>> GOOD AFTERNOON. I'M DELIGHTED TO INTRODUCE PETER PALESE FROM MOUNT SINAI IN NEW YORK THANKING YOU TO COME HERE ON A RAINY DAY. PETER EARNED A MASTERS AND Ph.D. AT VIENNA UNIVERSITY IN AUSTRIA, POSTDOCTORAL TRAINING AT THE ROCHE INSTITUTE, JOINING MOUNT SINAI IN 1971, PROFESSOR IN 1978, CHAIR IN 1987. ON A MORE PERSONAL NOTE I MET PETER IN 1990 I WHEN I JOINED THE LAB AT A POSTDOC AND COLLABORATED ON A PROJECT TO INFLUENZA B AND A GENES. PETER HAS BEEN HELPFUL IN MY CAREER, SO THANK YOU. HE RECEIVED THE HOWARD TAYLOR RICK ET CETERETTA AWARD, THE EUROPEAN VIROLOGY AWARD IN 2010, AND THE PASSTEUR AWARD IN 2012. IN 1998 HE WAS ELECTED FELLOW OF THE AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE, AND IN 2000 FELLOW OF THE AMERICAN ACADEMY OF MICROBIOLOGY, AND MEMBER OF THE NATIONAL ACADEMIES OF SCIENCES. IN 2012, HE WAS ELECTED TO THE INSTITUTE OF MEDICINE OF THE NATIONAL ACADEMIES OF SCIENCES, IN 2005 HE SERVED AS THE PRESIDENT OF THE AMERICAN SOCIETY FOR VIROLOGY. DR. PALESE'S RESERB HAS BEE RESEARCH HAS B EEN IN THE AREA OF INFLUENZA VIRUS. HE IDENTIFIEDED THE FUNCTION OF SEVERAL VIRUS GENES AND DEFINED MECHANISM OF NEURAMINIDASE INHIBITORS. PIONEER IN THE FIELD OF RNA INHIBITORS, WITH THE STUDY OF VIRAL GENE, PATHOGENESTY AND DEVELOPMENT OF VACCINES. MOST RECENTLY, HE HAS TURNED HIS ATTENTION TO THE TOPIC OF TODAY'S LECTURE, WHICH IS "TOWARD A UNIVERAL INFLUENZA VIRUS VACCINE?." PLEASE JOIN ME IN WELCOMING OUR SPEAKER FOR TODAY'S LECTURE. PETER? [APPLAUSE] >> THANK YOU VERY MUCH FOR THE NICE INTRODUCTION. I'M HAPPY TO SEE SO MANY FRIENDS HERE. LAST TIME I WAS ACTUALLY IN THIS ROOM WAS WHEN GEORGE BUSH GAVE A LECTURE, AND HE GOT A STANDING OVATION AT THE END. SO I HOPE YOU WILL BE EQUALLY TOLERANT TO ME. [ LAUGHTER ] CAN YOU MAKE AN INFLUENZA VIRUS VACCINE? WE'LL BE SPECIFIC ABOUT THIS ONE TOPIC WITH A SHORT INTRODUCTION. THIS IS THE VIRUS, IT'S AN RNA CONTAINING VIRUS, RNA IN THE CENTER, CENTER RNA'S AROUND IT, AND THEN WE HAVE A LIPID MEMBRANE AND WHAT IS REALLY RECOGNIZED BY THE IMMUNE SYSTEM ARE THE SURFACE GLYCOPROTEINS, HEMAGGLUTININ AND NEURAMINIDASE AND THEY COME IN DIFFERENT SUBTYPES AND WE'LL TALK ABOUT THIS A LITTLE BIT MORE AS WE GO INTO THE SUBJECT. WHAT IS IMPORTANT TO KNOW THE RNA SEGMENTS ARE LIKE MINI CHROMOSOMES, IF YOU HAVE TWO INFECT THE SAME CELLS ONE WITH GET AN EXCHANGES AND VIRUSES COME OUT AND YOU CAN SEE THAT THERE'S A LOT OF GENETIC EXCHANGE POSSIBLE, AND THAT IS GIVING RISE TO WHAT WE REFER TO AS PANDEMIC INFLUENZA VIRUSES AND WE'LL ALSO DISCUSS THIS IN MORE DETAIL. NOW, WHAT IS THE SUBJECT OF TODAY'S TALK IS THAT WE BELIEVE WE CAN MAKE BETTER VACCINES. THE PRESENT VACCINES AGAINST INFLUENZA ARE PRETTY GOOD. BUT THEY HAVE TO BE GIVEN EVERY YEAR AND THERE'S SOME OTHER PROBLEMS ASSOCIATED WITH THEM. WE WANT TO MAKE A VACCINE WHICH CAN LAST FOR 20 YEARS OR EVEN A LIFETIME. AND THE PRINCIPLE OF THIS IS THE HEMAGGLUTININ IN ONE OF THESE, THE MAIN GLYCOPROTEIN, THE HEMAGGLUTININ HAS A CONSERVED DOMAIN BUT A HEAD WHICH IS MUCH MORE VARIABLE. AND ACTUALLY THE HEAD IS TEN TIMES MORE VARIABLE THAN THE STALK, SO WE WANT TO MAKE A VACCINE BASICALLY WHERE THE IMMUNE SYSTEM IS DIRECTED AGAINST THE STALK. HEMAGGLUTININ AND NEURAMINIDASE. THINS THE PRINCIPLE WE WANT TO MAKE. WHEN THE REGULAR IS GIVEN, IT'S AGAINST THE HEAD. WE WANT IT AGAINST THE SUBDOMINANT STALK OF THE HEMAGGLUTININ AND THE IMMUNOSUBDOMINANT NEURAMINIDASE. JUST A LITTLE BIT MORE BACKGROUND. WE HAVE MANY DIFFERENT SUBTYPES OF THE HEMAGGLUTININ. THERE'S ACTUALLY NOW A TOTAL OF 18 DIFFERENT SUBTYPES AND THEY ARE VERY DIFFERENT. IF YOU LOOK HERE AT THIS SCALE, THIS IS 4% DOWN HERE, AND SO YOU CAN -- IF YOU ADD THE DISTANCES, WE CAN REALLY CLEARLY SEE THAT SOME OF THESE HEMAGGLUTININS HAVE A SEQUENCE CONSERVATION OF 40, 50%. SO THESE 18 DIFFERENT HEMAGGLUTININS FALL INTO TWO MAJOR GROUPS. GROUP 1 ON THE TOP AND GROUP 2 ON THE BOTTOM. BUT WHAT IS VERY IMPORTANT IS THAT SINCE 1918, WE ONLY HAD H 1, H 2, H 3. THESE ARE THE ONES WHICH ARE BOXED IN RED, AND SO THE H 1 AND H 2 BELONG TO THE GROUP 1. HEMAGGLUTININ, H 3 BELONGS TO GROUP 2. THAT WILL BECOME IMPORTANT AS WE GO ON. THEN JUST SORT OF TO GIVE YOU A