>>WELCOME. I WOULD LIKE TO THANK, FIRST, ALL OF YOU WHO HAVE TRAVELED FROM NEAR AND FAR AND MAKE THIS SYMPOSIUM POSSIBLE AND IT IS IMMUNO BIOLOGY REUNION. ALSO, MY APOLOGIES TO ALL WHO WANTED TO BE PRESENT IN-PERSON BUT WE COULD NOT ACCOMMODATE DUE TO THE DISTANCE REGULATIONS AND I HOPE THAT YOU HAVE TUNED IN AND YOU ARE WATCHING US. SO, NEXT FOUR MINUTES, I WILL TROY TO GIVE YOU A BRIEF HISTORY OF IMMUNO BIOLOGY SECTION. SO, IT STARTED AT HARVARD WHEN ALAN OPENED HIS LAB IN THE DEPARTMENT OF PATHOLOGY AND MEDICINE IN 1977. THREE YEARS LATER, HE WAS RECRUITED TO THE LAB OF DISEASES AND IMMUNO BIOLOGY SECTION WAS BORN. OVER THE TIME, HE MOVED FOUR TIMES. ALMOST IN A DECADE INTERVALS. BUT WHAT IS IMPORTANT IS THAT DURING EACH DECADE, A NEW GROUP OF STUDENTS AND POSTDOCS CAME TO THE LAB. SO ALAN TRAINED 75 GRADUATES AND 60 OF THEM ARE POSTDOCS. SO, WE CAME TO BE TAUGHT SCIENCE AND GOT -- WE WERE TAUGHT CUTTING-EDGE TECHNOLOGY OR HOW TO WRITE THE BEST ABSTRACT OR HOW TO BEST PRESENT YOUR DATA. WE GOT MUCH MORE THAN THAT. ARRIVAL OF NEW PERSON IN THE LABS SOMEHOW ALWAYS COINCIDES WITH ALAN AND STEPHANIE FINDING A TV, BIKE, SOFAS, A SET OF DISHES THAT THEY DON'T NEED ANYMORE. SO, OUR NEW, EMPTY ROOM OR APARTMENT, THE FIRST PIECE WAS THE PIECE FROM THEIR HOME. MANY CAME ALL AROUND THE WORLD. FROM ALL CORNERS OF THE GLOBE. THEY CAME WITH A DIFFERENT BACKGROUND, WITH DIFFERENT CULTURAL EXPERIENCES AND ALAN TAUGHT US TO SPEAK ENGLISH. HE TAUGHT EVEN ENGLISH-SPEAKING PEOPLE ENGLISH. HE TOOK INTEREST IN US. SO, HE LEARNED AND HE IS NOW PROFICIENT IN PORTUGUESE, SPANISH, GERMAN, FRENCH, SPECIALTY SWEDISH. [LAUGHTER] THERE'S STILL SOME ROOM FOR IMPROVEMENT IN SLAVIC LANGUAGES. [LAUGHTER] THAT'S A MEME. SO OVER THESE 45 YEARS, THE LAB TAZ BEEN WORKING ON THE LAB HAS BEEN WORKING ON NINE DIFFERENT PATHOGENS THAT CAUSE GLOBAL DISEASES OF IMPORTANCE AND IF CAN BE TRACED TO EGYPTIANS MEMBER HE'S TO SARS-CoV-2, THAT WAS NOT KNOWN THROW YEARS AGO. TO THE LAB HAS BEEN WORKING ON DIFFERENT PROJECTS AND DIFFERENT TOPICS FROM PARASITE VACCINE ANTIGENS TO -- [LAUGHTER] NO. YEAH, I KNOW WHY YOU ARE LAUGHING NOW. LET'S GO SERIOUS. SERIOUS MATTER IS THERAPY TO INFECTIOUS DISEASES. AND HAS PUBLISHED OVER 560 RESEARCH PUBLICATIONS. WHAT IS MORE IMPORTANT IN ADDITION TO THIS IMPRESSIVE IMPRINT LEGACY IS A LIVE LEGACY OF THE LAB. HERE IS A LIST OF SOME SENIOR POSITIONS HELD BY EX MEMBERS OF THE LAB, THAT INCLUDES THREE TENURE NIAID INVESTIGATORS, EIGHT TENURE PROFESSOR, DIRECTORS OF RESEARCH INSTITUTES, IN INDUSTRY AND THE REGULATORY PROGRAMS EXECUTORS AND DIRECTOR OF PRIVATE FOUNDATIONS. THAT IS PRETTY IMPRESSIVE. SO THE LEGACY IS ACTUALLY WHAT IS VERY IMPORTANT AS I WAS SAYING AND ALAN TAKES PRIDE IN SUCCESS OF EVERY AND EACH OF US. HE REFERS TO US USUALLY AS A SCIENTIFIC CHILDREN AND TRAINEES OF US AS HIS SCIENTIFIC GRANDCHILDREN. SO THAT IS -- I KNOW THAT IMMUNO BIOLOGY SECTIONS WILL BE CLOSING THIS SUMMER, BUT IT WILL NOT PERISH. FOR ONE, ALAN IS NOT RETIRING AND ALL OF US WILL CARRY ON AND KEEP GROWING IMMUNO BIOLOGY SECTION. THANK YOU. [APPLAUSE] SO, Dr. FAUCI WAS NOT ABLE TO COME IN-PERSON, BUT HE DID TAPE A MESSAGE THAT WE WILL BE WATCHING NOW. >>THANK YOU Dr. JANKOVIC. GREETINGS. THANK YOU ALL FOR JOINING US IN WHAT PROMISES TO BE AN EXCITING AND I AM INFORMATIVE SYMPOSIUM ON THE INTERFACE BETWEEN IMMUNITY AND INFECTION. OVER THE NEXT TWO DAYS, YOU WILL HEAR NOT ONLY GREAT SCIENCE, BUT ALSO PERSPECTIVES FROM MANY COLLEAGUES WHOSE RESEARCH WAS INSPIRED BY AND OFTEN UNDERTAKEN COLLABORATIVELY WITH ALAN. I CAN TELL YOU WITHOUT LIBRARY PER BLEE HE HAS BEEN THE MOST INFLUENTIAL SCIENCE. YOU WILL HEAR ABOUT HIS SCIENTIFIC ACCOMPLISHMENTS IN RESEARCH AS WELL AS HIS IMPORTANT EXPANSION OF THE LAB INTO TUBERCULOSIS RESEARCH. HOWEVER, I WANT TO RECOGNIZE ALAN AS A LEADER WHO HAS HELPED SHAPE SCIENCE AND THE SCIENCE WITHIN NIAID NATIONALLY AND GLOBALLY. FROM THE TIME HE ARRIVED HERE IN 1980 FROM HARVARD, HE HAS WORKED HARD TO PROMOTE COLLABORATIVE AND COLLEGIAL RESEARCH, UNSELFISHLY MENTORING AND SUPPORTING THE WORK OF COLLEAGUES WHEN HE COULD. WITHIN NIAID AND ACROSS AND OUTSIDE OF NIH, INCLUDING THE NIH OXFORD CAMBRIDGE SCHOLARS PROGRAM, THE NIH GRADUATE PARTNERSHIP PROGRAM AMONG OTHERS. ALAN HAS ALWAYS BEEN A TRIPLE THREAT WITH CUTTING-EDGE SCIENCE, HUMANISM AND FOSTERING A SUPPORTIVE ENVIRONMENT. THIS UNBEATABLE COMBINATION HAS CATALYZED BOTH THE STELLAR OUTPUT OF HIS LAB, AND A DISTINGUISHED CAREER OF THOSE WHO WERE TRAINED IN IT AND