Articles, Blog

Epidemiology and Virology of the 1918 Flu Pandemic

October 23, 2019


>>IT’S A PLEASURE TO MODERATE THIS NEXT SESSION. MY NAME IS RALPH BARIC, A PROFESSOR IN EPIDEMIOLOGY IN THE GILLINGS SCHOOL OF PUBLIC HEALTH. THIS SESSION WILL FOCUS ON THE EPIDEMIOLOGY AND VIROLOGY OF THE 1918 FLU PANDEMIC VIRUS. WE’RE ACTUALLY QUITE FORTUNATE TO HAVE THE NEXT FOUR SPEAKERS, ACTUALLY THREE IN THIS SESSION, ONE IN THE NEXT, WHO ARE CLEARLY RANKED AMONG THE TOP FLU VIROLOGISTS IN THE WORLD. THEY WOULD BE ON ANYONE’S TOP TEN LIST, I BELIEVE. THE SESSION WILL FOCUS ON THE PATHOGENESIS OF THE 1918 FLU VIRUS, TALK ABOUT RECENT DEVELOPMENTS IN VACCINES AND THE CURRENT STATUS OF ANTI-THERAPEUTIC COMPOUNDS USED TO TREAT VIRUS INFECTIONS, NOT ONLY CONTEMPORARY FLU STRAINS BUT AVIAN AND OTHER PANDEMIC FLUS THAT MIGHT EMERGE IN THE FUTURE. WE ASK YOU HOLD YOUR QUESTIONS TO THE END, AND THEN WILL HAVE THE SPEAKERS COME BACK TO THE TABLE WHERE THEY WILL TAKE QUESTIONS FROM THE AUDIENCE. IT IS HARD TO ACTUALLY DO JUSTICE TO EACH OF THE THREE SPEAKERS IN THIS SESSION. I HAVE HAD THE PRIVILEGE TO ACTUALLY COLLABORATE AND WORK WITH ALL OF THEM. THEY ARE GREAT COLLEAGUES AND WONDERFUL FRIENDS. THE FIRST SPEAKER AND Dr. ADOLFO GARCIA-SASTRE, USING SEQUENCING INFORMATION AND DATABASES, RESURRECTED THE VIRUS IN THE 21ST CENTURY. HAVING HAD ACCESS TO THAT VIRUS ALLOWED US TO BEGIN TO UNDERSTAND ITS PATHOGENESIS, TRANSMISSION, SUSCEPTIBILITY TO VACCINES AND THERAPEUTICS SO HE IS GOING TO TALK TO US ABOUT THE VIRUS AND HOW IT CAUSES DISEASE. ADOLFO IS THE FISHBERG PROFESSOR OF MICROBIOLOGY, MEDICINE AND INFECTIOUS DISEASES, THE DIRECTOR OF THE GLOBAL HEALTH OF THE ICAHN SCHOOL AT MOUNT SINAI IN NEW YORK. HE HAS STUDIED INFLUENZA VIRUS AND OTHER STRANDED R-VIRUSES AS WELL AS OTHER VIRUSES THAT APPEARED IN HUMAN POPULATIONS FOR AT LEAST 20 YEARS. HE HAS MADE SIMILAR CONTRIBUTIONS THAT ARE REFERENCED THROUGHOUT THE LITERATURE. HE IS CURRENTLY THE DIRECTOR OF THE CENTER OF RESEARCH ON PATHOGENESIS WHICH IS ONE OF THE FIVE NIH FUNDED CENTERS ON RESEARCH AND SURVEILLANCE. OUR NEXT SPEAKER IS DOCTOR BARNEY GRAHAM. BARNEY IS THE DEPUTY DIRECTOR OF THE VACCINE RESEARCH INSTITUTE AT NATIONAL INSTITUTE OF HEALTH AT INFECTIOUS DISEASES AT NIH, THE CHIEF OF PATHOVIRAL GENESIS LIBRARY AND OVERSEES MUCH OF RESEARCH THAT FOCUSES ON NEW, VACCINES AS WELL AS HIGHLY PATHOGENIC RESPIRATORY VIRUSES AND OTHER VIRUSES. HIS RESEARCH INCLUDES VACCINE DEVELOPMENT FOR HIV, RESPIRATORY VIRUSES, CORONA VIRUSES AND OTHER EPIDEMIC VIRUSES THAT HAVE EMERGED IN THE LAST DECADE. HE IS AN IMMUNOLOGIST, VIROLOGIST AND CLINICAL PHYSICIAN AND I LOOK FORWARD TO HEARING THE LATEST UPDATES ON THE DEVELOPMENT OF NEW FLU VACCINE STRATEGIES TO CONTROL THIS IMPORTANT HUMAN PATHOGEN. OUR FINAL SPEAKER IS Dr. FREDERICK HAYDEN WHO IS DIRECTOR OF AND PROFESSOR EMERITUS OF MEDICINE AND PATHOLOGY AT THE UNIVERSITY OF VIRGINIA SCHOOL OF MEDICINE. HE HAS STUDIES RESPIRATORY AND CLINICAL PATHOGENESIS FOR MANY YEARS AND HAS BEEN AT THE FOREFRONT OF DEVELOPING AND TESTING FLU THERAPEUTICS USED IN HUMAN POPULATIONS, SO HE WILL GIVE US UPDATES NOT ONLY ON THE CURRENT LIST OF ANTIVIRALS THAT ARE AVAILABLE FOR TREATMENT OF INFLUENZA VIRAL INFECTION BUT ALSO WILL GIVE US AN UPDATE ON THINGS IN THE PIPELINE THAT WILL PROVIDE HOPE FOR FUTURE GENERATIONS. SO WITH THAT, ADOLFO, PLEASE LEAD US OFF. WELCOME. [APPLAUSE]>>THANKS A LOT, THANKS FOR HAVING ME HERE. THIS IS A REALLY NICE MIXTURE BETWEEN PUBLIC HEALTH AND SCIENTISTS AND I AM ENJOYING THE FIRST TALKS OF THIS MORNING. SO IF YOU LIKE MY TALK THAT WAS ALMOST GIVEN BY RALPH, ALMOST EXACTLY WHAT I WAS GOING TO SAY — [LAUGHTER]>>SO I WAS INVOLVED ON THE TEAM THAT RECONSTRUCTED