FEELING ABOUT THE NEURAMINIDASE, WE HAVE 9 SUBTUBE SUBTYPES -- 10 SUBTYPES AT THIS POINT, 2 APPEARS OVER THE LAST 100 YEARS. NEVERTHELESS, THE BIG PROBLEM WITH INFLUENZA VIRUSES IS THEY ARE CHANGING. THERE IS A LOT OF VARIABILITY. IF WE TALK ABOUT MEASLES VIRUS VACCINE, MUMPS VIRUS VACCINE, WE'RE USING THE SAME VACCINE WHICH WAS DEVELOPED IN THE '50s. SO THE VIRUS HAS NOW CHANGED. OTHERWISE THE VACCINE WOULDN'T WORK TODAY. WITH INFLUENZA, THIS IS DIFFERENT. WE HAVE THE CONTINUING CHANGE FROM YEAR TO YEAR. WE HAVE ANTIGENIC DRIFT, OR EVEN WE CAN HAVE AN ANTIGENIC SHIFT, WHICH IS -- A NEW RNA AFTER THIS REASSORTMENT APPEARS AND WE HAVE THESE PANDEMIC STRAINS WHICH ARE ASSOCIATED WITH MUCH MORE DISEASE AND MUCH MORE MORBIDITY AND MORTALITY. THIS IS SORT OF A RATHER COMPLICATED SLIDE WHICH SHOWS THE DIFFERENT INFLUENZA VIRUSES OVER THE LAST MORE OR LESS 100 YEARS, AND WE HAVE INFLUENZA A VIRUS AND INFLUENZA B, ALSO INFLUENZA C IS USUALLY NOT CASINCAUSING A LOT OF DISEASE. WHAT WE CAN SEE ON THE LEFT IS THE 1918, THAT WAS BASED ON JEFFREY'S SEQUENCE WHICH HE OBTAINED BY VERY ELEGANTLY SEQUENCING MATERIAL FROM PATIENTS WHO DIED AT THAT TIME, AND IN COLLABORATION WE WERE ABLE TO ACTUALLY RECONSTRUCT IN THE LABORATORY SUCH A VIRUS, SO THE VIRUS IS EXTINCT BUT THE H1N1 FROM 1918 HAS GIVEN US A LOT OF INFORMATION WHY THE PANDEMIC VIRUS WAS SO BAD AND THIS IS SORT OF A HALLMARK OF MORE THAN 50, UP TO 100 MILLION PEOPLE DIED IN THIS 1918-1919 PANDEMIC. THEN WE SEE THE VIRUS WAS PRESENT IN H1N1. REMEMBER H 1 BELONGS TO ONE OF THE HEMAGGLUTININS, AND THE VIRUS CHANGES CONTINUOUSLY, THAT'S WHY I HAVE SORT OF CHANGED THE COLOR HERE IN TERMS OF FROM STARTING 1918, WE HAVE THE FIRST VIRUSES, HUMAN VIRUSES GOING BACK TO 1933, AND THEN IN 1957 WE SUDDENLY GOT AN H 2, A NEW PANDEMIC STRAIN, THAT WAS WITH US ONLY 11 YEARS. AGAIN, YOU SEE THE COLOR CHANGE, WHICH MEANS THAT AFTER 2 OR 3 YEARS THE VIRUS HAS SUFFICIENTLY CHANGED ANTIGENIC DRIFT SO WE CAN GET DISEASE AGAIN AND WE CAN ACTUALLY REALLY REINFLEC REINFECT AND GET THE DISEASE. THIS IS ACCENTUATED IF WE GET A NEW PANDEMIC STRAIN AS IN 1968, AN H3,N 2, THE SUBTYPE IS STILL WITH US IN 2014. WE HAD A REAPPEARANCE IN 1977 OF THE 1950 STRAIN, WITH US UNTIL 2009, CHANGING AND THAT IS SHOWN IN THIS COLOR CHANGE. THEN WE HAD A PANDEMIC VIRUS OF THE SAME H1N1, SUBTYPE AS WE HAD THIS SEASON WHAT WE REFER TO AS A SEASONAL H1N1, BUT THE HEAD OF THIS PANDEMIC H1 WAS VERY DIFFERENT, AND WE'LL SEE RIGHT THERE WHY THIS IS OF IMPORTANCE. WE BELIEVE ONE OF THE VERY GOOD EXPLANATIONS WHY THE PANDEMIC H1N1, THAT LED TO THE EXTINCTION OF THE OLD H1N1, WAS BECAUSE SOME STALK-SPECIFIC ANTIBODIES WERE INTRODUCED. WE BELIEVE THE EXTINCTION OF THE REGULAR H1N1 IN 2009 HAD TO DO WITH THE INDUCTION OF ANTI-BODIES WHICH WE'RE RECOGNIZING THE STALK OF THE OLD SEASONAL H1N1. NOW, I MENTIONED THE PANDEMIC OF 1919, 1918, AND THAT ALWAYS IS SORT OF IN THE BACK OF PEOPLE'S MINDS, WHEN WE TALK ABOUT INFLUENZA, I THINK THIS IS A VERY DIFFERENT STORY. TODAY ONE IS THAT WE HAVE ANTIBIOTICS SO IT'S A SUPERINFECTION BY BACTERIA WOULDN'T BE SO BAD BUT ALSO WHAT WE'VE LEARNED, THESE ARE STUDIES IN THE LABS, WE FIND THE VIRUS IS ACTUALLY SENSITIVE TO DRUGS, AND MORE IMPORTANTLY, WE ALL HAVE SOME ANTIBODIES NOW AGAINST THESE H1 VIRUSES SO THAT EVEN IF SUCH A VIRUS WOULD BE FOUND IN HUMANS, IT WOULD NOT HAVE THE SAME IMPACT AS THE 1918 H1N1. SO IT'S NOT ONLY THAT -- THIS VIRUS WAS VERY VIRULENT, BY ALL MEANS, AND ALL THE EXPERIMENTS WE CAN DO IN THE LAB, THIS WAS A REAL VIRULENT VIRUS BUT IT ALSO DEPENDS ON THE IMMUNE EXPERIENCE IN THE HOST AND CLEARLY IN THE 1918 MUST HAVE BEEN A VERY DIFFERENT SITUATION. SO I THINK BY LEARNING ABOUT WHAT THE 1918 VIRUS WAS WE'RE IN A MUCH BETTER POSITION NOW TO BE PREPARED IF SOMETHING ELSE OF THAT ILK WOULD HAPPEN. IN ADDITION TO THE H 1, H 2, H 3, THE ONLY THREE SUBTYPES WE'VE SEEN AND EXPERIENCED AS HUMANS IN THE 1919 PERIOD ON, THERE ARE MANY AVIAN INFLUENCES WHICH HAS BEEN IDENTIFIED AND THEN JUST LAST YEAR IN CHINA, ALL IN CHINA, HALF THE POPULATION OF THE WORLD LIVES IN THIS AREA, WE HAD H 10 AND H 7 AND 9, H 6 AND 1, ALL IN 2014. YOU CAN SEE THERE WERE H10 