IT IS A PLEASURE TO INTRODUCE SHANE CROTTY, REMARKABLY AS AN UNDERGRAD HE COMBINED THIS TRAINING AND WRITING AHEAD OF THE CURVE, A BIOGRAPHY OF DAVID BALTIMORE. IF YOU GOOGLE SEARCH FOR CROTTY, AND BALTIMORE AND CSPAN YOU WILL FIND A VERY YOUNG SHANE'S 2001 APEERPANCE ON BOOK TV TALKING ALL ABOUT HIS BOOK FOR 45 MINUTES. SHANE NEXT DID A Ph.D. WITH ALSO FANTASTIC PRODUCT OF DAVID BALTIMORE NAMED [INDISCERNIBLE] STUDY AT UCSF, STUDYING POLIO VIRUS, STUDYING THE LONG STANDING PROBLEM ON HOW THE 1 OF THE FIRST ANTIVIRAL DRUGS WORKS. AND SHANE SHOWED THIS WORKS MOSTLY IN VIVO BY INDUCING ERROR CATASTROPHE IN RNA VIRUSES INCLUDING THE POLIO VIRUS WE'RE STUDYING. SHANE ALSO SHOWED 1 OF MY FAVORITE PAPERS OF HIS THAT QUITE REMARKABLY POLIO VIRUS IS ABLE TO ESCAPE THE ANTIVIRAL EFFECTS OF--1 OF MY FAVORITE DRUGS WHICH TARGETS THE HOST SECRETORY PATHWAY, PEOPLE SAY THAT TARGETING THE HOST PATHWAY, YOU GET AROUND THE PROBLEM OF VIRAL AVOIDANCE ON THE HOST AND SHANE SHOWED BRILLIANTLY IN THIS PAPER WHICH I DON'T THINK A LOT OF PEOPLE KNOW ABOUT, THIS IS JUST IS NOT TRUE, VIRUSES ARE VERY, VERY CLEVER. I HAD THE PLEASURE OF FIRST MEETING SHANE WHEN HE APPLIED FOR A POST DOC IN MY LAB. HE MIGHT REMEMBER MEETING DR. HATCHER IN HIS BATH ROBE WHEN WE KNOCKED ON THE WRONG DOOR LOOKING FOR HIS SISTER WHO WAS LIVING ON CAMPUS AT NIH. IT WAS CLEAR TO ME AT THE TIME THAT SHANE WAS FULLY CAPABLE OF BEING A PI AND I THINK I MADE THAT CLEAR TO SHANE. I HOPE I DID, THE PROMISE HE SHOWED WAS JUST ABSOLUTELY REMARKABLE. HE CHOSE WISELY CHOSE ON DO A SHORT AND HIGHLY PRODUCTIVE POST DOC NOD IN MY LAB, WHERE IN JUST 2 YEARS HE BECAME AN EXPERT IN VIRAL IMMUNOLOGY, FOCUSING MORE OR LESS ON THE GENERATION OF MEMORY B-CELLS BUT I'M SURE HE GOT GREAT EXPOSURE TO T-CELLS WHICH [INDISCERNIBLE] LAB IS ALSO FAMOUS FOR. SHANE STARTED HIS LAB IN 2003 AND BASICALLY BECAME A WORLD LEADER IN ADAPTIVE ANTIVIRAL UMPIRESUNITY, PHOBEUSING ON T-CELL BIOLOGY, PARTICULARLY IN THE CONTEXT OF HIV AND VACCINATION. WHEN COVID CAME ALONG, SHANE ALONG WITH HIS CLOSE COLLEAGUE, ALONG WITH ALEX [INDISCERNIBLE] PUBLISHED CLASSIC STUDIES OF HUMAN BNT RESPONSES TO SARS-COV-2. MANY OF YOU KNOW SHANE FROM HIS NUMBER OF APPEARANCES ON THIS WEEK IN VIROLOGY, IT'S A FANTASTIC PODCAST FROM [INDISCERNIBLE]. IF YOU DON'T KNOW IT, JUST SEARCH FOR TWIV, AND CROTTY AND SHANE'S APPEARANCES ARE JUST FANTASTIC. HOW MANY TIMES HAVE YOU BEEN ON THERE, 4, 5? MANY. A LOT. IN ANY EVENT THIS IS A FANTASTIC FORUM FOR VIRUSES AND VIRAL IMMUNOLOGY AND SHANE IT'S A GREAT PLEASURE TO WELCOME YOU VIRTUALLY ANYWAY TO NIH AND I HOPE TO SEE YOU SOMETIME IN THE NEAR FUTURE IN PERSON. >>THANKS FOR THAT INTRO, I LOVE HOW YOU MENTIONED THE VIRUS PAPER. THAT LITERALLY WAS A BET OVER A BEER WITH ANOTHER GRAD STUDENT THAT THEY WERE THE CELL BIOLOGIST IN THE LAB AND DOING GREAT WORK AND THEY JUST SAID IT'S IMPOSSIBLE FOR THE VIRUS TO ESCAPE A CELLULAR PROTEIN REQUIREMENT AND BET A BEER ON IT. [LAUGHTER] AND IT WAS DETERMINED AND TESTED OUT. I WON'T BE TALKING ABOUT THAT FOR THE REST OF THE DAY. THANKS FOR HAVING ME AND THIS IS A GREAT FORUM AND A FANTASTIC GROUP OF RESEARCHERS TO TALK TO. I WILL TALK ABOUT PROJECTEDS WE'VE DONE IN THE CONTEXT OF COVID, AND SOME RELATED PROJECTS ACTUALLY THAT REALLY GET AT INFORMING, SHOWING OW COVID VACCINE STUDIES AND COVID IMMUNOLOGY INFORMS OTHER PROGRAMS. I'M GOING TO START HERE WITH SOMETHING YOU ALL KNOW WELL BUT IT IS CERTAINLY INCREDIBLY IMPORTANT AND OUR UNDERSTANDING OF COVID AND OBVIOUSLY THE COVID VACCINES HAVE REALLY BEEN HUGE THINGS OVER THE PAST COUPLE OF YEARS. OUR WORK IN THIS AREA REALLY STARTED WITH ALEX [INDISCERNIBLE] EARLY IN 2020 WHEN THERE WAS A HUGE AMOUNT OF SPHERE AND UNCERTAINTY ABOUT MOST EVERYTHING RELATED TO COVID-19 AND INCLUDING WHETHER THIS WAS A DISEASE THAT WAS A CONSEQUENCE OF A FACT THAT PEOPLE JUST DIDN'T MAKE IMMUNE RESPONSES OR ADAPTIVE IMMUNE RESPONSES TO THIS VIRUS AND/OR THAT PEOPLE MADE VERY UNUSUAL ADAPTIVE IMMUNE RESPONSES TO THIS VIRUS AND THOSE WERE THINGS THAT WE THOUGHT WE COULD POTENTIALLY HELP OUT WITH IN UNDERSTANDING AND THAT, SUCH UNDERSTANDING MAY BE HELPFUL FOR VACCINE PROGRAMS, WHICH VACCINE PROGRAMS WERE ALREADY UNDERWAY INCLUDING AMAZING EFFORTS BY THE VRC AND MODEL --MODERNA AND THEE LOTS OF CONCERNS ABOUT WHETHER SPIKE ALONE WAS A TARGET AND AGAIN, WHETHER THERE WAS SOME SORT OF IMMUNO DIVERGENT RESPONSES RELATED TO THIS VIRUS THAT MIGHT BE PROBLEMATIC FOR THIS EXAMPLE, FOR VACCINES OR IF WE WERE JUST GOING TO THE WRONG TYPE OF IMMUNE RESPONSES FOR THIS TYPE OF IMMUNITY. SO THAT WAS A SIMPLE TOPIC OF THIS PAPER FROM APRIL, MAY, FROM MAY 2020 WHERE WE SAID, WELL, WHAT ARE THE ADAPTIVE IMMUNE RESPONSES LOOK LIKE IN PEOPLE WITH AVERAGE CASES OF COVID-19. SO PEOPLE HAVE GOT INFECTED, DID NOT REQUIRE HOSPITALIZATION BUT CLEARED THE VIRUS AND PRESUMABLY MADE A SUCCESSFUL ADAPTIVE IMMUNE RESPONSE THAT CLEARED THE VIRUS, WHAT DID THAT IMMUNE RESPONSE LOOK LIKE AND IN BRIEF, WE SAW LIKE MULTIPLE LABS HAD SEEN AT THAT POINT THAT ESSENTIALLY ALL OF THOSE INDIVIDUALS MADE ANTIBODY RESPONSES DESPITE BUT NOTABLY THEY ALSO MADE CD4 T-CELL RESPONSES AND CD-4 T-CELL RESPONSES TO VIRAL PROTEINS AS WELL AS MANY PEOPLE HAD DETECTABLE REACTIONS TO THE RESPONSES AND THE FUNCTIONALITY OF THIS CELLS LOOK LIKE 1 WOULD EXPECT FOR A QUOTE-UNQUOTE OR JUST AN UNKNOWN NEW RESIRATTORY VIRAL INFECTION, THE FUNCTIONALITIES LOOK LIKE THE TYPES OF THINGS YOU WOULD GET IN AN ANTIVIRAL RESPONSE AND IN ADDITION SPIKE WAS AN IMMUNO DOMINANT TARGET OF THOSE RESPONSES INDICATING THAT SPHIEK WAS INDEED A REASONABLE CHOICE IF YOU WERE GOING WITH A SINGLE COMPONENT VACCINE AND THESE STUDIES HAD A VARIETY OF RAMIFICATIONS, CERTAINLY 1 OF THEM WAS--IT WAS REALLY INTERPRETED BY A NUMBER OF VACCINE MANUFACTURES THAT ESSENTIALLY THEY WERE ON THE RIGHT PATH AND THAT THEY COULD FOCUS THEIR EFFORTS INSTEAD OF TRYING MANY DIFFERENT VACCINE STRATEGIES, WHERE I THINK I COULD FOCUS ON SPIKE ONLY VACCINES AND TRY AND ILLICIT CONVENTIONAL CD4 RESPONSES IF THEIR PLATFORMS ALLOWED. THE SECOND PROGRAM WE EXPLORED INTENSIVELY, IN THE FALL OF 2020 THERE WAS LOTS OF FEAR AND UNCERTAINTY ABOUT WHETHER PEOPLE WOULD BE PROTECTED FROM REINFECTION, WERE CORONA VIRUSES SPECIAL IN SOME WAY AND THAT PEOPLE JUST DON'T DEVELOP IMMUNE MEMORY AND THAT WAS 1 OF THE COMMON TALKS POINTSA THE TIME AND WE FELT WE COULD CONTRIBUTE TO THAT, NOT IN TERMS OF DIRECTLY MEASURING REINFECTION RATES, RIGHT? THAT WOULD COME FROM OBVIOUSLY EPIDEMIOLOGICAL STUDIES BUT BY DIRECT MEASUREMENTS OF IMMUNE MEMORY TO A PRIMARY VIRAL INFECTION IN HUMANS AND SEE WHAT IT WOULD LOOK LIKE AND THAT WAS PUBLISHED IN THIS STUDY WHICH WAS POSTED ONLINE AROUND THANKSGIVING 2020. WE HAD A LAB PARTY RECENTLY WHERE SOMEBODY MADE A VERY NERDY CAKE FOR THE LAB THAT HAD LAB PAPER QR CODES PRINTED ALL