INFLUENCED BY IT. THE GENERATIONS OF SCIENTISTS THAT ALAN MEN FORWARD TO AND TOD IS WHO IS WHO. INCLUDING SOME OF YOU TODAY. THIS IS NOT JUST AN AMERICAN GROUP OF SCHOLARS, BUT A TRULY GLOBAL GROUP WORKING IN THE THE VERY PLACES WHERE THESE IMPORTANT DISEASES KILL AND DISABLE. AS A SCIENTIST MENTOR, IT SHOULD NOT BE SURPRISING THAT ALAN WAS AWARDED THE PRESTIGIOUS BAILEY K. IN TROPICAL MEDICINE AND HYGIENE, THE NIH DIRECTOR MENTORING AWARD AND THE UNITED STATES PUBLIC-HEALTH SERVICE SUPERIOR SERVICE AWARD AMONG OTHERS. I TAKE PARTICULAR NOTE OF ALAN'S LONGSTANDING COMMITMENT TO SUPPORTING WOMEN SCIENTISTS, WHICH BEGAN DECADES AGO. AT A TIME WHEN FEW WOMEN SCIENTISTS WERE ALLOWED TO BREAK THROUGH THE GLASS CEILING. IT IS A HIGH TRIBUTE IN 2015 THE ASSOCIATION OF WOMEN AND SCIENCE AWARDED ALAN THE EXCELLENCE IN MENTORING AWARD. PERHAPS THIS DOGGED CAREER-LONG SUPPORT OF WOMEN SCIENTISTS COMES FROM HAVING THE RIGHT GENES. ALAN'S MOTHER, ROSLYN, WAS A PH.D MICROBIOLOGIST, HIS MOTHER'S COUSIN AND BEST FRIEND TRUDEAU' BELL ELION A CO RECIPIENT OF THE 1988 NOBEL PRIZE OF MEDICINE FOR NEW APPROACHES TO RATIONAL DRUG DESIGN AND HIS AUNT A RENOWNED PEDIATRIC VAX ENOLOGIST AND OF COURSE, ALAN HAD THE VERY GOOD SENSE TO EXTEND THIS STREAK OF ACCOMPLISHMENT BY MARRYING OUR COLLEAGUE, STEFANI JAMES. ALAN, YOU HAVE PROVEN ONCE AND FOR ALL THAT THE VERY BEST SCIENTISTS ARE THOSE WHO REALIZE THAT WOMEN ARE AMONG THE VERY BEST SCIENTISTS. WE ARE HERE TO THANK I FOR YOUR LASTING ACCOMPLISHMENTS, AND FOR SHARING YOURSELF WITH THE MANY GREAT SCIENTISTS YOU HAVE MEN FORWARD TO AND INFLUENCED OVER MORE THAN FOUR DECADES AT NIAID. WE ARE PARTICULARLY GRATEFUL THAT INSTEAD OF RETIRING, YOU ARE STAIR ON TO WORK WITH ANOTHER GENERATION OF SCIENTISTS, THEY TOO NO DOUBT, WILL CARRY ON YOUR TRADITION OF EXCELLENCE. THANK YOU FOR YOUR SERVICE AND DEDICATION AND MAY THE STANDARDS YOU HAVE SET INSPIRE US LONG INTO THE FUTURE. >>GOOD MORNING, IT'S VERY NICE TO BE HERE AND SEE SO MANY FAMILIAR FACES AND I ACTUALLY I THINK I CAN RECOGNIZE MOST OF YOU IN SPITE OF ALL THE TIME, LONG TIME IN ALL THE PHENOTYPIC CHANGE OVER THE YEARS. FIRST I'D LIKE TO THANK DRAGANA FOR ORGANIZING THIS GREAT TWO DAYS OF SYMPOSIUM. I THINK SHE DID A GREAT JOB AND REAL SWS HOW ALAN WAS IMPORTANT AS A MENTOR AND LEADER OF THE FIELD. IT'S AN INCREDIBLE PROGRAM. I ASK DRAGANA TO SQUEEZE FEW WORDS ABOUT ALAN DURING THIS INITIAL SESSION. DON'T GET SCARED, ALAN, I WILL ONLY SAY GOOD THINGS. I THINK OF COURSE I DON'T NEED TO REPEAT ALL OF HIS ROLE ON MENTORSHIP AND LEADERSHIP IN IMMUNOLOGY OF INFECTIOUS DISEASE BUT HE HE HAD A VERY IMPORTANT RESPECT OF TRAINING BRAZILIAN SCIENTISTS. I COUNTED, OVER THE YEARS, HE HAD ABOUT 12 POSTDOCS AND I THINK ALL OF THEM WENT BACK TO BRAZIL AND REALLY MADE A REALLY IMPORTANT LEADERSHIP IN BRAZIL AND ALSO ALAN HELPED TO LEARN VERY IMPORTANT INTERNATIONAL SCIENTIFIC MEETINGS IN BRAZIL. REALLY, HE HAD A VERY IMPORTANT ROLE IN DEVELOPING THIS AREA AND FOR THAT, ELLA WARDED, I THINK 10 YEARS AGO, AS A MEMBER OF THE BRAZILIAN ACADEMY OF SCIENCE AND THEN HE GOT THIS FAMOUS PRIZE FOR THE SCIENTIST THAT DRINK MOST AT A DISEASE MEETING. [LAUGHTER] SO HE HAS THIS -- WHEN BACK IN THOSE DAYS WHEN HE HAS THIS ANIMAL PARTY INSIDE OF HIM. [LAUGHTER] SO, ANYWAY, I CAME TO ALAN'S LAB, I WAS DOING MY PH.D IN OF COURSE THE TH1 AND TH2 DICHOTOMY WAS FLOURISHING AND WAS A VERY IMPORTANT. I GOT VERY EXCITED. I READ A PAPER WITH ALAN IN ALAN'S LAB SHOWING THIS TH1 AND TH2 AND I THOUGHT THAT'S THE PLACE TO BE AND WHEN I ARRIVED, THERE WAS ALL THESE WORK FROM DNX AND A LOT OF PEOPLE -- PEOPLE THAT DID THE WORK LIKE BOB COULD HAVE MAN, TIM HOFMANN, AND THEY ARE ALL HERE AND THEN GEORGE AND ALAN HAS ALL THOSE GREAT MODELING FOR PARA CIVIC DISEASES THAT WERE GREAT MODELS TO STUDY, T CELLS, INNATE IMMUNITY SO I FELT LIKE I WAS IN THE RIGHT PLACE AT THE RIGHT TYPE. ALTHOUGH, LOOKING BACK, IT WAS ALWAYS THE RIGHT TIME IN ALAN'S LAB. SO, HE NEEDED TO HAVE A TH1 PARASITE SO HE HAD TH2 PARASITE THAT WAS SIS TOE AND A PARASITE THAT THE HOST CONTROLLED THE DEVELOPMENT OF T CELLS AND NOW HE HAD TO HAVE A TH1 PARASITE AND I WAS WANTING TO BE THE FIRST ONE TO WORK ON HIS LAB AND WE ACTUALLY HAVE VERY -- WE STUDIED THE ROLE OF IL12 AND I WAS EXPECTED IT WAS A KEY CYTOKINE FOR IN DUCING TH1 BUT IT WAS A BITTEN COU BIT INCOUNTE BECAUSE THE PARASITES USE IO10 TO EVADE IMMUNE RESPONSE AND ESTABLISH AND WE FOUND THAT IO10 IN DEED MAKES THE MOUSE MORE RESILIENT TO INFECTION AND A LOT OF