VIRUSES LIKE H10N7, AND H7N2 BEFORE THAT. WE HAVE OCCASIONALLY, AND THIS I THINK HAS TO DO WITH THE INCREASED SURVEILLANCE WHICH WE HAVE HAD OVER THE LAST ON ONE ONE TO TWO DECADES, WE SEE SOME OF THESE VIRUSES ACTUALLY CAN JUMP OR CAN JUMP FROM AVIAN POPULATIONS, MOSTLY CHICKEN, INTO HUMAN. PARTICULARLY H5N1 HAS CAUSED PROBLEMS STARTING, FIRST APPEARS IN 1987 AND SINCE 2003. HOWEVER, NONE OF THESE DIFFERENT AVIAN VIRUSES WITH THESE DIFFERENT HEMAGGLUTININS AGAINST WHICH WE HUMANS DO NOT HAVE ANY KIND OF COMMUNITY AND PROTECTION, NEVERTHELESS THESE VIRUSES DO NOT HAVE THE CAPABILITY, SO FAR, TO REALLY JUMP FROM ONE HUMAN TO THE OTHER. IN MOST INSTANCES, IT IS LIKELY THAT ONLY A LARGE DOSE OF INFECTIOUS PARTICLES HAS BEEN INHALED BY PEOPLE, ANIMAL HANDLERS, CHICKEN MARKETS, OR HAD TO DO WITH CHICKENS IN RURAL AREAS. IN THIS CASE, I THINK ALL OF THESE VIRUSES, EVEN THOUGH THEY HAVE BEEN FOUND IN HUMANS, I THINK THERE IS NOT THE GREATEST LIKELIHOOD THAT THESE WILL BE THE NEXT PANDEMIC STRAINS. WE HAVE SORT OF ASKED THE QUESTION, WHY IS THE -- WHY DO THESE VIRUSES CHANGE SO DRAMATICALLY IN TERMS OF ANTIGENIC DRIFT, OBSERVABLE IN HUMANS, AND NOT, FOR EXAMPLE, WITH OTHER VIRUSES, AND WE DID -- THIS IS WORK BY A POSTDOC FELLOW, NICK HEATON, WHO DID AN INTERESTING ANALYSIS, THIS IS ONE SLIDE TO SHOW US WHY WE BELIEVE THAT THE HEMAGGLUTININ IS SUCH A UNIQUE MOLECULE. WHAT HE DID WAS ON THE LEFT SIDE WE SEE A MONOMER AND ONE CAN SORT OF DIVIDE INTO GLOBULA AND INTO A STALK DOMAIN, WHAT HE DID IS HE MUTEAGENIZED, INSERTED INTO EVERY POSITION OF 1800 NUCLEOTIDES, 50 NUCLEOTIDE LONG PIECE, AND THEN ASKED THE QUESTION, CAN WE RESCUE VIRUSES, REACH STABLE, AND IT TURNS OUT, VERY INTERESTING, THE BOTTOM LINE, BASICALLY ALL THE MUTATIONS WERE JUST IN THE HEAD, AND THIS CAN BE SHOWN HERE, SO THIS IS THE GLOBULAR HEAD, THIS IS THE STALK HERE, AND THE STALK IS GREEN HERE, SO THE FIRST 52 AMINO ACIDS, THEN GOING FROM 277 TO 565. AND SO WE'RE LOOKING HERE TO INPUT, WE CAN SHOW ALL THE COLORS, IN ESSENCE WE WERE ABLE TO SHOW THAT MUCOGENESIS WORKS, ALMOST EVERY NUCLEOTIDE POSITION WAS CHANGED BY INTRODUCTION OF 50 NUCLEOTIDES, AND THEN AFTER THE RESCUE, WE PUT THIS IN EGGS, AND WE LOOK HERE, WHAT WAS THE MAJORITY OR WHAT CAME OUT? WHAT WAS RESCUABLE? WHAT WAS RESCUED? IF YOU LOOK HERE CAREFULLY, WE HAVE THIS BLUE HERE, AND THE SEQUENCE FALLS RIGHT INTO HERE, IN THE GLOBULAR HEAD, AND THEN WE HAVE A LIGHT BLUE HERE, NOT THAT MUCH, ANOTHER BLUE, THIS BLUE HERE, AND YELLOW, RED, AND THIS YELLOW HERE, AND ALL OF THIS, ALL THE WAY DOWN HERE, IF YOU LOOK, GO BACK HERE, THESE ARE ALL IN THE GLOBULAR HEAD, THESE ARE ALL 15 NUCLEOTIDES, A ROBUST MUCOGENESIS, THE HEAD IS MORE TOLERANT, FLEXIBLE IN TERMS OF MUCOGENESIS AND THE STALK DOESN'T ALLOW US TO MAKE THE MUTATIONS. THIS IS WHY WE BELIEVE THE STALK IS MORE CONSERVED AS COMPARED TO THE HEAD. THE STALK IS INVOLVED IN FUSION, AND IT HAS TO SORT OF FUNCTION -- IT'S MORE IMPORTANT THERE ARE CHANGES. THIS IS A VERY RECENT EXPERIMENT THAT JUST CAME OUT, IT ALLOWED US TO SAY WHY WE BELIEVE THAT THERE'S SO MUCH GENETIC VARIATION IN THE GLOBULAR HEAD. IF YOU LOOK DOWN HERE THERE'S ANOTHER BLUE ONE, BUT IF YOU LOOK DOWN HERE, THIS IS INSERTION OUTSIDE OF THE CODING SEQUENCE, THIS IS REALLY NOT REFLECTED. THE INTRODUCTION OF 50 NUCLEOTIDES PAST. THIS IS QUITE INTERESTING RESULT, AND WE'RE DOING EXPERIMENTS RIGHT NOW WITH MEASLES VIRUS BECAUSE WE CANNOT SELECT SUCH TRANSPOSAL MUTANTS BECAUSE THEY WON'T TOLERATE. OTHERWISE I WOULD FEEL WE SHOULD BE ABLE, THE VIRUS WOULD BE ABLE TO TOLERATE CHANGES. SO WE SUGGEST THE HEAD DOMAIN OF THE HEMAGGLUTININ IS UNIQUELY TOLERANT TO MUTATIONS, THIS IS AN EXPLANATION FOR THE RAPID ANTIGENIC DRIFT AND ALSO AT THE SAME TIME EXPLAINS WHY WE DON'T SEE THIS WITH OTHER VIRUSES YOU ABOUT THOSE DATA WE DON'T HAVE YET. OKAY, SO LET'S TALK SPECIFICALLY ABOUT THE VACCINES WHICH WE HAVE. AGAIN, WE HAVE PRETTY GOOD VACCINES AVAILABLE RIGHT NOW, THEY FALL INTO TWO CATEGORIES, KILLED, MEANING THEY ARE FORMALDEHYDE TREATED VIRUS, AND THEN SPLIT TO MAKE IT LESS REACTOGENIC AND LIVE ATTENUATED, USUALLY GIVEN THROUGH THE NOSE. WE HAVE THE TWO FLAVORS OF