OVER IT AND I WAS LIKE, IT WAS REALLY UNEXPECTED, AND THEY'RE LIKE OH YEAH, THIS IS A THING NOW, YOU DO IT WITH YOUR SLIDESSA THE SEMINAR AND YOU HAVE THE QR CODE IN SEMINAR AND SO THERE YOU GO AND I'M TRYING IT OUT AND IT'S IN QR CODE. A CRITICAL PROBLEM WITH IMMUNE MEMORY GOING BACK TO MY DAYS IN [INDISCERNIBLE] LAB AND MANY OTHER LABS BEFORE IT SHANKAR IT IS QUITE HARD TO PREDICT IMMUNE MEMORY AND SO A FOCUS FOR US WAS TO WAIT UNTIL 6-8 MONTHS POST INFECTION SO WE CAN GET A SENSE OF THE CONNECTED IMMUNE MEMORY AND REALLY TAKING THE APPROACH WE'VE TAKEN IN ALMOST ALL OF OUR STUDIES IS REALLY A WHOLISTIC LOOK AT THE ADAPTIVE IMMUNE RESPONSES. CERTAINLY MEASURING ANTIBODIES AND ALSO MEASURING THE OTHER MAJOR COMPONENTS OF ADANTE FOWLERRIVE IMMUNITY WHICH WOULD BE MEMORY B-CELLS, MEMORY CD4S AND MEMORY CD8S AND ACROSS MULTIPETRESSABLE ANTIGENS AND WHEN WE DID THAT, WE SAW THAT PEOPLE DID DEVELOP ENHANCED MEMORY AFTER INFECTION AND BY WAITING UNTIL APPROXIMATELY 8 MONTHS POST INFECTION, WE BEING SEE WHAT THE KINETETTICS OF THOSE RESPONSES ARE AND ON THE LEFT HERE WE HAVE ANTIBODY TITERS AND THESE WERE REPORTED BY MULTIPLE LABS AT THE TIME WHICH ARE THAT ANTIBODIES ARE RELATIVELY LOW AND AFFECT PEOPLE BUT SOMETHING LIKE 90-98% OF PEOPLE DO BECOME ANTIBODY POSITIVE AND THOSE TITERS DROP OVER A RELATIVELY SHORT TIME FRAME BUT BY 6 MONTHS ARE REALLY RATHER STABLE AND INDEED THEIR DATA IN THE LITERATURE NOW OUT TO A YEAR AND A HALF OR 2 YEARS SEEING THESE INFECTION INDUCED ANTIBODY TITERS ARE TABLELY MAINTAINED AND WHAT WE WERE ADDING TO THAT LITERATURE WAS REALLY THE CELLULAR ASPECTS, THE MEMORY B-CELLS, THE CD8 T-CELLS AND CD4 T-CELLINGS AND PROBABLY THE MOST SURPRISING FINDING WAS THAT PEOPLE HAVE MORE MEMORY B-CELLS POST INFECTION AND AT 1 MONTH POST INFECTION TO FOLLOW THE OPPOSITE KINETICS OF THE ANTIBODY RESPONSE AND WE SAW THIS FOR SPIKE RBD AND NUCLEOCAPSID AND THIS IS REPRODUCED BY MULTIPLE LABS. AND THAT THE KINETICS AND T-CELL MEMORY WAS DIFFERENT FROM THE KINETICS OF MEMORY B-CELLS AND OF SERUM ANTIBODIES SO THESE DIFFERENT COMPONENTS OF IMMUNE MEMORY DO NOT ALL FOLLOW THE SAME KINETICS AND THAT FOR CD8S ABOUT 70% OF PEOPLE HAD DETECTABLE RESPONSES EARLY, ABOUT 50% AT 8 MONTHS, BUT OF THE DETECTABLE RESPONSES THE HALF LIFE OF THOSE RESPONSES WAS ABOUT 125 DAYS, WHICH ACTUALLY MATCHED WHAT [INDISCERNIBLE] HAD DEFINED EARLIER FOR THE YELLOW FEVER VIRUS VACCINE WHICH IS A RESPONSE TO INFECTION LOOKED LIKE IT WOULD BE QUITE GOOD IN TERMS OF THE QUALITY OF DURATION EMPLOY THE CD4 RESPONSE TO THIS VIRUS ARE QUITE ROBUST, ALMOST A HUNDRED% OF PEOPLE WERE POSITIVE INITIALLY AND 98% OF PEOPLE POSITIVE AT 6-8 MONTHS WITH THE A SIMILAR HALF LIFE SUGGESTING THAT THE CD4 AND 8 MEMORY WOULD BE LONG LASTING. THERE WAS BT ANY BENCHMARK IN THE LITERATURE FOR LOOKING AT KINETICS OF CD4 T-CELL MEMORY TO A PRIMARY HUMAN INFECTION. SO THIS IS ACTUALLY NOW A PRIMARY REFERENCE POINT FOR THE FIELD FOR WHAT IMMUNE MEMORY LOOKS LIKE IN THE CD4 T-CELL COMPARTMENT TO A VIRAL INFECTION AND IN FACT, OVERALL, THIS WAS A VERY LARGE STUDY IN TERMS OF LOOKING AT MEMORY TO ALL OF THESE COMPONENTS AND PROBABLY THE LARGEST STUDY BEFORE THAT TO AN ACUTE INFECTION WAS PROBABLY ABOUT 3 OR 4 INDIVIDUALS AND IN THIS STUDY WE HAD 188. I WILL SAY JULIE [INDISCERNIBLE] AND [INDISCERNIBLE] HAD A SIMILAR STUDY PUBLISHED I THINK 6 MONTHS LATER AND I THINK THEY DID A NICER JOB OF CALCULATING HALF LIVES, WHERE INITIALLY THE HALF LIVES LOOK LIKE THIS AND THEY PROBABLY PLATEAU OR NEARLY PLATEAU THAT THE HALF LIVES PROBABLY BECOME MORE LIKE A YEAR AT THESE LATER TIME POINTS AND IT WOULD BE GOOD TO SEE FURTHER FOLLOW UP ON THOSE, BUT ALTOGETHER, THE RESULTS INDICATED THERE WAS SUBSTANTIAL IMMUNE MEMORY AFTER THIS INFECTION, AND IT WAS PROBABLY SUFFICIENT IMMUNE MEMORY TO PROVIDE PROTECTION AT LEAST FROM SERIOUS SARS-COV-2 INFECTIONS FOR YEARS INTO THE FUTURE, THIS IS BEFORE THE EXISTENCE OF VARIANTS WHICH IS DIRECTLY LOOKING AT IMMUNE MEMORY AND IN FACT LOOKING AT EPIDEMIOLOGICAL STUDIES HAVE SUPPORTED THAT OVER AND OVER AGAIN AT THIS POINT. AND JOHN MENTIONED WHAT WOULD HAPPEN IF YOU GOOGLED MY NAME AND CSPAN IN BALTIMORE. IF YOU GOOGLE MY NAME AND CSPAN, YOU WILL FIND DIFFERENT HITS INVOLVING INTERESTING DEBATES RELATING INTERPRETING TO THIS DATA WHICH INCLUDED SENATOR RAND PAUL AND TONY FAUCI AMONGST OTHERS BUT THAT'S A TOPIC FOR ANOTHER DAY. WITH ALL OF THAT ESSENTIALLY AS BACKGROUND, WE THEN MOVE TO UNDERSTAND IMMUNE MEMORY TO COVID-19 VACCINES. AND FOR NOW I WILL TALK THROUGH THE WORK WE PUBLISHED EARLIER THIS YEAR WHICH WAS THE FIRST PUBLICATION TO LOOK AT HEAD-TO-HEAD COMPARISON TO IMMUNE MEMORY TO 4 COVID-19 VACCINES, INCLUDING A PROTEIN VACCINE TO LOOK AT IMMUNE MEMORY OUT AT LEAST 6 MONTHS WHERE WE FEEL LIKE WE REALLY DO HAVE TO WAIT THAT LONG TO GET A SENSE OF WHAT MEMORY'S LOOKING LIKE AND TO DO SO AGAIN IN A RELATIVELY WHOLISTIC WAY OF LOOKING AT ALL 3 MAJOR BRANCHES OF ADAPTIVE IMMUNITY, THE HUMORAL IMMUNITY AS WELL AS CD4 AND 8 T-CELLS AND WE FELT LIKE THESE WERE KEY COMPARISONS TO DO FOR UNDERSTANDING WHY ARE THERE SIMILAR OR DIFFERENT OUTCOMES WITH DIFFERENT COVID-19 VACCINE PLATFORMS AND ALSO FOR LOOKING INTO THE FUTURE, LOOKING TO THE FUTURE FOR VACCINE DEVELOPMENT TO OTHER PATHOGENS. ARE SOME OF THESE PLATFORMS MORE SUITED TO SOME PATHOGENS THAN OTHERS FOR EXAMPLE, OR IS--OR RNA VACCINES ESSENTIALLY MIRACULOUS, THAT THEY DID SOMETHING PHENOMENAL THAT REALLY CONVENTIONAL OR MORE CONVENTIONAL VACCINE TECHNOLOGIES COULD NOT. THOSE WERE STILL OPEN QUESTIONS FROM AN IMMUNOLOGICAL PERSPECTIVE BECAUSE IT IS HARD TO RECRUIT PEOPLE WHOF GOTTEN EACH OF THESE DIFFERENT VACCINES AND PARTICULARLY FROM A T-CELL SIDE, YOU HAVE TO BE DOING THOSE ASSAYS IN THE SAME LAB FOR THEM TO BE COMPARABLE AND IN ADDITION, WE HAD LOCAL INFECTED INDIVIDUALS SO THAT WE CAN COMPARE VACCINE ILLICITTED IMMUNE MEMORY. SO WHAT DID ALL OF THAT LOOK LIKE? HERE'S THE WAY WE STRUCTURED THE FIGURURES, WE DID ANTIBODIES, AND THEN EACH OF THESE OTHER IMMUNE COMPONENTS AND IN HERE WE'RE SHOWING THE RESPONSES TO EACH OF THE VACCINES SO MODEL CITIZEN BERNEA, PFIZER, J& J AND NOVAVAX, AND AT THE DOSES AND TIMING AND USE IN THE UNITED STATES AND I'M SORRY, I SHOULD HAVE BEEN CLEAR--FOR ANYBODY WHO'S NOT FAMILIAR WITH THE NOVAVAX VACCINE, NOVAVAX IS A RECOMBIN ANT SPIKE PROTEIN WITH [INDISCERNIBLE] ADJUVANT AND IT SERVES AS A USEFUL COMPARISON HERE BECAUSE THERE'S A HUNDRED YEARS MORE EXPERIENCE WITH PROTEIN BASED VACCINES THAN RNA VACCINES AND SO, PROVIDES A VALUABLE COMPARISON. ONE, CAVEAT TO THIS IS THAT WE WERE NOT ABLE TO RECRUIT THESE INDIVIDUALS IMMEDIATELY AFTER IMMUNIZATION, SO WE ONLY HAVE THE MEMORY TIME POINTS FOR THE NOVAVAX INDIVIDUALS. T1 WOULD BE PREVACCINATION. T2 IS AFTER A FIRST VACCINATION IN THE CONTEXT OF THE RNA VACCINES, T3 AND 2 WEEKS AFTER