THE 1918 INFLUENZA VIRUS AND WOULD LIKE TO TELL THIS STORY ABOUT WHY WE DID THAT AND WHAT WE HAVE LEARNED FROM THIS VIRUS AND WHY PARTICULARLY WE FELT IT WAS IMPORTANT, THE WORK THAT WE DID AT THIS TIME AND HOW IT MAY TEACH US HOW TO FIGHT AGAINST THE INFLUENZA VIRUS. SO I HAVE SOME SLIDES, PROBABLY YOU ARE FAMILIAR WITH SOME OF THEM AS WELL AS THIS ONE. SHOWS THE LIFE EXPECTANCY IN THE LAST 100 YEARS THAT HAS BEEN INCREASED DUE TO MODERN MEDICINE, ANTIBIOTICS, HYGIENE TECHNIQUES, ET CETERA, ET CETERA, AND THE BIG DIP HAPPENED IN 1918 AND IT WAS CAUSED BY THE 1918 EPIDEMIC, NOT THE GREAT WAR GOING ON AT THIS TIME. THE GREAT WAR WAS A CONTRIBUTION TO THE LIFE EXPECTANCY, NOT ONLY IN THE UNITED STATES BUT THE WHOLE WORLD. THE OTHER IS A PICTURE FROM THE SMITHSONIAN. THIS IS A PICTURE OF A LITTLE VIRUS IN ALASKA, THE SURVIVALS OF 1918 WHERE THE VIRUS ORIGINATED REMOTELY IN ALASKA AND WHAT YOU CAN SEE FROM THE PICTURE IS THERE WERE NO ADULTS IN THE PICTURE BECAUSE ALL OF THE ADULTS DIED IN 1918. IT WAS A VERY DRAMATIC INFECTION IN HEALTHY ADULTS, CAUSED 24 PERCENT DEATHS BUT IN REMOTE AREAS LIKE THIS VILLAGE IN ALASKA WHERE THERE WAS NO IMMUNITY AGAINST THE INFLUENZA VIRUS, IT WAS MORE DEVASTATING IN THOSE AREAS AND IN THIS CASE, ALL THE ADULTS DIED. NOW, WHY STUDY A VIRUS THAT WAS 100 YEARS OLD? WELL, THIS VIRUS CONTAINS THE TERMINALS RESPONSIBLE FOR ITS SUCCESS AND WE DON’T HAVE TOO MANY EXAMPLES OF INFLUENZA VIRUS IN HUMANS. WE HAD THREE SAMPLES AND THEY WERE THERE FROM 1918 WAITING TO BE RECOVERED. NOW, THE INFORMATION WE HAVE FROM THIS VIRUS, WHAT STARTED THE PANDEMIC, IT WAS NOT UNDERSTOOD WHEN WE STARTED RECONSTRUCTION OF THE VIRUS AND THOUGHT WE WOULD UNRAVEL SOME NEW VIROLOGY THAT WE DIDN’T UNDERSTAND BEFORE FOR VIRUS IN HUMANS. AND THE 1918 VIRUS, IT MAY HAVE BEEN A SINGLE EPISODE BUT THIS ONE DOES NOT EXCLUDE THE POSSIBILITY THAT IT CONTAINED SIMILAR CHARACTERISTICS THAT COULD COME IN THE FUTURE AS A PANDEMIC VIRUS SO WE THINK BY STUDYING THE VIRUS IN 1918, WE WILL HAVE MORE INFORMATION ON WHAT MAKES IT A LIVING VIRUS IN HUMANS AND HOW WE CAN FIGHT AGAINST THIS VIRUS. SO THE PROBLEM YOU KNOW IN 1918, IT WAS KNOWN THAT INFLUENZA WAS CAUSED BY A VIRUS AND THERE WAS NO TECHNIQUE TO ISOLATE THE VIRUS AND THEN THE VIRUS BECAME EXTINCT. THE SAMPLE FROM THE VIRUS CAME FROM HUMANS BUT DUE TO EVOLUTION, THE VIRTUAL LENS OF THIS VIRUS CHANGED AND BECAME LESS VIRULENT. NOW, THE VIRUS WASN’T AVAILABLE BUT WAS AVAILABLE ON SOME SURFACE OF TISSUE WHERE THE VIRUS WAS NOT THERE, THE INFECTIOUS VIRUS BUT THERE WAS STILL GENETIC MATERIAL, DISINTEGRATED BUT IT WAS STILL THERE AND SO THE INDIVIDUAL AT NIH GOT ACCESS TO MATERIALS LIKE THAT AND HAS PRODUCED TWO TYPES OF SAMPLES OF THE 1918 VIRUS, COLLECTED THUS FAR FROM DATA BANKING OF TISSUES OF THOSE WHO DIED OF KNOWN DISEASES AND GOING ON FOR MANY YEARS. THE OTHER WAS THE BODY THAT WAS BURIED IN ONE OF THESE REMOTE ALASKA VILLAGES, RECOGNITION OF A PERSON WHO DIED IN 1918. I SHOWED YOU THE PICTURE BEFORE, MOST OF THE ADULTS DIED IN THIS VILLAGE, THE ARMY CAME AND BURIED THE BODY UNDER THE PERMA FROST LINE SO THE BODIES KEPT THE VIRUS IN THEM, THE DURATION OF THE TISSUE FOR THE INFECTIOUS VIRUS WAS NOT THERE BUT GOT THE TISSUES FOR PERFORMING SEQUENCING. SO WE START WITH PATHOLOGICAL SPECIMENS OF PEOPLE WHO DIED IN 1918 OF THE VIRUS INFECTION AND THEN THROUGH THE SEQUENCING, WE ARE STILL NOT VERY SOPHISTICATED WITH SEQUENCING TECHNIQUES LIKE THE ONES WE HAVE NOW, BASICALLY LITTLE BY LITTLE GETTING THE SEQUENCE OF THE WHOLE VIRUS AND ONCE YOU HAVE THE SEQUENCE, YOU CAN RECONSTRUCT THE GENES AND DNA AND INTRODUCE THE TECHNIQUE AS A VIRUS TO ASK QUESTIONS ABOUT HOW THE VIRUS LOOKS LIKE. AND FOR EXAMPLE, A