THE NEXT YEARS, THERE WAS A LOT OF STUDIES OF MECHANISMS THAT ARE INVOLVED ON IL12 INDUCTION, IL10 AND THAT WAS TWO FINDINGS THAT WERE VERY IMPORTANT FOR THE FUTURE RESEARCH ON TOXO. SO SAYING THAT, I GUESS THE SPEAKERS OF THIS SECTION WILL REALLY TALK ABOUT ALL THESE MECHANISMS AND THE ROLE OF DIFFERENT CYTOKINES THAT ARE INVOLVED ON CONTROL OF INFECTION AND SO THE WONDERLAND OF MOUSE IMMUNOLOGY TO TOXO INSPECTION SO WITH THAT INVITE CHRIS HUNTER TO BE THE FIRST SPEAKER AND DRAGANA ASKED ME TO KEEP YOU GUYS ON TIME. [APPLAUSE] SO, MY JOB HERE, I THINK, I'M NOT QUITE SURE WHAT MY JOB IS. YOU SAID EVERYTHING I WANTED TO SAY AND RICARDO SAID THE SAME THING SO AS THIS MEETING -- I THINK I'LL MANAGE. I HAVE SOME REALLY FUNNY STORIES ABOUT ALAN THAT I WAS GOING TO TELL BUT I THOUGHT ABOUT THEM AND I SAID THEM OUT LOUD AND THEY ALWAYS REFLECTED BADLY ON ME. [LAUGHTER] SO I'M NOT GOING TO TELL THOSE STORIES. SO, MOST OF YOU WILL REMEMBER THE FIRST TIME YOU METAL AN AND ALAN DOESN'T REMEMBER THE FIRST TIME HE MET ME. I HAVE 1990 AT BIOLOGY PARTISAN COURSE. ALAN LECTURED AND TALKED ABOUT TH1 AND TH2 WHICH I THOUGHT AT THE TIME SOUNDED RIDICULOUS. YOU KNOW, AS A YOUNG POSTDOC AND THIS IS THE FIRST TIME I HEARD THAT AND THE NEXT TIME, I WAS DOING A POSTDOC WITH JACK RAMMINGTON ON THE WEST COAST AND I STARTED TO WORK ON IL12 AND ALAN LAID OUT A SERIES OF STUDIES FROM RICARDOER AND I CAN AND ALAN'S LAB AND ALL THE WORK THAT I HAD BEEN DOING BUT HERE IT WASN'T PAPERS AND IN TALKS AND A YEAR TO TWO YEARS AHEAD OF EVERYTHING THAT I'VE BEEN DOING, I REALLY DEVASTATED ME SO BUT IT KIND OF TAUGHT ME A LOT. IT TAUGHT ME ABOUT RESILIENCE AND HOW TO MOVE A FIELD FORWARD. I METAL AN AT A CONFERENCE AND AS I TRANSITIONED TO BEING AN ASSISTANT PROFESSOR, I REALLY THOUGHT MY CAREER HAD GONE UP IN FLAMES. I NEED COUNCILING AND ALAN MAY OR MAY NOT REMEMBER THIS AND I CAME TO TALK TO ALAN ABOUT THIS AND THE LPD, YOU GUYS KNOW THIS BUT FROM THE OUTSIDE, THIS WAS REALLY THE THE WORLD NEXUS AND I WAS COMING DOWN AND I CAME TO SEE ALAN'S OFFICE AND IT'S A SHI TTY OFFICE. I KNOW SOMEONE ELSE IS GOING TO SAY THAT AND I'M THINKING, THIS -- THIS IS THE LPD AND CLEARLY, THERE'S ONLY ONE OFFICE THAT WAS IN THIS WORSE AND THAT WAS DAVID SACKS OFFICE. [LAUGHTER] >> AND I'M SITTING THERE AND THANK YOU DRAGANA FOR SHOWING THAT CARTOON MANY OF IT'S ON ALAN'S DOOR, PUBLISHED PUT PERISHED. I THOUGHT IT WAS PUBLISHED OR PERISHED. I WASN'T A SON OF CHER, I COME FROM A COMPETITIVE PERSPECTIVE AND ALAN TOOK THE TIME TO GIVE ME ADVICE AND MENTOR ME THROUGH SO IT WAS A TOUGH TIME AS I STARTED MY LAB. HE HAS FOR GONE OF COURSE. I DIDN'T FORGET IT. AND I'M LOOKING AROUND TODAY THINKING, IT WAS A LITTLE BIT OF A BACK WATER FOR SOME PEOPLE. AND THEN WITH GEORGE YO AND ANNE AND IMMUNOLOGISTS I WAS INTERESTED IN THE PARASITIC SYSTEMS AND IT WASN'T COUNTING TCRs BUT OUTCOMES OF INFECTION AND ALAN WAS REALLY SORT OF KEY FOR THAT. THE LAST COMMENT I WANT TO MAKE IS JUST REALLY THE SECTION ON TOX' PLASMA, ALAN HAS HAD A TERRIFIC TRACK RECORD IN PATHOGENS BUT FROM MY PER SPECIFIC I HAVE THE GROUPS OF PEOPLE THAT WORKED ON ALAN'S LAB, THE POSTDOCS, ALL OF THE POSTDOCS REALLY MOVED THIS FIELD FORWARD AND ALAN WAS THE COM ANALITY SO IT'S HARD TO SAY RICARDO GETS THE CREDIT. ALAN IS THE DRIVING FORCE FOR THIS COMMUNITY AND I THINK HE IS REALLY CREATED A PRODUCTIVE COMMUNITY AND INVOLVED IN MENTORING, NOT JUST HIS OWN TRAINEES BUT EVERYBODY ELSE WITHIN THE COMMUNITY AND I'VE APPRECIATED THAT AND WITH THAT I THINK IT'S BEST TO HAND IT OVER TO THE SCIENTISTS AND TO THANK DRAGANA FOR DOING ANY OF THIS. WELL DONE, DRAGANA. WITH THAT, I THINK I'M DONE. [APPLAUSE] [OFF MIC] >> SO, THANK YOU, DRAGANA FOR ORGANIZING THIS. IT'S FANTASTIC TO BE HERE. SO, FOR THOSE OF YOU, I'M ERIC DENKERS, I SPENT TIME AT CORNELL UNIVERSITY AND KIND OF NEEDED A CHANGE AND WANTED TO MOVE ON SO I MOVED TO THE UNIVERSITY OF NEW MEXICO, FIVE AND A HALF YEARS AGO. AND SO I'M WITH THE CENTER FOR EVOLUTIONARY AND THEORETICAL IMMUNOLOGY WITHIN THE DEPARTMENT OF BIOLOGY AND SO FOR THOSE OF THAW DO KNOW, I STUDY TOX' TOXA PLASMA SO I HAD TO SHOW SOME PICTURES. HERE ARE A FEW PICTURES WHEN I WAS A POSTDOC IN ALAN'S LABS WHICH WERE 1990 TO 1995. A LOT OF THESE PICTURES HERE, WE WERE IN BUILDING FOUR BACK IN THOSE DAYS AND THIS IS THE INFAMOUS SLOT. THIS IS WHERE POSTDOCS HAD THEIR OFFICES. THERE WERE THREE OF US. YOU CAN HERE IS ME RIGHT THERE. THERE'S ALAN. TOM WIN, RICARDO, PHIL AND GEORGE AND -- THAT'S JEFF