THE SECOND IMMUNIZATION, T4 WOULD BE 3 MONTHS OUT AND T5 WOULD BE 6 MONTHS OUT APPROXIMATELY FOR EACH OF THESE, OKAY? AND THEN THE THICK LINES IN EACH CASE WHICH YOU CAN FOCUS ON ARE THE GMEs OF THE ENTIRE COHORT, AND THEN WE HAVE CONVENIENT REFERENCE OVER HERE FOR HOW DOES THIS ALL COMPARE TO INFECTED INDIVIDUALS. SO INFECTED INDIVIDUALS ARE BEING SHOWN IN YELLOW WITH THE SAME COLOR CODES FOR THE VACCINES OVER HERE. SO IN TERMS OF ANTIBODY RESPONSES THIS WORK WAS DONE IN THE GROUP BY POST DOCTORAL FELLOWS ZALI ZHANG, AND ASSISTANT PROFESSOR AT UCSD GEN DAN AND WE CAPITULATED THAT THE VACCINES ARE FANTASTIC AT ILLICITTING HIGH ANTIBODY TITERS QUICKLY AND CONSISTENTLY ACROSS INDIVIDUALS BUT WITH A PRETTY DRAMATIC DECAY OVER A 6 MONTH PERIOD OF TIME AND TO ME THE BEST DAT I'VE SEEN IS THAT THOSE REALLY CONTINUED TO DECAY FOR 10 MONTHS AFTER 2 DOSES OF RNA VACCINE. AND ACTUALLY OUT AT 6 MONTHS THE NOVAVAX PROTEIN HAS NEUTRALIZING ANTIBODY TITERS QUITE COMPARABLE TO MODEL CITIZEN DERNA AND PFIZER WHICH ARE SIGNIFICANTLY HIGHER THAN WHAT 1 GETS AFTER INFECTION OR AFTER A SINGLE DOSE J& J. NOW NOTE ABLE I WAS INCREEINGING ABOUT THE J& J VACCINE AND OTHER GROUPS HAVE NOTED THIS AS WELL IS THAT AT A MONTH POST VACCINATION, IT'S REALLY RELATIVELY POOR NEUTRALIZING ANTIBODY TITERS, IT'S ONLY ABOUT 50% OF PEOPLE HAVE TITERS THAT ARE ABOVE 20 BUT INTRIGUINGLY THOSE TITERS INCREASE OVER TIME AND AS A COHORT PEOPLE HAVE HIGHER AN AVERAGE NEUTRALIZING TITERS AT 1 MONTH THAN THEY DID AT 6 MONTHS. WHICH IS THE TREND YOU GET WITH RNA VACCINES INDICATING THESE VACCINE PLATFORMS ARE ENGAGING DIFFERENT IMMUNOLOGICAL MECHANISMS OF ACTION. IF WE LOOK AT CD4 T-CELL RESPONSES WE SEE THAT THE RNA VACCINES ARE VERY GOOD AT ELUCIDATING ROBUST RESPONSES BOTH ACUTE AND OUT AT MEMORY TIME POINTS. SIX MONTHS BOTH VACCINATION, THERE'S REALLY LESS THAN A 2 FOLD DECLINE IN THE MAGNITUDE OF THE SPIKE SPECIFIC CD4 T-CELLS WHICH IS REALLY QUITE IMPRESSIVE IN MY OPINION FOR THE R NA VACCINES AND IN FACT, THEY RECEIVED HIGHER CD4 RESPONSES THAN INFECTION. NOW INFECTION ALSO ILLICITS T-CELL RESPONSES TO OTHER ANTIGENS, RIGHT? BESIDES SPIKE, BUT STILL IF YOU SPECIFICALLY FOCUS ON SPIKE, THESE ARE STILL PRETTY IMPRESSIVE RESPONSES AND REALLY THE RNA VACCINE RESPONSES ARE IN LINE WITH THE NOVA VAX AND IT CAN DO WELL AT ILLICITTING RESPONSES IN COMPARISON TO RNA AND IN OUR HANDS WITH THESE TYPES OF ASSAYS ACTUALLY J& J WAS THE LOWEST OF THE RESPONDERS WHICH REALLY CONTRAST WITH INITIAL EXPECTATIONS THAT A VIRAL VECTOR WOULD BE THE BEST AT ILLICITTING T-CELL RESPONSES SO AT LEAST WITH CD4S, WE DID NOT FIND THAT TO BE THE CASE BY ANY OF THESE METRICS ALTHOUGH IT CERTAINLY DID ILLICIT CD4 RESPONSES AT 4 AND 6 MONTHS. WE DID BREAK THIS DOWN INTO INDIVIDUAL CYTOKINES AND I WILL HEIGHT LIGHT WE WERE INTRIGUED THAT THERE ARE A NUMBER OF GRANZYME B CD4 CELLS ILLICITTED BY EACH OF THESE 4 VACCINE PLATFORMS IN HUMANS WHICH WASN'T NECESSARILY WHAT WE EXPECTED TO SEE. OF KEY IMPORTANCE, IS THE ABILITY OF THESE VACCINES TO ILLICIT TFH CELLS, THEY ARE THE SUBSET OF CD4s THAT ARE REQUIRED FOR HELPING B-CELLS AND THEY'RE REQUIRED FOR THE MAJORITY OF NEUTRALIZING ANTIBODY RESPONSES TO SARS-COV-2 AND THEY'RE CERTAINLY REQUIRED FOR ALL OF THE OMICRON ANTIBODIES HAVE YOU CIRCULATING IN YOUR BLOOD WHICH ARE THE SOURCE OF--WHICH ARE THE PRODUCT OF GERMINAL CENTERS AND AFFINITY MATURATION WHICH WE WILL COME BACK TO LATER. SO THESE CELLS ARE CRITICAL. THEY'RE NOT GENERALLY AT HIGH FREQUENCIES IN THE BLOOD BUT THEY ARE DETECTABLE AND SO WE'RE ABLE TO SEE THAT THE RNA VACCINES GENERATE THESE AND GENERATED THEM WELL, GENERATED THEM EVEN A BIT BETTER THAN J& J AND IN LINE WITH NOVA VAX, AND I WOULD ACTUALLY CONCLUDE THAT I THINK THAT THE POTENCY OF RNA VACCINES FOR ILLICITTING NEUTRALIZING ANTIBODIES AND BEING SO HAWAIILY PROTECTIVE SO QUICKLY REALLY COMES DOWN TO THEIR ABILITY, THEIR IMPRESSIVE ABILITY TO PRIME CD4 T-CELL RESPONSES AND DRIVE TFH CELL DIFFERENTIATION AND I THINK WE'RE SHOWING THAT HERE. IN ADDITION TO GENERATING THOSE TFH CELLS, THESE VACCINES ARE GENERATING MEMORY TFH CELLS WHICH IS IMPORTANT FOR BOOSTER RESPONSES AND SUBSEQUENT TIME POINTS. FOR CD8 T-CELLS. THIS IS WHERE THE LITERATURE WAS A LOT MORE MIXED IN GOING BACK TO ACTUALLY EARLY PUBLICATIONS THAT WE'RE OBSERVING VERY SIMILAR IMMUNE RESPONSES BETWEEN THE PFIZER, AND MODERNA VACCINE EXCEPT IN CD8S WHERE PFIZER REPORTED RELATIVELY STRONG CD8 RESPONSES AND CONTINUED WORKING WITH MODERNA GENERALLY REPORTED LOW TO UNDETECTABLE CD8 T-CELL RESPONSES. WE AND OTHER VS WORKED ON THIS IN A VARIETY OF WAYS, IN PARTICULAR IN THIS CONSORTIUM, IT WAS JOSE MATEUS, WORKING WITH [INDISCERNIBLE]. AND SO A LOT OF THIS PARTICULAR PROJECT WAS ALEX SETTE, MYSELF AND DANIEL [INDISCERNIBLE] A REALLY FANTASTIC ASSISTANT RESEARCH PROFESSOR HERE, AND MATEUS HANDLING THE BRUNT OF THE WORK, WHERE WHAT WE SEE WITH ESSENTIALLY AN OVERNIGHT EPITHELIAL RACELLULAR CYTOKINE STAINING AND THE FOLLOWING THAT ACTUALLY MODERNA AND PFIZER VACCINES ARE QUITE GOOD AT ILLICITTING CD8 T-CELL RESPONSES TO THE EXTENT THEY ARE DETECTABLE IN 85-90% OF INDIVIDUALS AFTER IMMUNIZATION AND ARE STILL DETECTABLE AND 2/3RDS OF INDIVIDUAL ADDED 6 MONTHS NOT AS GREAT AS I WOULD LIKE BUT CERTAINLY PRESENT IN THE MAJORITY OF INDIVIDUALS AND IN FACT, THOSE RESPONSES ARE HIGHER OR AS HIGH THAN THE J& J VACCINE. WHAT OTHER GROUPS HAVE FOUND IS THAT IF INSTEAD YOU USE A SHORT-TERM STIMULATION ONLY A 6 HOUR STIMULATION INSTEAD OF AN OVERNIGHT STIMULATION, WHAT YOU SEE IS RESPONSES TO AN ADEN O VACCINE IN HUMANS AND NOT TO THE RNA VACCINES SO THIS COMES IN DOWN GENERAL ASSAYS 1 USES, AND IN ASSESSING T-CELLS, ICS IS REALLY MORE IN LINE WITH WHAT YOU'RE INTERESTED IN BECAUSE DO YOU WANT TO KNOW ARE THE CELL SAYS THERE GOOD, THEY DID SEE A REINFECTION WOULD THEY RECALL, WHEREAS A 6 HOUR ASSAY IS TELLING YOU THE CAPACITY FOR IMMEDIATE EFFECTOR FUMPLEGZS WHICH IS VERY HIGH, WHETHER CD8 T-CELL RESPONSE WAS ILLICITTED OR NOT. WE DO SEE A LITTLE INDICATION OF THE CD8 RESPONSE AFTER THE IN, OVAVAX, WE HAVE REPRODUCED THIS IN A COHORT, 25 INDIVIDUALS HAVE SOME DETECTABLE RESPONSE, I WILL NOT FOCUS ON THAT NOW BAH WE DO THINK THAT'S INTERESTING BECAUSE IT'S UNCONVENTIONAL FOR A PROTEIN VACCINE. AND THE FINAL IMMUNE MEMORY COMPONENT WE LOOKED AT ACROSS THESE 4 VACCINES HEAD-TO-HEAD WERE THE MEMORY B-CELLS WHICH WERE SPIKE AND RBD, WE FOUND ALL 4 DO ILLICIT MEMORY B-CELLS AND THOSE DO INCREASE BETWEEN 3 MONTHS AND 6 MONTHS POST VACCINATION AND I WILL COME BACK TO THIS AND IT IS INTERESTING THAT THESE GO UP, WHEREAS THE ANTIBODY TITERS ARE CLEARLY GOING DOWN AND THE T-CELL RESPONSES CERTAINLY AREN'T GOING UP AND THIS IS SIMILAR TO WHAT WE SEE IN THE CONTEXT OF INFECTION AND COMPARING ACROSS THE 4 VACCINES OTHERS AGAIN THEY'RE DOING A FANTASTIC JOB OF GENERATING