VACCINES, KILLED, ADMINISTERED BY NEEDLE, IM, OR WE HAVE BY SYRINGE A LIVE VIRUS VACCINE, CODE ADAPTED. THAT'S WHAT WE HAVE RIGHT NOW. AND AS I MENTIONED BEFORE, WE HAVE VACCINES WHICH HAVE TO BE CHANGED ACTUALLY EVERY YEAR, AND THIS IS A -- NOT COMPLICATED, JUST LOOK AT THE COLORS. WE HAVE THE VACCINE FORMULATIONS OF THE VACCINES WHICH WERE USED FROM 2000 TO 2010, AND RED MEANS THE VACCINE, THIS PARTICULAR COMPONENT, WE HAD AN H 1 COMPONENT, H 3 COMPONENT, B COMPONENT. RED MEANS THE STRAIN WAS CHANGED FROM THE PREVIOUS YEAR, THE FIRST ONE HERE, TWO COMPONENTS WERE CHANGED, IN THIS ONE ALL THREE WERE CHANGED, 2008-2009, ONE YEAR, 2003-2004 IS IDENTICAL TO THE PREVIOUS YEAR, 2002-2003. THIS IS COMPLICATED. WE SOMETIMES DON'T HIT THE RIGHT VACCINE STRAIN, AND IT IS QUITE CUMBERSOME TO PREPARE THIS EVERY YEAR. ALSO IN A CASE OF A PANDEMIC, AND THIS IS WITH THE C.D.C. DATA, ONE CAN SEE THAT WE WERE TOO LATE IN 2009 WHEN THIS PANDEMIC OCCURRED, AND THE RED SHOWS WHEN WE HAD THE MAXIMUM INFLUENZA ACTIVITY IN THE COUNTRY, AND THE BLUE ONE WAS WHEN THE VACCINE WAS SHIPPED OUT, IT WASN'T EVEN DISTRIBUTED YET AND BOTH LIVE AND KILLED TAKES TEN DAYS UNTIL THEY ARE EFFECTIVE. YOU CAN SEE THAT WE WERE MUCH TOO LATE. FORTUNATELY THE PANDEMIC 2009 H1N1 VIRUS WAS NOT VERY HIGHLY VIRULENT. OTHERWISE WE WOULD HAVE HAD A REAL PROBLEM BECAUSE THE VACCINE -- I MEAN BASICALLY WE WOULD HAVE TO SHIF SHIFT UNTIL IT WORKED, ANOTHER FOUR OR FIVE WEEKS, WE WOULD HAVE BEEN TOO LATE. THAT'S THE PROBLEM WITH OUR CURRENT VACCINE. IT TAKES TOO LONG AND I CAN EASILY DISCUSS THIS IN MORE DETAIL, WHY IT TAKES SO LONG BUT WE HAVE TO LOOK AT THE STRAINS WHICH ARE CIRCULATING IN THE SOUTHERN HEMISPHERE, IN JULY AND AUGUST AND THEN WE MAKE UP OUR MINDS, THE C.D.C. MAKES RECOMMENDATIONS, THE F.D.A. ACCEPTS THE -- DEFINES WHAT THE NEW STRANGE SHOULD BE AND IT HAS TO BE CHANGED, PARTICULARLY WITH RESPECT TO THE KILLED ONE IT IS NOT GOOD. WITH RESPECT TO THE PANDEMIC, WE WERE TOO LATE. ALSO THIS WAS PARTICULARLY LAST YEAR WHERE WE HAD MOSTLY AN NH 3 YEARS, THE VACCINE STRAIN DIDN'T REALLY MATCH WHAT WAS GOING AROUND, AND THE HOSPITALIZATIONS, MORE THAN 20% OF ALL THE HOSPITALIZATIONS IN THE LAST TWO WEEKS OF JANUARY WERE ASSOCIATED WITH INFLUENZA INFECTIONS. THERE ARE QUESTIONS CAN WE IMPROVE AND THE QUESTION IS HOW CAN WE DO BETTER. THAT'S THE SORT OF CONCEPT OF A UNIVERSAL INFLUENZA VIRUS VACCINE. THE IDEA IS NOT A NEW ONE. PEOPLE TRIED BASICALLY IN THE PAST TO LOOK AND COULD B CONSERVE, WE HAVE SERVE OF THE UNIVERSAL APPROACHES IN THE PAST, ONE WAS THE M2, VERY MUCH CONSERVED, A SMALL PROTEIN, AND ALSO THE NEURAMINIDASE, PEOPLE LOOKED AT THE NEURAMINIDASE, THAT'S A VERY IMPORTANT COMPONENT OF A UNIVERSAL INFLUENZA VIRUS VACCINE, IT TENDS TO BUILD UNIVERSAL CONVENIENCE ON CTL EPITOPES AND PASSIVE ADMINISTRATION OF ANTIBODIES, BUT THAT OBVIOUSLY WOULD NOT BE FEASIBLE IN TERMS OF A VACCINE FOR EVERYONE. IT'S SORT OF IMPOSSIBLE FOR PEOPLE IN INTENSIVE CARE UNITS, ET CETERA. WHAT I WANT TO CONCENTRATE ON IS TO THE CONCEPT OF REDIRECTING THE IMMUNE RESPONSE AND THE HIGHLY CONSERVED NEURAMINIDASE. IN TERMS OF HEMAGGLUTININ TALK-SPECIFIC ANTIBODIES, WE'VE IDENTIFIED HUMAN, WILSON, WE GROUP AROUND CRUSELLE, IDENTIFIED HUMAN MONOCLONOL ANTIBODIES BINDING NOT TO THE HEAD OF THE HEMAGGLUTININ, WHICH VARIES A LOT, BUT RATHER TO THE STALK. AND THEN WE HAVE ALSO IDENTIFIED OR DESIGNED A VACCINE APPROACH IN MICE WHERE WE VACCINATED WITH AN H 3 FIRST, ONE STALK AND ONE HEAD, AND THEN WE VACCINATED SAME MOUSE WITH BASICALLY THE SAME STALK AND DIFFERENT HEAD, AND THE THIRD TIME WITH THE SAME STALK AND A DIFFERENT HEAD, AND WHAT WE GOT OUT WAS MOSTLY ANTIBODIES AGAINST THE CONSERVED STALK, THAT WAS AN EXPERIMENT IN MICE. AND SUCH SPECIFICITY INTENSE THE STALK CAN BE EASILY SEEN IF ONE DOES A MOUSE EXPERIMENT VACCINATING IN THE WAY I DESCRIBED IT. NOW, THE MOST IMPORTANT, HOWEVER, IS WHAT DOES NATURE DO. WE BELIEVE THAT NATURE USES THIS MECHANISM OF DIRECTING ANTIBODIES AGAINST THE STALK AND THIS WAS THE MECHANISM WHICH EXPLAINS THE ELIMINATION OF THE EARLIER STRAIN IN TERMS O TERMS OF THE H1N1, SEASONA SEASONAL H1N1, BY THE PANDEMIC ONE IN 2009. THIS IS