HERE. THIS IS RICARDO. PHIL, CHRIS AND GEORGE WERE REMARKING THIS MORNING THAT RICARDO HAS NOT CHANGED AT ALL. A LITTLE COMPLIMENT THERE, RICARDO. AND SOME OTHER PEOPLE. HERE IS DRAGANA. I DON'T WANT TO POINT EVERYONE OUT BECAUSE I'LL BE HERE BEFORE. ISABEL OSWALD. HERE IS ANOTHER PICTURE FROM AROUND THE SAME TIME. A MORE FORMAL PICTURE OF THE LAB AND OF COURSE, ALAN HAD THESE GREAT PARTIES AT HIS HOUSE WHERE THE WINE FLOWED AND THERE WAS A LOT OF MUSIC AND SONG AND DANCE. THIS IS OUR MONICA WHO WAS A GREAT SINGER FROM ARGENTINA. >> SHE COULDN'T COME. >> TOO BAD. SO, ANYWAY, LET'S MOVE ON. I WANT TO TELL YOU ABOUT RESEARCH GOING ON IN MY LAB THE LAST YEAR OR TWO. SO, I WORK ON TOXOPLASMA AND AS YOU KNOW, IT WAS FIRST DESCRIBED IN 1908 I BELIEVE, AND SO FOR THE FIRST 80 YEARS AFTER ITS DESCRIPTION IT WAS A BACK QUOTAS PARASITE. THERE WAS SOME RESEARCH BUT NOT A LOT AND IT WAS ONLY IN THE 1980s WITH THE EMERGENCE OF HIV/AIDS THAT TOXOPLASMA WAS APPRECIATED FOR BEING AN IMPORTANT OPPORTUNISTIC PATHOGEN AND THAT IS WHERE REALLY RESEARCH STARTED TO SORT OF BUILD AND IT COINCIDED WITH THE COMING ON-LINE OF MOUSE GENE KNOCK OUT TECHNOLOGY IN COMBINATION WITH THE ABILITY OF REALLY TO CULTIVATE TOXOPLASMA AND AN EXPLOSION OF RESEARCH. IN MY MIND, LATE 1980s AND EARLY 1990s IT'S WHEN IN MY MIND, WHERE MAUD ARE NOT DAY TOXO PLASMA RESEARCH WAS BORN. AND OF COURSE, ALAN'S LAB WAS REALLY RIGHT THERE AT THE BEGINNING AND IT CONTINUED OVER SEVERAL DECADES TO BE PROMINENT IN TOXOPLASMA RESEARCH OF THE OVER THE YEARS, WE CAME UP WITH THIS GENERAL MODEL FOR THE IMMUNE RESPONSE TO TOXO PLASMA AND A LOT OF PEOPLE IN THIS ROOM WERE INVOLVED IN COMING UP WITH THIS SORT OF DOGMA OF THE IMMUNE RESPONSE TO TOX OWE PLASMA AND N PARTICULAR CELLS ARE IMPORTANT IN THE EARLY RESPONSE AND DENDRID I CAN CELLS MAKE IL12 AND IT'S BELIEVED TO BE THE KEY CYTOKINE THAT TRIGGERS INTER FEWER ON GAMMA AND IT COMES INITIALLY IN NK CELLS AND LATER FROM CD4 AND CROSS CD-8 CELLS AND MORE RECENTLY NUTRFILS IS AN IMPORTANT SOURCE OF SO IT'S REALLY THOUGHT TO BE THE CENTRAL CYTOKINE IN RESISTANCE TO TOXOPLASMA AND IT ACTUALLY IS MECHANISMS BUT ONE OF THE IMPORTANT MECHANISMS IS THROUGH TRIGGERING MICROBE IN MONOCYTES AND AS I SAID, I COULD GO THROUGH AND MENTION ALL THE NAMES OF THE PEOPLE IN THIS ROOM WHO CONTRIBUTED TO THIS MODEL BUT I DON'T WANT TO BECAUSE IT WOULD TAKE FOREVER. SO WHEN DRAG AN ASKED ME TO GIVE THIS TALK, I FELT SOME PANIC BECAUSE I DIDN'T KNOW WHAT ON EARTH I WAS GOING TO TALK ABOUT. WAS I SUPPOSED TO TALK ABOUT SORT OF SUMMARIZE MY CAREER SINCE I LEFT ALAN'S LAB? THAT WOULD TAKE A LONG TIME AND IT WOULD BE PRETTY BORING. I THOUGHT, MAYBE SO SHOULD I TALK ABOUT A RECENT PAPER I PUBLISHED. BUT THAT ALSO SOUNDED BORING. I'VE DECIDED TO PRESENT SOMETHING TO YOU, MORE ALONG THE LINES OF A WORK IN PROGRESS. THIS IS A PROJECT THAT WE'VE BEEN INVOLVED WITH OVER THE LAST YEAR-AND-A-HALF OR TWO YEARS AND SO I WANT TO PRESENT TO YOU SOME DATA FROM A GRADUATE ATT GRAD SE DOUGHTERY. THIS IS ONE OF YOUR SCIENTIFIC CHILDREN. YOU MAY HAVE MET HER IN SANTA IF HE THREE YEARS AGO. WE'RE GETTING READY TO PUBLISH THIS DATA AND IT SORT OF REVEALS NEW ASPECTS, I THINK, OF THE IMMUNE RESPONSE TO TOXOPLASMA. SO WE'VE BEEN WORKING WITH PARASITE STRAIN CALLED CPS. ANYONE THAT WORKS ON TOXO Mrs. TOX PLASMAWILL KNOW THIS PLAIN. THIS IS A URACIL AUXOTROPH IT WAS CREATED IN DARTMOUTH MEDICAL SCHOOL SOME 20 YEARS AGO YOU ACTUALLY. IT'S BEEN A VERY USEFUL TRAIN TO USE AND SO THE THING ABOUT THIS STRAIN, AS I SAID, IT'S A URACIL AUXOTROPH AND WHEN YOU INOCULATE MICE IT'S A STRONG PROTECTIVE IMMUNE RESPONSE AND YET THE PARASITE ITSELF IS NOT THOUGHT TO PERSIST SO YOU INOCULATE MICE, THE PARASITE STICKS AROUND AND IT'S GONE AND THEN IT TRIGGERED A NICE, PROTECTIVE IMMUNE RESPONSE THAT YOU CAN STUDY. TWO YEARS AGO WE LOGGED AT THIS AND WE HAD THIS IDEA THAT CPS SHOULD BE A GREAT WAY TO LOOK FOR EVIDENCE OF TRAINED IMMUNITY DURING TOXOPLASMA INSPECTION SO THE IDEA WOULD BE THAT YOU WOULD TAKE, FOR EXAMPLE, RAG 1 KNOCK OUT MICE SO NO ADAPTIVE IMMUNE SYSTEM, LET'S INOCULATE THEM WITH CPS AND COME BACK A MONTH LATER AND LOOK FOR EVIDENCE FOR A PROTECTIVE IMMUNE RESPONSE, AN INNATE PROTECTION. SO WE DID THAT EXPERIMENT, AND -- GO FORWARD. AND WE GOT THIS BIG SURPRISE. SO, WHEN WE INOCULATED THE MICE, WHAT WE FOUND, SO HERE WILD TYPE MICE INOCULATED WITH CPS THEY'RE PERFECTLY LINE BUT WHEN WE INOCULATED THE RAG 1 KNOCK OUT MICE THEY WERE SUSCEPTIBLE TO THIS PARASITE STRAIN. CHECK