B-CELL MEMORY, IT WILL BE ADEN O VIRAL VECTOR OR EVEN THE PROTEIN VACCINE. AND HOW DOES ALL THIS CONNECT TO PROTECT EVALUATION PROCESS IMMUNITY? AND I'LL COMMENT JUST A BIT ON THIS, HAVING TRAINED AS A VIROLOGIST IT'S ALWAYS CRITICAL TO THINK OF PROTECTIVE IMMUNITY IN THE CONTEXT OF THE ANATOMY AND PATHOGENESIS OF THE PARTICULAR VIRAL INFECTION THAT YOU ARE TRYING TO DEAL WITH AND IN THE CASE OF SARS COV-2 THIS IS CLEARLY A VIRUS THAT IS RAPID AT REPLICATING NAISAL CAVITIES AND ORAL CAVITIES AND VERY RAPID AT CAUSING ASYMPTOMATIC TO COLD LIKE DISEASE AND IT'S A RATHER DIFFICULT VIRUS TO STOP QUICKLY BUT IN TERMS OF PATHOGENESIS IT'S A RELATIVELY SLOW INFECTION IN THE LUNG AND INFECTION IN THE LUNG IS LOW WITH HOSPITALIZATIONS, FREQUENTLY 2 WEEKS POST INFECTION OR SYMPTOM ONSET AND THIS PROTECTIVE IMMUNITY NEEDS TO KEEP THESE DIFFERENTIAL CONCEPTS OF PROTECTION, I. E. PROTECTION FROM INFECTION, VERSUS PROTECTION FROM SEVERE DISEASE IN MIND AS WELL AS THE DIFFERENT TISSUES IN WHICH THOSE ARE HAPPENING. IN BRIEF MY PERSONAL INTERPRETATION OF THE LITERATURE IS AS FOLLOWS. FIRST OF ALL THE SIMPLEST OPTION FOR ANY VACCINE IS HIGH LASTING NEUTRALIZING ASPECT BODIES, AND SARS-COV-2 IS PRIMARY FOR THIS, I MEAN THIS IS--THIS GENERAL TOPIC IS LITERALLY WHAT I FOUNDED MY LAB BASED ON 19 YEARS AGO WAS GENERATING NEUTRALIZING ANTIBODY RESPONSES IS KEY FOR MOST VACCINES AND WHAT WE NEEDED TO BETTER UNDERSTAND IS THE FUNDAMENTAL IMMUNOLOGICAL PRINCIPLES UNDERLYING HOW DO YOU GENERATE THOSE RESPONSES AND HOW DO YOU GET THEM TO BE DURABLE. BUT GIVEN THAT IT WAS CLEAR THAT INFECTION ITSELF DIDN'T ELICIT PARTICULARLY HIGH NEUTRALIZING ANTIBODIES AND THERE WAS LOTS OF CONCERN ABOUT DURABLE ANTIBODIES, WE AND OTHERS RAISE THE ISSUE OF LIF YOU CAN'T GENERATE HIGH ENOUGH LEVELS OF NEUTRALIZING ANTIBODIES OR IF THEY'RE NOT LONG LASTING, ARE THERE OTHER POTENTIAL MECHANISMS OF PROTECTIVE IMMUNITY AGAINST COVID-19 AND ADDITIONAL LAYERS OF ANTIBODIES. AND IT HAS BEEN HARD TO MAINTAIN ANTIBODIES LONG LASTING EITHER IN THE CONTEXT OF VACCINE BEING USED OR IN THE CONTEXT OF VARIANTS AND THE VITRATTURE HAS VARIOUS LINES OF EVIDENCE POINTING TO CONTRIBUTION OF T-CELLS AGAINST COVID-19. I WON'T GO THROUGH ALL OF THOSE IN THE INTEREST OF TIME WE CAN TALK ABOUT THEM IN THE Q&A. OVER ALL MY PERSPECTIVE OVERALL WHICH IS THE SAME BACK IN 2020 IS THAT IT'S REASONABLE TO CONSIDER THAT HOSPITALIZATION LEVEL COVID-19 IS PREVENTED BY ANY DESCENT COMBINATION OF ANTIBODIES, MEMORY B-CELLS, CD4S AND CD8S. I HAVE FOUND THAT SOMETIMES PEOPLE HEAR THIS IN RATHER DIFFERENT WAYS SO I WILL SAY IT AGAIN A LITTLE BIT DIFFERENTLY, I AM NOT SAYING THAT T-CELLS, THAT ALL THE PROTECTIVE IMMUNITY AGAINST COVID-19 IS T-CELLS, SO T-CELLS IS THE MAJORITY OF IT, WHAT I AM SAYING IS THAT THE AVAILABILITY DATA POINT TO CONTRIBUTIONS OF T-CELLS AGAINST COVID-19 AND IN PARTICULARLY IN THE CONTEXT OF MORE SEVERE DISEASE, IT'S REASONABLE TO CONSIDER THAT THEY DO CONTRIBUTE AS WELL AS MEMORY B-CELLS CONTRIBUTING. AND I LIKE TO THINK OF THIS IN A LAYERED DEFENSES TYPE OF MODEL WHERE YOU WOULD LOVE TO HAVE THE VIRUS STOPPED BY NEUTRALIZING ANTIBODIES BUT IF YOU DON'T HAVE ENOUGH, DO YOU HAVE ADDITIONAL LAYERS OF THE DEFENSE THAT CAN HELP PREVENT SEVERE COVID-19 FROM OCCURRING. AND REALLY IF YOU CAN CONSIDER LAYERED DEFENSES, WE NEED TO MOVE BEYOND JUST THINKING ABOUT CIRCULATING IMMUNITY WHICH IS EVERYTHING THAT I'VE TALKED ABOUT SO FAR, RIGHT, WERE ALL MEASUREMENTS IN THE BLOOD TO ALSO CONSIDERING LOCAL IMMUNITY, TISSUE RESIDENT MEMORY WHICH WOULD BE MORE FRONT LINE DEFENSES AND SO THERE HAVE BEEN PLENTY OF QUESTIONS ABOUT, WELL, DID INFECTION ILLICIT TISSUE RESIDENT MEMORY OR DO THE VACCINES ILLICIT TISSUE RESIDENT MEMORY, THOSE ARE MUCH HARDER QUESTIONS TO ASK BECAUSE YOU HAVE TO ACCESS HUMAN TISSUE TO DO SO. AND SO THOSE ANSWERS HAVE FOLLOWED BEHIND THE MUCH MORE COMMON MEASUREMENTS IN THE CIRCULATION. AND 1 OF THE FIRST STUDIES TO SHOW OR TO ASSESS WHETHER OR NOT THERE WAS TISSUE RESIDENT MEMORY AFTER SARS-COV-2 INFECTION WHICH IS THIS 1 LED BY THE WONDERFUL DONNA FAR BERNEA DEET AND MOST OF YOU WILL KNOW DONNA FAR FARBER HAS DONE AMAZING WORK MAKING USE OF ORGAN DONOR PROGRAMS IN THE NEW YORK METROPOLITAN AREA TO DIRECTLY LOOK AT HUMAN IMMUNE MEMORY IN TISSUES, IN COMPARISON TO BLOOD. AND SO IN THIS PARTICULAR STUDY WHAT BECAME AVAILABLE JUST THROUGH THAT--JUST AN EXTENT RANDOM ENROLLMENT PROCESS WAS THERE WERE 4 INDIVIDUALS QUHO DIED FROM COVID UNRELATED CAUSES AND HAD UNREMARK CALVERT CASINGS OF COVID-19 AT SOME PREVIOUS TIME POINTS AND WE COMPARED WHETHER THOSE INDIVIDUALS, THOSE 4 INDIVIDUALS WHO HAD A HISTORY OF COVID-19 DID THEY HAVE DETEBTIBLE B-CELL OR T-CELL MEMORY IN THEIR LUNGS AND THE LUNG DRAINING LYMPHNODES. SO THOSE INDIVIDUALS ARE SHOWN IN RED HERE. AND THE SHORT ANSWERS WERE YES, THEY HAD TISSUE RESIDENT MEMORY CD4 T-CELLS AND TISSUE MEMORY CD8 T-CELLS IN LUNGS AND IF ANYTHING, THERE WERE A HIGHER FREQUENCY OF VIRUS SPECIFIC CD8 IN THE LUNGS THAN IN THE BLOOD, AND IT IS NOTABLE HERE BECAUSE AGAIN, THESE ARE PEOPLE WITH UNREMARKABLE CASES OF COVID-19 AND THERE STILL WAS ENOUGH ANTIGEN IN THE LUNGS RIGHT TO RESULT IN SIGNIFICANT AMOUNTS OF RESIDENT MEMORY T-CELLS. WHAT ALEX IN MY GROUP WAS RESPONSIBLE FOR WAS ASKING THE QUESTION WERE THERE RESIDENT MEMORY B-CELLS WHICH IS A MUCH LESS STUDIED AREA, PARTICULARLY IN HUMANS SO WE WERE THE FIRST TO DEMONSTRATE THAT THERE ARE INDEED ROBUST AMOUNTS OF RESIDENT MEMORY B-CELLS IN THE LUNG, SPIKE SPECIFIC AND RBD SPECIFIC RESIDENT MEMORY B-CELLS IN THE LUNGS OF PEOPLE WHO HAVE HAD AGAIN UNREMARKABLE CASES OF COVID-19 COMPARED TO NEGATIVE CONTROLS. THESE PEOPLE WERE ALL ENROLLED BEFORE THE VACCINES WERE AVAILABLE SO WE KNOW THESE ARE ALL INFECTION INDUCED EVENTS, OKAY. AND WE DID ALSO SEE MEMORY IN THE LUNG DRAINING LYMPHNODES, THAT'S WHAT LLN STANDS FOR AS WELL AS GUT TRAINING LYMPHNODES WHICH IS AN IMPORTANT AND INTERESTING TOPIC. OTHER LABS HAVE ALSO PUBLISHED THAT MEMORY T-CELLS ARE DETECTABLE IN LUNGS OF INVESTED INDIVIDUALS AND MORE RECENTLY THERE'S BEEN NICE DATA AVAILABLE ON NAISAL, NAISAL PHARYNGIAL MEMORY CELLS, BY UPON J. SUN AND [INDISCERNIBLE] AMONG RPGHTS DEMONSTRATING THERE CLEARLY ARE CDEFICIENCY, 8 AND CD4 MEMORY T-CELLS IN THESE TISSUES AFTER INFECTION AND NOT AFTER VACCINATION WITH R NA VACCINES WHICH I THINK LARGELY MATCHES OUR EXPECTATION BUT IS IMPORTANT TO SEE AND ANYTHINGLY IT LOOKS LIKE THESE MAY BE RELATIVELY DURABLE RESPONSES WHICH WARRANTS MORE INFORMATION BUT AT LEAST FOR THE MOMENT IT DOESN'T LOOK LIKE THEY JUST SHOW UP FOR 2 MONTHS AND IN THIS PERSON WHO IS INFECTED OR IN HYBRID IMMUNITY IF YOU'VE BEEN INFEBTED OR VACCINATED AND THEN VACCINATED AND HAVE A BREAK THROUGH INFECTION, YOU END UP WITH HYBRID IMMUNITY WHERE YOU GET TISSUE RESIDENT MEMORY CELLS, REALLY AS