>>HELLO, WELCOME EVERYONE TO
OUR LATEST LECTURE IN THE COVID
INTEREST GROUP.
IT IS REALLY A GREAT PLEASURE TO
INTRODUCE OUR SPEAKER, DR. ALI
ELLEBEDY, WHO IS A FACULTY IN
THE DEPARTMENT OF PATHOLOGY AND
IMMUNOLOGY AND CO-DIRECTOR FOR
THE CENTER FOR VACCINES AND
IMMUNITY AT THE WASHINGTON
UNIVERSITY SCHOOL OF MEDICINE IN
ST. LOUIS.
STARTING WITH HIS GRADUATE WORK
AT THE UNIVERSITY OF TENNESSEE
AND POST DOCTORAL WORK AT EMRY,
DR. ELLEBEDY HAS BEEN AT THE
FOREFRONT OF ELUCIDATING
MECHANISMS OF SUCCESSFUL
RESPONSING TO IMMUNIZATION.
FIRST FOCUSING ON INFLUENZA, HE
DESCRIBED AS HIS FIRST AND REAL
LOVE.
MORE RECENTLY ON VACCINATION FOR
SARS-COV-2.
IN HIS OWN LAB AT WASHINGTON
UNIVERSITY WHERE HE HAS BEEN
SINCE 2017, ACTUALLY, VERY
RECENT, STARTING HIS LAB, DR.
ELLEBEDY'S WORK HAS BEEN NOTABLE
FOR HIS GROUNDBREAKING STUDIES
OF IMMUNE RESPONSES TO
VACCINATIONS.
NOT ONLY IN THE BLOOD WHERE MOST
STUDIES HAVE BEEN CARRIED OUT,
BUT ALSO IN THE TISSUE BY TAKING
THE BRAVE STEP IN SERIAL LIMP
NODE BIOPSIES AND BONE MARROW
BUY -- BIOPSIES.
THIS IS AN AMAZING STEP IN, I
THINK, HUMAN IMMUNOLOGY AND
SERVES AS A MODEL FOR ALL OF US.
TOGETHER HIS WORK HAS HELPED TO
DESIGN SCLAR AND MOLECULAR
MECHANISMS, HELPED US UNDERSTAND
THE HETEROGENEITY OF MEMORY B
CELL RESPONSES AND DERNLING THE
RULES FOR ELICITING LARGELY
NEUTRAL B CELL RESPONSES AGAINST
PATHOGENS SUCH AS INFLEW ENDSA
AND SARS-COV-2.
DR. ELLEBEDY'S WORK IS AT THE
FOREFRONT OF TECHNOLOGIES USING
TECHNIQUES THAT WOULDN'T HAVE
BEEN POSSIBLE JUST A FEW YEARS
AGO AND I THINK IT IS REALLY A
TESTIMONY TO HOW THIS FIELD HAS
MOVED TO HELP US UNDERSTAND WHY
VACCINES WORK.
WE LOOK FORWARD TO HEARING HIS
EXCITING FINDINGS.
SO WELCOME, DR. ELLEBEDY.
>>THANK YOU SO MUCH FOR THE
VERY, VERY KIND INTRODUCTION.
AND THANK YOU FOR THE
INVITATION.
I'M REALLY HAPPY AND THRILLED TO
BE SHARING OUR WORK WITH THIS
GROUP.
YEAH.
IT IS -- I'M GOING TO GO
THROUGH -- I SEE THAT WE HAVE
ALREADY THE DISCLOSURE, SO I
WILL SKIP THE DISCLOSURE PART.
I'M GOING TO TELL YOU THE STORY
OF WHAT I HAVE LEARNED SO FAR IN
THE LAST FEW YEARS AND ONE OF
THE THINGS YOU GET ASKED WHEN
YOU APPLY FOR A POSITION TO
START YOUR OWN LAB, TELL US WHAT
YOU ARE GOING TO DO IN THE NEXT
FIVE YEARS.
I LOOK BACK AND I ACTUALLY
LOOKED BACK AT MY PLAN, WHATEVER
THE PLAN WAS AT THE TIME, AND,
OF COURSE, IT HAS NOTHING TO DO
WITH WHAT I HAVE DONE IN THE
LAST FIVE YEARS OR WHAT WE HAVE
LEARNED IN THE LAST FIVE YEARS.
THERE ARE THINGS YOU CANNOT
REALLY PLAN FOR.
REALLY, EVERYTHING CHANGED.
AS YOU WILL SEE, EVERYTHING
CHANGED QUICKLY AND WE HAD TO
ADAPT TO BE ABLE TO LEARN MORE
IN THE ONGOING STUDIES.
I KNOW WE ARE ALL EXPECTING
SARS-COV-2 AND MY FIRST SLIDES
ARE ABOUT A DIFFERENT VIRUS.
ABOUT THE -- IT IS AN EPIDEMIC
THAT HAD BEEN TALKED ABOUT AND
WROTE ABOUT A LOT IN THE MEASLES
EPIDEMIC IN THE FARE ISLANDS IN
1846.
THESE ARE A GROUP OF ISLANDS
THAT ARE SELF-GOVERNING ISLANDS
IN THE ATLANTIC OCEAN BETWEEN
ICELAND AND NORWAY.
THE REASON I'M BRINGING THIS UP,
VERY INTERESTINGLY AT THE TIME,
STARTING FROM 1,500, THE DANISH
DYNASTIES OR WHOEVER WAS RULING
AT THE TIME WAS REALLY CONCERNED
ABOUT THE POPULATION FOR MANY,
MANY OBVIOUS REASONS.
SO THEY STARTED MONITORING THE
OUTBREAKS AND AT THE TIME THEY
SENT A DANISH EPIDEMIOLOGYIST.
THERE WAS A HUGE OUTBREAK.
BUT ONE OF THE FAMOUS
EPIDEMIOLOGIST, WHAT HE WROTE
WHEN HE INVESTIGATING THAT
OUTBREAK, I LIKED SOME OF THE
WRITING.
HE MADE FEW OBSERVATIONS THAT
REMAIN TRUE TO NOW.
IT REALLY TELLS YOU HOW
INSIGHTFUL IT CAN BE CAREFULLY
DONE EPIDEMIOLOGY FOR THE
STUDIES.
THE FIRST VERSION IS VIRGIN
POPULATIONS OR NAIVE
POPULATIONS.
WHEN THERE IS AN OUTBREAK WITH A
VIRUS, EVERYONE GETS INFECTED.
THE REASON HE SAID NEARLY ALL
WOULD BE INFECTED IS THERE WAS
AN OUTBREAK IN 1781 DOCUMENTED
IN THE ISLANDS.
IT WAS A MEASLES OUTBREAK.
PEOPLE WHO ARE 65 OR OLDER DID
NOT GET AFFECTED BY THE 1846
OUTBREAK.
AT THAT TIME IT WAS AN
INTERESTING OBSERVATION ABOUT
EXPERIENCING OUTBREAK BEFORE CAN
PROTECT YOU.
AND THEN HE MADE THAT
OBSERVATION THAT NATURAL
INFECTION CAN GIVE YOU LIFE-LONG
IMMUNITY.
WELL, I GUESS WE WILL GO
STRAIGHT TO THE FAROE ISLANDS,
I WAS SHOWING WHERE THEY ARE AND
THE EPIDEMIOLOGIST FROM DENMARK.
I SAID THE INSIGHT OF BEING
EXPOSED TO AN EPIDEMIC HELPED
PEOPLE.
THE VIRUS DIDN'T CHANGE BUT YOU
GET THIS LIFE-LONG IMMUNITY.
THAT IS BASICALLY IT.
I'M GLAD WE TAUGHT THIS.
THIS LIFE-LONG IMMUNITY COME
FROM TWO MAIN POPULATION OF B
CELL POPULATION.
THE BONE MARROW PLASMA CELLS,
THE ANTIBODY SECRETING CELLS.
IF THOSE AREN'T ENOUGH IN TERMS
OF PROVIDING PROTECTION, WE HAVE
CIRCULATING B-CELLS.
BOTH OF THESE POPULATION COME
FROM REACTION.
THIS IS A REACTION THAT
HAPPENED, A VERY UNIQUE ACTION,
I WILL DISCUSS IN DETAIL IN THE
NEXT SLIDE.
THIS REACTION IS CRITICAL FOR
THE GENERATION OF THOSE TWO
POPULATIONS.
ESPECIALLY HIGH AGENERAL FI, OR
HIGH BINDING AFFINITY POPULATION
OF THOSE CELLS.
SO YOU CAN THINK OF ALL OF OUR
SUCCESSFUL VACCINATIONS IN THE
LAST FEW HUNDRED YEARS.