PRE-2009, AND WE ARE HAVING A STALK HERE AND A HEAD, AND THE HEAD IS ALWAYS MORE IMMUNODOMINANT, PROBABLY -- POSSIBLY BECAUSE IT'S SORT OF REALLY ON THE OUTSIDE, THAT'S WHAT THE IMMUNE SYSTEM SEES FIRST. AND ONE GETS VACCINATE AND MAKES ANTIBODIES AGAINST THE STALK, AGAINST THE HEAD, NOT AGAINST THE STALK. WHAT HAPPENED IN 2009 WAS THAT WE HAD AGAIN EXPOSURE TO THE PRE-2009 SEASONAL H1N1 AND THEN THE PANDEMIC H1N1 CAME ABOUT WHICH HAD 20% DIFFERENCE IN THE HEAD AND MORE OR LESS CONSERVED STALK, AND WHAT WE BELIEVE HAPPENED WAS THAT THIS WAS SORT OF -- WE WERE NAIVE AGAINST THE NEW HEAD BUT WE HAD THEN A RECALL AND THEN IDENTIFICATION OF THE B MEMORY CELLS AGAINST THE STALK, WE FOUND THAT THE AVERAGE OF PEOPLE WHO WERE INFECTED WITH THIS NEW PANDEMIC H1N1 SHOWED INCREASED STALK OF 15 TIMES. WE BELIEVE THIS CONTRIBUTED TO THE ELIMINATION OF THE SEASONAL H1N1 BY THE POPPING UP OF THIS PANDEMIC ONE, AND THIS IS MOSTLY BY PICA, A GIFTED Ph.D. STUDENT. HOW ARE WE GOING TO MAKE THIS VACCINE? AND WE ACTUALLY HAD TO DEVELOP ASSAYS TO MEASURE THE STALK ANTIBODIES, THOSE ARE TO MAKE ANTIGENS. WHAT WE FOUND, JUST REMEMBER WE LOOK AGAIN HERE AT A MONOMER OF THE HEMAGGLUTININ, A HEAD, A STALK DOMAIN, AND SO THE EDGE IS DIFFERENT, A CHIMERIC H6, A DIFFERENT SUBTYPE. REMEMBER THE SECOND SLIDE WHERE WE SHOWED THE DIFFERENT HEMAGGLUTININ SUBTYPES, SO WE MAKE NOW A CHIMERIC STALK OF THE HEMAGGLUTININ HEMAGGLUTININ,, WHERE THE HE AD IS AGH 6, THE STALK IS H 1. THIS IS H 9, THIS WOULD BE A STALK AGAIN FROM THE H 1, IT'S A CHIMERIC H 9, SLASH 1, A HEAD FROM H 9 SUBTYPE INFLUENCE HEMAGGLUTININ AND A STALK FROM SUBTYPE 1, AND IN TERMS OF THE LINEAR SEQUENCES, AGAIN, WE HAVE THE 52 AND 277, EXCHANGE INTO AN H6, H9, USING DIFFERENT STALKS SO THIS WOULD BE A PR8 STALK, THIS IS THE CAL09, IN THE PRESENT 2014 VACCINE, AND THEN H 1, H 2, NEURAMINIDASE, A NOT ONLY IN TERMS OF THE HEAD FROM GROUP 1 BUT ALSO GROUP 2. WE HAVE MIXED -- A HEAD FROM GROUP 2, FOR EXAMPLE, H7, WITH A STALK FROM GROUP 1, PR8, WHICH IS 1934, OR THE 2014 VACCINE COMPONENT AND DIFFERENT AMINODASE YOU CAN SEE THE DIRECTION WE'RE DOING, WE HAVE HERE NOW, FOR EXAMPLE, AN H5 WITH A DIFFERENT -- H5 IS GROUP ONE, WITH A GROUP TWO STALK, AND HERE WE HAVE A GROUP 2 HEAD WITH GROUP 2 STALKS. SO WHAT IS REMARKABLE IS THAT THESE VIRUSES GREW VERY WELL, SO WE HAVE, FOR EXAMPLE, HERE, THOSE WHO HAVE DEALT WITH HEMAGGLUTININ INHIBITION TITLES, THE RED BLOOD CELL IS EGG IN, THESNEGATIVEAND THESE ARE POSITIVE, THE VIRUS HAS AN H5 HEAD, PR8 STALK, IN THIS CASE A CAL09 TALK. THIS IS GOOD, THOSE WHO ARE FLOCKED VIRUSES WILL LOVE THAT BECAUSE THIS IS PR8, WHICH IS SORT OF THE HOUSE ANIMAL, WHICH EVERYONE IS USING IN THE LABORATORY, IT GOES WELL, THIS IS ARE THE FLOX, THE CHIMERIC, FOR EXAMPLE, IS ALMOST INDISTINGUISHABLE IN TERMS OF ITS ABILITY TO GROW ONE BUT ALSO TO FORM FLOX. WHAT I'VE SAID SO FAR, A BRIEF SUMMARY, WE HAVE DEVELOPED ANALYTIC TOOLS, THESE VIRUSES, IF THEY HAVE AN IRRELEVANT HEAD, AND NEURAMINIDASE, THEY WILL ONLY BIND IF WE HAVE ANTIBODIES AGAINST THE STALK. AND ALSO, WE CAN USE NOT ONLY TO DETECT AND QUANTIFY BUT ALSO WE CAN HAVE VARIATIONS WHICH CAN BE USED TO NEUTRALIZE VIRUSES FROM DIFFERENT HEADS, CONTAINING DIFFERENT HEADS, AND I WOULD SHOW SEVERAL DATA IN TERMS OF -- UNFORTUNATELY NOT HUMANS, IN TERMS OF ANIMALS. I WILL SHOW DATA IN TERMS OF MICE AND IN TERMS OF FERRETS. OKAY. AND THEN ALSO I THINK THAT'S A VERY IMPORTANT ASPECT IS THAT WE HUMANS DO MAKE THESE STALK ANTIBODIES, AND THE INCREASED STALK ANTIBODIES, WITH THE PANDEMIC H1N1 SERVES AS AN EXPLANATION WHY THE PREVIOUS H1N1 VIRUSES HAVE DISAPPEARED FROM THE CIRCULATION. AND SO NOW THE NEXT COUPLE OF SLIDES WILL BE IN TERMS OF WHAT HAVE WE DONE TO SHOW THAT THIS CAN WORK, SO WHAT THE PRINCIPLE IS WE HAVE ALL PRE-EXISTING COMMUNITY, FOR EXAMPLE IN H1N1, WE HAVE NOT AS MUCH STALK ANTIBODIES WE COME WITH THE VACCINE, IDENTICAL TO THE PRESENT VACCINE EXCEPT WITH AN IRRELEVANT DIFFERENT HEAD, THE SAME IN 2014 H1, BOOSTING THIS SO THAT WE WOULD AMPLIFY THE B-CELLS, B MEMORY CELLS AGAINST THE STALK AND THE SECOND TIME WITH A CHIMERIC, IN THIS CASE H6, IT COULD BE H7, HBECAUS9 BECAUSE WE DON'T HAVE