VERY STRONG CIRCULATING MEMORY GENERATED BY VACCINATION. THERE WAS 1 ADDITIONAL PIECE OF INFORMATION BY THESE ORGAN DO DONORS THAT SURPRISED US AND THAT WAS WHEN WE LOOKED IN THE LUNG DRAINING LYMPHNODES WE COULD SEE GERMINAL CENTERS THAT IN AND OF ITSELF IS UNSURPRISING, PEOPLE DO GET OBVIOUSLY EXPOSED TO MANY THINGS, BUT WE HAD THE CAPACITY WITH FLUORESCENT PROBE STAINING TO DEMONSTRATE THERE WERE SARS-COV-2 SPIKE SPECIFIC GERMINAL CENTER B-CELLS IN THESE LUNG DRAINING LYMPHNODES AND THE PREVIOUS PROBABLY FOR 3 OUT OF 4 OF THESE INDIVIDUALS AND THEY'RE PREVIOUSLY RECORDED INFECTIONS WERE 6 MONTHS OR MORE IN THE PAST AND SO THIS SUGGESTS THAT THERE WERE ONGOING GERMINAL CENTERS FOR AT LEAST 6 MONTHS AFTER AGAIN AN UNREMARKABLE CASE OF COVID-19 IN THESE LUNG DRAINING LYMPHNODES. AND SO NOW, WITH THAT, I'LL PIVOT TO THESE CONNECTIONS BETWEEN WHAT WE'VE BEEN OBSERVING IN THE CONTEXT OF COVID-19 AND COVID-19 VACCINES AND WHAT WE'VE BEEN WORKING ON I WOULD SAY MORE BROADLY IN VACCINE IMMUNOLOGY, PARTICULARLY USING--WORKING ON HIV VACCINES BOTH BECAUSE THE WORLD NEEDS AN HIV VACCINE AND BECAUSE IT'S A RATHER FANTASTIC MODEL SYSTEM FOR VACCINE IMMUNOLOGY BECAUSE OF THE MANY TOOLS WE HAVE AVAILABLE. WE--I MENTIONED TF8 CELLS BEING IMPORTANT BEING ESSENTIAL FOR T-CELL AND B-CELLS, AND WE'VE BEEN STUDYING TFH CELLS FOR A NUMBER OF YEARS NOW AND AFTER WE AND OTHERS HOPE TO ESTABLISH THAT TF8 STYLES WERE REAL AND IMPORTANT IN 2009, THERE HAVE BEEN A LOT OF GREAT ADVANCES IN THAT FIELD BY MANY GROUPS INCLUDING PAM [INDISCERNIBLE] GROUP AT NIH AND MANY OTHERS AND BETWEEN THE ENTIRE WORK OF THE FEELTD WE HAVE LEARNED MANY THINGS ABOUT TFH CELLS, AND THEY HELP THESE CELLS EARLY IN IMMUNE RESPONSES AS EARLY AS THE FIRST FEW DAYS, BUT ARE THEN ABSOLUTELY CRITICAL IN GERMINAL CENTERS FOR THE ENTIRE PROCESS OF GERM LINE CENTERS INCLUDING THE PRODUCTS, THE PRODUCTS THEMSELVES. GERMINAL CENTERS ARE ABSOLUTELY AMAZING IMMUNOLOGICAL EVENTS AND STRUCTURES. THESE ARE LITERALLY REALTIME EVOLUTION WHERE ANY EVOLUTIONARY PROCESS HAS TO HAVE GENERATIONS ISSUES RIGHT? SO THAT'S CELL DIVISION, MUTATIONS, WHICH ARE SOMATIC HYPERMUTATIONS THAT OCCUR, IN THE CENTERS AND A SELECTIVE PRESSURE OR A LIMITING RESOURCE AND IN THIS CASE, THE LIMITING RESOURCE IS ACTUALLY GENERALLY THE TFH CELLS AND THE TFH CELLS ARE LIMITED IN NUMBER AND PROVOID A LIMITED AMOUNT OF HELP TO THE B-CELLS AND THEY SIGNAL THE B-CELLS TO UNDERGO MUTATION AND PROLIFERATION AND THEN AMONGST THOSE EVENTS, AFTER A PSYCHOEVENT, THOSE B-CELLS HAVE TO COME BACK, INTERACT WITH THE TFH CELLS AGAIN AND OVERALL IN THE POP IELINGS, THERE'S A TENDENCY FOR THE B-CELLS TO HAVE THE HIGHEST AFFINITY TO GET THE MOST HELP FROM THE TFH CELLS AND THEIR ADDITIONAL SIGNALS, THAT GO ON, BEYOND DRIVING THIS EVOLUTIONARY PROCESS, CAN THEN DRIVE FURTHER DIFFERENTIATION TO GENERATE PLASMA CELLS OUT OF THAT GERMINAL CENTER B-CELLS OR MEMORY B-CELLS. AND IT'S VERY CLEAR FROM BEAUTIFUL WORK FROM A NUMBER OF LABS THAT REALLY ESSENTIALLY, ALL--AS I WAS SAYING EARLIER ALL OF YOUR OMICRON NEUTRALIZING ANTIBODIES IN THE BLOOD ARE COMING FROM THIS PROCESS WHERE A B-CELL THAT STARTED WITH THE ABILITY TO RECOGNIZE THE ANCESTRAL STRAIN PRESENT IN YOUR ORIGINAL MODERNA OR PFIZER OR OTHER VACCINE, THOSE B-CELLS UNDERWENT EXTENSIVE MUTATION AND SELECTION AND A NUMBER OF THEM BECAME CAPABLE OF BINDING TO OMICRON, WITH HIGH AFENNITY EVEN THOUGH THEY'VE NEVER SEEN OMICRON, AND MAKING ANTIBODIES THAT COULD DO SO. OR IN FACT, IN GENERAL, THAT DIDN'T HAPPEN DIRECTLY AFTER THE FIRST OR SECOND IMMUNIZATION, WHAT WAS HAPPENING AFTER THE FIRST AND SECOND IMMUNIZATION FOR THE GENERATION OF MEMORY B-CELLS THAT HAD THE POTENTIAL TO BIND OMICRON AND REALLY 1 KEY ASPECT OF GERMINAL CENTERS THAT'S BECOMING MORE AND MORE APPRECIATED IS THAT THEY REALLY ARE A SOURCE OF GENERATING B-CELL DIVERSITY FROM A TELEIO LOGICAL PERSPECTIVE OF THE IMMUNE SYSTEM RECOGNIZING THAT THE PRESENCE OF 1 VIRUS SLOWING UP NOW MAY MEAN THAT THEY WOULD BE RELATED VIRUSES SHOWING UP IN THE FUTURE AND THUS MAKING A DIVERSITY OF MEMORY B-CELLS, MIGHT BE QUITE HELPFUL IN RECOGNIZING VIRAL VARIANTS IN THE FUTURE AND INDEED THAT'S EXACTLY WHAT GETS SEEN. WE HAVE BEEN TRYING TO UNDERSTAND THOSE GERMINAL CENTER PROCESSES FROM OTHER PERSPECTIVES. THE SIMPLEST MODEL OF A GERMINAL CENTER HAS ALWAYS BEEN THAT IT'S WHERE YOU HAVE RAPID EVOLUTION AND SELECTION OF THE BEST B-CELLS AND THE BEST B-CELLS WIN OUT AND WHAT WE SAID WAS THAT DIDN'T REALLY MATCH OUR OBSERVATIONS IN SOME OF THE HIV VACCINE FIELDS WHERE IT'S BEEN VERY DIFFICULT TO GENERATE NEUTRALIZING ANTIBODIES, SO, WHY WOULD YOU HAVE SITUATIONS WHERE YOU WOULD IMMUNIZE WITH AN ANTIGEN THAT HAD NEUTRALIZING ANTIBODY EPITOPES ON IT BUT YOU WOULDN'T SEE THOSE NEUTRALIZING ANTIBODIES COME OUT. AND WE CONSIDERED THAT IMMUNO DOMINANCE MIGHT BE A REASON FOR THAT PROBLEM. THAT WAS CERTAINLY IN THE HIV FIELD THAT WAS CONTROVERSIAL AS WELL AS SOME OTHER FIELDS AND SO WE TRIED TO LAYOUT SOME CLEAR CONCEPTS THAT WERE TESTABLE THAT WOULD ADDRESS THOSE B-CELLS AND IMMUNO DOMINANCE EXISTS AND IF IT DID WHAT PARAMETERS DOES IT ABIDE BY AND SO 1 MODEL WAS THAT YOU CAN GET IMMUNO RESPONSES BASED ON INFINITY DISPARITY, AND IF YOU HAVE FOR EXAMPLE, EASY EPITOPES ON AN ANTIGEN, LET'S CALL THESE THE NONNEUTRALLIZING EPITOPES, IF THESE HAVE A BETTER AFFINITY THAN THE B-CELLS THAT RECOGNIZE DIFFICULT NEUTRALIZING EPITOPES THEN PERHAPS THE B-CELLS THAT RECOGNIZE THE NONNEUTRALLIZING EPITOPES, WIN OUT IN COMPETITION SIMPLY BUZZ THEY START WITH A BETTER AFFINITY ON AVERAGE. OR YOU COULD HAVE A NUMERICAL DISPARITY WHERE MAYBE THERE ARE JUST MANY MORE B-CELLS THAT CAN RECOGNIZE NONNEUTRALLIZING EPITELOMERES, THAN NEUTRALIZING EPITOPES AND THAT NUMERICAL DISPARITY MIGHT BE SO LARGE THAT YOU FAILED TO GET NEUTRALIZING ANTIBODY RESPONSES BECAUSE THEY GET NIEWMERICALLY OUTCOMPETED. THESE CAN HAPPEN THROUGH MULTIPLE MECHANISMS, 1ICISMEM CONCEPT IS GLYCANS SO VIRUSES LIKE HIV, THIS IS THE ENVELOPE SHOWN WITH GLYCANS SHOWN IN BLUE, THESE ARE SELF-GLYCANS SO HUMAN B-CELLS THAT DEVELOP IN HUMAN B-CELLS THAT HAVE HIGH AFFINITY FOR CELL GLYCANS GET ELIMINATED BECAUSE THEY'RE AUTOREACTIVE SO YOU INTRINSICALLY HAVE LOWER AFFINITY B-CELLS AND RARE B-CELLS IF THE EPITOPES ARE GOING TO INVOLVE A SELF-GLYCAN COMPONENT. ADDITIONALLY, IT COULD BE THAT LIMITED T-CELL HELP COULD HELP IN B-CELL IMMUNE DOMINANCE BECAUSE AFTER AN INITIAL PRIMING EVENT THERE ARE VERY FEW CD4 T-CELLS IN THE FIRST FEW DAYS AND YET WE KNOW THAT MOST IMMUNE RESPONSES B-CELLS REALLY ONLY UNDER GO 1 CELL DIVISION BEFORE THEY SEE T-CELL HELP AND THEN ALL SUBSEQUENT CELL DIVISIONS ARE T-CELL DEPENDENT IN A MAJORITY OF CASES AND IN THAT SCENARIO WITH A LIMITED NUMBER, IT'S PRETTY EASY TO IMAGINE THAT SOME B-CELLS MAYBE JUST THE MOST COMMON B-CELLS WOULD GET HELP AND