THINKING ABOUT SMALLPOX, POLIO,
YELLOW FEVER, TETANUS,
DIPHTHERIA, MMR, SAVING MILLIONS
AND MILLIONS OF LIVES.
WHILE WE HAVEN'T REALLY STUDIED
THE IMMUNE RESPONSE -- OR AT
LEAST THE GERMINAL RESPONSE WE
CAN ASSUME THEY HAVE A ROBUST
RESPONSE.
WE HAVE OBSERVED THESE
POPULATIONS TO SURVIVE FOR A
LONG TIME.
GERMINAL CENTER REACTION IS A
CRITICAL REACTION FOR THE
MEMORY.
THIS IS OVERSIMPLIFIED, IF YOU
GET IMMUNIZED, YOU WILL SEE WHY
I THINK THIS IS OVERSIMPLIFIED
BECAUSE IN HUMANS WE HAVE MORE
THAN ONE LYNCH NODE DRAINING.
ASSUMING THERE IS A DRAINAGE,
THE B-CELLS WILL REACT TO
PLASMA, THOSE ARE SHORT LIVED
AND THOSE DON'T PROVIDE THE
FIRST WAVE OF PROTECTIVE
ANTIBODIES.
THEY ARE NOT AFFINITY MATURE,
WHICH HAPPENS IN THE GERMINAL
SYSTEM.
SOME OF THE RESPONDING BCELLS,
THEY WILL HAVE REACTION IN THE
CORTEX, IN THE CENTER OF THE
B-CELL FOLLICLES.
B-CELLS WILL UNDERGO MULTIPLE
ROUNDS OF PROLIFERATION,
REGULATE THE IDEAS, ALLOWED TO
INTRODUCE MUTATIONS IN THE BCR.
THOSE SHOW THE BINDING AFFINITY
TO THE ANTI GERKS ENS,
SPECIALIZED CELLS IN THE
DRAINING LIMP NODES.
SO NOW YOU HAVE DETAILED, IF YOU
GO CLOSER, THERE ARE TWO
DIFFERENT ZONES.
ONE ZONE REALLY SPECIALIZED FOR
THE AMPLIFICATION OF THE B-CELLS
AND INTRODUCTION OF THE B-CELL
RECEPTOR MUTATIONS.
EVERYTHING HAPPENED HERE WITH
THE HELP OF A VERY CONTROLLED
AND HELP FROM T-CELLS.
RESPONDING B-CELLS, OR AT LEAST
THE B-CELLS EXITING, SO THEY CAN
COME OUT AS CIRCULATING MEMORY
B-CELLS OR LONG LIVE PLASMA
CELLS THAT WILL MIGRATE TO THE
BOTTOM NOW.
THIS IS THE REACTION, WHAT WE
LEARNED FROM ANIMAL MODELS,
ACTUALLY, IN THE LAST 50 TO
MAYBE 60 TO 70 YEARS SINCE WE
STARTED LEARNING IN RABBITS AND
SMALLER RATS AND MICE.
WE REALLY LEARNED MOST OF THESE
DETAILS FROM THESE REALLY
IMPORTANT EXPERIMENTS IN ANIMAL
MODELS.
BUT THEN IN HUMANS, HOW CAN
WE -- GIVEN HOW CRITICAL THIS
REACTION IS AND GIVEN THESE
CELLS DO NOT CIRCULATE IN BLOOD,
HAVING THESE CELLS CONFINED TO
THIS MICROATOMIC STRUCTURES IN
THE DRAINING INFLUENCE, HOW CAN
WE KNOW, FOR EXAMPLE, IN THE
CASE OF INFLUENZA VACCINE, HOW
CAN WE IMPROVE THE VACCINE IF WE
DON'T HAVE A GENERAL REACTION.
IN MY TALK I WILL TELL YOU MY
JOURNEY OF HOW I STARTED ASKING
THESE IMPORTANT QUESTIONS.
THIS IS WHAT DROVE REALLY THE
TOOLS AND TECHNIQUES THAT WE
TRIED TO ANSWER THESE QUESTIONS.
I WILL START FIRST WITH THE
PLASMA -- THIS IS MY POST DOC I
MENTIONED.
HE DID THIS STUDY 15 YEARS AGO,
PUBLISHED VERY SIMPLE ONE,
LOOKING AT HEALTHY YOUNG
DIFFICULTS.
THE REL INFLUENZA VACCINE.
THEY NOTICED THIS POPULATION OF
COMING VERY QUICKLY PLASMABLAST,
ANTIBODIES ARE RESPONDING TO THE
VACCINE AND HE MADE THE NOTE
THESE ARE COMING FROM MEMORY
CELLS.
FIRST EVERY ONE OF US HAS A POOL
OF MEMORY B-CELLS FOR INFLUENZA.
IF YOU PUT THIS ON PLATES COATED
WITH THE ANTIGEN, YOU WILL SEE
THEY ARE MOSTLY SECRETING.
THEY COME QUICKLY.
THAT IS A FAST KIND.
AND THEY HAVE ACCUMULATED LARGE
NUMBER OF MUTATIONS, RELATIVELY.
SO THAT MEANS THEY ARE MOST
LIKELY CAME FROM MEMORY CELLS.
THESE MEMORY SELLS WENT THROUGH
INTERACTION IN THE PAST AND
ACCUMULATED THESE MUTATIONS.
THESE MUTATIONS HAPPEN AS I
DISCUSSED.
SO WHAT I WANT YOU TO LEARN FROM
SO FAR IS THAT WE HAD, BY
LOOKING AT BLOOD, WE CAN LEARN A
LOT OF THINGS.
WE CAN ISOLATE THE RESPONDING
PLASMABLAST.
WE CAN LEARN A LITTLE BIT ABOUT
THEIR ORIGIN.
WE CAN LOOK AT THE IGG, THE
ISOTYPE THEY ARE SPEAKING.
WE COULDN'T LEARN IF WE CAN
GENER
GENERATE
[ INDISCERNIBLE ]
CAN WE ACTUALLY LOOKED A OTHER
CELLS IN THE BLOOD, THAT ARE NOT
DIFFERENTIATING THROUGH
PLASMABLAST.
I CALL THEM ACTIVATED B-CELLS AT
THE TIME.
THOSE ARE THE CELLS YOU CAN
DEFINE THEM AS 71 HIGH CELLS,
THEY ARE PROLIFERATING BUT DON'T
DIFFERENTIATE.
THESE ARE ALL DETAILED IN THIS
PAPER.
IF YOU DID THE SEQUENCE ANALYSIS
OF THE RESPONDING B-CELLS, THE
LEVEL OF MUTATIONS DIDN'T CHANGE
MUCH AND THIS IS JUST LOOKING AT
AVERAGE MUTATION FOR THE SAME
EXACT CLONE THAT ARE RESPONDING
TO A CHANGE OVER TIME.
AT THE TIME IT WAS INTRIGUING TO
ME.
I WAS FINISHING MY POST DOC
FOLLOWSHIP, IF WE HAVE A GOOD
RESPONSE WHY DON'T WE HAVE A
GERMINAL CENTER.
SO THIS REALLY, THAT QUESTION
REMAINED WITH ME AS I MOVED FROM
ATLANTA TO ST. LOUIS AND I
THOUGHT THE FIRST QUESTION I
WANTED TO ASK IS CAN WE STUDY
THE GERMAL CENTER IN VACCINE.
WE KNOW THAT IT IS A SUB --
VACCINE, BUT DOESN'T INDUCE A
GERMINAL CENTER REACTION.
IF IT DOESN'T, WE NEED TO
IMPROVE, IF IT DOES, HOW MANY?
AT THE TIME A LAB PUBLISHED A
COUPLE OF PAPERS OF SAMPLING
LYNCH NODES FINDING -- SO THIS
IS ONE OF THE THINGS THAT REALLY
BEING MORE OR LESS NAIVE ABOUT
HOW THINGS GO, I JUST WENT TO
OUR GEOLOGIST AT WASH U, IN
2017, ONE WEEK AFTER I STARTED
BY LAB.
I WALKED INTO THE RADIOLOGIST
OFFICE AND SAID I SAW THIS PAPER
THAT IT CAN DO THIS TECHNIQUE
AND I READ YOU DO THIS DYING --
DIAGNO
DIAGNOSTICALLY.
I WOULD LIKE YOUR HELP IN
SAMPLING THE NODES.
OBVIOUSLY, IT TOOK SOME TIME TO
CONVINCE THAT THIS IS AN
IMPORTANT QUESTION, BUT AT THE
SAME TIME I LEARNED THAT HUMANS
HAVE MULTIPLE LYNCH NODES.
IT TOOK US THE FIRST YEAR TO
LEARN HOW TO DO THIS TECHNIQUE
AND HOW TO ACTUALLY SAMPLE
RELIABLELY THE GENE.
