE CHANGES IN
NON-HUMAN PRIMATE DEMAND CAN
OCCUR UNPREDICTABLY, THE
SITUATION TODAY IS NOT SO
DIFFERENT FROM WHAT IT WAS WHEN
COLONIES WERE FIRST ESTABLISHED.
IN THIS CASE HIGH DEMAND IS
BEING DRIVEN BY THE COVID-19
PUBLIC HEALTH CRISIS AND THE
DECISION TO CEASE EXPORTING,
EXPANSION REQUIRES LONG LEAD
TIMES, 3 TO 6 YEARS BEFORE
ADDITIONAL RESEARCH AGED ANIMALS
BECOME AVAILABLE.
THIS HIGH DEMAND AND LOW
AVAILABILITY IS FUELING
COMPETITION FOR A LIMITED POOL
OF NON-HUMAN PRIMATES, RESULTING
IN SUBSTANTIAL INCREASE IN COST
FOR MOST IN DEMAND SPECIES.
UNFORTUNATELY THIS CAN ALSO FEED
UNSUSTAINABLE ILLEGAL TRADE IN
THESE ANIMALS.
NEXT SLIDE.
FOR THESE REASONS WE SUPPORT THE
RENEWAL OF THE NIAID SPF MACAQUE
BREEDING COLONIES TO ENSURE A
RELIABLE DOMESTIC SUPPLY OF HIGH
QUALITY PRIMATES.
THIS SOLICITATION TYPE MECHANISM
AND DURATION REMAIN UNCHANGED
FROM THE PREVIOUS RENEWAL.
THANK YOU FOR YOUR ATTENTION, BE
HAPPY TO TAKE QUESTIONS AT THIS
TIME.
>>THANKS FOR THE PRESENTATION.
ARE THROWS COLONIES OWNED BY NIH
OR OWNED BY THEIR CONTRACTOR?
>>THE ANIMALS ARE OWNED BY
NIAID, AND WE, YEAH, HAVE A
CONTRACT FOR THEIR CARE AND
BREEDING.
>>GREAT.
THANK YOU.
>>INTERESTING QUESTION,
DIFFICULT TO MOVE THESE ANIMALS
BUT WE HAVE THAT CAPABILITY IF A
NEW CONTRACTOR IS CHOSEN THROUGH
COMPETITOR PROCESS.
AND IT'S BEEN AN EXTRAORDINARILY
VALUABLE RESOURCE FOR US
CONSIDERING WHAT WE COULDN'T
POSSIBLY HAVE ANTICIPATED ABOUT
THE COST OF ANIMALS AND QUALITY
AND ASSURANCES WE HAVE THROUGH
OWNING THIS COLONY.
>>TOTALLY UNDERSTAND.
IT'S AWESOME YOU'RE EXPANDING
COLONIES.
SOUND GREAT.
>>I'M CURIOUS, MAYBE THIS IS AN
ASIDE, BUT DO YOU HAVE TO GET
INVOLVED IN MONITORING THE
ILLEGAL TRADE AND MAYBE EVEN
STEMMING IT SINCE IT'S SOMETHING
YOU MENTIONED?
>>I DON'T THINK SO BECAUSE WE
HAVE OUR OWN SUPPLY HERE BUT IT
IS VERY DIFFICULT FOR PEOPLE WHO
ARE TRYING TO PURCHASE THEM
COMMERCIALLY, SOMETIMES TO KNOW.
>>WHAT WOULD HAPPEN IF A
RESEARCHER MAKES AN INNOCENT
MISTAKE PURCHASING AN ILLEGAL
ANIMAL?
>>I DON'T KNOW.
THAT'S A GOOD QUESTION.
I THINK THERE HAVE BEEN SOME
THAT HAVE BEEN HELD AT CERTAIN
FACILITY
FACILITIES FOR A WHILE BECAUSE
THEY WERE SUSPECTED TO HAVE BEEN
ILLEGALLY ACQUIRED.
>>YEAH.
>>WE DON'T ACTUALLY RECEIVE
THOSE ANIMALS.
>>THE SITUATION WE'VE
THANKFULLY AVOIDED BY HAVING
THIS COLONY AND BEING ABLE TO
MEET THE NEEDS WITH THE ANIMALS
READY FOR RESEARCH.