MANY OTHERS WOULD NOT AND THEN THESE CELLS WOULD START EXPONENTIALLY DIVIDING, RIGHT WHICH WHICH MAKES IT VERY UNLIKELY THAT THESE WOULD EVER BE ABLE TO CATCH UP. SO WE PUT THESE OUT THERE AND WE TESTED THESE IDEAS AND IN BRIEF AND IN THE INTEREST OF TIME WE FIRST SHOWED THAT IN HUMAN NAIVE B-CELLS, THIS DEFINITELY OCCURS, YOU CAN HAVE SIGNIFICANT DIFFERENCES IN AFFINITY BETWEEN NAIVE B-CELLS, AND 2 DIFFERENT EPITOPE TAUPES AND YOU CAN HAVE LARGE DIFFERENCES IN FREQUENCIES, SO FOR A BROADLY NEUTRALIZING EPITOPE, THE VRC 1 CLASS WHICH IS THE VRC IS VERY, VERY FAMILIAR WITH, WE FOUND THOSE ARE FREQUENT AT HIGH AFFINITY AT 1 AND A MILLION TO THE HUMAN B-CELLS WHERE IN AN EASY EPITOPE THERE MIGHT BE 1 AND 5000. THAT IS A REALEE BIG IN FREQUENCIES FROM A COMPETITIVE STANCE. SO WE THEN TESTED THESE IN DIFFERENT MOUSE MODELS, USING PHYSIOLOGICALLY RELEVANT CONDITIONS AND AGAIN, IN BRIEF, WE DEMONSTRATED ALL 3 OF THESE MECHANISMS OF IMOWN O DOMINANCE DO CONSIDER. AND CAN RESULT IN ESSENTIALLY NO NEUTRALIZING ANTIBODIES DEVELOPING IN SOME CASES AND THE RESPONSE TO THAT WAS ESSENTIALLY AT THE OTHER GROUPS HAVE CONFIRMED THOSE OUTCOMES, THE RESPONSE TO THAT TO SOME DEGREE WAS, OKAY, FINE IMMUNO DOMINANCE IS A REAL PROBLEM, NOW WE'RE JUST SAD. AND THE KEY THING THERE IS, FREQUENTLY TO SOLVE A PROBLEM, YOU HAVE TO FIRST UNDERSTAND WHAT THE PROBLEM WAS, SO KNOWING THAT IMMUNO DOMINANCE WAS AN ISSUE, WE DID A SERIES OF STUDIES TO TRY AND SEE IF IMMUNO DOMINANCE COULD BE MODULATED BY DIFFERENT IMMUNOLOGICAL EVENTS AND WAWE HAVE FOUND IS THAT BY CHANGING THE KINETICS OF THE DELIVERY OF ANTIGEN, YOU CAN HAVE MASSIVE IMPACTS ON IMMUNO DOMINANCE WHERE A CONVENTIONAL INJECTION OF HIV ENVELOPE INTO MONKEYS GENERALLY GIVES ONLY ABOUT 50% OF THE MONKEYS MAKE NEUTRALIZING ANTIBODIES APPROXIMATE AND IT'S ONLY ABOUT AFTER 6 MONTHS OF IMMUNIZATIONS BUT IF WE CHANGE THAT TO A SLOW DELIVERY SUCH THAT THE EXACT SAME AMOUNT OF PROTEIN AND ADJUVANT IS DELIVERED, BUT IT'S DELIVERED OVER--IT'S SPREAD OUT OVER 14 DAYS OR 12 DAYS INSTEAD OF A SINGLE INJECTION, THOSE ANIMALS NOW MAKE NEUTRALIZING EABT BODIES AND THEY MAKE THEM FASTER AND OUR MEASUREMENT TO THE B-CELL RESPONSES TESTED 2 IDEAS, 1 WOULD BE THIS HAPPENS BECAUSE SOMATIC HYPERMUTATION IS BETTER AND YOU JUST HAVE MORE AFFINITY MATURATION, MORE AFFINITY AND THAT'S WHY YOU END UP WITH NEUTRALIZING ANTIBODIES OR IT WAS AN IMMUNO DOMINANCE PROBLEM OF WHICH B-CELLS GOT RECRUITED AND IT TURNED OUT THAT THEY ALL POINTED TO THE IMMUNO DOMINANCE PROBLEM AND UNDER CONVENTIONAL IMMUNIZATION, THERE ARE LOW TO NO HIV B-CELLS THAT GET RECRUITED INTO THE GERM LINE CENTER RESPONSE AT ALL AND THEN THEY GET OUTCOME PETEED QUICKLY. BUT IN THE SLOW DELIVERY CONTEXT WHERE WE HAVE A RESPONSE WE HAVE MUCH BETTER RECRUITMENT OF THE RARER AND LOWER AFFINITY, HIV NEUTRALIZING ANTIBODY CAPACITY B-CELLS AND THEN THAT THOSE HAVE MORE TIME AND MORE ABILITY TO BE KEPT IN A GERMINAL CENTER AND NOT BE OUTCOME PETEED AND ATTAIN ADDITIONAL AFFINITY. AND SOME OF THAT WE THINK HAS TO DO WITH ANTIGEN ANTIBODY COMPLEX FORMATION AND BETTER RETENTION OF ANTIGEN AND WE CAN TRACK ALL OF THAT USING LYMPHNODE FINE NEEDLE ASPIRATES AND IN THE INTEREST OF TIME, I WILL JUMP TO THE FINAL PIECE; OKAY WHICH IS 1 OF THE THINGS WE OBSERVED IN ALL THOSE STUDIES CHES THAT THE GERMINAL CENTER COULD BEHAVE DIFFERENTLY THAN THE SIMPLE MODELS ORIGINALLY IN PLACE AND A GERMINAL CENTER COULD BE A MUCH MORE INCLUSIVE ENVIRONMENT AND ALSO COULD EXIST FOR A SIGNIFICANTLY LONGER PERIOD OF TIME THAN WE THOUGHT. WE THOUGHT GERMINAL CENTERS AM PEAK AT 2 OR 3 WESTBOUNDS BASED ON MOUSE MODELS AND OTHER DATA BUT WE HAD BEEN SEEING GERMINAL CENTERS STILL AT 8 WEEKS AND THAT WAS LONGER THAN WE EXPECTED AND SO WE SET UP A STUDY WHICH WE JUST RECENTLY PUBLISHED TO ASK, WELL, HOW LONG CAN THESE GERMINAL CENTERS LAST AFTER A PRIMING IMMUNIZATION AND IT USED TO BE ABOUT ESSENTIALLY NOTHING HAPPENED AFTER A PRIMING IMMUNIZATION BECAUSE YOU COULDN'T MEASURE ANYTHING IN THE BLOOD AND WHAT YOU SHOWED IS THAT CERTAINLY THERE ARE THINGS IN GOING ON IN THE LYMPHNODE AND MAYBE LOTS OF THINGS ARE GOING ON IN THE LYMPHNODE. SO, WE DID A STUDY THAT WAS AN EXERCISE IN PATIENTS SO HERE I'M SHOWING YOU LYMPHNODE SAMPLES FROM THE SAME ANIMALS, LONGITUDINALLY TRACKED OVER TIME AND LOOKING AT THE GERMINAL B-CELLS AND AT 10 WEEKS POST VACCINATION, WE SAW LOTS OF GERMINAL B-CELLS THAT WERE ENVELOPE SPECIFIC STILL AND AT 4 MONTHS, POST VACCINATION, THOSE GERMINAL CENTERS WERE STILL THERE, AND AT 6 MONTHS, THOSE GERMINAL CENTERS WERE STILL ONGOING EVEN THOUGH WE PROVIDED NO NEW ANTIGEN IN THE SYSTEM SINCE DAY 12. OKAY? SO I DON'T--THIS IS JUST FANTASTIC. THIS WAS REALLY AMAZING THESE GERMINAL CENTERS WERE GOING ON THIS LONG AND MY POST DOC SAID THIS IS TERINCREASE IN BODY, THIS EXPERIMENT'S GOING ON FOREVER, AND SO WE DID END IT AT THAT POINT. AND FOR COMPARISON, WE IMMUNIZED ANIMALS WITH HIV ENVELOPE WITH ALLUM AND WE GOT A PEAK AT ABOUT WEEK 3 AND THEN THEY WERE GONE BY WEEK 10 BUT IF WE DID THIS SLOW DELIVERY IMMUNIZATION, AGAIN JUST OVER A COURSE OF 12 DAYS WITH AN ADJUVANT WE REALLY LIKE WHICH WE CAN DISCUSS, WE SEE AT WEEK TEN-DOLLAR'S OVER A HUNDRED FOLD DIFFERENCE IN THE GERMINAL CENTER RESPONSE BETWEEN THESE 2, IMMUNIZATION APPROACHES AND A HUNDRED FOLD DIFFERENCE IN B-CELLS IS A MASSIVE DIFFERENCE IN THE AMOUNT OF EVOLUTIONARY SPACE THAT CAN BE EXPLORED BECAUSE THEY'RE DIVIDING APPROXIMATELY ONCE EVERY 4-6 HOURS AT LEAST BASED ON MOUSE MODELS AND HERE'S THE QUANTIFICATION OF THE DATA OUT TO 6 MONTHS. SO 1 CRITICAL QUESTION WAS ARE THESE ACTUALLY REAL GERMINAL CENTERS, ARE THESE FUNCTIONAL? SO WE DID SINGLE CELL TRANSCRIPTOMICS AND WE CAN SEE THAT THE GERMINAL CENTER B-CELLS AT WEEK 3 LOOK SIMILAR TO THE GERMINAL CENTER B-CELLS IN WEEK 29 INCLUDING PROLIFERATION MARKERS AND SOMATIC HYPERMUTATION, SO, TRANSCRIPT ORDER OF MICRONSICS THESE B-CELLS LOOK VERY EQUIVALENT BETWEEN EARLY AND LATE. THEY ARE STILL FUNCTIONAL, THEY ARE STILL DIVIDING AND WE SHOW THAT THEY ACONTINUE TO ACCUMULATE FUNCTIONS SO THEY CONTINUE TO ACCUMULATE SOMATIC HYPERPUTATION FOR 6 FULL MONTHS. BUT REALLY TO SUMMARIZE IT ALL IN A SINGLE SLIDE, WE COULD TRACK LYNNERAGES OVER 6 MONTHS IN NISSAN MALS, SINGLE B-CELL LINEAGES BY GOING BACK INTO THESE GERMINAL CENTERS OVER AND OVER AGAIN AND WE COULD SEE THERE WERE LINEAGES THAT STAYED IN THESE GERMINAL CENTERS FOR 6 MONTHS AND THAT THEY WOULD UNDERGO AFFINITY MATURATION. HERE'S 1 THAT'S PRETTY STRIKING EXAMPLE OF ESSENTIALLY UNDETECTABLE AFFINITY AT TIME POINTS AND NOW A THOUSAND FOLD IMPROVEMENTS IN AFFINITY BY 6 MONTHS LATER. THAT DOES HAVE PRACTICAL SMLICATIONS BECAUSE WE CAN ACTUALLY SEE FOR THE FIRST TIME EVER, THAT THESE ANIMALS ACTUALLY DEVELOPED AUTOLOGOUS BEFORE, THIS INDICATING THE B-CELLS WERE FUNCTIONAL BEFORE BOOSTER. SO