THIS IS AN ULTRA SOUND GUIDED
AUXILIARY NODE.
WE SAMPLE THE CORTEX REASON.
WE DO NOT APPLY SUCTION SO WE
TRY TO KEEP THE STRUCTURE OF THE
NODE INTACT SO WE CAN DO THIS
MULTIPLE TIMES.
SO 2017-18 WAS THE YEAR WE
LEARNED HOW TO DO THIS.
SO WE DIDN'T REALLY GENERATE ANY
MEANINGFUL DATA IN THAT TIME.
IN 2018-19, NOW WE HAVE BEEN
REALLY SET UP TO DO THIS, WE
LEARNED WE CAN ACTUALLY PREDICT
WHICH NODES WILL BE RESPONDING
TO VACCINE.
THESE ARE THE LATERAL CLUSTER,
THROUGH TRIAL OR ERROR, THOSE
ARE THE NODES WE SHOULD SAMPLE
BECAUSE THEY ARE NOT DRAINING
THE UPPER ARM.
YOU CAN SEE ONE OF THE NODES WE
CORRECTLY PREDICTED WOULD
RESPOND TO THE VACCINE, THE
CORTEX IS VERY THIN, BUT OVER
TIME AT VACCINATION, THE CORTEX
GETS MUCH THICKER.
THAT IS A GOOD SIGN THERE MUST
BE GOING ON.
IF YOU LOOK AT THE SAME TIME
POINTS, WEEK ONE IS BLAST MA --
PLASMABLAST ARE WORKING.
THESE ARE B-CELLS RESPONDING TO
SOME STIMULATION OR MEMORY
CELLS.
WEEK TWO WE STARTED SEEING THE
GERMINAL CENTER REACTION, SOME
POUND TO THE HA, THE MAIN
PROTEIN IN THE INFLUENZA
VACCINE.
THIS IS THE FIRST DETECTION OF
B-CELLS AFTER VACCINATION.
ALLOWED US TO ANSWER A SIMPLE
QUESTION.
REMEMBER, I MENTIONED
PLASMABLAST ALMOST COME FROM THE
MEMORY B-CELL COMPARTMENT
BECAUSE THE VACCINE IS NOT -- IT
RELIES ON STEM LATING THE MEMORY
B-CELLS.
WE SEE MULTIMILLION TIME POINTS
OF THE SAME CLONES, RESPONDING
TO VACCINATION WITH THE
INFLUENZA CAN BECOME PART OF THE
GC OR A COMPLETELY DIFFERENT
NODE OR WE HAVE A MIX.
TO ANSWER THIS QUESTION WE
SINGLE SOURCED PLASMABLAST FROM
BLOOD.
WE DID THE SAME FOR THE GC
B-CELLS.
TOTAL GC.
ONCE YOU HAVE THE ANTIBODIES YOU
CAN CONFIRM ITS BINDING.
ONCE YOU HAVE IT CONFIRMED TO
THE VACCINE, YOU NOW HAVE THIS
REARRANGED FROM THAT PARTICULAR
CLONE AS YOUR I.D. OR
FINGERPRINT, YOU CAN TRACK THE
EVOLUTION.
THIS IS WHAT WE DID HERE.
THIS IS A SINGLE CELL ANALYSIS
OF ONE INDIVIDUAL, B-CELLS OR
LIMP NODE B-CELLS.
WE TRACKED THAT OVER TIME.
YOU CAN HAVE THE TRANSCRIPTION
TO DIVIDE B-CELLS PLASMABLAST,
MEMORY AND SAME IN THE LIMP NODE
BUT YOU ADD THE GERMINAL CENTER
IN BLUE.
THESE AT BASELINE WERE ANGI
SPECIFIC.
THAT BRC COMPLIED WITH THAT.
FIRST WEEK EVERYTHING HERE IN
THE PLASMABLAST IS SPECIFIC
BECAUSE THAT IS WHAT WE KNEW.
THE EXPECTATION AND THOSE ARE
COMING FROM MEMORY, THOSE ARE
HIGH LEVEL OF MUTATIONS.
THEY ARE FROM BLOOD, ACTIVATED
B-CELLS WHICH IS THE WORK I DID
WITH RAFY AS A POST DOC.
THOSE BECOME RESTING CELLS AND
SLOWLY BECOME OUR MEMORY CELLS.
THE LYMPH NODE IN THE GERMINAL
YOU CAN SEE THEM FOR TWO MONTHS.
SOME CLONES I COLORED IN BLUE
THAT ARE RECORDING IN THE
PLASMABLAST.
ANYONE WE SHARED I COLORED
PURPLE.
THERE ARE SOME CLONES GOING BOTH
WAYS, THE MEMORY SELLS ARE THE
MAJOR PRECURSOR FOR THE
PLASMABLAST, THOSE CELLS IN BLUE
ARE COMING FROM NAIVE.
THEY WILL HAVE LOWER MUTATIONS
IN THAT CASE.
WE DID THE ANALYSIS FROM THREE
INDIVIDUALS AND IN THAT CASE,
THIS IS LOOKING AT MUTATION
LEVEL AND PLASMABLAST OR SHARED
PLASMABLAST AND GCs ARE
SIGNIFICANTLY HIGHER MUTATED
CELLS.
WE MADE THE CORRESPONDING MONO
CLO
CLONES.
WE CAN GENERATE B-CELLS TO THE
REACTION AND WHY THIS IS
IMPORTANT.
IN THE CASE OF RAPIDLY INVOLVING
VIRUSS OR ANTIGENS YOU GET
EXPOSED TO REPEATEDLY, YOU WANT
TO UPDATE YOUR REP -- R
REPRETOIRE.
IF YOU HAVE A ROBUST YOU CAN
ENGAGE THE CELLS LATER IN THE
RESPONSE.
AND THEY ARE TARGETING NOVT
ISOTOPES AND WE SHOWED THAT IN
THIS PAPER.
SO THIS IS NOW JUST TO PUT YOU
IN THE CONTEXT OF THE TIME AND
THE SYSTEM, PROFESSOR, TWO AND A
HALF YEARS AND I WAS SO EXCITED.
WE HAVE OUR FIRST BIG PAPER AND
WE ARE REALLY EXCITED AND WE CAN
ADDRESS QUESTIONS ABOUT GERMINAL
CENTER DYNAMICS IN HUMANS.
I WANTED TO LOOK AT THE BONE
MARROW PLASMA CELLS.
THAT IS WHERE THE ULTIMATE
TARGET OF HAVING THIS I ENLISTED
SOME HELP WITH ONCOLOGISTS.
WE CAN DO A VACCINE STUDY TO
SAMPLE LYMP NODE, BLOOD AND THIS
IS SO EXCITING.
WE ARE VERY READY TO GO.
OF COURSE, AS YOU ALL KNOW THIS
HAPPENED AND I GOT THIS CALL IN
MARCH 2020, THIS WAS A VERY,
VERY DEPRESSING WEEK WHERE WE
HAD EVERYTHING HAS TO BE
STOPPED.
WE HAVE TO SWITCH.
NO WORK ON INFLUENZA ANYMORE.
ALL OF MY DREAMS, ALL OF MY
PLANS SO EXCITED TO START, BUT
THEN I REALIZED RIGHT AWAY THIS
IS A GOOD OPPORTUNITY BECAUSE
OBVIOUSLY INFECTION WITH THE
SARS-COV-2 AS A NOVEL VIRUS IN A
WAY, BUT THE VACCINATION PART.
THIS IS A VACCINE PLATFORM NEVER
TESTED IN HUMANS.
IF PEOPLE HAVEN'T BEEN INFECTED,
THIS COULD BE CLOSER TO A
PRIMARY RESPONSE I NEVER GET A
CHANCE TO STUDY IN HUMANS
BECAUSE INFLUENZA MODELS IS
ALWAYS REALIZING THE --
RESPONSE.
WE HAVE THE SAME QUESTION WITH
THE SARS-COV-2 VACCINE.
PFIZER LOOKING AT PEOPLE GETTING
TWO DOSES.
IN DECEMBER 2020, LIKE MANY
OTHER GROUPS AROUND THE WORLD.
QUESTION RECRUITED INDIVIDUALS.
THE ONLY DIFFERENCE IS WE
SAMPLED BLOOD AND LYMPH NODE
OVER TIME THEN WE ASKED FOR
BLOOD FOR LOOKING AT THE
PLASMABLAST.
THERE ARE SOME RESPONSES
DETECTED AT THREE WEEKS.
THIS IS PROTEIN IGG.
ONE WEEK AFTER SECOND DOSE
EVERYONE RESPONDED WELL.
YOU CAN SEE SOMETHING VERY
SIMPLE AS TWO DOSES NEEDED TO
GET REALLY HIGH, BUT IF YOU HAVE
BEEN INFECTED WITH THE VIRUS,
ONE DOSE IS ENOUGH BECAUSE YOU
HAVE MEMORY CELLS FROM THE VIRUS
INFECTION.