IT OUT. THEY DIE WITHIN TWO TO THROW WEEKS AFTER INOCULATION. THIS IS IP INOCULATION BY THE WAY. AND IF YOU LOOK IN THE CAVITY OF THESE NICE OVER HERE I BELIEVE THAT THIS IS DAY 10 AFTER INSPECTION. YOU CAN SEE THE WILD TIME MICE CONTROL THE INFECTION AND PARASITES ARE GOING AWAY BUT IN THE RAG 1 KNOCK OUT MICE, WE'RE SEEING AN EXPANSION OF THIS PARASITE STRAIN. IT WAS UNEXPECTED, WE WERE SURPRISED. SO KIND OF OUR FIRST THOUGHT WAS, OH MY GOD, WE SELECTED FOR A ROW VERT ANT PARASITE OR MAYBE WORSE, OUR PARASITES, OUR CPS PARASITES ARE CONTAMINATED WITH WILD TYPE PARASITES. THAT'S NOT THE CASE BECAUSE IF YOU TAKE A PARASITES FROM THE CAVITY OF THESE MICE AND PUT THEM BACK INTO FIBER BLAST IN VITRO, YOU CAN SEE THAT THEY STILL REQUIRE URACIL FOR THIS IN VITRO GROWTH. SO WE THOUGHT THAT WAS PRETTY INTERESTING AND WE THOUGHT IT WOULD BE WORTH OF PURSUING SEEING WHAT IS GOING ON IN THESE MICE. SO, RAG 1 DEPENDENT CELL SEEMS TO BE INVOLVED IN RESISTANCE. YOU ARE IS SIL -- YOU'RE A SIL - WE'RE ASKING WHAT IS GOING ON IN THE CAVITY AFTER INFECTION WITH THESE PARROT SIGHTS SO YOU CAN SEE UP HERE, B1B CELLS NOT MUCH GOING ON AND RED IS CELL NUMBER AND BLUE IS TOTAL PERCENT IN THE PARA TOE KNEEL CAVITY AND THERE'S A BLIP OF ACTIVITY BETWEEN DAY FOUR AND SIX. AND WE ALSO SEE A FAIRLY PROMINENT INCREASE IN CD8 CELL NUMBER IN THE PERITONEAL CAVITY SO WE SUSPECT T CELLS WERE INVOLVED HERE AND IT MAY NOT BE TOO SURPRISING. AND SO WE GOT SOME KNOCK OUT MICE. THESE WERE HORRIBLY EXPENSIVE EXPERIMENTS TO DO, BY THE WAY. WE GOT KNOCK OUT MICE AND YOU CAN SEE HERE ARE THE RAG 1 KNOCK OUT MICE. THEY ALL DIE. WILD TYPE MICE SURVIVE. HERE ARE GAMMA T GAM DELTA MICED THEY'RE FINE AND THIS IS SUPPOSED TO SAY MUMT SO THEY'RE ALSO RESISTANT AND ON THE OTHER HAND, AS WE KIND OF SUSPECTED WHEN WE INOCULATED TCR BETA KNOCK OUT MICE, THEY HAVE THE SAME PHENOTYPE AS A RAG 1 KNOCK OUT MICE. SO CLEARLY IT'S T CELLS THAT ARE INVOLVED HERE. WE ALSO DID ANTIBODY DEPLETION EXPERIMENTS. IT WORKS WELL AS WE ALL KNOW AND SO WHEN WE DO ANTI- CD4 AND ANTI-CD8 DEPLETION THEY DIE. IN THIS EXPERIMENT, WE DID -- WE TREATED THE MICE WITH DEPLETING ANTIBODIES BUT WE DIDN'T START THE DEPLETION UNTIL DAY 6 AFTER INOCULATION. WHEN WE SEE THAT INCREASE IN T CELLS IN THE PARA TENILLE CAVITY SO IF WE WAIT UNTIL DAY 6 WE STILL SEE A SUN ACCEPT TA BILL TEE AND W WE -- THEY'RE INVOLVED IN RESISTANCE. AND IF YOU LOOK AT THE PHENOTYPE OF THESE CELLS COMING INTO THE PERITONEAL CAVITY, THIS IS WHAT YOU SEE. SO CD4, CD8. THIS IS STAINING -- CAN'T READ THIS HERE. THIS WOULD BE CD44 AND ON THE Y AXIS AND THIS IS CD62 ON THE X AXIS. HERE IT IS DOWN HERE. SO MOST OF THE CELLS THAT WE'RE SEEING HAVE AN EFFECT TO BE/MEMORY. THESE ARE THE CD8 AND CD4. IF WE LOOK FORWARD. IF WE LOOK AT CD8 CELLS THEY'RE EXPRESSING CD69 AND THEY'RE EXPRESSING TBED SO THEY'RE LOOKING LIKE EFFECTOR CELLS. SO, THERE'S A REALLY SURPRISING RESULT FOR US, ON THIS SLIDE HERE. SO WE WENT BACK TO OUR SUSCEPTIBLITY STUDIES AND WE GOT KNOCK-OUT MICE AND SO WHAT WE CAN SEE IS THAT THIS IS THIS PROTECTIVE RESPONSE DOES NOT REQUIRE MYD88. THAT IS MAYBE NOT TOO SURPRISING, MY LAB HAS DONE QUITE A BIT OF WORK ON MYD88 INDEPENDENT IMMUNITY BUT THE REAL SURPRISING RESULT WE FOUND IS THAT IL12 KNOCK OUT MICE CAN SURVIVE CPS INFECT INPERFECTLY . NOT REQUIRED. AND OVER HERE, SAME TYPES OF EXPERIMENTS, HERE WE ASK WHAT THE REQUIREMENT WAS FOR IN TER FEWER ON GAMMA AND IN THE ABSENCE, SO THEY'RE ONLY PARTIALLY SUSCEPTIBLE SO THEY DO NOT PINOT COPY THE CTR BETA KNOCK OUT MICE SO THIS IS AN IL12 UNDER PEND PROTECTIVE RESPONSE AND IT'S ONLY PARTIALLY REQUIRES INTERFERON GAMMA. A FEW YEARS AGO, THERE WAS A LOT OF INTEREST IN INFLAMMATORY MONOCYTES THAT EXPRESSED THE MARKER GR1. SO, THIS IS WORK THAT WAS MAINLY DONE BY DAVID CIBLY AT WASHU SO HE SHOWED RESISTANCE OF TOXOPLASMA REQUIRED INFLAMMATORY MONOCYTES AND HE SHOWED THIS DURING MUCOSAL INFECTION AND DURING IP INFECTION. SO, WE HAD TO WONDER INFLAMMATORY MONOCYTES WERE INVOLVED IN THIS PROTECTIVE RESPONSE. THE ANSWER IS, AS FAR AS WE CAN TELL, IS THAT THIS IS A INDEPENDENT OF THE PRESENCE OF SO HERE IS THE DEPLEAION AND THIS IS AND THE MICE ARE PERFECTLY FINE. SO AT THIS POINT, WE'RE IN A QUANTITY. THESE MICE ARE CLEARLY RESISTANT AND IT'S T-CELL DEPENDENT BUT WE'RE STRUGGLING WITH WHAT ON EARTH COULD BE INVOLVED HERE AND WHAT IS THE MECHANISM OF RESISTANCE? ONE THING WE SAW VERY EARLY ON IN THE FIRST EXPERIMENTS IS IF YOU FOCUS ON MARCO FACTS IN THE PERITONEAL IF