SINGLE GENE THAT IS INCLUDED FROM 1918 THAT IS TYPICAL NOW OF HUMAN INFLUENZA VIRUS. SO THE FIRST STEP IS TO TAKE THE VIRUS AND KEEP IT IN HIGH CONTAINMENT AND JUST FROM LOOKING AT IT, ONE COULDN’T SAY WHAT WAS THE DETERMINANCE OF THE VIRTUAL LENS OF THIS VIRUS. THERE WAS NOTHING KNOWN AT THAT TIME ABOUT THE SEQUENCE OF THE VIRUS THAT MAY TELL US WHY IT WAS SO VIRULENT. SO WHAT IS THE VIRTUAL LENS OF THE VIRUS? WE INFECT MICE AND IF DO YOU IT WITH A HUMAN, HNO1 — [INDISCERNIBLE] — YOU DON’T GET FATALITY IN MICE SO THIS IS MICE, 50, YOU NEED MORE THAN 6 PFU IN THE MICE FROM 1991. NOW, WE INTRODUCED JUST TWO GENES FROM 1918, NOW YOU GET THE VIRUS THAT IS ABLE TO BE LIVING IN MICE WITH 10 TO THE 5TH MORE OR LESS IS REQUIRED TO GET PATHOLOGY IN MICE. WITH MORE GENES, YOU DON’T CHANGE SO MUCH THE LETHALOGY OF THE MICE BUT YOU INCREASE AROUND ONE AND A HALF — [INDISCERNIBLE] SO THE FIRST THING WE LEARN FROM THESE STUDIES THAT THE VIRULENCE OF THIS VIRUS IS MUTAGENIC. THERE IS NOT A SINGLE GENE RESPONSIBLE FOR THE VIRUS, WHICH GIVES US A LITTLE MORE HOPE THAT IT WOULD BE VERY DIFFICULT TO GET A VIRUS LIKE THAT BECAUSE YOU NEED TO COMBINE MULTIPLE VIRULENCE MARKERS IN ORDER TO COME UP WITH A VIRUS LIKE THAT. SO IT IS NOT A SINGLE DETERMINANT. THEN IF WE INJECT MICE, INFECT MICE TO 10 TO THE 6TH PFU, ALL OF THEM SURVIVE, THE ONE WITH THE VIRUS FROM 1918, THEY ALL DIED AND IF YOU GET TO THE 1918, THEY DIE MORE QUICKLY FROM THE BASE POLYMERS. THEN WE GET THE VIRUS FROM 1991, THIS VIRUS IS NOT ABLE TO KILL MICE SO THIS IS IMPORTANT AS WE LOOK TO WHAT HAPPENED TO THE REPLICATION IN THE ANIMAL. THE ONE THAT IS NOT LETHAL GROWS TO 1006 UNITS FOR THE LIFE OF THE ANIMALS. THIS ONE AROUND 10 TO THE 6TH, THIS ONE A LITTLE MORE LETHAL, 10 TO THE 8TH AND THIS ONE, NOT LETHAL, 10 TO THE 5, SO REPLICATION LOOKS LIKE IT AFFECTS THE VIRULENCE AND IF IT IS ABLE TO REPLICATE MORE IN THIS ANIMAL, YOU GET LETHALITY. NOW, IN COLLABORATION, WE LOOKED TO THE HOST IMMUNE RESPONSE HAPPENING IN THIS ANIMAL AND WE DID THAT IN A GENERAL WAY BY LOOKING JUST TO THE WHOLE RESPONSE WITH HOW MANY GENES ARE COMING UP AND DOWN WHICH IS SEEN HERE IN THIS PLOT. SO BASICALLY WHAT I WANTED TO SEE HERE IS THE CHANGE IN GENUS EXPRESSION FOR THE LENGTH OF MICE INFECTED BY THE CONTROLLED VIRUS OR THE COMPLETE 1918 — [INDISCERNIBLE] AND THE MOST INTERESTING THING TO SO SEE IS IN THIS PART, THE ONE INFECTED WITH IT FROM 1918 AND DIFFERENT MICE, WHAT YOU SEE IS MORE COLOR. MORE COLOR MEANS MORE CHANGES AND MORE CHANGES MEAN THERE IS A LOT OF INFLAMMATION IN THIS ANIMAL. SO THE MORE RESPONSE, THE MORE INFLAMMATION. NOW, MOST OF THIS CHANGE IS HAPPENING WITH GENES, MICROPHAGES OR T-CELLS, THESE CHANGES CORRELATE WITH THE ABILITY OF THE VIRUS TO REPLICATE IN THE MICE. THE HIGHER YOU REPLICATE, THE MORE CHANGES YOU GET. AND WHAT HAPPENS WITH THE REPLICATION FROM WHAT IT APPEARS TO BE, DEFINITELY A RELATION OF THE EXPRESSION OF THE CHANGES THAT GOES RIGHT TO THE ANIMAL PATHOLOGY AND THAT IS WHAT WE THINK IS KILLING THIS ANIMAL. NOW, THE AUDIENCE WILL HEAR THIS PRESENTATION A LITTLE LATER, HE CONSTRUCTED THE VIRUS AND FOUND VERY SIMILAR THINGS IN MACAQUES AND FOUND THAT THE VIRUS IS EVEN ABLE TO KILL A MACAQUE DUE TO AN INVASIVE RESPONSE. NOW, WITH THE GROUP, WE USED A FERRET MODEL AND THE REASON WE USE FERRETS, THEY LOOK A LOT LIKE HUMANS, AS YOU CAN SEE. [LAUGHTER]>>THEY BEHAVE LIKE HUMANS WHEN IT COMES TO INFECTIONS. THEY ARE NOT NECESSARILY SUSCEPTIBLE TO THE INFLUENZA VIRUS INFECTION, SO WE GIVE THEM THE INFECTION AND THEY EXHIBIT SIMILAR SYMPTOMS TO INFLUENZA, VARIOUS INFECTIONS, RUNNY NOSE, FEVER. AND RESPOND TO MEDICINE. SO WE PUT 1918 VIRUS IN FERRETS, USED THREE FERRETS WHERE THE VIRUS WAS