THIS IS THE DATA THAT HAS BEEN
SHOWN A THOUSAND TIMES NOW BY
MANY, MANY DIFFERENT GROUPS.
WHAT ABOUT THE GERMINAL CENTER?
AT THE TIME, INFLUENZA VACCINE
IN HUMANS IN THE U.S. IS REALLY
POORLY MANAGED VACCINE IF IT
WASN'T JUST RELYING ON MEMORY
CELLS.
THE LYMPH NODE RESPONSE WAS MORE
ROBUST HERE.
ROBUST IS MUCH BIGGER NODE, MUCH
MORE ACTIVE, REACTIVE NODES.
I GOT THE PHONE CALL YOU NEED TO
COME AND SEE THIS BECAUSE THIS
IS SOMETHING WE HAVEN'T SEEN
WITH FLU BEFORE.
JUST TO GIVE YOU THE EXAMPLE,
THIS IS OUR LYMPH NODE RESPONSE
TO -- ISOTOPE B-CELLS.
THIS IS BEFORE VACCINATION.
B-CELL IS THE MAIN
TRANSSCRIPTURE FACTOR.
YOU CAN TELL THERE IS AN
INCREASE.
THE SAME POPULATION ONE WEEK
AFTER THE SECOND PFIZER
IMMUNIZATION, WHICH IS BASICALLY
THE TWO DOSES THAT WERE THREE
WEEKS APART, ONE WEEK AFTER THAT
SECOND DOSE, THIS IS THE
POPULATION OF THE LYMPH NODES.
THIS IS OUR FIRST VOLUNTEERS.
WE ARE LOOKING AT SOMETHING
COMPLETELY DIFFERENT.
PEOPLE FELT THERE IS A VERY
ROBUST RESPONSE AND SOME PEOPLE
SAID THE NODES WERE REALLY, YOU
CAN FEEL THEM, I GUESS.
THAT ACTUALLY -- WE PUBLISHED
THIS AND SAID WE ARE A ROBUST
RESPONSE.
HOW LONG WILL THAT RESPONSE
LAST?
WE HAD 15 PEOPLE PARTICIPATING
IN THIS.
NONE OF THEM WAS INFECTED AND
NONE OF THEM WAS INFECTED DURING
THE PERIOD OF THE STUDY SO
EVERYTHING HERE IS VACCINE
INDUCED.
WE TRACKED THE SAME NODES SIX
TIMES FOR SIX MONTHS AFTER
SECONDS IMMUNIZATION.
HOW LONG THAT GERMINAL CENTER
CAN LAST?
THIS IS AN EXAMPLE WHERE A
REALLY SURPRISING FINDING FOR US
AT THE TIME THAT FOR SIX MONTHS,
10 OUT OF THE 15, HAD A REALLY
ROBUST GERMINAL CENTER GOING ON.
THIS IS GATED IN GC B-CELLS, 80%
TO THE SPIKE PROTEIN.
THIS IS A DRIVEN RESPONSE.
AND THE SUMMARY DATA SO THE
DETAILS ARE REALLY OUT THERE.
WE HAVE SOMETHING WE HAVE NEVER
EXPERIENCED IN ANY ANIMAL THAT
YOU CAN HAVE THE TIME.
SUCH A ROBUST AND PERSISTENT
GERMINAL CENTER REACTION IN
HUMANS.
BUT THEN ANOTHER THING WE
NOTICED IS EVERY TIME WE SEE THE
GERMINAL CENTER WE SEE LYMPH
NODE PLASMA CENTERS.
YOU CAN DETECT PLASMA BLAST
EARLY IN BLOOD, BUT PLASMA CELLS
IN THE LYMPH NODE, I ASSUME,
THESE ARE COMING OUT OF THE GC.
THAT IS ONE OF THE OUTCOMES OF
THE GCs.
WE DETECT THESE AS 38 HIGH,
POSITIVE.
SOME OF THEM BIND TO THE SPIKE.
THE VCR GETTING DOWN REGULATED.
WE POSTULATED THOSE CELLS ARE
LIKELY GOING TO BECOME THE ONES
THAT EXIST THE GC AND GO THROUGH
THE LYMPH NODE.
AT THE TIME WE DIDN'T HAVE SOLID
DATA FOR THAT.
WHAT HAPPENS IN THE BONE MARROW?
11 OUT OF THE 15, GIVE US ANS
ASPIRATE, WE ISOLATED THE BONE
MARROW CELLS AND THOSE, THINK OF
THEM AS CONTINUOUSLY ANTIBODIES
SECRETING MACHINES.
YOU DON'T NEED TO STIMULATE
THEM, PUT THEM OVERNIGHT ON
PLATES WITH THE ANTIGEN.
EVERY SINGLE PERSON IN THOSE 11
HAD A REALLY, LOTS OF PLASMA
CELLS AGAINST INFLUENZA.
1 IN 100 OF OUR PLASMA CELLS ARE
SPECIFIC TO INFLUENZA ANTIGEN,
MUCH LESS IN TETANUS DIPHTHERIA,
FOR NOSE 9 OF 11, VERY LOW, BUT
DETECTABLE SPIKE SPECIFIC PLASMA
CELLS.
WE THOUGHT THIS IS GREAT.
WE CAN DETECT THE PLASMA.
IT IS MUCH LOWER THAN INFLUENZA,
UH BUT IT IS THERE.
CAN WE, IF YOU LOOK AT BLOOD,
YOU WILL SEE THIS IS LIKE
EVERYONE ELSE YOU HAVE A VERY
NICE INCREASE EARLY ON, BUT THEN
YOU HAVE THIS DECAY WHICH GOT
SOME PEOPLE CONCERNED AFTER THE
PEAK RESPONSES.
WE THOUGHT THIS IS NORMAL
BECAUSE THIS WILL BE THE PLASMA
PLUS DRIVEN ANTIBODIES AND THESE
PLASMABLASTS WILL DIE, BUT LATER
YOU HAVE GERMINAL CENTER AND
BONE MARROW CELLS.
THESE ANTIBODIES SHOULD BE MORE
MATURE.
BETWEEN THESE TWO TIME POINTS
THERE IS ROBUST GERMINAL CENTER
REACTION.
TO ENSURE THIS IS THE CASE, SO
WE PICKED EIGHT INDIVIDUALS THAT
GIVE US THE THREE TIME POINTS,
BLOOD, LYMPH NODE AND BONE
MARROW AND SEQUENCE THE GERMINAL
CENTER FROM THE LYMPH NODE AND
THE LYMPH NODE PLASMA CELLS.
WE MADE THE CORRESPONDING
MONOBJECTION CLONAL ANTIBODIES
AND TESTED THAT.
THAT GAVE US MORE THAN 1,500
CLONES COMBINED BETWEEN THE
EIGHT INDIVIDUALS BINDING TO THE
SPIKE PROTEIN.
NOW WE HAVE MORE THAN 15,000
FINGERPRINT FROM EIGHT
INDIVIDUALS WE CAN FOLLOW OVER
TIME.
DOES THIS ROBUST GERMINAL CENTER
REACTION GIVE YOU THIS
ENHANCEMENT OF YOUR CAPACITY TO
ACTUALLY NEUTRALIZE THE VIRUS.
WHAT IS THE POINT OF INVESTING,
OF HAVING THIS ROBUST GERMINAL
CENTER REACTION.
TO GO BACK TO THE SEQUENCING
DATA.
EVERYTHING I'M GOING TO SHOW YOU
IS ANGO SPECIFIC.
WE TESTED THESE MODELS AND ASKED
THE SAME QUESTION, IF WE LOOK AT
THE PLASMABLAST IN BLOOD EARLY
ON, HOW MUCH OF THAT RESPONSE
OVERLAPS WITH THE GC?
IN THE EIGHT INDIVIDUALS WE SEE
VARIABLE FRACTIONS.
EVERYONE SHOWED SOME OVERLAP.
THIS IS CONFIRMING WHAT WE SAW
WITH INFLUENZA.
YOU CAN HAVE MEMORY CELLS IN THE
GC.
IN THE PLASMABLAST AND --
BUT IMPORTANTLY, WE CAN TRACK IF
A SAME CLONE BECAME A
PLASMABLAST.
A CRONE WILL PRORIFF PROLIFERATE
AND WE CAN TRACK THE SAME CLONE.
IF IT IS A PLASMA BLAST IS THE
LEVEL OF MUTATIONS.
PLASMA BLAST DETECTED ONE WEEK
AFTER.
GC HAS ALMOST THE SAME MUTATION.