>>ANY OTHER COMMENTS OR
QUESTIONS ABOUT THIS OR ANY OF
THE INITIATIVES JUST PRESENTED?
>>OKAY, MERCY, LET'S GO ON.
YOU HAVE THE FINAL ONE.
>>YES, I WILL NOT TAKE LONG.
THANKS, BRIAN.
I'M MERCY, AND I'M PROGRAM
OFFICER AT THE BASIC IMMUNOLOGY
BRANCH.
NEXT SLIDE PLEASE.
I'LL BE TALKING TO YOU ABOUT THE
RENEWAL OF THE PROGRAM ON
MATERNAL FETAL IMMUNITY, AND THE
MAIN OBJECTIVES ARE TO DETERMINE
THE ROLES AND INTERACTIONS OF
HUMAN IMMUNE CELLS AT THE
MATERNAL FETAL INTERFACE AND TO
ELUCIDATE EFFECTS OF VACCINATION
IN PREGNANCY ON INTERFACE CELLS
AND DEVELOPING FETAL IMMUNE
SYSTEM, INITIATED IN FISCAL YEAR
2019, AND CURRENTLY 11 AWARDS, 9
FROM DAIT, 2 DMID, AROUND 53
PUBLICATIONS.
NEXT SLIDE PLEASE.
SO, THE MATERNAL-FETAL INTERFACE
UNDERGOES DYNAMIC IMMUNOLOGIC
MODELING AND CHANGES DURING
PREGNANCY.
AND SO THREE IMMUNE EVENTS ARE
CRUCIAL TO SUCCESSFUL PREGNANCY
OUTCOMES, AND FIRST IT'S THE
EMBRYO IMPLANTATION, EARLY
TRANSPLANTATION THAT ARE
IMPORTANT, EARLY EVENTS ARE
COUNTER-BALANCED BY STATE OF
TOLERANCE INDUCED BY VARIOUS
FACTORS WHICH INCLUDE
RECRUITMENT AND PROLIFERATION OF
DIFFERENT REGULATORY CELLS AND
EXPRESSION OF VARIOUS REGULATORY
MOLECULES.
A SECOND PHASE LASTS UNTIL
PARTURITION IS INDUCED,
INCREASED PRODUCTION OF
PRO-INFLAMMATORY CYTOKINES.
SOME IMPORTANT INSIGHTS WERE
DISCOVERED BY INVESTIGATORS IN
THE PROGRAM.
SEVERAL QUESTIONS REMAIN AND WE
WOULD LIKE TO BUILD ON THE
FOUNDATION ESTABLISHED UNDER
FIRST INITIATIVE INCLUDING
LOOKING AT HEALTHY UTERINE
IMMUNOLOGIC CHANGES AT INTERFACE
CONTRIBUTING TO RECURRENT
IMPLANTATION FAILURE AND
RECURRENT PREGNANCY LOSS.
THERE'S A NEED FOR TOOLS THAT
REFLECT THE MATERNAL-FETAL
INTERFACE, AND MODELS THAT
APROXIMATE THE INTRAUTERINE
ENVIRONMENT DURING PREGNANCY,
EARLY PREGNANCY, AND THERE'S A
LACK OF REAL HUMAN MODELS,
RELIANCE ON MICE INHIBITS
ABILITY TO MAKE PROGRESS,
LOOKING FOR PRIMARY CELL
ISOLATION, ORGANIZED
MICROPHYSIOLOGIC SYSTEMS AND THE
LIKE AND WANT TO BE ABLE TO
INVESTIGATE THE VARIOUS CHANGES
THAT WILL ALLOW CONSTRUCTION OF
IMMUNE LANDSCAPE DURING
PREGNANCY AND COVID-19 ALSO HAS
BEEN SHOWN TO BE MORE SEVERE IN
PREGNANT WOMEN, AND HIGHLIGHTS
THE NEED FOR ATTENTION TO UNIQUE
STATE OF
NEXT SLIDE PLEASE.
I'D LIKE TO HIGHLIGHT A FEW
ACCOMPLISHMENTS.