YOU LOCK AT DAY 10 AFTER CPS INFECTION WE HAVE A LOT OF MACRO PAGES IN THE KNOCK OUT MICE AND IF YOU TAKE THEM AND STAIN FOR CPS AND IN THIS EXPERIMENT ABOUT A THIRD OF THOSE CELLS ARE POSITIVE. WE'RE NOT SEEING MANY MACRO FAGES AND REALLY VERY FEW OF THEM, SOMEWHERE AROUND 12% IN THIS EXPERIMENT, AND IN SOME SENSE IT'S COMING FULL CIRCLE FOR ME BECAUSE WHEN I WAS A POSTDOC IN ALAN'S LAB, I SPENT TWO YEARS DOING CHROME YUM RELEASE ASSAYS AND LOOKING AT CTL ACTIVITY TURNING TOXOPLASMA INFECTION SO IT'S INTERESTING TO COME BACK TO DIRECT KILLING OF T CELLS AFTER SO MANY YEARS. WE LOCKED AT THAT AND SO WE CAME UP WITH THIS PROTOCOL TO MEASURE CYTO TOXICITY AND SO WHAT THE PROTOCOL CLAIRE AND I CAME UP WITH IS THAT WE WOULD DECIDE TO GENERATE MACROPHAGES AND THEN AT WHATEVER IT IS DAY FIVE AFTER CULTURE WE TAKE OUR MACROPHAGES AND SPLIT THEM INTO TWO POPULATION AND ONE POPULATION WE INFECT WITH PARASITES WITH TOXOPLASMA AND WE LABEL THIS POPULATION CFSE AND THEN OTHER POPULATION IS LABELED WITH CELL TRACE VI LOT AND WE TAKE THEM AND MIX THEM UP IN A TUBE AND THEN WE ADD IN PERITONEAL CELLS FROM CPS INFECTED MICE AND GIVE THEM SIX HOURS TO INCUBATE AND COLLECT THE CELLS FOR ANALYSIS SO THE IDEA IS TO LOOK AT WHAT IS HAPPENING TO THE INFECTED MACROPHAGES RELATIVE TO THE MACROPHAGES THAT ARE NOT INFECTED OR EXPOSED TO PARASI PARASITES. AND HERE IS OUR STRATEGY AND WE'RE STARTING OUT WITH OUR PLOT AND AND SO HERE THESE WOULD BE THE EFFECTER CELLS FROM WE CAN SEPARATE THEM FROM POSITIVE CELLS AND THE CTC POSITIVE CELLS SO WE COLLECT ON THESE CELLS AND FOR CELLS AND DO A LIVE DEAD CELL AND WE HAVE THE CELLS FROM CTV POSITIVE CELLS AND IF YOU LOOK CLOSELY AT THESE PERCENTAGES, THEY ADD UP TO 100 AND IT'S COMPLETELY WHAT YOU WOULD EXPECT. WHAT HAPPENS TO THE RATIO OF CFS POSITIVE CELLS TO CTV POSITIVE CELLS. THAT'S THE QUESTION HERE. OK, SO, HERE IS ONE EXPERIMENT AND SO THE TOP PANEL SHOWS TARGET CELLS ALONE SO THERE'S NO EFFECTER CELLS HERE AND JUST TAKING THE LABEL CELLS AND LETTING THEM INCUBATE FOR SIX HOURS AND SO HERE ARE THE PERCENTAGE HERE SO 63% OR CTS AND CFE POSITIVE IF YOU GATE ON THESE CELLS, ABOUT A QUARTER OF THEM ARE INFECTED AND IN PARTICULAR, IF YOU LOOK AT THE CFSE POSITIVE CELLS THERE'S A DECREASE FROM 37% DOWN TO 23% AND IF YOU GATE ON THESE CELLS, AND ASK HOW MANY ARE INFECTED IF THE NUMBERS DROPPED TO ABOUT 11%, IF YOU TRADE IN EFFECTOR CELLS, THE NUMBER OF TARGET CELLS DECREASES REALLY PRETTY DRAMATICALLY IN THIS EXPERIMENT AND WE CAN DO THE SAME THING NOW. EVERYTHING IS IN THE SAME TUBE INTERNALLY CONTROLLED. AND SO WE CAN LOOK AT NON INFECTED TARGETS AND THESE WOULD BE CFSE POSITIVE CELLS THAT DON'T HAVE ANY PARASITES RELATIVE TO CTV POSITIVE MACROPHAGES AND HERE YOU CAN SEE THAT THERE'S REALLY NOT MUCH OF A EVIDENCE FOR A DISAPPEARANCE OF THESE CELLS AND SO TO GET BACK TO THIS MODEL, SOP WHAT WE THINK WE'VE GOT HERE IS SOMETHING REALLY INTERESTING THAT MAYBE PEOPLE HAVEN'T APPRECIATED BEFORE. SO WE HAVE A PROTECTIVE RESPONSE, DOESN'T REQUIRE IL12, DOESN'T REQUIRE TLR SIGNALING AND SEEMS TO DEPEND ON CD8 AND CD4T CELLS WE NEED TO TEASE APART THE CONTRIBUTIONS TO BOTH TO THIS PROTECTIVE RESPONSE AND DOESN'T REQUIRE INFLAMMATORY MONOCYTES. AND INTERFERE AN GAMMA IS ONLY PARTIALLY REQUIRED. OUR WORKING HYPOTHESIS RIGHT NOW IS PROTECTION IS DEPENDENT UPON T-CELL ACTIVITY AND WE STILL NODE TO WORK ON AND IT MIGHT BE INTERESTING IS TO FIGURE OUT THE MAC REQUIREMENTS FOR THIS RESPONSE AND ALSO THE SPECIFIC INVOLVEMENT OF CD4s IN THIS KILLING RESPONSE. SO, I'M GOING TO LEAVE IT THERE. SO THIS IS MY CURRENT LAB. WE HAVE A SMALL LAB BUT WE'RE GETTING ALONG WELL WHICH IS NICE AND WE WORK WELL TOGETHER AND WE HAVE A GO AHEAD TIME TOGETHER AND AS I SAID, THIS IS WORK THAT WAS MOSTLY DONE BY CLAIRE DOUGHTERY. A SECOND YEAR GRADUATE STUDENT IN THE LAB WITH HELP FROM LINDSEY SCHNEIDER A POSTDOC, JESSICA ORTEGA IS A BEGINNING GRADUATE STUDENT AND ALSO SOME TALENTED UNDERGRADUATES. THANK YOU. [APPLAUSE] >> THANK YOU. >> I WAS STRUCK BY THE FACT THAT IN THE INTER FEWER AN GAMMA KNOCK OUT MICE, IT LOOKED AS THOUGH 60% OF THEM DIED QUICKLY AND THE OTHER 40% THERE WAS A LONG PLATEAU OF SURVIVAL, LONG-TERM SURVIVAL AS IF THERE WAS SOME HETEROGENEITY IN THE MICE AND ONLY A LITTLE MORE THAN HALF OF THEM WERE SUSCEPTIBLE. IS THERE ANYWAY THAT YOU CAN LOOK FOR HETEROGENEITY IN THOSE MICE? >> IT MIGHT GIVE YOU A CLUE TO WHAT IS GOING ON? >> I SUPPOSE YOU COULD. YOU COULD TAKE -- YOU COULD BASICALLY JUST ASK WHAT IS GOING