IF YOU CHECK THIS OVER TIME, YOU
SEE THE SAME CLONE IN THE
GERMINAL CENTER EMANATED MUCH
MORE SIGNIFICANT MUTATIONS.
HAVING THAT GERMINAL CENTER IS
GREAT BECAUSE IT IS WHAT WE
SHOULD EXPECT.
LYMPH NODE PLASMA KROLLS, I
MEENGSED THEY ARE LIKELY COMING
FROM THE GC.
IF THIS IS THE CASE THEY SHOULD
BE IDENTICAL AND MUTATION RATES
SHOULD NOT HAVE THAT DYNAMICS.
IF WE ARE COMPARING THEM TO GC B
CELLS THEY SHOULD BE ON THE SAME
LEVEL.
WHAT YOU SEE HERE IS THE
MUTATION RATE.
WHAT ABOUT THE BONE MARROW
PLASMA CELLS.
ARE THE CELLS THAT ARE SPIKE
SPECIFIC, ARE THEY COMING FROM
HIGH LEVEL OF MUTATIONS OR LOW
LEVEL?
SO WE DID THIS ANALYSIS
COMPARING PLASMA BLAST, LYMPH
NODE AND BONE MARROW AT THE
SIX-MONTH TIME POINT.
THAT IS WHAT WE SEE HERE, A
RESPONDING PLASMA BLAST,
MUTATIONS OVER TIME AND
STRIKINGLY BONE MARROW PLASMA
CELLS HAVE ALMOST THE SAME
MUTATIONS AT THE LYMPH NODE AT
THE SAME TIME POINT.
THIS GIVES US THIS IDEA THAT,
WELL, ACTUALLY, IT IS LIKELY
THOSE LYMPH NODE PLASMA CELLS
ARE SHIPPED TO THE BONE MARROW,
WHICH IS GREAT.
THE CELLS THAT ARE COMING OUT
HERE, THE ANTIBODIES PRODUCED BY
THOSE CELLS SHOULD BE HIGHER
QUALITY.
WE HAVE ALL THE SEQUENCES AND WE
KNOW THERE ARE ANGI SPECIFIC.
WE CAN EXPRESS THE ANTIBODIES AS
A PLASMA BLAST.
WE CAN EXPRESS IT AS A BONE
MARROW PLASMA CELLS ADDING THE
MUTATIONS NATURALLY ACCUMULATED.
THE FIRST ANTIBODY BINDING
AFFINITY.
OR BY NEUTRALIZATION.
YOU CAN SEE IN BOTH CASES
ANTIBODY THAT BECAME STRONGER
BINDING FOR THE MOST PARTS IT
BECAME BETTER NEUTRALIZERS.
AFTER THESE TWO DOSES, THIS IS
THE FIRST TWO DOSES WERKS HAD A
REALLY ROBUST RESPONSE.
MOST OF THE EARLY ANTIBODIES
WERE DRAWN FROM SHORT LIVED
PLASMA BLAST.
AFTER A ROBUST GERMINAL CENTERS
WHERE B CELLS ARE RESPONDING
ALLOWED THEM TO BECOME LATER
LIKELY THE BONE MARROW CELLS.
THE ANTIBODY WILL BE LESS IN
MAGNITUDE, BUT HIGHER IN
QUALITY.
WE PUBLISHED THIS YEAR LAST WEEK
AND OBVIOUSLY, WE ALL KNOW WHAT
HAPPENED.
THE VIRUS DIDN'T -- AND WE GOT
TO LEARN MORE LESSONS FROM THE
FAROE ISLANDS.
A PREPRINT CAME OUT IN DECEMBER
2021 ABOUT A MEETING IN HEALTH
CARE WORKERS IN THESE ISLANDS OF
PEOPLE, 33 HEALTH CARE WORKERS
HAD THIS SOCIAL GATHERING.
ALL OF THEM HAVE BEEN TRIPLE
VACCINATED WITH THE PFIZER
VACCINE, TRIPLE VACCINATED AND
BOOSTED.
ALL OF THEM TESTED NEGATIVE
BEFORE THAT SOCIAL GATHERING.
WE KNOW WHAT HAPPENED.
OMICRON CAME IN.
OUR VACCINE DIDN'T BLOCK THE
INFECTION, EVEN THREE DOSES
AFTER ALL OF THESE RESPONSES.
I'M NOT GOING TO DISCUSS WHY
THIS IS THE CASE BECAUSE THERE
ARE SO MANY REASONS.
I'M NOT GOING TO STUDY THAT, BUT
THERE'S MANY REASONS WHY WE CAN
HAVE A GOOD IMMUNE RESPONSE BUT
ONCE THE VIRUS ANTIGENETICALLY
CHANGED.
OMCRON HAS SIGNIFICANT
MUTATIONS.
MODERNA MADE THE BIVALUE LENT.
MODERNA ASKED CAN WE SEE IF
HAVING A BIVALENT VACCINE WILL
HELP IN REDUCING NAIVE B-CELLS?
IF YOU BOOST PEOPLE THAT HAVE
ALREADY BEEN IMMUNIZED WITH MRNA
VACCINE, THEY HAVE THIS REALLY
NICE MEMORY B-CELL COMPARTMENT,
IF YOU BOOST THEM WITH A VACCINE
WITH NOVEL SPIKE, CAN WE GET A
RESPONSE?
WE RECRUITED NINE PEOPLE, GAVE
THEM THE BIVALENT VACCINE AND
EVERYBODY RESPONDED ROBUSTLY TO
THE PLASMA BLAST.
HOW ABOUT THE GERMINAL CENTER.
EIGHT WEEKS, THIS IS TWO MONTHS.
REMEMBER, MY EXPERIENCE WITH
INFLUENZA IS THE GERMINAL CENTER
IS WHERE IT SHOULD BE.
WE HAVE THIS WINDOW TO INDUCE
NAIVE B-CELLS.
LET'S INVESTIGATOR -- LET'S GO
FOR THE GERMINAL CENTER.
AND WE MADE SURE THAT NONE OF
THE INDIVIDUALS GOT INFECTED.
WE LOOKED AT THE RESPONSE
AGAINST SARS-COV-2 AND IT WAS A
FLATLINE.
AFTER THE NINE PEOPLE, FEW
RESPONDED, WELL, WE ALMOST SAW
RESPONSES IN EVERYONE EXCEPT
THIS PERSON.
FIVE IN THE MIDDLE HERE
RESPONDED VERY ROBUSTLY AGAINST
THE SPIKE PROTEIN.
GC B-CELLS, THE SPIKE PROTEIN.
WE DON'T KNOW HOW MANY OF THOSE,
WITH THE ORIGINAL SPIKE.
WE DON'T KNOW HOW MANY OF THOSE
COULD BE NAIVE OR COULD BE
COMING AGAINST THE OMICRON
SPIKE.
WE WENT BACK TO THE GC B-CELLS
FROM THE FIVE INDIVIDUALS AND
SINGLE CELL SORTED EVERY SINGLE
ONE.
AND THIS IS WHAT I'M SHOWING YOU
HERE.
WE LOOKED AT THE LYMPH NODES,
AND THE 10X, SINGLE CELL
SEQUENCE OF B 10X ANALYSIS OF
THOSE RESPONDING B-CELLS.
WE CAN LOOK AT THE GERMINAL
CENTER.
TOOK EVERY ONE OF THEM, MADE THE
CORRESPONDING ANTIBODIES, TESTED
AGAINST THE SPIKES AND MAKE SURE
THEY ARE ALL BINDING TO THE
SPIKE.
BUT WE TESTED THEM AGAINST BOTH
SPIKES, ARE THEY BINDING TO THE
ORIGINAL SPIKE OR BINDING TO THE
OMICRON SPIKE OR BINDING TO
BOTH?
WE EXPECT A LOT TO BE CROSS
REACTIVE BECAUSE THOSE PEOPLE
ARE IMMUNE.
WE DIDN'T GET A SINGLE YAENT
BODY THAT BOUND TO B1, OMICRON,
BUT NOT TO THE ORIGINAL.
THIS IS NOT ABOUT DETECTION.
WE SEQUENCED EVERY CELL AND MADE
THEM MONOCLONAL.
IT IS NOT THAT WE COULDN'T
DETECT THEM BECAUSE WE DIDN'T GO
DEEP ENOUGH.
WE WENT AS DEEP AS WE COULD GO
IN THIS FIVE INDIVIDUALS AND
COULDN'T DETECT A SINGLE B1
SPECIFIC.
SO I THOUGHT MAYBE WE WENT TOO
EARLY.
HERE EIGHT WEEKS IS MAYBE TOO
EARLY.
AND ALSO THE OTHER THING IS THIS
VACCINE HAS A 50% ORIGINAL
STRAIN MRNA SO MAYBE THAT
BIASSED THE RESPONSE TO THE
CROSS REACTIVE ANTIBODIES WE SEE
HERE, THE 71% HERE.