IMMUNOLOGICALLY THIS IS REALLY FASCINATING BECAUSE BACK TO THE ENVELOPE CALCULATIONS WOULD BE THAT THAT'S ABOUT 750 CELL DIVISIONS THAT THESE GERMINAL CENTER B-CELLS ARE UNDER GOING OVER 6 MONTHS. SO I THINK THAT MEANS WE DON'T REALLY UNDERSTAND THE ABILITY OF THESE CELLS TO PROLIFERATE THE WAY WE THOUGHT AND THAT THESE GERMINAL CENTERS ARE FUNCTIONAL THEY CAN HAPPEN IN RESPONSE TO A PRIMING IMMUNIZATION AND THAT ESSENTIALLY PATIENTS IN A VACCINE STRATEGY CAN HAVE LOTS OF BENEFITS WHEN YOU'RE TRYING TO GENERATE RESPONSES TO DIFFICULT TARGETS, AND THIS WAS LED BY A FANTASTIC INSTRUCTOR JMPLET H. LEE WHO IS CAMERA SHY, AN AMAZING SCIENTIST AND HARRY SUTTON, DID SOME OF THE WORK, AND JUST TO TIE IT BACK TO THE COVID WORK, WE'RE NOT SHOWING ANYTHING NEW HERE IN OUR EXPERIMENTAL DATA, I THINK THE DAILY BASIS AT IS CONSISTENT WITH GERMINAL CENTERS IN SARS-COV-2 INFECTED INDIVIDUALS 6 MONTHS LATER SUGGESTING THAT IS A NORMAL BIOLOGICAL PROCESS AND I THINK [INDISCERNIBLE] HA BETA THAT GERMINAL CENTERS ARE DETECTABLE IN PEOPLE 6 MONTHS LATER WE THINK YOU DON'T HAVE TO INVOKE RNA EXPRESSION FOR 6 MONTHS TO EXPLAIN THOSE RESULTS BUT THEY COULD EXPLAIN WHY MEMORY B-CELLS ARE INCREASING OVER A 6 MONTH PERIOD. IT MAY ALSO BE TRUE FOR ADEN O VIRAL VECTORS BUT THESE KINETICS ARE DIFFERENT FROM THESE KINETICS SUGGESTING THAT THESE DIFFERENT PROCESSES IN THE GERMINAL CENTER, RIGHT AND LUNG GERMINAL CENTERS MAY HAPPEN IN DIFFERENT WAYS WITH DIFFERENT PLATFORMS SO EVERYTHING I SHOWED ABOUT THE HIV STUFF IS COLLABORATIVE SO I DID JUSTMENT TO HIGHLIGHT THAT THAT'S OUR CHILD CONSORTIUM LED BY DENNIS BURREDON, ALL OF THE ANTIGENS ARE PROVIDED BY BILL [INDISCERNIBLE] AND MOST OF OUR WORK IS IN CLOSE COLLABORATION BILL AND ADAPTED RUE AND WITH THAT I WILL STOP AND TAKE ANY QUESTIONS, I KNOW I WENT A SMIJ LONG, YEAH, THANKS IF ARE YOUR ATTENTION. >>GREAT, THAT WAS WONDERFUL SHANE AND I DO WANT TO EMPHASIZE FOR PEOPLE IN THE AUDIENCE THAT PART OF THE REASON WHY SHANE AND ALEX AND OTHERS ON THE TEAM THERE HAVE BEEN ABLE TO REALLY UNCOVER THESE AND STUDY THESE IS JUST BECAUSE OF THE FUNDAMENT AMILLIO WORK ON TFH AND GERMINAL CENTER BIOLOGY THAT HE DESCRIBED AT THE END AND WE HAVE A LOT OF QUESTIONS, I WILL TRY TO GET THROUGH THEM. THERE WERE A COUPLE OF QUESTIONS YOU PROBABLY--SOME ADDRESSED AT THE END ON WHY--WHAT IS IT ABOUT MRNA VACCINES THAT YOU THINK THAT GIVES THIS LONG RESPONSE PARTICULARLY. IS IT THAT THEY'RE BETTER AT TFH CELLS OR CD4 CELL RESPONSES AND WHY IS IT OR WHY DO YOU THINK? >>IT'S DEFINITELY--THEY ARE FANTASTIC AT PRIMING CDFOUR RESPONSES AND FOR EXAMPLE, RBD ALONE IS QUITE SMALL AND DIDN'T HAVE GREAT T-CELL EPITOPES IN IT AND YET BIOENTECH USING RBD WAS ABLE TO ILLICIT QUITE ROBUST CD4 RESPONSE AND THAT'S 1 KEY PIECE OF DATA FROM ME DEMONSTRATING THAT. AND IN TERMS OF KINETICS, WE THINK THAT RNA VACCINES ARE PROBABLY MIMICKING THESE TYPES OF KINETICS WHERE THERE IS ANTIGEN EXPRESSION FOR AT LEAST A PERIOD OF A COUPLE OF WEEKS AND THAT THAT IS INVOKING THESE TYPES OF MECHANISMS THAT WE'RE TALKING ABOUT THAT THEN ALLOW FOR SUSTAINED GERMINAL CENTERS AND I THINK REALLY WHAT WE'RE DEALING WITH HERE ARE THE IMMUNE SYSTEMS EVOLVE FOR MILLIONS OF YEARS TO TRY AND RECOGNIZE PATHOGENS SO IT'S NOT--LET'S SEE IF I CAN--IT'S REALLY USED TO NOT BEING EXPOSED TO A BOLUS IMMUNIZATION, RIGHT? BUT INSTEAD SEEING IT OVER TIME MECHANISMS GET INVOKED AND I WILL--GET INVOKED AND I THINK RNA RACK SEENS ARE EXPRESSING ANTIGEN OVER SOME PERIOD OF TIME. SCOTT BOYD HAS DATA ON HA AND THAT THOSE RESULTS IN THE ABILITY TO SUPER THESE LONGER GERMINAL CENTERS IN OUR CASE, THEY WILL SHOW, THE RESULTS IN A DRAMATIC ANTIGEN CAPTURE AND IT'S A BEAUTIFUL IMAGES FROM DALE URBAN'S LAB WHERE WE LOOK AT WHOLE CLARIFIED LYMPHNODES IN THE MONKEYS WITH A FLUORESCENT ANTIGEN 2 KAYS AFTER CONVENTIONAL SYRINGE. THIS IS A VIDEO, THERE ARE BITS OF ANTIGEN THERE COMPARED TO SLOW DELIVERY OF THE EXACT SAME TOTAL AMOUNT OF ANTIGEN ASK THERE'S ENORMOUS DIFFERENCE IN THE ANTIGEN TO BE CAPTURED AND GO TO GERMINAL CENTERS IF YOU GET THE IMMUNE SYSTEM AHEAD START AND SOME ADDITIONAL TIME. YEAH, GO AHEAD, PAM. >>OH, NO, I WAS REMINDED TO TELL PEOPLE THAT THE CME CODE IS 44465 AND JUST A LOT OF QUESTIONS COMING IN. SO 1 FROM AMANDA HENNING, ARE YOU CONCERNED AT ALL ABOUT INCREASED RISK OF LYMPHOMA DEVELOPMENT WITH LONG GERMINAL CENTER ACTIVITY AND AWLINGS FROM STEPHEN [INDISCERNIBLE] 2 PEOPLE WITH THE SAME QUESTION. >>HMM, YOU KNOW THAT GOT BUMPY THERE SO I WILL PAUSE, CAN YOU ASK THE QUESTION AGAIN? >>YES, SORRY. ARE YOU CONCERNED ABOUT A POSSIBLE RISK OF INCREASE DEVELOPMENT OR AUTOIMMUNITY WITH MORE PROLONGED GERMINAL CENTER ACTIVITY. >>YEAH, WE ABSOLUTELY HAVE TO KEEP THAT IN MIND. CLEARLY THESE RESPONSES CAN GO ON DRAMATICALLY LONGER THAN WE WERE GENUINELY THINKING AND AGAIN, I THINK WHAT WE'RE DOING IS ACTUAL NORMAL BIOLOGY AND I THINK THIS IS PRETTY COMMON IF NOT NORMAL EVEN AFTER VIRAL INFECTIONS AND OTHER EXPOSURES, AGAIN GOING BACK TO SEEING THESE LUNG GERMINAL CENTERS AFTER REMARKABLE CASES OF COVID-19. I THINK THIS IS PROBABLY A PRETTY NORMAL BACKGROUND EVENT IN HUMANS AND THAT VACCINES MAY BE ABLE TO ENGAGE THIS IN A BETTER WAY SO THAT AGAIN, YOU GET THINGS LIKE OMICRON RECOGNITION FROM A VACCINE THAT DOESN'T ACTUALLY INCLUDE OMICRON SEQUENCES BUT YEAH, IT'S A GOOD POINT. >>OKAY. ANOTHER QUESTION, MOST OF YOUR WORK, THIS IS FROM [INDISCERNIBLE] ON SARS-COV-2 INDICATES THAT THE IMMUNE RESPONSE TO THE VIRUS IS EXPECTED AND EXPECTED WITH TB INTERACTIONS BUT THIS INFECTION'S BEEN UNLIKE ANY INFECTION WE HAVE. DO YOU THINK IT'S IMMUNOLOGICALLY DIFFERENT OR IS IT THE VIRUS THAT'S DIFFERENT? >>I THINK IT'S MORE THAN 1 THING. ANY SITUATION WHERE YOU HAVE A PROBLEM THAT'S BECOME AS VAST AS COVID-19. IT'S NOT JUST 1 THING. I ALWAYS--SO, I SAID FOR QUITE A WHILE LOOK THIS VIRUS' SUPER POWER IS THAT IT'S INCREDIBLY GOOD AT EVADING INNATE IMMUNITY. IT'S VERY SILENT ANYBODY THAT CAN HAVE CAUSE AND EFFECT OF DISEASE HAS TO HAVE SOME GOOD IMMUNE EVASION TRICK AND THIS VIRUS IS UNUSUALLY GOOD AT BEING SILENT TO THE INNATE IMMUNITY. RIGHT? THAT'S WHY YOU HAVE ASYMPTOMATIC AND PRESYMPTOMATIC AND PEOPLE ARE TRANSMITTING LARGE AMOUNTS OF VIRUS WITHOUT KNOWING THEY'RE SICK, THAT'S BECAUSE THIS VIRUS AVOIDS TRIGGERING INNATE IMMUNITY FOR I LONG TIME AND THAT DOES HAVE IMPLICATIONS ON THE ADAPTIVE IMMUNE RESPONSE AND THE AABILITY OF THE ADAPTIVE IMMUNE RESPONSE TO REPLY AND IN A PAPER I DIDN'T TALK ABOUT, YOU KNOW WE TRIED--WE COMPARED HOSPITALIZED VERSUS NONHOSPITALLIZED PATIENTS AND OUR INTERPRETATION OF THE DATA WAS THAT IT'S PROBABLY PEOPLE WHO HAVE A RELATIVELY LATE AND SLOW INNATE IMMUNE RESPONSE THAT ARE THE PEOPLE ARE AT HIGHEST RISK FOR HOSPITALIZATION BECAUSE WHAT WE'RE SEEING IS PEOPLE IN THE HOSPITAL WITHOUT DETECTABLE CD4 OR 8 T-CELL RESPONSES SUGGESTING THAT