FIRST ONE IS FROM JUDY
LIEBERMAN'S LAB AT BOSTON
CHILDREN'S HOSPITAL, HERE SHE'S
SHOWN HUMAN DECIDUAL NK CELLS
EXPRESS PEPTIDE, TRANSFERRED VIA
NANOTUBES TO KILL EXTRACELLULAR
LISTERIA WITHOUT KILLING THE
TROPHOBLASTS, AND THESE
NANOTUBES ORIGINATE FROM THE NK
CELL AND TRANSFER IN AN
ACTIN-DEPENDENT MANNER.
CAROLINE COIN'S GROUP SHOWED
MID-GESTATION PERIOD INDUCES
INFLAMMASOME SIGNALING, SHE
SHOWED THAT THESE TROPHOBLASTS
SECRETE INFLAMMASOME-ASSOCIATED
CYTOKINES LIKE IL-1 BETA, AND
THEN SHOW THIS -- IDENTIFY
INFLAMMASOME AS IMPORTANT
MECHANISM BY WHICH HUMAN
PLACENTA REGULATES SYSTEMIC AND
LOCAL IMMUNITY DURING PREGNANCY
TO DEFEND AGAINST INFECTION.
NEXT SLIDE PLEASE.
AND THEN NEXT STUDY, WHILE
SEVERE CORONAVIRUS DISEASE IS
ASSOCIATED WITH IMMUNE
ACTIVATION OF MATERNAL-FETAL
INTERFACE, RESPONSES TO
ASYMPTOMATIC OR MILD INFECTION
DURING PREGNANCIES REMAIN
UNKNOWN.
THE LAB LOOKED AT RESPONSES TO
ASYMPTOMATIC OR MILD COVID-19
DURING PREGNANCY, AND REPORT
ATTENUATION OF ANTIGEN
PRESENTATION AND TYPE I
INTERFERON SIGNALING AND THEN
ALSO REMODELING OF LANDSCAPE OF
THE INTERFACE, AND WHICH HAS A
POTENTIAL FOR LONG-TERM ADVERSE
OUTCOMES FOR THE OFFSPRING.
AND THEN LASTLY, A LAB IN
CINCINNATI SHOWED PREGNANCY --
DURING PRESS NANCY THERE'S
DEACETYLATION WHICH EXPANDS
PROTECTION TO EXTRACELLULAR
PATHOGENS, HERE THEY DESCRIBE
PREGNANCY SPECIFIC MODIFICATION
OF MATERNAL ANTIBODIES WHICH
THEN ENHANCE PROTECTION AGAINST
LISTERIA INFECTION AND SUGGESTS
THE DEACETYLATION THEN ENABLES
CD22 BINDING AND INHIBITION OF
SUPPRESS IVE B CELL SUBSET.
NEXT SLIDE PLEASE.
SO, THE SCOPE OF THE PROPOSED
PROGRAM, THE RENEWAL, IS TO
INCLUDE MECHANISMS OF INDUCTION,
ACTIVATION OF VARIOUS LEUKOCYTE
RESPONSES AND MAINTENANCE OF
HOMEOSTASIS AT MATERNAL FETAL
INTERFACE AND WILL LOOK AT THE
EFFECT OF PERTURBATION, WHETHER
INFECTIOUS OR NON-INFECTIOUS ON
IMMUNE REGULATORY MECHANISMS AND
ARCHITECTURE AND FUNCTION AT THE
INTERFACE, AND WILL ALSO
DETERMINE IMMUNE CONTRIBUTIONS
TO KEY OBSTETRICAL DISEASES IN
PREGNANCY AND DEFINE REPERTOIRE
OF IMMUNE CELLS AND PROTEINS
DURING GESTATION, ALSO TO
ENCOURAGE DEVELOPMENT OF EX VIVO
SYSTEMS THAT WE CAPITULATE THE
INTERFACE.
SO, POTENTIAL PARTNERS NIEHS,
NICHD, NIDA, AND WE WANT TO
CONTINUE WITH R01 MECHANISM.
AND WITH THAT WE'LL END THE
PRESENTATION AND I'M HAPPY TO
TAKE ANY QUESTIONS.
>>LOOKS LIKE CREATIVE RESEARCH
AND FINDINGS THERE, VERY
EXCITING.
>>YES, A LOT OF NEW THINGS THAT
HAVE COME UP OUT OF THE PROGRAM.
IT'S GRATIFYING.