ON IN THE PERITONEAL CAVITY IN TERMS OF CELLS RECRUITED THERE. I DON'T KNOW -- I'M NOT SURE HOW MUCH SIGNIFICANCE I WOULD ATTACH TO THAT RESULT. IT'S VARIABLE FROM EXPERIMENT TO EXPERIMENT HOW MUCH SUSCEPTIBLE YOU GET FROM THE INTERFERON GAMMA KNOCK OUTS SO THE INTER FEWER ON GAMMA IS PARTIALLY REQUIRED NOT TOTALLY REQUIRED BUT THE SPECIFIC CONNETICS OF WHAT THE MICE DIE CAN VARY FROM EXPERIMENT TO EXPERIMENT. IT ON THE EXCLUDE TLR3 OR EVEN TLR4 SIGNALING THROUGH TRIFF AND IF INTER FEWER ON CAM IS ONLY PARTIALLY REQUIRED. >> WE HAVE NOT DONE THAT EXPERIMENT. GOOD IDEA. THESE KNOCK OUT OUTS ARE EXPENSE TO DO. >> ERIC, YOU ACTUALLY MAY REMEMBER YOU AND I DID SOME EXPERIMENTS ON THOSE CHROME YUM >> WHEN WE WERE LOOKING AT CROSS PRESENTATION SO THAT COMES BACK TO THIS WHICH IS WHETHER THIS PARTICULAR STRAIN IS MORE PRONE TO GET OUT OF HAVE ACCESS TO THE CLASS 1 PRESENTATION PATHWAY AND TRYING TO PUT MANY DIFFERENT THINGS TOGETHER AND I KNOW YOU SAID IT WAS IL12 INDEPENDENT BUT IT COULD STILL BE DC1 DEPENDENT AND HAVE YOU TESTED BA3 KNOCK OUT MICE. >> WE HAVE NOT TESTED THOSE MICE BUT IN TERMS OF CROSS PRESENTATION, YEAH, I DEFINITELY REMEMBER THOSE EXPERIMENTS SO ONE QUESTION WE HAD WHETHER WE WOULD SEE EVIDENCE FOR KILLING OF NO NON INFECTED BYSTANDER CE. THERE WAS A LITTLE BUT IT WASN'T DRAMATIC. >> DOES THE INITIAL PRIMING INVOLVE A DENDRID I CAN CELL POPULATION THAT IS THE EFFECT AND JUST KILLED THE EFFECTIVE MACROPHAGES SO AND THE BAT F-3 KNOCK OUT MODELS MIGHT BE A GOOD WAY. >> I GROW, THANK YOU FOR THE SUGGESTION. >> I HAVE A QUICK QUESTION ABOUT THE STRAIN AND DO HAVE YOU ANY IDEA WHAT MAKES IT GROW IN HIGHLY -- >> YEAH, RIGHT. SO THAT'S KIND OF LIKE IT KEPT ME UP A LOT AT NIGHT WONDERING WHY ON EARTH THESE MICE WHY DYING. THE ONLY SORT OF THING I CAN COME UP WITH IS MICE HAVE THEIR OWN URACIL, RIGHT, IT'S NOT A URACIL FREE ENVIRONMENT SO WE'RE WORKING HYPOTHESIS IS THAT URACIL LEVELS ARE ENOUGH TO ALLOW SOME PERSISTENCE AND PROLIFERATION OF THE CPS STRAIN AND IN WILD TYPE MICE THERE'S A EFFECTER RESPONSE THAT COMES IN AND CONTROLS THE INFECTION BUT WHEN YOU TAKE THE T CELLS, YOU HAVE UNCONTROLLED INFECTION. THAT'S WHAT I THINK. GO WITH THAT? >> YEAH. I GUESS. >> YEAH. >> THANK YOU, A LOT. IT'S NEVER TOO LATE, ERIC IS A PROFESSOR AT NEW MEXICO UNIVERSITY AND I THINK WE'RE STARTING THE POSTDOC IN ALAN'S LAB ALMOST AT THE SAME TIME AND I'M REALLY VERY IMPRESSED WITH HIS WORK. THE NEXT SPEAKER IS FELIX AT THE UNIVERSITY OF ROCHESTER AND FELIX, WE CALL THE PROFALIN GUY AND THERE'S FOR DECADES PEOPLE TRYING TO PURIFY A TOXO COMPONENT THAT EFFECT HE WAS THE ONE THAT DID IT. >> THANK YOU, RICHARD AND BEFORE I START, I ABSOLUTELY HAVE TO TAKE A PICTURE OF THE MOST AMAZING SCIENTIST IN THIS ROOM BECAUSE I REALIZE THAT THIS IS THE PLACE WHERE TO BE TO TAKE THIS PICTURE. THANK YOU, DRAGANA FOR PUTTING THIS TOGETHER. EVEN THOUGH I WILL START THIS STORY WITH PROVE OWE LYNN THE THINK ABOUT THIS NO MATTER WHAT YOU GO AFTER THAT PEOPLE STILL REMEMBER YOU FOR ONE THING, RIGHT. [LAUGHTER] SO, IN REALITY, I CAME TO ALAN CHERYSE LASHER'S LAB AT I PERFE. IT LANDED IN THE TEXTBOOK. HE DID A SIMPLE AND PROBABLY VERY ELEGANT EXPERIMENT WHEN HE INFECTED DEFICIENT MICE AND HE OBSERVED THE LACK OF OR INTER FEAR AN GAM AND THEY WERE SUSCEPTIBLE TO THE PARASITE. SIMPLE AND H ELEGANT E EXPERIME. THE QUESTION WAS SIMPLE WHEN I JOINED THE LAB, WHAT TRIGGERS THE ACTIVATION SO THE IDEA IS THERE'S ALREADY KNOWLEDGE THAT THERE'S A SOLUBLE PROTEIN THAT TRIGGER RESPONSE VERY A PATHWAY SO BASICALLY IT'S SIMPLE, JUST GO AHEAD AND ISOLATE THE MOLECULE AND IDENTIFY THE RECEPTORS, RIGHT, THIS IS THE QUESTION AND THIS IS THE APPROACH SO IDEA AND I WAS LUCKY TIMES WERE BEHIND YOU WAS GROWING AND SOLUBLE AND AND YOU TAUGHT ME AND DO IT AND THEN AND TEST FRACTION ON DENDRID I CAN CELLS AND TEST THEIR ABILITY TO TRIGGER AND WE HAVE AND THE LAB WAS SURE AND AND AND IT WOULD BE EASIER BUT THERE'S A REASON BY I'M SHOWING THIS BECAUSE I'VE NEVER TOLD AND THIS STORY. DURING THE FIRST WEEK, AT THIS CAME USUALLY BEEN 15 MINUTES AND I STARTED DOING AND I TURNED ON THE MACHINE IN A MINUTE, FOUR MEETERS OR MORE THE WASHING SOLUTIONS WERE ON THE FLOOR AND IT WAS COMPLETE MESS AND CAN YOU IMAGINE THE FIRST WEEK IN THE LAB AND YOU CREATE A MAJOR MESS IN ALAN SHER'S LAB AND I PANICKED AND IT GETS WORSE. WHAT HAPPENED NEXT, I COMPLETELY KNOCKED DOWN AND BROKE THE FULL LITER CYLINDER SO THAT WAS A COMPLETE DISASTER AND ESPECIALLY IN EVERYONE WHO MET PAT KNOWS ABOUT THAT