I REACH OUT TO MODERNA AND I
SAID YOU HAVE THE MONO VALENT
VACCINE.
SO IF I COMPARE THAT AND INSTEAD
OF LOOKING AT EIGHT WEEKS LOOK
AT FOUR MONTHS AFTER BOOST, I
CAN SEE IF THERE IS A CHANCE OF
GETTING THE NAIVE RESPONSE TO BE
ENGAGED.
I COMPARED THAT TO PEOPLE WHO
GOT LIKE MOST OF US THE ORIGINAL
VACCINE AS A BOOSTER
IMMUNIZATION.
SO WE HAVE THE EIGHT PEOPLE THAT
WE GOT THE MONO VALENT AND WE
PICKED THREE PEOPLE FROM EACH
AND SORTED THEIR MEMORY CELLS AT
FOUR MONTHS' TIME POINT.
WE LOOKED AT TOTAL MEMORY
WHATEVER BOUND TO EITHER SPIKE,
SINGLE CELL THEM AND ASKED THE
SAME QUESTION, CAN ANY OF THE
ANTIBODIES BIND TO THE OMICRON?
WELL, IF YOU HAVE BEEN ONLY
IMMUNIZED WITH THE ORIGINAL
STRAIN, SO FOR EXAMPLE, TWO
PFIZER, TWO MODERNA EARLY ON AND
GOT THE THIRD, THE MONOCLONAL IS
COMING OUT.
YOU SHOULDN'T EXPECT ANYTHING TO
BE OMICRON SPECIFIC AND THAT IS
WHAT WE SEE HERE.
IF YOU HAVE BEEN EXPOSED TO THE
ORIGINAL STREAM, PFIZER OR
MODERNA, YOU HAVE 2/3, BUT 1/3
SPECIFIC TO ORIGINAL.
AS YOU WOULD EXPECT, THERE IS
NOTHING HERE SPECIFIC TO THE
OMICRON BY ITSELF.
BUT NOW I PREDICTED THAT WE HAVE
WAY MORE ANTIBODIES AGAINST BA1
AND I WAS WRONG.
NONE OF THE ANTIBODIES -- SO IF
YOU HAVE BEEN BOOSTED WITH MONO
VALENT BA1 AND WAIT FOUR MONTHS,
THERE IS STILL SIGNIFICANT CROSS
REACTIVE ANTIBODIES, BUT NONE
BOUND TO THE BA1.
I THOUGHT THIS IS VERY DIFFERENT
FROM WHAT WE EXPECT, FROM WHAT I
EXPECTED.
SO I THOUGHT MAYBE BECAUSE WE
ARE LOOKING AT MEMORY, WE ARE
NOT GOING DEEP ENOUGH.
TO GO DEEPER IN THIS COMPARTMENT
AND DETECT ANY BA1 SPECIFIC, I
SORTED THE BA1 SPECIFIC MEMORY
CELLS.
THIS IS VERY TARGETED TO BA1.
FROM THOSE CELLS, THE ONE THAT
TENDED TO NOT BIND FROM THE
WASHINGTON STRAIN.
WE MADE THE MONOCLONALS AND CAN
WE SEE THE BA1 CELLS?
AFTER DOING THAT, SIX
MONOCLONALS THAT ONLY BOUND TO
BA1 NOT THE WA1.
THEY SHOULD HAVE SLIGHTLY LOWER
MUTATIONS AND INDEED HAVE LOWER
MUTATIONS COMPARED TO THE CROSS
REACTIVE ONES.
ONE COULD ASK, WHY DO WE CARE?
WHY THE INSISTENCE OF SHOWING WE
HAVE THESE NAIVE RESPONSES?
BECAUSE THOSE SIX MONOCLONALS,
RESIST BA1.
IT IS NOT A LUXURY.
YOU ACTUALLY INCREASE YOUR
ABILITY TO ADD TO YOUR
NEUTRALIZING BA SPOBSS TO THE
ORIGINAL STRAIN.
THE FACT BECAUSE THEY ARE
NEUTRALIZING, WE WERE ABLE TO
MAP WHERE THEY BIND.
THIS IS WITH THE VSV, NOT THE
REAL VIRUS, PSEUDOTYPE.
THE VSV WERE PSEUDOTYPE WITH BA1
AND WE GENERATED, WE SEQUENCED
FROM THIS NEUTRALIZATION AND WE
WERE ABLE TO SHOW THE RESIDUES
THEY ARE LIKELY RECOGNIZING ARE
ONLY SPECIFIC TO BA1.
SO AFTER ALL OF THAT, WHAT WE
LEARNED IS THE BA1 RESPONSE OR
THE OMICRON RESPONSE CAN ENGAGE
IS VERY IMMUNOGENI THAT
GENERATED MANY B-CELLS.
A BOOSTER ENGAGED THE MEMORY
B-CELLS AND BROUGHT THEM BABLG
TO THE GERMINAL CENTER REACTION.
WE SHOWED WITH INFLUENZA AND
SHOWED WITH THE BA1 VACCINE.
CHANGING THE VACCINE CAN ALLOW
ENGAGEMENT OF BA CELLS.
THIS ENGAGEMENT CAN BE NOT VERY
EFFICIENT.
IF YOU ARE CLOSELY RELATED
SEQUENCE WISE TO THE ORIGINAL
STRAIN, THIS WILL ALLOW A LOT OF
MEMORY CELLS TO BE ENGAGED.
AND THEN ALSO WE KNEW FROM OUR
PREVIOUS WORK WITH RAFY, ONE
DOSE MIGHT NOT BE ENOUGH TO GET
A DETECTABLE RESPONSE.
ONE DOSE LIKELY MULTIPLE
RESPONSE TO OMICRON WILL
INCREASE AND MORE DETECTABLE IN
THE NYE VEEF RESPONSES.
ANOTHER SURPRISE WE LEARNED IN
OUR STUDY, WE DIDN'T PREDICT AT
ALL, IS GERMINAL RESPONSE IN
HUMAN CANS BE PERSISTENT AND
LONGER THAN ANY PREDICTION FROM
SMALL ANIMAL MODELS.
WE WERE THINKING THIS IS
SOMETHING SPECIFIC TO THE MRNA
PLATFORM, BUT WE DON'T THINK
THIS IS THE CASE.
WE THINK THIS IS REALLY
SOMETHING THAT RELIES ON THE
CONTEXT WHERE THE VACCINE WAS
GIVEN OR WHO GETS THE VACCINE
AND THE PREEXISTING IMMUNITY.
THIS WORK COMES FROM -- LAB AS
WELL.
THE PREEXISTS BASELINE SITS
THERE AT THE ROBUSTNESS AND
DURATION OF THE RESPONSE.
LE INDUCTION OF PERSISTENT GC IS
WHAT KEEPS IT GOING.
WE REALLY DON'T KNOW.
WE WOULD LIKE TO SHOW THAT.
AND ALSO THIS BRINGS ME TO
SOMETHING THAT IS NOT IDEAL.
YOU ONLY LOOK AT CELL
SUSPENSION, SO YOU CANNOT GET
ANY INFORMATION ABOUT
PERSISTENCE.
OVER THE LAST YEAR WE OPTIMIZED
OUR TOOLS INSTEAD OF GETTING
ASPIRATES, WE GET ASPIRATED PLUS
CORE WITH THESE.
I WILL SHOW YOU DATA WE JUST GOT
THE LAST COUPLE OF WEEKS, AFTER
SOME TRIALS AND ERRORS WE CAN
GET CORE BIOPSIES.
WE CAN STAIN FOR THE GERMINAL
CENTER.
THIS WAS VERY EXCITING FOR US TO
BE ABLE TO SEE THAT WITHIN THE
B-CELL FOLLICLES WE CAN SEE A
RESPONDING GC FROM SOMEONE
IMMUNIZED WITH MRNA VACCINE.
THE NEXT QUESTION FOR US IS CAN
WE SEE HOW LONG THE ANTEGEN CAN
PERSIST AND HOPEFULLY THIS WILL
BE COMING NEXT.
I MENTIONED A LOT OF
EXPERIMENTS, A LOT OF WORK I
DID.
I DIDN'T GENERATE ANY SINGLE
PIECE OF DATA.
I AM REPRESENTING THE TERRIFIC
TEAM THAT DID ALL THE WORK IN MY
LAB.
OBVIOUSLY, YOU CAN ASSUME THERE
IS A LOT OF COLLABORATORS THAT
ARE BRILLIANT AND TERRIFIC, NOT
ONLY TAKING CARE OF PATIENTS,
BUT HELPING US WITH THESE
STUDIES.
A LOT OF COORDINATORS THAT WERE
VERY KRCRITICAL.
INSIDE AND OUTSIDE OF WSU.