INOCULATED BUT IN ADDITION, WE USED THREE CONTACT FERRETS WHERE THEY WERE NOT INFECTED BUT THEY WERE PUT IN CLOSE PROXIMITY TO THE INFECTED FERRETS ONE DAY AFTER THE INFECTION, NOT ALLOWING CONTACT BUT TRANSMISSION BY DROPLETS AND LOOKING AT THE ABILITY OF THE FERRETS TO TRANSMIT AND WHAT TYPE WILL BE IN THE FERRET THAT WAS PRESENT NOT FROM EXPERIMENTAL IMMUNIZATION BUT FROM CONTACT AND AGAIN, TWO OUT OF THREE FERRETS DIE OF THE ONES THAT WERE INFECTED AND THE CONTACT FERRETS, THE VIRUS WAS TRANSMITTED TO THE CONTACTS AND QUITE SEVERE WITH ONE ANIMAL OUT OF THREE DYING IN THIS GROUP. AND THIS CORRELATED ALSO WITH THE ILLUSTRATION OF IMMUNE CELLS IN THE FERRETS THAT WERE INJECTED WITH THE 1918 VIRUS AND IN THE CONTACT FERRETS, YOU SEE FEWER CELLS. NOW, I TOLD YOU ABOUT SOME OF THE GENES RESPONSIBLE FOR THAT BUT IN ORDER TO SEE WHAT ACTUALLY ARE THE SPECIFIC GENES RESPONSIBLE FOR THE MAXIMAL REPLICATION OF THE VIRUS OF 2018, AGAIN, WE SUBSTITUTED EVERY SINGLE GENE OF THE 1918 VIRUS WITH ONE GENE OF THE HN101. SO WE GET A VIRUS THAT WAS IDENTICAL TO 1918 EXCEPT ONE OF THE GENES THAT WE NOW HAVE FROM THE HN101 FROM 1991. AND THEN WE LOOK AT THEIR ABILITY TO REPLICATE THE HUMAN CELL AND AS YOU CAN SEE, MOST OF THEM REPLICATE WELL EXCEPT FOR THIS TREE HERE AND ARE THIS INCLUDES THE MICRO– [INDISCERNIBLE] AND SO AGAIN THE ABILITY TO REPLICATE TO HIGH LEVELS AND REQUIRES H1N1 TO GET IT TO KILL THE MICE. SO AGAIN, CORRELATING HIGH LEVELS OF REPLICATION WITH ABILITY TO — OR LETHALITY IN THE MICE. NOW, HE NOT ONLY SEQUENCED ONE OF THE GENES BUT HE ALSO SEQUENCED FROM TWO PEOPLE WHO DIED IN 1918, ONE FROM SOUTH CAROLINA, ONE FROM NEW YORK AND TWO LONDON SAMPLES HE WAS ABLE TO OBTAIN FROM TOMBS IN LONDON WHERE THE BODIES WERE BURIED IN IT LEATHER CASES. AND IT WAS IDENTICAL IN OTHER CASES EXCEPT FOR ONE CHANGE HERE FROM B TO G, SO THERE WERE TWO DIFFERENT EXAMPLES FROM 1918 FROM THESE TWO SEQUENCES AND WHAT IS INTERESTING IS THIS HAS BEEN KNOWN TO PLAY A ROLE IN THE HEMOGLUTINEN. SO IT IS ABLE TO PLAY A ROLL IN TWO LINKAGES — [INDISCERNIBLE] THEY HAVE TWO VIRUSES THAT BIND TO 21 AND THE HUMAN ONE HERE, THEY BIND TO 26. NOW, WE WOULD PREDICT THAT BY THIS CHANGE WITH SPECIFICITY AND EVEN IF YOU INTRODUCE A NEW CHANGE HERE, YOU WOULD CHANGE COMPLETELY THE RECEPTOR SPECIFICITY. SO WE THEN USED THE ONE WE USED BEFORE, THE SUBMISSION FROM SOUTH CAROLINA, THEN CONSTRUCTED THE VIRUS WITH A SINGLE CHANGE AND THE VIRUS THAT CONTAINS THE TWO CHANGES. AND THEN WE LOOK FOR ABILITY TO BIND TO THE RECEPTORS, 23 AND 26. SO THEY INTRODUCE IT MAINLY BY INTRODUCING ON THE 23. THE HUMAN SAMPLES FROM MOSCOW OR SOUTH CAROLINA IN 1918 BY THE RECEPTOR. IN NEW YORK, CHANGED FOR SPECIFICITY — [INDISCERNIBLE] AND THIS IS INTERESTING BECAUSE IN ADDITION TO THE RECEPTOR TO CREATE 26 IN HUMANS, 23 IS MORE ABUNDANT IN THE LOWER RESPIRATORY TRACT OF PEOPLE WHERE INFECTION IS MORE SEVERE IF THE VIRUS GOES THROUGH. SO THEN WE USED THE LARGEST OF THE EFFECTS AGAINST FERRETS. THE PREVIOUS VIRUS FROM 1918, NOW WITH THIS, THEY DIE, THE FERRET, BUT THIS ONE DOES NOT PROCEED AND THAT WAS ONE OF THE FIRST EVIDENCE FOR TRANSMISSION OF THE VIRUS AND LATER CONFIRMED WITH THE H5N1 VIRUS THAT THEY NEED TO HAVE THE RIGHT RECEPTORS TO BE ABLE TO TRANSMIT TO SOMEONE. NOW, THE OTHER DESIGN OF STUDYING THE VIRUS IS WE NEED TO BE SURE WE CAN FIGHT SOMETHING LIKE THAT. SO ARE THE ANTIVIRALS EFFECTIVE AGAINST THE 1918 FLU VIRUSES? WE USED A LITTLE CHALLENGE WITH THE INHIBITORS, THE ANIMALS SURVIVE AND IF WE USE AMANTADINE, WE SEE IT WAS ABLE TO INHIBIT THE VIRUS. SO IF WE CONTROL INFLAMMATION, CAN WE CONTROL THE VIRUS SO THESE ARE THE EFFECTS YOU GET AND WITH THE MAIN INFLAMMATORY CELLS GOING THERE, YOU DON’T