IMMUNITY AGAINST THESE DIFFERENT HEADS, AND IN EACH CASE HERE WE HOPE THAT WE GET ENOUGH OF THIS ANTI-STALK AND ANTIBODY ENHANCED IN TERMS OF A PROTECTIVE IMMUNE RESPONSE. SO IF WE GO NOW TO DATA WHICH WE HAVE IN THE MOUSE, WE HAVE TO -- MICE ARE NAIVE, THEY HAVE NEVER BEEN INACCOUNT IFED. INFECTED. WE'RE PRIMING THEM WITH A CHIMERIC H9/1 DNA, NO NEURAMINIDASE BECAUSE WE WANT TO STUDY THE EFFECT OF THE STALK IN THE MOUSE. WE BOOST WITH THE AND THE ONLY COMMON DENOMINATOR IS THE STALK IN EACH CASE AND WE CHALLENGE WITH DIFFERENT VIRUSES, SO WE PRIME HERE, WE GET SOME ANTI-BODIES AGAINST THE HEAD, SOME AGAINST THE STALK, AND THEN WE BOOST, WE SHOULD GET AGAIN SOME ANTIBODIES AGAINST THE HEAD BUT MORE ANTIBODIES AGAINST THE STALK, AND THE SECOND BOOST ACTUALLY WE GIVE YET A DIFFERENT HEAD AND WE GET HERE EVEN MORE STALK ANTIBODIES. AND THEN WE ARE CHALLENGING, AND THIS IS NOW VERY IMPORTANT. SO WE HAVE HERE -- THE STALK HERE WAS AGAINST THE 1934 VIRUS, AND WE USED THE 1947, OR CHALLENGED THE 2009, OR THE COMPLETELY DIFFERENT SUBTYPE, H5N1 AND H6, ALL THE ANIMALS HAVE SEEN PROTEIN, NOT ANYTHING ELSE. WHAT WE CAN VERY NICELY SEE IS THAT THE VACCINATION WITH THESE CHIMERIC HA STALKS ALLOW PROTECTION, IT JUST SHOWS THE 2009, SO WE USE THE 1934 STALK, AND WE HAVE HERE MICE, WE HAVE STARTING AN INITIAL WAIT. IF WE HAVE NOVATION AFTER NO VACCINATION AFTER 7 TO 10 DAYS, THE MICE ARE RED. DEAD. IN TERMS OF HOMOLOGOUS VACCINATION, THAT SHOULD WORK TOO BUT MORE IMPORTANTLY VACCINE BASED ON A MUCH EARLIER 1934 STALK, ALSO WORKS VERY WELL. IF WE LOOK HERE, AT THE SURVIVAL, ALL THE ANIMALS SURVIVED, YOU CAN CLEARLY SEE THIS HERE. AGAIN, WHAT WE'VE BEEN ABLE TO SHOW IS THAT A STALK IMMUNIZATION, WITH THE STALK FROM 1934, PROTECTS AGAINST IN THE HOUSE AGAINST THE CHALLENGE OF A 2009 VIRUS, AND IT IS BASICALLY AS GOOD AS THE HOMOLOGOUS VACCINATION. AND MORE IMPORTANTLY, WE CAN ALSO CHALLENGE NOW WITH THE DIFFERENT SUBTYPE VIRUS. WE USE AN H5 OR H6, THESE ARE THE SAME ANIMALS, ANIMALS HAVING BEEN IMMUNIZED IN THE SAME WAY. WE FIRST THE DNA PRIMING AND THEN BOOSTING HERE WITH THESE TWO DIFFERENT CHIMERIC PROTEINS, AND THE INCREASE IN TITER, WE CAN MEASURE IN TERMS OF THE ASSAY AND ONE SLIDE HERE SO THAT WE HAVE HERE, WE MEASURE THE STALK ANTI-BODY WHICH IS VERY GOOD, AND MORE IMPORTANTLY WE CAN ALSO PASSIVE TRANSFER OF SERUM PROTECTS FROM VIRAL CHALLENGE. SO THIS IS CLEARLY TELLING US SOMETHING ABOUT THE MECHANISM, THAT IT IS MEDIATED BY ANTIBODY. WE CAN ALSO DO THE SAME THING WITH FERRETS, WHICH WE'VE DONE WITH MICE AND IN THAT CASE AGAIN THE CHIMERIC HA-VACCINATED ANIMALS, YOU CAN SEE THERE'S QUITE A DROP IN, FOR EXAMPLE, OLFACTORY, NASAL, LUNG TITERS, ET CETERA. WE CAN DEMONSTRATE THE MEDIATION OR MECHANISM IS THE SAME AS WE SAW IN THE MOUSE. WE CAN SHOW THE VACCINATION OF THE FERRETS INDUCES A BROADLY REACTIVE ANTIBODY, THE RED CURVE, THIS ONE I THINK IS IS -- OKAY, THIS WILL BE THE 1918. THIS WOULD BE I THINK SOUTH CAROLINA. NO, CAN'T BE. THIS WOULD BE AN H2, THIS WOULD BE AN H17, ONLY A BAD VIRUS, AND NO VIRUS IS ISOLATED. SO THIS IS JUST BASED ON THE SEQUENCE. WE CAN CLEARLY SEE THAT ALSO THE FERRET MAKES THESE ANTIBODIES DIRECTED AGAINST THE STALK OF THE HEMAGGLUTININ AND PROVIDES PROTECTION. AND THEN OBVIOUSLY IF YOU REMEMBER, WE HAVE GROUP 1 AND GROUP 2 HEMAGGLUTININS, AND WE HAVE THESE H3, THAT'S THE ONLY ONE SO FAR, H7, THAT'S SORT OF ONE OF THE VIRUSES WHICH HAVE POPPED UP IN CHINA, AND THAT BELONGS TO THE SAME GROUP. SO WHAT WE DID IS WE DID A VACCINATION NOW, WE TESTED FIRST WHERE THERE IS ANY EVIDENCE THAT THE GROUP 1 VACCINATED ANIMALS ARE PROTECTED AGAINST THE GROUP 2 VIRUS, AND THAT IS NOT THE CASE, WHICH WILL BE ON THE LEFT SIDE SO WE HAVE HERE ANIMALS WHICH WE VACCINATED THE SAME WAY, IDENTICAL WHAT I SAID BEFORE, NAMELY AGAINST THE GROUP 1 STALK, THEN BOOSTED WITH CHIMERIC H6 WITH THE GROUP 1 STALK, HERE IS THE GROUP 1 STALK. WHEN WE LOOK AT THIS IN TERMS OF THE ANTIBODIES AGAINST THE GROUP 2 STALK WE DON'T SEE ANYTHING. MORE IMPORTANTLY IN A CHALLENGE EXPERIMENT, AGAIN, THE H3 VIRUS IS NOT -- IS A CHALLENGE VIRUS, IT IS UNFORTUNATELY KILLING ALL THESE ANIMALS. SO THAT MEANS FOR A TRULY