>>OKAY.
IF WE HAVE NO OTHER QUESTIONS OR
COMMENTS, ON THIS OR ANY OF THE
OTHER CONCEPT CLEARANCES
PRESENTED, WE SHOULD MOVE TO EN
BLOC MOTION TO PROCEED AND A
SECOND.
>>THESE ARE IMPRESSIVE
PRESENTATIONS.
ACCOMPLISHMENTS ARE AWESOME.
I MOVE FOR A VOTE,
RECOMMENDATION FOR ALL FOUR.
>>I SECOND.
>>GREAT, THANK YOU.
REMINDER THAT THE CONCEPT
CLEARANCES WE DIDN'T DISCUSS,
THOSE THAT I MENTIONED BRIEFLY
FOR THE SMALL BUSINESS CONTRACTS
AND THE ODD SET INFORMATICS
PROPOSALS ARE IN THE ELECTRONIC
COUNCIL BOOK AND WE NEED YOUR
APPROVAL TO GO THROUGH WITH
THOSE, NO NEED FOR DISCUSSION.
YVETTE, HAVE WE COVERED
EVERYTHING BEFORE WE ADJOURN?
>>WE HAVE.
I WANT TO REMIND COUNCIL MEMBERS
TO VOTE ON THE FOUR CONCEPT
CLEARANCES THEY HEARD AND FOUR
DATE FYIS AND TWO ODD SETS,
EVERYTHING IS IN ECB TO APPROVE
I WOULD APPRECIATE THAT.
>>TERRIFIC.
THANK YOU TO THE SPEAKERS TODAY
AND EVERYBODY ELSE WHO JOINED US
AND COUNCIL MEMBERS, WE HOPE
THAT COME JUNE WE'LL SEE YOU ALL
IN PERSON.
>>OKAY.
VERY GOOD.
>>WE ANTICIPATE THAT I WILL
NEED TO BE AVAILABLE IN JUNE OR
PROBABLY NOT, RIGHT?
>>THIS SHOULD BE YOUR LAST
COUNCIL ROUND.
I THINK THEY EXTENDED YOU, AND
LET ME SEE.
I BELIEVE THIS IS YOUR LAST
ROUND.
>>ALL RIGHT.
>>APPRECIATE IT THOUGH.
>>OH, WELL, THEN A SPECIAL
THANKS TO BOTH OF YOU FOR YEARS
OF SERVICE AND FOR THE
EXTENSION.
WE'VE REALLY ENJOYED HAVING BOTH
OF YOU ON THE COUNCIL
SUBCOMMITTEE AND YOUR COMMITMENT
AND CONTRIBUTIONS HAVE BEEN
TERRIFIC.
ORDINARILY I THINK IF IT WOULD
HAVE BEEN IN PERSON IT WOULD
HAVE BEEN A PHOTO WITH DR. FAUCI
OR IN THIS CASE YOU.
>>I THINK AT THE LAST ONE OUR
FINAL ONE, RIGHT?
THAT'S WHY I WAS ASKING ABOUT
THE EXTENSION BECAUSE I DON'T
THINK THEY WERE SO OBVIOUS.
BUT ANYWAY, IT'S BEEN FUN, MOST
HAS BEEN.
>>YEAH.
>>A PLEASURE.
THANK YOU SO MUCH.
THANKS TO ALL THE PEOPLE IN THE
OFFICE THERE.
ALL THE FOLKS THAT PRESENTED
THIS AMAZING INFORMATION.
>>GREAT.
WELL, THANK YOU AGAIN.
AND WE'LL ADJOURN HERE AND LOOK
FORWARD TO SEEING PEOPLE IN
JUNE.
>>THANK YOU.
BYE-BYE NOW.
CAPTION TEST, CAPTION TEST.
IS NOT A GOOD TARGET FOR T CELLS
HERE, SPIKE IS A GOOD TARGET
VACCINE MADE OF SPIKE ONLY,
WHICH WAS ORIGINALLY BEING
DEVELOPED.
AT THE SAME TIME WE SAW T-CELL
RESPONSES CD4 AND CD8 RESPONSE
AGAINST A NUMBER OF SARS-COV-2
ANTIGENS, RAISING THE PROSPECT
IF A BROADER SPECTRUM OF T