REDUCE MORTALITY, YOU INCREASE MORTALITY SO ALTHOUGH INFLAMMATION IS A GOOD WAY TO GET IN THESE VIRUS, FOR TREATMENT OF THE VIRUS, YOU CANNOT JUST REDUCE INFLAMMATION BECAUSE IF YOU DO THAT, YOU GET MORE CELL GROWTH. NOW, HOW ABOUT VACCINES? WE USE A VACCINE THAT IS BASED ON THE NA VIRUS FROM 1918, IT DOES NOT WORK BUT EVEN VACCINES FAR AWAY, THEY ARE PARTIALLY PROTECTIVE AND AS A YOU GET CLOSER TO THE 1918, YOU GET SOME RESPONSE SO AT LEAST WE FOE THAT THE VIRUS WE HAVE WORK ON THE 1918 VIRUS. SO WHAT DO WE KNOW NOW? THE 1918 VIRUS IS THE ONLY KNOWN HUMAN INFLUENZA VIRUS LETHAL TO MIGHT, FERRET AND MACAQUES. , SINGULAR AM AINO AS A SID CHANGE IN HA CHANGES IN FERRET BUT THE GOOD NEWS, THE H1 VIRUS RESPONDS TO VACCINES. SO IF IT WOULD BE THE EXACT 1918 VIRUS, THE GOOD NEWS IS THE NEW HAN101 VIRUS, IT IS NOT HIKE THE VIRUSES WE HAVE NOW. AND THE VACCINES WE HAVE ARE NOT ONLY INHIBITING THE VIRUS FROM THE 1989, IT IS EFFECTIVE AGAINST 1918 SO IF WE HAVE SOME BIG VIRUS TO BE A PANDEMIC, IT WILL REQUIRE SOMETHING DIFFERENT THAN THE ONE FROM 1918. [INDISCERNIBLE] THANK YOU. [APPLAUSE]>>GOOD MORNING, MY PLEASURE TO BE HERE. I THANK RALPH FOR INVITING ME TO THE ORGANIZERS FOR ALLOWING TO US SHARE SOME OF OUR WORK. I AM GOING TO GO THROUGH WHAT SOME OF THE OPTIONS ARE FOR UNIVERSAL FLU IMMUNIZATION AND TRYING TO FIND VACCINES THAT ARE BETTER THAN THE ONES WE ALREADY HAVE. SO I WORK AT THE VACCINE RESEARCH CENTER ON THE CAMPUS IN BETHESDA NIH. WE’RE IN THIS BUILDING 40 HERE AND THESE ARE THE PRINCIPAL INVESTIGATORS AND PROGRAM HEADS THAT I WORK WITH EVERY DAY. VRC WAS FOUNDED TO WORK ON HIV INFECTION AND DEVELOP AN AIDS VACCINE BUT IN THE MEANTIME, WE HAVE TAKEN ON A NUMBER OF THESE OTHER PROJECTS, MANY VIRUSES THAT YOU HEARD OF COMING OVER THE LAST FOUR OR FIVE YEARS. MUCH OF THE TECHNOLOGY THAT WE’RE USING TO ADDRESS THESE OTHER VIRUS INFECTIONS HAS COME AS A RESULT OF FUNDING FOR HIV AND AIDS SO JUST AS THE 1918 FLU EPIDEMIC I THINK STIMULATED FUNDING AND INVESTMENT, FOR INSTANCE, BY THE ROCKEFELLER FOUNDATION, I THINK THE AIDS EPIDEMIC HAS STIMULATED INVESTMENT AND VIROLOGY AND IMMUNOLOGY THAT HAS RESULTED IN SOME OF THESE PROJECTS THAT I WILL BRIEFLY TELL YOU ABOUT. WE USE A NUMBER OF DIFFERENT DELIVERY PLATFORMS DEMONSTRATED HERE, EITHER GENE-BASED DELIVERY OF A VACCINE ANTIGEN, MAKING VIRUS-LIKE PARTICLES OR PROTEIN BASED VACCINES EITHER AS SOLUBLE PROTEINS OR MOUNTED ON NANOPARTICLES. I WILL TELL YOU STORIES TODAY ABOUT A FLU VACCINES BASED ON AN ANTIHISTAMENIC ARTICLE AND WE ALSO HAVE A PROCESS WHERE WE CAN DO IT. WE HAVE A PILOT PLANT THAT CAN MAKE GMP PRODUCT FOR USE IN HUMAN TRIALS. WE HAVE OUR OWN PHASE 1 CLINIC DO, A GOOD LABORATORY PROCESS FOR SAMPLE ANALYSIS BUT AS WE ENTER INTO PHASE 1 AND 2, WE NEED PARTNERS. THE REASON WE NEED A UNIVERSAL FLU VACCINE AND I THINK YOU HAVE HEARD THIS BEFORE, WE’RE STILL USING 70-YEAR-OLD TECHNOLOGY. WE STILL GROW THE VIRUS IN EGGS, INVACUATE THEM IN THE SAME WAY AND IN GOOD YEARS, IT CAN BE 60 PERCENT EFFECTIVE, IN BAD YEARS, AS LOW AS 10 PERCENT EFFECTIVE. I THINK IT IS BETTER TO SAY IT IS BETTER THAN NOT TO GET THE FLU VACCINE BUT PEOPLE ARGUE IT IS ONLY MARGINALLY BETTER IN SOME YEARS. WE HAVE TO REMAKE IT EVERY YEAR, WE HAVE TO HAVE TWO MEETINGS A YEAR TO PREPARE FOR SOUTHERN AND NORTHERN HEMISPHERE OUTBREAKS AND GENERALLY NOT GOING TO BE EFFECTIVE ABOUT THE PANDEMIC STRAIN. SO WHAT ARE WE GOING TO DO ABOUT THAT? THE REASON IT IS SO DIFFICULT IS THE ANTIGENIC AND GENETIC VARIATION OF THE VIRUSES FROM YEAR TO YEAR EITHER THROUGH DRIFT WHICH YOU JUST HEARD DESCRIBED OR THROUGH SHIFT WHERE YOU GET A WHOLE NEW GENE LEVEL IN THE VIRUS. THIS IS ONLY SEEN IN A FEW OTHER VIRUSES LIKE HIV AND HEPATITIS