FUNDING, YOU CANNOT DO THIS
WITHOUT GENEROUS VOLUNTEERS.
I'M HAPPY TO TAKE QUESTIONS.
>>THANK YOU, ALI, VERY
FASCINATING TALK.
BEFORE WE START, THE CMA CODE
FOR TODAY'S TALK IS 44480.
SO THE FIRST QUESTION IS YOUR
STUDIES HAVE SHOWNED THAT GC
REACTIONS ARE MAINTAINED FOR A
LONG TIME, SIX MONTHS OR LONGER,
ALLOWING INFECTIONAL
VACCINATION.
OTHER STUDIES HAVE SHOWN THERE
IS A WANING OF NEUTRALIZING
ANTIBODIES OR VACCINE EFFICACY
TWO TO MONTHS FOLLOWING
VACCINATION.
HOW DO YOU RECONCILE THOSE TWO
FINDINGS THAT LONG LASTING LYMPH
NODE B-CELL ACTIVITY WITH THE
ANTIBODIES RESPONSE AND VACCINE
EFFICACY?
>>THAT IS A VERY IMPORTANT AND
VERY COMMON QUESTION I GET ABOUT
THIS.
SO WE HAVE TO DISTINGUISH A
COUPLE OF POINTS.
WANING IS SOMETHING I'M NOT
MEASURING.
THE VIRUS IS DIFFERENT, CHANGED
SIGNIFICANTLY IN THE CASE OF
OMICRON.
IN CASE OF DELTA THE VIRUS
INTRINSICALLY BECAME MORE
INFECTIOUS.
SO THERE ARE OTHER FACTORS THAT
DECIDES WHAT THE OUTCOME OF
EXPOSURE, IN TERMS OF
SYMPTOMATIC.
IT IS VERY DIFFERENT THINGS.
WHAT I'M LOOKING AT IS DETECTION
OF ANTIBODIES.
NOW, YOUR FIRST QUESTION, WHICH
IS A VERY IMPORTANT QUESTION, SO
MRNA VACCINES, THIS IS THE
THEORETICAL FRAMEWORK I'M
CURRENTLY THINKING ABOUT, MRNA
VACCINES DELIVER A LARGE DOSE OF
PROTEIN EARLY ON.
THAT DRIVES A LOT OF PLASMA
BLAST RESPONSE.
ALMOST DISPROPORTIONATE
PLASMABLAST RESPONSE.
WE GET THIS NICE SPIKE EARLY ON
IN ANTIBODIES YOU CAN SEE A VERY
NICE INCREASE.
AND ONCE THESE CELLS, NOT ALL OF
THOSE CLONES, RESPONDING CLONES,
SO ONCE THOSE CELLS START TO
DIE, YOU CAN SEE THIS, AGAIN,
DRAMATIC DECAY, WHICH I THINK
GOT PEOPLE CONCERNED.
WHAT I THINK IS IF YOU JUST LOOK
IN TERMS OF -- UNFORTUNATELY, WE
DIDN'T HAVE A CHANCE TO HAVE A
THREE, FOUR-YEAR-LONG STUDY TO
SEE THERE IS A PLATEAU OF THE
ANTIBODY RESPONSE YOU CAN DETECT
AFTER SIX TO SEVENTH RESPONSE.
YOU CAN SEE THE ANTIBODY
REACHING PLATEAU, NO MORE DECAY,
BUT AT THE TIME, IF YOU GOT
INFECT OR BOOSTED YOU COULDN'T
SEE THAT FOR A LONG PERIOD, BUT
IT WAS THERE.
SO I THINK THERE ARE INDUCTION
OF PLASMA BLASTS.
BUT IF YOU COMPARE IN THE SHORT
TIME PERIOD.
I KNOW SIX MONTHS DOESN'T SOUND
LIKE A SHORT TIME, BUT IF YOU
COMPARE WITH SUCCESSFUL
VACCINES, SMALLPOX AND HVC, 10
YEARS ARE FLATLINES.
THAT IS YEARS COMPARED TO SIX
MONTHS.
YEAH, IF YOU DO THE HALF LIFE
DECAY, IT WOULD LOOK REALLY
DRAMATICALLY BAD FOR MRNA
VACCINATION.
I DON'T THINK THERE IS SOMETHING
ISO INHERENTLY IN HUMANS THAT
MRNA CANNOT DELIVER A LONG TERM
ANTIBODY.
>>TO FOLLOW UP ON THAT PART,
SINCE YOU MOSTLY, AT LEAST
SHOWED MRNA BASED VACCINES,
RIGHT, HAVE YOU LOOKED AT FOR
COVID MRNA AS A PROTEIN OR ADD
FIVE BASE VACCINES.
DO THE GC RESPONSES DIFFER?
>>YEAH.
WE DID THE SMALL TRIAL, AS YOU
MIGHT RECALL DURING THE TIME
THERE WAS PAUSE IN USING IT.
WE COULDN'T REALLY, IDEALLY, WE
WOULD HAVE LOVED TO DO THAT
STUDY HEAD TO HEAD, MRNA VERSUS
NOVA VAX AND JNJ BASED.
RIGHT NOW IT IS VERY HARD
BECAUSE OF THE DIVERSE AND
HETEROGENEITY IN THE POPULATION.
IT IS HARD TO DO IT HEAD TO
HEAD.
UNFORTUNATELY, WE DIDN'T HAVE
THE OPPORTUNITY.
WE ARE DOING IT NOW BUT NOT WITH
SARS-COV-2.
INFLUENZA MRNA AND REGULAR.
THAT HOPEFULLY WILL BE COMING
OUT SOON.
>>IN THAT CONTEST, FOR THE FLU,
HAVE YOU LOOKED AT AT
[ INDISCERNIBLE ]
>>GREAT QUESTION.
ONE OF THE STUDIES I STARTED
BEFORE THE PANDEMIC AND ONE OF
THE STUDIES WHY I HATED TO
PANDEMIC, IT GOT CUT.
WE WERE COMPARING THE FLU VIRUS
AND THE REGULAR.
AND THAT IS THE SAME
FORMULATION.
I WOULD LOVE TO GO BACK AND DO
THAT STUDY AGAIN.
WE ARE NOW COMPARING MRNA WHICH
ARE DIFFERENT, BUT COMPARING TO
THE CONVENTIONAL.
ADDING MRNA TO THAT WOULD BE
GREAT.
>>IN THIS CONTEST OF VACCINES,
HAVE YOU LOOKED AT ELDERLY
POPULATION TO SEE IF RESPONSE IN
GCs OR LYMPH NODE DIFFER?
>>YEAH.
AGAIN, ANOTHER GREAT QUESTION.
IN THE BOOSTER STUDIES, THIS IS
THE DATA THAT -- OH, HERE IT IS.
THREE OUT OF THOSE FIVE
INDIVIDUALS WERE 65 AND ABOVE.
WHICH IS REALLY, FOR ME, ANOTHER
FASCINATING ASPECT THAT WE CAN
SEE SUCH A ROBUST RESPONSE.
BUT WE DON'T HAVE NUMBERS REALLY
TO BE ALLOWED TO SAY SOMETHING
DEFINITIVELY.
WE HAVE A STUDY THAT IS PLANNING
TO ENROLL HOPEFULLY IN THE FALL
WHERE WE ARE DIRECTLY ASKING
THAT.
SAME VACCINE.
MRNA VERSUS CONVENTIONAL, 65
ABOVE FOR OLDER INDIVIDUALS OR
18-50 FOR YOUNG ADULTS.
>>SO A QUESTION ON THIS SLIDE,
SINCE YOU ARE SHOWING THE FNA,
DID YOU LOOK AT BONE MARROW AND
IF YOU SEE CHANGE IN BOOSTING
EFFECTS
>>WE DID.
I HAVE THAT IN EXTRA SLIDES.
I DIDN'T KNOW IF I WILL GO OVER
TIME.
SO BONE MARROW IS HERE.
SO THIS IS THE BONE MARROW LEVEL
AGAINST THE SPIKE.
IF YOU HAVE NEVER BEEN INFECTED
OR VACCINATED FOR SARS-COV-2,
THESE PEOPLE ARE HARD TO FIND.
YOU HAVE ALMOST BASICALLY AN
UNDETECTABLE BONE MARROW AGAINST
THE SPIKE PROTEIN.
IF YOU HAVE BEEN INFECTED ONCE
AND NEVER BEEN VACCINATED, THIS
IS ONE IN 1,000, YOU HAVE SPIKE
SPECIFIC, NOTHING COMPARED TO
FLU.
IF YOU HAVE BEEN VACCINATEDMAN
WITH TWO DOSES AND NOT INFECTED
YOU ALMOST HAVE THE SAME LEVEL
AS INFECTED.