UNIVERSAL VACCINE WE HAVE TO HAVE A GROUP 1 STALK, AS WELL AS A GROUP 2 STALK. AND WE HAVE SHOWN THIS NOW AGAIN, I ONLY SHOW THE DATA IN TERMS OF THE MOUSE, WHERE WE HAVE HERE THE PRIME HERE, NOW WITH THE GROUP 2 STALK, SO H3, H4 HEAD, GROUP 2 STALK AGAIN, DIFFERENT HEAD, GROUP 2 STALK, A DIFFERENT HEAD. AND THEN WE CHALLENGE WITH THE PHILIPPINE STRAIN, H3, 1968, H 3, ALSO H7 AND 9, I WOULDN'T SHOW YOU THE DATA UNLESS I CAN SHOW YOU NOW WE GET PROTECTION IN MICE WHICH HAVE BEEN IMMUNIZED WITH AN H3 STALK, WHICH IS GROUP 2 STALK. AND WE GET COMPLETE PROTECTION ALSO AGAINST THE DIFFERENT SUBTYPE, IN THIS CASE THE H7. ALL ANIMALS ARE PROTECTED IN TERMS OF SURVIVAL HERE. AND SO THE LAST SLIDE IS JUST A SUMMARY OF WHAT WE HAVE DONE IN TERMS OF UNDERSTANDING THE MECHANISM, SO THE STALK-SPECIFIC ANTIBODIES CAN INHIBIT HEMAGGLUTININ, MEDIATED FUSION, AND THERE ARE SOME ASSAYS WHICH CAN BE USED. WE CAN ALSO SHOW THAT CLEAVAGE, AND THIS IS NOT ONLY FROM OUR DATA, BUT ALSO IAN WILSON, AND THAT THE TRIPSON MEDIATED CLEAVAGE CAN BE INHIBITED. WE SHOWED THE STALK-SPECIFIC ANTIBODIES ENGAGE RESENTERS, MACROPHAGES, K-CELLS, ONE OF THE MOST IMPORTANT MECHANISMS BY WHICH THE STALK ANTIBODIES WORK, THEY ACTIVATE COMPLEMENT AND ACTUALLY ARE SIMILAR, THEY ACTUALLY ALSO INHIBIT VIRAL EGRESS. WE'RE SPENDING TIME TO REALLY UNDERSTAND WHAT THESE ANTIBODIES ARE DOING AND WHAT THE MECHANISM IS. THE LAST SLIDE IS HERE WHAT OUR CONCLUSIONS, WE HAVE IN A VACCINE APPROACH HERE, WHICH WE HOPE WILL REDUCE THE IMMUNODOMINANCE OF THE HEAD OF THE HEMAGGLUTININ, AND ENHANCE THE IMMUNOGENICITY, THROUGH CHIMERIC HEMAGGLUTININ CONSTRUCTS, NOT INDUCING AS MUCH OF AN ANTIBODY, THE REDIRECT IMMUNE RESPONSE TO THE TALK AS WELL AS NEURAMINIDASE. THESE ARE ALL ANIMAL DATA ONLY THAT WE CAN PROTECT AGAINST THE CHALLENGE WITH HETEROLOGOUS, VARIANTS OF THE SAME SUBTYPE, BUT ALSO GOING FROM ONE SUBTYPE TO THE OTHER, AS LONG AS IT IS IN GROUP 1, WITH GROUP 1 STALK, SATAN MALVACCINATE ANIMALS. THE ANTIBODY IS MEDIATED, WE CAN SHOW GOOD PROTECTION AGAINST POTENTIAL PANDEMIC VIRUS SUCH AS H5N1 AND H7N 9. LET ME CLOSE HERE AND I'M HAPPY TO TAKE SOME QUESTIONS. [APPLAUSE] NO QUESTIONS? OKAY. >> WE HAVE TIME FOR QUESTIONS, THERE ARE TWO MICROPHONES BACK THERE. I WOULD LIKE TO REMIND YOU THAT THERE'S A RECEPTION IN THE LIBRARY RIGHT AFTER THIS. >> YES, THANK YOU, DR. PALESE. THAT'S WONDERFUL. DO WE KNOW OR CAN WE SAY, CAN WE IMAGINE A TIME LINE TO PHASE 1, PHASE 1 TRIAL? >> I WAS TOLD EARLY ON I SHOULD NEVER GIVE ANY TIME LINES. >> OKAY. OKAY. FAIR ENOUGH. >> NO, BUT IT'S AN EFFORT WHICH IS DRIVEN BY NIH AND I'M HOPEFULLY THIS WILL GET INTO PEOPLE VERY SOON. >> OKAY, THANK YOU. >> BUT THIS IS NOT YET A VACCINE. >> SO IF IT IS A UNIVERSAL VACCINE DO WE NEED VACCINATION EVERY YEAR? HOW LONG DOES IT COVER? >> OKAY, OKAY. SO CLEARLY THE IDEA OF A UNIVERSAL VEEP VACCINE IS THAT WE DON'T HAVE TO VACCINATE EVERY YEAR. SO WE WOULD HOPE THAT BASED ON THE CHANGE WHICH WE'VE SEEN IN THE STALK WE WOULD LOVE TO SUGGEST IT WOULD LAST 20 YEARS, OR EVEN FOR A LIFETIME. IN THE ANIMAL, WE CAN HAVE EVEN A LONGER PERIOD. I POINTED THIS OUT, 1934 VERSUS 2009, THAT'S AN EVEN LONGER ONE. BUT THE UNIVERSAL VACCINE APPROACH IS THAT WE HOME WOULD NEED ONE, HOPEFULLY AT LEAST 20 YEARS, AND MAYBE A WHOLE LIFETIME. AND WE NEED TWO DOSES, FOR THOSE PHYSICIANS HERE, THEY ARE USING IT ALL THE TIME. TO GIVE TWO VACCINE DOSES IS NOT REALLY THE END OF THE WORLD, YOU KNOW? >> FROM THE CLINICAL TIME IT'S STARTING, OR WILL BE STARTING? >> I MEAN, IT HAS TO GO FIRST PHASE 1, PHASE 2, PHASE 3. YES. >> VERY EXCITING. I WAS JUST WONDERING, FOR THE CELL BIOLOGY OF THE NEUTRALIZATION, DO YOU KNOW WHETHER IT HAS TO TAKE PLACE BEFORE BINDING AND ENTRY OR IS IT POSSIBLE WHEN THE STALK IS MORE EXPOSED DURING THE -- IN THE ENDOSOME, ART CERTIFICATION THERE MIGHT BE TRAVELING OF ANTI-BODIES. >>> WE WANT TO INDUCE A POLYCLONLA ANTIBODY, TOWARDS THE STALK, INCLUDING ANTIBODIES WHICH RECOGNIZE ON THE VIRUS ITSELF, SUCH ANTIBODIES CAN BIND TO THREE-QUARTERS, TO THE HEMAGGLUTININS OF THREE-OF THEQUARTERS OF THE HEMAGGLUTININ ON THE VIRUS ITSELF, WE WOULD ARGUE SOME ARE ON THE OUTSIDE BUT WHEN TAKEN IN IT