C BUT A MAJOR PROBLEM FOR FLU. ANOTHER PROBLEM WE FACE IN IMMUNIZING FOR INFLUENZA IS PREEXISTING IMMUNITY AND I WILL GET INTO SOME T-CELL TECHNOLOGY IN THE END SHOWING HOW IT IS A PROBLEM FOR INDUCING NEW RESPONSES BUT ALSO GIVES US AN OPPORTUNITY TO IDENTIFY RESPONSES THAT COULD BE MORE EFFECTIVE IN THE FUTURE. AND THEN THE OTHER PROBLEM WE HAVE IS THAT FLU IS USUALLY MOST SEVERE AT THE EXTREMES OF AGE SO IMMUNIZING YOUNG INFANTS AND IMMUNIZING THE ELDERLY HAVE THEIR OWN PROBLEMS. SO THERE IS MANY TARGETS. THIS IS A CARTOON OF A FLU VIRUS WITH ITS GENE SEGMENTS INSIDE A LIPID ENVELOPE AND THE SURFACE PROTEINS INCLUDE THE HEMOGLUTINATE. I WILL FOCUS ON THAT MOLECULE TODAY AND GIVE YOU SOME EXAMPLES ON NEW TECHNOLOGIES AND HOW THEY MAY BE USEFUL. SO AS YOU HEARD, WE’RE FACING NOT ONLY FLU PANDEMIC THREATS BUT FROM MANY OTHER THINGS BECAUSE OF THE WAY WE DISRUPTED OUR ECOLOGY AND THE QUESTION IS CAN WE TURN VACCINE DEVELOPMENT INTO MORE OF AN ENGINEERING EXERCISE BY TAKING ADVANTAGES OF SOME OF OUR NEW TECHNOLOGIES TO DEVELOP DIFFERENT AND BETTER APPROACHES THAN THE TRADITIONAL APPROACH OF EMPIRICAL TRIAL AND ERROR TYPE OF APPROACH TO VACCINE DEVELOPMENT. AND I FEEL LIKE I AM A LITTLE BIT AT A RISK HERE OF BECOMING A CONTRACTURE OF THE OUTBREAK NARRATIVE BECAUSE I AM GOING TO GET INTO SOME TECHNICAL DETAILS. SO I AM GOING TO BE THE TECHNOLOGIST WHO THINKS WE MAY HAVE SOLUTIONS HERE. JUST TO MAKE IT A LITTLE MORE PERSONAL, LET ME TELL YOU A STORY OF HOW I AM CONNECTED TO THE 1918 EPIDEMIC. MY GRANDFATHER WAS AN OSTEOPATH IN KANSAS WHICH IS NOT TOO FAR FROM HASKELL COUNTY WHERE JOHN BARRY AT LEAST THINKS THE 1918 FLU EMERGED AND HIS FIRST WIFE DIED IN 1918. SO IF HE HAD NOT MARRIED MY GRANDMOTHER A FEW YEARS LATER, I WOULDN’T BE HERE TALKING ABOUT 1918 INFLUENZA. SO I HAVE MIXED FEELINGS ABOUT THE 1918 FLU OUTBREAK. [LAUGHTER] SO I AM GOING TO GIVE YOU ARE AN EXAMPLE OF HOW STRUCTURAL BIOLOGY HAS INFORMED OUR ABILITY TO NOW MAKE NEW TYPES OF VACCINES THAT HAVE MORE SPECIFICITY. I AM GOING TO TELL YOU A LITTLE BIT ABOUT HIGH SEQUENCING WHICH CAN IDENTIFY NOT ONLY THE VIRUSES THAT EMERGE BUT THE PROCESS THAT CAN INFORM OUR TARGET FOR VACCINE RESPONSES AND THEN HOW RAPID ISOLATION OF HUMAN BODIES THAT HAS REALLY JUST HAPPENED THE LAST EIGHT TO NINE YEARS HAS CHANGED EVERYTHING IN HOW WE THINK ABOUT VACCINE DESIGN. SO RSV IS A CHILDHOOD VIRUS PRIMARILY. IT INFECTS YOUNG INFANTS WHICH HAPPENS ABOUT TWO, THREE MONTHS OF AGE, INFECTS THE AIRWAYS, CAUSES LOTS OF INFLAMMATION AND NOT AS LETHAL AS INFLUENZA BUT IN THE ELDERLY ADULTS, IT HAS MORE AFFECTABILITY THAN THE OTHERS AND IS THE LEADING CAUSE OF DEATH FOR CHILDREN UNDER 5 GLOBALLY. SO IT HAS A FUSION PROTEIN THAT IS VERY SIMILAR TO THE HUMAN INFLUENZA, A CLASS 1 FUSION PROTEIN AND YOU CAN SEE IF YOU DID CRYOMICROBIOLOGY ON THE SURFACE, THERE IS A LONG AND SHORT OF THIS F PROTEIN AND FOR THE LAST 40 TO 50 YEARS, WE HAVE BEEN USING THIS PROTEIN FORM AS A VACCINE. ABOUT FIVE YEARS AGO, WE WERE ABLE TO SOLVE THE STRUCTURE OF THIS PREFUSION FORM BEFORE IT REORGANIZES AND REARRANGES. AND IT REVEALED SOME NEUTRALIZATION SENSITIVE SITES ON THE PROTEIN THAT HAD NOT BEEN KNOWN BEFORE WHEN WE WERE JUST USING THIS PROTEIN AND SO BY IDENTIFYING THE STRUCTURE OF THE FUNCTIONAL FORM OF THIS PROTEIN BEFORE THE REARRANGEMENT OCCURS AND IDENTIFYING NEUTRALIZATION SENSITIVE SITES, MAKING A VACCINE, WE SHOW IT IS MORE ANTIGENIC AND PRODUCING RESPONSES THAT ARE 16 TO 25 FOLD HIGHER THAN BASELINE INSTEAD OF 2 TO 3 BASELINE FROM FAILED PREVIOUS EFFORTS, AND WE THINK IT IS ONE OF THE FIRST CLINICAL PROOF OF CONCEPT FOR STRUCTURE VACCINE DESIGN KNOWING THE ATOMIC LEVEL OF THE PROTEIN AND UNDERSTANDING THE EXACT TARGET OF