IF YOU HAVE BEEN VACCINATED AND
BOOSTED, THIS IS YOUR LEVEL.
SO, YES, BOOSTING NOT ONLY GETS
YOU A GERMINAL CENTER REACTION
IT CAN INCREASE FREQUENCY OF
BONE MARROW PLASMA CELLS.
THIS IS A YEAR AND A HALF OLD.
RIGHT NOW MANY PEOPLE GOT NOT
ONLY TRIPLE IMMUNIZATION, BUT
GOT INFECTED, INFECTED TWICE,
FOUR IMMUNIZATION.
WE ARE VERY CLOSE TO INFLUENZA
FOR SARS-COV-2.
>>IN TERMS OF GC, THE QUESTION
COMES IS THERE A RELATIONSHIP OF
DURATION OF GC ANDEAN YAN AT
EACH TIME POINT.
DO SOME OF THE B-CELLS MORE
MATURE TEND TO LEAVE EARLY
COMPARED TO OTHERS IN THE
OPPORTUNITY IN DIFFERENT
INDIVIDUALS IS DIFFERENT OR
EVERYBODY THE SAME?
>>YES.
ALL THE QUESTIONS ARE BEAUTIFUL
TODAY.
IT IS HARD -- IT IS A TOUGH ONE.
IF WE DO SEE, OBVIOUSLY, FROM
THE LEVEL HERE OF MUTATION OVER
TIME, WE DO SEE, SORRY, I'M
MOVING THE SLIDES TOO FAST BUT
I'M TRYING TO GET TO THE SLIDE
THAT -- OH, HERE IT IS.
SO GC OVER TIME, THESE ARE THE
LYMPH NODE PLASMA CELLS I SHOWED
YOU EARLIER, SAME LEVEL OF GC
B-CELLS.
WE MADE MONOCLONAL FROM THE SAME
CLONE OVER TIME.
SOMETIMES YOU CAN DETECT THE
GRADUAL INCREASE IN BINDING
AFFINITY.
WHETHER THIS AFFECTS THE OVERALL
SERUM RESPONSE, WE HAVEN'T
LOOKED AT THAT.
TO BE HONEST, I DOUBT WE WILL BE
ABLE TO DETECT THAT IN A GRADUAL
FASHION.
I THINK WE CAN DETECT IT FROM
ZERO, BASELINE VERSUS SIX
MONTHS.
TO SEE THAT IMPACT OVERALL IN
SERUM, THESE HAVE TO
DIFFERENTIATE FROM THE -- CELLS
AND HAS TO BE A SIGNIFICANT
FRACTION OF POLY CLONE
ALRESPONSE.
>>TO FOLLOW ON THAT, WE COMBINE
TWO QUESTIONS.
IN THE LYMPH NODE OF POST
VACCINATED SUBJECTS, DO YOU
DETECT ATYPICAL B-CELLS AND HAVE
YOU LOOKED AT AUTOANTIBODIES
RESPONSES IN THE LYMPH NODE
CELL?
>>I WILL START WITH THE SECOND
ONE.
WE DID NOT TEST THAT.
WE DID NOT, I MEAN, WE HAVE THE
MONOCLONALS.
WE DID NOT -- WE DON'T DO THIS
ROUTINELY IN OUR LAB SO WE DID
NOT TEST -- I DON'T KNOW WHAT
WILL BE THE DEFINITION.
I GUESS BINDING TO SOME OF THE
ANTIGEN STRAND OF DNA.
WE HAVEN'T DONE THAT SO I DON'T
KNOW.
THE FIRST QUESTION ABOUT -- THE
FIRST QUESTION WAS -- CAN YOU
REMIND ME, ACTUALLY?
>>IN YOUR LYMPH NODE STUDIES
ARE THERE VACCINATION FOLLOWING
INFECTION, HAVE YOU DETECTED ANY
ATYPICAL MEMORY --
>>OH, YES.
THE DEFINITION IS THE -- I'M
ASSUMING.
WE DID SEE THOSE CELLS.
I CAN'T REMEMBER MAINLY IN BLOOD
OR LYMPH NODES, WE CAN DETECT
THEM, WE CAN SEE THEM.
THE FREQUENCY IS NOT AS HIGH.
ONE OF THE THINGS WE ARE TRYING
TO LEARN AND WE DON'T HAVE A
DEFINITIVE ANSWER IS ARE THOSE
CELLS REALLY COMING OUT FROM THE
GC OR ARE THESE CELLS GOING INTO
THE GC.
IT IS HARD FOR US TO TELL WHERE
ARE THESE CELLS COMING FROM OR
ARE THESE CELLS GOING TO THE GC.
ARE THESE PHENOTYPE ACQUIRED
BEFORE OR AFTER THE GC.
WE CAN SEE THEM, BUT WE DON'T
HAVE A DEFINITIVE ANSWER YET.
>>A LOT OF QUESTIONS.
THE LAST TWO QUESTIONS, WE YOU
COMPARE HOMOLOGOUS OR
HETEROLOGOUS, ARE THEY THE SAME?
>>WE DIDN'T DETECT A DIFFERENCE
THERE.
IT IS NOT THE SAME.
IT LOOKED THE SAME.
>>SO THE LAST QUESTION IS BASED
ON WHAT YOU HAVE SHOWN IN TERMS
OF OKAY KRON VACCINATION AND
SMALL NUMBER OF B-CELLS SPECIFIC
FOR BA1 ONLY.
WHAT ARE THE IMPLICATIONS OF
THAT FINDING?
LET'S SAY DOWN THE ROAD WE WANT
TO HAVE THE NEWLY EMERGING
VARIANTS, THAT WILL TEND TO
GENERATE MOST ANTIBODIES THAT
ARE CROSS REACTIVE OR MORE
SPECIFIC FOR THE PROTOTYPE WA1
STRAIN.
HOW DO WE OVERCOME THAT?
HOW DO WE HAVE VACCINES
GENERATING MORE ANTIBODIES
AGAINST THE EMERGING STRAINS?
>>SO THE VERY FIRST PART, THE
VERY FIRST THING I LEARNED IS WE
DON'T REALLY NEED THE ORIGINAL
STRAIN IN THE VACCINE ANYMORE.
OMICRON BY ITSELF, IT IS VERY
CLEAR, CAN REENGAGE MEMORY CELLS
EFFICIENTLY.
AT LEAST AS A BOOSTER, NOT AS A
PRIMING.
AS A BOOSTER IMMUNIZATION, WE
DON'T NEED THE BIVALENT AND IN
MY OPINION WE CAN GO WITH THE --
WE SEED A VARIANT ANTIGENICALLY
DIFFERENT.
OMICRON IS 97.5% SIMILAR TO THE
ORIGINAL IN TERMS OF HEMOLOGY.
IF WE THINK WE NEEDED TO SEE A
SIGNIFICANT ENGAGEMENT, I THINK
WE NEED EITHER TWO DOSES OR
SOMETHING THAT IS SIGNIFICANTLY
DIFFERENT.
AND THE SIGNIFICANTLY DIFFERENT
PART IS HARD TO MEASURE, BUT
OBVIOUSLY, OUR EXPERIENCE WITH
THE 97.5% WHICH IS OBVIOUSLY
DIFFERENT STILL NOT DIFFERENT
ENOUGH FOR ONE DOSE TO DO THE
JOB.
>>YOU BRIEFLY MENTIONED ABOUT
YOU ARE LOOKING AT OTHER
PLATFORMS.
>>YES.
>>IN TERMS OF BOOSTING, IS A
PLATFORM, LET'S SAY IF YOU ARE
INITIALLY PLANNED WITH MRNA, DO
YOU GET A BETTER BOOSTER WITH
PROTEIN-BASED?
>>UNFORTUNATELY, WE DON'T HAVE
THAT.
I MEAN IT WILL BE VERY -- I'M
VERY CURIOUS TO DO THESE STUDIES
BECAUSE IT WILL BE GREAT TO SEE
HOW THE EARLY DYNAMICS AND
ANTIGEN CLEARANCE WILL BE
DIFFERENCE BETWEEN THE COMMON
PROTEIN OR MRNA.
HOW THAT AFFECTS THE OVERALL
RESPONSE.
UNFORTUNATELY, WE DON'T HAVE
ACCESS TO THESE VACCINES.
I WOULD LOVE TO DO THESE
STUDIES.
IT IS AN IMPORTANT QUESTION FOR
ALL OF US TO LEARN.
>>THANK YOU SO MUCH, ALI.
IT WAS A FASCINATING TALK.
WE HAVE GONE OVER THE TIME, WE
LOOK FORWARD TO DISCUSSING MORE.
THANK YOU VERY MUCH.
>>THANK YOU FOR HAVING ME.
I ENJOYED ALL THE QUESTIONS.
THANK YOU SO MUCH.