YOUR ADAPTIVE IMMUNE RESPONSE DOESN'T JUST START UNTIL THE INNATE IMMUNE RESPONSE, RIGHT? YOU NEED THAT INNATE IMMUNE SIGNAL SO THIS DELAY IN THE INNATE HAS THIS DOWN STREAM LARGE IMPACT ON THE WAY IN THE ADAPTIVE WHICH THE OLDER YOU ARE, IS MORE LIKELY TO HAPPEN AND CAN PROBABLY BE A MAJOR CAUSE OF MUCH OF THOSE PROBLEMS. CERTAINLY NOW I THINK WE ALL FINALLY RECOGNIZE THAT THE OTHER SUPER POWER OF THIS VIRUS IS VARIANTS AND IT'S ABILITY TO MUTATE AND IT'S INCREDIBLY FRUSTRATING. I THINK SOME OF THAT WASN'T APPARENT EARLY ON BECAUSE IT DOESN'T SHOW THAT MUCH DIVERSITY BUT IT'S SHOWN THAT IF IT DOES--GIVEN THE ABILITY TO IBT--INTEGRATE FECT OVER A BILLION PEOPLE, RIGHT? YOU ROLL THE DICE THAT MANY TIMES WITH SEQUENCE EVOLUTION OF THE VIRUS AND IT CAN COME UP WITH WAYS TO EVADE QUITE A VARIETY OF NEUTRALIZING ANTIBODIES. >>I THINK THIS IS A RELATED QUESTION BUT IT'S ON YOUR INTERPRETATION OF WHY THE VACCINES DON'T HAVE A SIGNIFICANT IMPACT ON TRANSMISSION AS OPPOSE TO SEVERE ILLNESS. >>THEY DO HAVE A SIGNIFICANT IMPACT ON TRANSMISSION, I MEAN TRANSMISSION IN THE EPIDEMIOLOGICAL STUDIES I'M FAMILIAR WITH, TRANSMISSION WAS REDUCED 50-75% IN VACCINATED COMPARED TO UNVACCINATED WHICH YOU KNOW WE START MULTIPLYING THAT ACROSS POPULATIONS, THAT IS SUBSTANTIAL. I DO THINK THE VIRUS, THERE'S BEEN PLENTY EVALUATION PROCESS EVIDENCE THAT THE VIRUS EVOLVED TO BE MOAGHT MORE INFECTIOUS AND HAVE HIGHER TITERS, IT IS HARDER AND HARDER AS TIME HAS GONE ON TO PREVENT TRANSMISSION IS BECAUSE IT'S BECOME AN EVEN FASTER VIRUS AT TRANSMITTING. FOR THE ANCESTRAL STRAIN IT WAS REASONABLE TO EXPECT SOME IMPACT ON TRANSMISSION BECAUSE THERE WAS A SEVERAL DAY WINDOW THERE FOR THAT TO HAPPEN AND THAT'S GOTTEN COMPRESSED MORE OVER TIME. >>SOMEONE'S ASKED, DO YOU KNOW ANYTHING ABOUT THE ENZYME PATHWAYS THAT CLEARS THE MODIFIED RNA IN THE COVID VACCINES? >>NO, IT'S A GREAT QUESTION. I MEAN, THE STANDARD STATEMENTS BY BOTH MODERNA AND PFIZER HAVE BEEN THAT THE RNA IS DETECTABLE 1 OR 2 DAYS POST IMMUNIZATION. THERE HASN'T BEEN GREAT LITERATURE PUBLISHED BY THEM, WELL THERE HASN'T BEEN LITERATURE PUBLISHED BY THEM ON THOSE TOPICS AND I THINK THE DATA FROM SCOTT BOYD WHERE HE SHOWS IN CORE LYMPHNODE BIOPSIES IN HUMANS DETECTABLE RNA 2 WEEKS AND 4 WEEKS POST IMMUNIZATION SUGGESTS AND PROTEIN SUGGESTS THERE MAY BE FUNCTIONAL RNA FOR AT LEAST THAT AMOUNT OF TIME AND AND MAY SUGGEST THAT THE DEGRADATION OF THE RNA, THE MODIFIED RNA IS SLOWER THAN INITIALLY THOUGHT, YEAH. BUT I DON'T KNOW WHAT THE PATHWAYS ARE. THATYA A GOOD QUESTION. >>I THINK IT REALLY HIGHLIGHTS THE CHALLENGES OF LOCAL TISSUE RESPONSES, RIGHT? THIS IS CLEARLY A VIRUS THAT DOES HAVE ORAL INFECTIONS AND SO YOU WOULD LOVE TO KNOW WHAT ARE THE B-CELL AND T-CELL RESPONSES IN THAT TISSUE, RIGHT? THAT'S A HARD QUESTION, THAT IS NOT A HUMAN SITE THAT'S EASY TO SAMPLE AND THERE IS SOME ABILITY TO START SAMPLING, NAISAL TISSUES AND SO I'M HOPING THERE'S MORE AND MORE DATA THAT GET ACCUMULATED FOR EVEN WITH THE KINETICS OF RESPONSES LOOK LIKE, THAT'S HARD TO DO THOUGH, RIGHT? YOU HAVE TO CAPTURE PEOPLE DURING ACUTE INFECTION, PREFERABLY SUPER EARLY DURING THE INFECTION SO YOU KNOW WHAT THE BASE LINE IS BUT WE--PEOPLE WOULDN'T SHOW UP UNTIL SYMPTOMATIC. SO THOSE ARE REALLY IMPORTANT QUESTIONS FOR US TO BED UNDERSTAND WHAT MECHANISMS OF PROTECTIVE IMMUNITY REALLY OCCUR IN THESE TISSUES BECAUSE I DO THINK IT'S QUITE HARD TO EXTRAPOLATE FROM ANIMAL MODELS, ANIMAL MODELS ARE AWESOME BUT YOU NEED SOME DIRECT EVIDENCE IN PEOPLE. I REALLY--I DO--I AM QUITE IMPRESSED BY THE HUMAN CHALLENGE MODELS BOTH FOR SARS-COV-2 AND FLU, FOR EXAMPLE IN HUMAN CHALLENGE MODELS IN FLU, INFECTED PEOPLE, PEOPLE WHO GOT INFECTED PEOPLE WHO HAD CD8 T-CELLS IN THEIR LUNG, THAT'S WHAT CORRELATED WITH LOWER OVERALL VIRAL LOAD DURING YOU KNOW WHEN THEY GOT CHECKED FOR VIRAL LOAD OVER 7 DAYS AND I JUST DON'T THINK YOU CAN FIND THOSE THINGS UNLESS YOU GET TISSUE SAMPLES WHERE YOU FIND [INDISCERNIBLE] HAD IS A HIGH BAR. IT'S ACTUALLY AMAZING, WHAT THE STUDIES ARE BEING DONE NOWADAYS. SO 1 PERSON WANTED TO ASK YOU TO SPEAK TO THE IDEA OF THE ORIGINAL ANTIGENIC SIN AND HOW COVID AND COVID VACCINES HAVE CAUSED THE FIELD TO UPDATE OR MODIFY THIS IDEA IF AT ALL. >>YEAH, WE DON'T--SO WE HAVE A STRONG INTEREST IN THAT TOPIC, WELL FIRST AS HUMAN BEINGS WE WANT TO BE PROTECTED AGAINST COVID BUT WE HAVE A STRONG INTEREST IN THE TOPIC IN THE CONTEXT OF HIV VACCINES AND SORT OF TRYING TO EXTEND UPON WHAT I TALKED ABOUT TODAY WHERE TO GENERATE BROADLY NEUTRALIZING ANTIBODIES WE HAVE A SHEPHERDING APPROACH AND THEN YOU'RE COMING BACK WITH A RELATED BUT DIFFERENT IMMUNO GENERATED AND THEN ANOTHER DIFFERENT BUT RELATED IMMUNOGEN SO YOU HAVE TO HAVE A RECALL RESPONSE THAT YOU CAN BUILD UPON AND HOW DO THINGS LIKE IMPRINTING OR CLINICAL GENETICSINAL [INDISCERNIBLE] IMPACT THAT. I THINK LIKE MOST OF THE FIELD, I TRIED TO MOVE AWAY FROM THE TERM ORIGINAL [INDISCERNIBLE] BECAUSE IT INVOKES QUITE A VARIETY OF EMOTIONAL RESPONSES FROM PEOPLE AND I THINK IMPLINTING IS A LITTLE MORE NEUTRAL. I CAN JUST COMMENT ON THE STATE OF THE LITERATURE WHICH IS THERE ARE BEAUTIFUL PREPRINTS FROM LAURA WALKER AND [INDISCERNIBLE] BETTY SHOWING THAT WHEN A PERSON SEES OMICRON OR ANOTHER FAIRLY DIVERGENT VARIANT, ALMOST ALL THE RESPONSE TO THAT IS MEMORY B-CELLS TO THE ORIGINAL AND WHILE EVERYBODY WAS ABLE TO DETECT NAIVE B-CELLS, THEY WERE LESS THAN 1 PERCENT OF THE RESPONSE AND SO THERE IS A VERY STRONG IMPRINTING. NOW I THINK IT'S EASY TO ELECTRIC AT THAT AS A PRETTY POSITIVE THING. THIS IS ESSENTIALLY WHAT I WAS TALKING ABOUT EARLIER OF IN THE GERMINAL CENTER RESPONSE IS, PART OF A GOAL OF THAT PROCESS IS TO GENERATE MEMORY B-CELLS, THAT HAVE DIVERSE RECOGNITION CAPACITY AND DON'T NECESSARILY RECOGNIZE THE CURRENT ANTIGEN THE BEST BUT MIGHT RECOGNIZE A RELATED ANTIGEN I. E. OMICRON MUCH BETTER THAN WHAT I NAIVE B-CELL COULD STARTING FROM SCRATCH SO THERE MIGHT BE MUTATIONS ON OMICRON THAT ARE AFFECTING AN EPITOPE BUT IF HAVE A B-CELL IT'S BINDING NOT JUST TO THE BINDING OF THE CENTER OF THE EPITOPE BUT BESIDE THE CELL, THE B-CELL CAN STILL HANG ON AND BE A GOOD RESPONDER TO THAT AND I THINK THAT'S WHAT REALLY THE FLU FIELD SEES IN A LOT OF ITS LITERATURE HAS JUST BEEN MORE DIFFICULT TO INTERPRET THE FLU LITERATURE BECAUSE PEOPLE HAVE SUCH INTENSE COMPLEX HISTORIES WITH FLU. YEAH, I GUESS I WILL STOP THERE. THOSE ARE MY MAIN THOUGHTS. >>YEAH, I THINK WE'RE GOING TO HAVE TO STOP BECAUSE OF THE TIME. >>COOL. >>THIS IS REALLY WONDERFUL AND YEAH, WE ARE--YOUR COMMENTS ON THE FLU AND THE HISTORY, IT DOES BRING HOME HOW WE ARE LIVING IN THE WORLD'S BASED EXPERIMENT AND WE ARE THE SUBJECTS. AND I THANK YOU FOR PROVIDING YOUR INSIGHT, SO, THANKS EVERYONE AND WE WILL SEE YOU IN 2 WEEKS. >>YEAH, THANKS IF YOUR TIME AND ALL THOSE EXCELLENT QUESTIONS.