NEUTRALIZING BODIES CAN REALLY CHANGE EVERYTHING. SO CAN WE APPLY THIS NEW TECHNOLOGY TO FLU? SO I AM SHOWING YOU ANOTHER CARTOON OF THE FLU VIRUS AND THE HEMAGGLUTININ MOLECULE, THIS IS ONE OF THE FIRST CRYSTALLIZED STRUCTURES EVER FORMED, HE HAD TO MAKE A STICK STRUCTURE AND IS STANDING BY IT SO WE HAVE KNOWN FOR A LONG TIME THE ATOMIC LEVEL STRUCTURE OF INFLUENZA. I WILL JUST GIVE YOU A PRESTRUCTURE TO TELL YOU WHAT I AM TALKING ABOUT. IT HAS A HEAD PROTEIN AND A HEAD AND STEM REGION, BLOCKED AND PROTECTED BY SUGAR MOLECULES THAT YOU SEE ON THE SURFACE AND MOST OF THE CHANGES YEAR TO YEAR HAPPEN ON THESE RED SPOTS ON THE HEAD BUT THERE ARE SITES OF VULNERABILITY, EITHER ONE IN THE STEM HERE OR ONE IN THE BINDING POCKET HERE AT THE TOP AND THERE ARE KNOWN BODIES THAT CAN BIND EITHER THE HEAD REGION ACROSS STRAINS OR THE STEM REGION ACROSS STRAINS AND THE QUESTION IS CAN WE MAKE VACCINES THAT CAN ILLICIT THESE BODIES THAT CAN CROSS REACT TO CROSS SUBTYPES AND MAYBE EVEN ACROSS GROUPS OF HEMAGGLUTININ MOLECULES. SO THERE IS LOTS OF WAYS TO MAKE THIS IN A VACCINE. YOU CAN EXPRESS IT IN A LOT OF VIRUSES. THE WAY WE MAKE THE CONVENTIONAL VACCINE, YOU GROW THE VIRUS IN EGGS, DISRUPT IT AND HARVEST WHATEVER PROTEINS ARE LEFT OVER AND THEN GIVE IT AS AN INACTIVATED PRODUCT WHICH IS MOSTLY HEMAGGLUTININ ANYONE. IT ALSO MAKES PARTICLES THAT DON’T HAVE THIS, YOU CAN GIVE IT AS A SOLUBLE PROTEIN AS A NORMAL MOLECULE OR MAYBE A CONERIC HEAD MOLECULE, I WILL DESCRIBE THAT IN A MINUTE, OR YOU CAN USE PROTEIN ENGINEERING OR DESIGN VACCINES THAT ARE BASED JUST ON THE FULL LENGTH HEMAGGLUTININ OR JUST THE STEM AND DISPLAY IT ON A NANO PARTICLE AND ASK CAN YOU DO BETTER THAN NATURAL IMMUNITY. SO I WILL FOCUS PRIMARILY ON THIS APPROACH AND GIVE YOU SOME EXAMPLES ON WHAT WE CAN DOTHE REASON IS IF YOU LOOK AT THE STEM STRUCTURE AND THE SEQUENCE OF THE HEMAGGLUTININ, YOU CAN DIVIDE THE MOLECULES, 18 OF THEM THAT BIRDS ALL CARRY, INTO TWO MAJOR GROUPS AND SEASONALLY, WE ARE EXPOSED TO THE H1N1 VIRUS, THE H1N2 VIRUS, THE 1918 WAS THIS H1. THEY ARE WORRIED ABOUT AVIAN FLU WHICH IS H5 OR H7 THAT SOMETIMES SHOWS UP IN CHINA THAT ARE LARGELY TRANSMITTED FROM AVIAN SOURCES. AND IF WE LOOK AT THE STEM REGION IN MORE DETAIL, YOU CAN SEE THAT THE BODIES THAT CAN NEUTRALIZE BASED ON STEM BINDING CAN OCCUPY TWO DIFFERENT TYPES OF FOOTPRINTS. ONE, THEY CAN GET THE GROUP 1 VIRUS, ONE THAT GET THE GROUP 1 VIRUSES AND SOME THAT CROSS BOTH BY SLIPPING IN BETWEEN THESE TWO SUGAR MOLECULES. SO IF YOU DON’T UNDERSTAND THAT GEOGRAPHY ON THE PROTEIN, YOU WON’T BE ABLE TO MAKE A VACCINE THAT CAN ILL ELICIT THOSE RESPONSES. NOW, AGAIN JUST TO ORIENT YOU, THIS IS A PLASMA CELL, THIS IS WHAT MAKES OUR BODIES, IT LIVES IN THE BONE MARROW. THESE ARE BODY MOLECULES AND THIS IS A FLU VIRUS SO ONE QUESTION IS IF YOU MAKE BODIES FROM THE STEM, CAN THEY ACCESS THE VIRUS AND GET TO THE RIGHT PLACE ON THE HEMAGGLUTININ AND DO SOMETHING TO BLOCK THE VIRUS. THIS STUDY BY AUDREY HARRIS A FEW YEARS SHOW THAT INDEED AN BODY THAT BINDS THE HEMAGGLUTININ IN HERE, YOU CAN SEE IT DEMONSTRATED HERE WITH THE ELECTROMICROSCOPY STUDY, THEY DO INDUCE THE STEM REGION SO IF WE CAN INDUCE THESE STEM BODIES, CAN WE MAKE A BETTER VACCINE FOR INFLUENZA? THOSE BODIES ARE NOT AS POTENT TO THE ONES ON THE HEAD BUT MUCH MORE CONCERNED BETWEEN STRAINS. SO HERE IS AN EXAMPLE OF HOW WE ADDRESS THIS. THIS IS A FULL HEMAGGLUTININ AND IF WE REMOVE THE HEAD AND DO SOME CIRCULAR PERMEATIONS TO TIE THE TOP OF THE HEAD TOGETHER, YOU CAN STABILIZE IT BY ADDING ADDITIONAL MOLECULES AND MUTATIONS FROM INSIDE THE PACT, HOLD IT TOGETHER FROM THE INSIDE, THEN YOU CAN MOUNT THOSE TRIMERS ON A SELF-ASSEMBLED MOLECULE CALLED PERTIN AND NOW