WOULD PREVENT FUSION, FOR EXAMPLE, AND ALSO SOME ANTIBODIES MAY BE WHERE IT BINDS TO THE VIRUS AND RECEPTOR BINDING WITH K-CELLS. MY ANSWER WOULD BE ALL OF THE ABOVE, WE HOPE. >> RIGHT. WE'VE ALWAYS BEEN BEMUSED BY THE FACT EVEN IF WE MAKE THE PEPTIDE AND ANTIBODY AGAINST THE FRAGMENT OF THE HEMAGGLUTININ THAT SHOULD BE BURIED, LIKE THE FUSION PEPTIDE, IT TURNS OUT TO BE NOT SUCH A BAD ANTIBODY FOR IMPACT PH7 VIRUS. >> YES, BUT WE HAVE SOME BREATHING GOING ON, A MILLISECOND, IT OPENS IT UP AND THEN IT CAN BIND THE ANTIBODY. WE WOULD ARGUE THIS HAPPENS HERE, THIS SHOULD HAPPEN WITH THOSE ANTIBODIES AS WELL. >> VERY INTERESTING. WHAT IS THE ANTI-BODY PROFILE AND DO THE ANIMALS DEVELOP ALLERGIES? THAT'S ONE QUESTION. THE OTHER QUESTION IS DO YOU HAVE ANY DIFFERENCES IN THE EFFECTIVENESS OF THESE VIRUSES TOWARDS AGING, IF IT IS DONE IN THE YOUNGER ANIMAL OR OLDER ANIMAL? >> OKAY, SO THE FIRST ONE, THE ANIMAL SYSTEM WE HAVE USED, OUR VACCINATION, WE HAVE ONLY DONE MOUSE, GUINEA PIG AND FERRET. THERE WE HAVE NOT SEEN ANY UNTOWARDS EFFECT WHATSOEVER. LITERALLY WE HAVE VACCINATED THOUSANDS OF MICE. AND BASED ON THE FACT, ALSO, THAT WE HUMANS DO MAKE THOSE ANTIBODIES, THAT WE DON'T -- IT'S NOT THAT WE DON'T MAKE ANY, PARTICULARLY IN THE CASE OF THE PANDEMIC H1N1 IN 2009, WE ACTUALLY MADE QUITE A LOT AND NOTHING HAS HAPPENED REALLY IN ESSENCE. SO I FEEL THIS IS PROBABLY NOT SO MUCH OF A PROBLEM. AND THE SECOND QUESTION WAS -- >> THE AGE. >> THE AGE, NO, WE HAVE NOT LOOKED AT THAT. OKAY. PURE ACADEMIC LABORATORY. >> THANK YOU. >> I JUST HAVE TO NEEDLE THE NIH A LITTLE BIT. YES? >> THANKS FOR A VERY NICE PRESENTATION. UNDERSTANDING THE STALK IS NOT IMMUNODOMINANT, THERE'S NATURALLY OCCURRING STEM ANTIBODY, WHY IS THAT PROTECTED AGAINST PANDEMIC INFLUENZA? >> OKAY, I WOULD ARGUE THAT WE ARE NOW MAKING IN THE POPULATION ACTUALLY SOME OF THE ANTIBODIES, STALK ANTIBODIES, WHICH ELIMINATE THE OLD ONE AND I WOULD PREDICT, AND I'M GOING OUT ON A LIMB, THIS H1N1 WILL NOT LAST LONG, BECAUSE WE WILL MAKE MORE OF THESE ANTIBODIES AND BASICALLY I WOULD ARGUE THIS H1N1 WILL NOT BE AROUND WITH US FOR MUCH LONGER. >> I HAVE A QUESTION FOR YOU. >> OKAY. SO THIS WOULD GO ALONG WITH THE IDEA THAT, YES, WE MAKE THE ANTIBODIES AND THEY SHOULD BE PROTECTED. AND IN A WAY, THE H2 WHERE I THINK THE SAME THING HAPPENED GOING FROM H1 TO H2, THE H2 ONLY LASTED 11ERS YAO. IF YO11 YEARS. GROUP 1 AND GROUP 2 CAN CO-CONSIST BUT WE'LL NEVER HAVE TWO GROUP 1 AND GROUP 2 AT THE SAME TIME. H1N1 WILL DIE OUT SOON. >> WE'VE HAD CLINICAL TRIALS, DO PEOPLE WHO RECEIVE THOSE VACCINES HAVE,, HAVE THEY MADE MORE STEM ANTIBODIES? >> YEAH. WE HAVE SOME DATA FROM NORWAY, WHERE WE CAN MEASURE THAT, AND WE HAVE ALSO SOME DATA FROM NORTHERN SCIENCES WHERE THERE'S A GOOD INCREASE IN ANTIBODIES. WHETHER IT'S ENOUGH, WE DON'T KNOW. WE WOULD HOPE BY HAVING TWO VACCINATIONS WITH DIFFERENT HEADS, THAT WE WOULD BE BETTER -- AND THE HEAD IS NOT UPRIGHT, IT'S H5 AND H7. WE WANT A 2014 STALK AND 2014 NEURAMINIDASE. >> HOW CONCERNED ARE YOU THAT YOU'LL DRAW ESCAPE MUTANTS IN THE STALK REGION? >> YEAH, NO, IT'S A VERY GOOD QUESTION. WHY WOULDN'T THAT STALK ALSO MUTATE ENOUGH AND, YES, TO ANSWER SUCH EVASION MUTANTS WILL HAPPEN. IT'S A POLYCLONAL RESPONSE WHICH IS MORE DIFFICULT, BUT WE WOULD HOPE THAT THESE STALK MUTANTS WOULD BE LESS FIT THAN THE WILDTYPE, AND THAT WE WOULD BE ABLE TO LAST AT LEAST 20 YEARS, HOPEFULLY LONGER. IF THAT HAPPENS WE WOULD NEED A NEW VACCINE WITH A NEW STALK. BUT WE WOULD HOPE THAT THIS IS -- THAT THE STALK IS NOT AS FLEXIBLE AND TOLERANT AS THE HEAD IS SO THE MUTATIONS WOULD BE TAXED BY A FITNESS PROBLEM AND WOULD NOT BE ABLE TO TRANSMIT AS EASILY SO THEY WOULD NOT SURVIVE AS EASILY. AND THE HOPE IS ALSO WE'RE REALLY BANKING ON THE NEURAMINIDASE ANTIBODIES AND STALK ANTIBODIES, TOGETHER THIS IS ABOUT 800 AMINO ACIDS. AND SO WE WOULD HOPE THAT THERE WOULD BE ENOUGH ANTIBODIES AGAINST THESE DIFFERENT EPITOPES SO EVEN ONE ESCAPE MUTANT WOULD HAPPEN, IT WOULDN'T CHANGE MUCH. BUT IT IS CLEARLY ONE OF THE CONCERNS. YES? >> ANY MORE QUESTIONS? IF NOT, I INVITE YOU TO JOIN US IN THE LIBRARY FOR A RECEPTION, AND THANK YOU, PETER. [APPLAUSE]