YOU CAN HAVE TRIMERS ON A STEM FROM THE MOLECULE. THAT IS APPROACH NUMBER ONE. APPROACH IS MORE DIFFICULT AND HAS TAKEN FIVE TO SIX YEARS TO ACHIEVE A MOLECULE THAT HAS THE RIGHT ATOMIC STRUCTURE. SO BY MAKING A VARIETY OF MUTATIONS, YOU CAN GET TO A PLACE WHERE FOR THE H2 VIRUSES, YOU CAN HAVE A SIMILAR TYPE OF VACCINE. NOW, I AM GOING TO GO BACK NOW TO WHAT I SAID ABOUT SEQUENCING AND BODY LINEAGES. WE HAVE DONE SOME STUDIES WITH H1 AND H7 ANTIGENS WHICH HUMANS DON’T HAVE SO MUCH IMMUNITY TO. AND IN A DEVICE THAT CAN MONITOR AND MEASURE PROTEINS ON INDIVIDUAL B CELLS GOING THROUGH A LASER DETECTION SYSTEM, YOU CAN ASK WHETHER — YOU CAN FIND THESE CELLS THAT REACT BETWEEN H5 AND H1 WITHIN GROUP 1 VIRUSES, THEY WOULD BE IN THIS UPPER RIGHT QUADRANT. EACH ONE OF THOSE DOTS REPRESENTS AN INDIVIDUAL B CELL SO YOU USE THE PROBES TO FIND THESE CELLS AND YOU CAN ALSO FIND THE CELLS THAT ACT BETWEEN H5 AND H3 FROM THESE PEOPLE WITH THE PANDEMIC STRAINS. WHAT WE FIND IS THERE ARE CERTAIN TYPE OF BODIES LINEAGES, THE HEAVY CHAIN SEQUENCE USED TO MAKE THE BODY, IT IS ALMOST ALL VH69, VERY HEAVILY BIASED TOWARDS THIS TYPE OF ANTIBODY. THE ONES THAT SEE CROSS GROUP RESPONSES ARE MUCH MORE DIVERSE AND COMPLEX BUT YOU CAN IDENTIFY ANTIBODIES AND EVEN IDENTIFY MOTIFS FROM THE BODIES THAT ANALYZE THE STEM IN A CROSS-REACTIVE WAY. SO WE NOW HAVE ABOUT A DOZEN OF THESE LINEAGES THAT WE KNOW AND FIND IN MORE THAN ONE GROUP. AND WE KNOW THESE THAT ARE MORE GROUP 1 ORIENTED, IMMUNIZING WITH A GROUP 2 VIRUS ARE MORE SIMILAR TO AN H3, WE INDUCE A NUMBER OF BODY LINEAGES THAT HAVE THE CAPACITY FOR CROSS GROUP NEUTRALIZATION AND ALSO CROSS SUBTIED IMMUNIZATION. SO WE HAVE MADE THESE TYPES OF PRODUCTS. WE HAVE SHOWN WE CAN PROTECT FERRETS. IF YOU MAKE AN H1 VACCINE, YOU CAN PROTECT AGAINST THE LETHAL H5 AS SHOWN HERE. WE MADE A GROUP 2 ON FERRETS THROUGH A NANOPARTICLE AND SINCE WE KNOW THE SEQUENCES OF THE BODIES WE’RE TRYING TO TARGET, WE CAN GO TO THE — [INDISCERNIBLE] — BEFORE IT MUTATES TO BECOME MORE EFFECTIVE AND WE CAN PUT THE GERM LINE VERSION OF THOSE IN A TISSUE FILTER AND ASK DO OUR ANTIGENS RECOGNIZE OR DO THOSE GERM LINE VERSIONS OF BODIES RECOGNIZE OUR VACCINE ANTIGENS AND INDEED WE CAN MAKE NOW, GROUP 2 VACCINES THAT CAN STIMULATE THE GERM LINE VERSIONS OF BODY LINEAGES WE KNOW HAVE THE CAPACITY TO BECOME BROADLY NEUTRALIZING BODIES. SO THESE TECHNOLOGIES HAVE CREATED A WHOLE NEW WAY OF DESIGNING, MEASURING AND TRACKING RESPONSES TO VACCINES. SO I JUST SAID THAT. WE ALSO, I WANT TO MAKE SURE IT WAS CLEAR THAT WE HAVE DEFINED THESE STRUCTURALLY DEFINED SITES OF VULNERABILITY AND SPECIFIC BODY LINEAGES THAT CAN NOW BE TARGETS FOR FLU VACCINES. WE DON’T KNOW IF WE CAN INDUCE THEM IN PEOPLE WHO ARE IMMUNE. IT MAY HAVE TO START BACK AT THE LEVEL OF THE NAIVE INFANT BUT WE WILL KNOW THAT IN THE NEXT FEW YEARS AND WE HAVE DESIGNS BASED ON FULL LENGTH HA, THE STEM, I HAVEN’T BEEN ABLE TO TELL YOU ABOUT ALL OF THOSE BUT YOU SEE THERE ARE LOTS OF NEW OPTIONS IN COMBINING TECHNOLOGIES TO GET TO THE — A BETTER FLU VACCINE AND JUST TO FINISH, I WANT TO TELL YOU HOW WE’RE TAKING ALL THESE TECHNOLOGIES BACK TO WORK ON HIV BECAUSE HIV IS PROBABLY THE MOST DIFFICULT VACCINE TARGET. IT HAS ALL THESE WAYS OF EVADING BODIES BUT BY MINING OUR WORK ON RSV AND FLU AND CORONAL VIRUSES, WE’RE LEARNING BETTER NOW HOW TO MAKE A BETTER HIV VACCINE. I AM GOING TO THANK THE PEOPLE WHO HELPED ME WITH THIS WORK FROM MY LAB AND THE VRC. [ LISTING NAMES ] AND I DON’T HAVE JEFF BOYNTON ON HERE BUT HE HELPED ME MAKE THESE MOVIES AND JASON MCLELLAN WAS INVOLVED IN THE ORIGINAL RSV STRUCTURAL DISCOVERY. THANK YOU VERY MUCH. [APPLAUSE]

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