AND SHE'S STRICT AND I KNEW SHE WOULD BE IN THE LAB IN 15 MINUTES SO I HAD TO CLEAN UP EVERYTHING AND MAKE IT BRAND NEW AND IT LOOKS LIKE NOTHING HAPPENS AND REMARKABLY, I EVEN IN THIS 15 MINUTES, I WAS ABLE EVEN THOUGH BORROW, I NEVER RETURNED IT -- THIS WAS THE BEGINNING OF THIS ADVENTURE, WHICH AFTER THAT IT BECAME MUCH BETTER. I HAVE TO TELL YOU THAT RELATIVELY QUICKLY WHY ISOLATED WHICH INDUCED INTO RESPONSE WHICH APPEARED TO BE PRO FILL VARIANT PROTEIN WHICH WE EFFICIENTLILY CALLED PRO FILL IN-LIKE PRE TEEN. THE REMARKABLE WHEN ICE WERE IMMUNIZED WITH STAT THEN A LOT OF CD40 CELLS RECOGNIZED PROFILIN AND DRAGANA CLONED THE CELLS AND HAD A LOT OF PROFILIN-SPECIFIC CD4 CELLS WHICH I DIDN'T KNOW THERE ARE SPECIFICITY, HOWEVER, I DIDN'T LEND IT IN THE TEXTBOOK BUT AS YOU CAN SEE IT IS EXTREMELY SUSCEPTIBLE TO INFECTION AND FOR US WHEN WE -- UNFORTUNATELY FOR ME BUT THIS IS WHAT WE'VE GOT WHEN WE INFECTED THE MICE THEY WERE MUCH MORE SUSCEPTIBLE TO THE INFECTION BUT THEY SHOW PHENOTYPE AND MANY OF THEM SURVIVED AT THAT TIME INFECTION AND THAT CERTAINLY BECAUSE THE BASIS FOR THE EXPERIMENTS AND WE WANTED TO UNDERSTAND A LITTLE BIT MORE HOW PROFILIN AND OTHER MOLECULES ARE RECOGNIZED. WITHIN A SHORT PERIOD OF TIME, IN MY LAB, I WAS SURPRISED TO FIND OUT THAT RECEPTORS 11 IS A RECEPTOR DESPITE AT THAT TIME, EVERYONE BELIEVED THAT ONLY SENSING TO RECEPTORS WAS THE AND THE SIGNALING FROM THE 11 DEPEND ON PROTEINS AND LATER ON, ALAN'S LAB AND MY LAB SHOWED THAT TLR11 FUNCTION IS A IN RECEPTORS AND AND FUNCTION IS A REQUIRED FOR DEPEND ANT INDUCTION OF IL-12. WITH THE CELLS THIS PROCESS HAPPENS IN DENDRID I CAN CELLS AND THERE WAS AS WELL AS FROM TE LAB IN THE CONTEXT OF MUCOSAL IMMUNOLOGY ESTABLISHED THAT THIS IS CR2 POSITIVE MONOCYTES. HOW THEY ACTIVATED AND HOW THEY RECRUITED TO THE SITE OF INFECTION IS NOT COMPLETELY UNDERSTOOD IN AND ONE POSSIBLE PATHWAY WHICH IT RECRUITMENT OF MONOCYTES TO THE SITE OF INFECTION AND WAS RECENTLY REVEALED AND WE OBSERVED THAT DURING TOXOPLASMA INFECTION NOT ONLY THERE'S THE RELEASE OF PROVE OWE LYNN AND PROTEIN BUT ALSO RELEASED THE PROTEIN WHICH IS CALLED S100A11 WHICH LEADS TO CCL11 AND MONOCYTES AND CELLS CONTRIBUTE TO INDEX AND IT'S JUST INTRODUCED US. OF COURSE THE CITATION IS MORE COMPLEX AND THIS IS A BEAUTIFUL SUMMARY FROM OUR CHRIS HUNTER RECENT REVIEW THAT IN ADDITION TO TL11 OF IL-12 THERE'S SOME CONTRIBUTION FROM RECEPTORS AND CONTRIBUTE OR TO THIS FIELD AND IN ADDITION AND WE SHOWED IS THAT AMPLIFY INTER FUR AN GAMMA INDEX AND MOST LIKELY VIA AND I INTRODUCED THE DAMP MEDIATED IMMUNITY TO AND ALL OF THESE PATHWAYS LEADS TO INTRA FUR ON GAMMA IN THE PARASITE IN THE INFECTED CELLS AND SELECTIVELY IN AND OF COURSE, OVER PRODUCTION OF INTER FUR AN GAMMA IS DETRIMENTAL AND BEAUTIFULLY ILLUSTRATED WITH INDEX OF I AM AN' PATHOLOGY IN TESTING AND RE AT NIH CAMPUS JASMINE INDICATED TO AND AND OUR LAB IDENTIFIED THIS THERE'S A PRECISE PATHOLOGY CHARACTERIZED AND DISAPPEARANCE AND IN THEIR DISAPPEARANCE LEADS TO UNCONTROLLED PROLIFERATION OF INDESTINNAL BACTERIA WHICH IS RESPONSIBLE FOR THE SEVERE. LOOKING BACK ON THE POSITIVE SIDE OF INTER FUR AN GAMMA IS A CRUCIAL SPECIFICALLY AND THERE ARE DIFFERENT FLAVORS OF BROADLY DIVIDED IN TWO GROUPS AND WELL CHARACTERIZED MACROPHAGE GENERATED FROM MONOCYTES AND INITIALLY FROM BONE MARROW BUT MAJORITY WAS SEEDED DURING OUR EMBRYONIC DEVELOPMENT AND THE CO TISSUE RESIDENT AND THEY HAVE DIFFERENT FLAVORS AND DIFFERENT NAMES AND THIS IS ILLUSTRATION FROM TOM VINCE AND OUR REVIEW WHERE YOU CAN SEE THIS TISSUE RESIDENT MACROPHAGE AND THE LEVER OF MIKE IT'S TWO GROUPS AND MA MACROPHAGES AND THEY'RE E CELLS AND PARA PAR AND THIS IS DIFFERENT WAYS TO DEFINE THIS DIFFERENT SUBSETS OF MACROPHAGES AND THEY EXPRESS HIGH LEVELS OF CELLS AND CERTAINLY ARE NOT CLASS 2 WHICH IS TIP I CANAL ASSOCIATED WITH SMALL PERITONEAL MACROPHAGES AND WE ASK WHAT IS THE DIFFERENT ROLE FOR THOSE MACROPHAGES SUBSETS AND THE NOW YOU ARE REQUIRED FAMILIAR THAT IN PERITONEAL CAVITY AND THE MACK OWE PAGES ARE THE DOMINANT CELL AND EVERYTHING CHANGES DURING GONDII INFECTION AND AND CLEARLY INDICATING THAT MACROPHAGES ARE LOST DURING TONS SO AND AND MACROPHAGE ACTIVATION AS WELL AS INDUCTION OF MAJOR CHANGES IN CELL COMPOSITION AND THE QUESTION WHETHER THERE'S LOSS OF MACRO PAGES IS MEDIATED BY INTERFERON GAMMA AND THEY HAVE A SIMPLE EXPERIMENT WHEN WE INFECTED MICE AND HERE WE CAN SEE IF WE BLOCK IT WE COMPLETELY MACRO PAGES AND CLEARLY INDICATING THAT INTERFERON GAMMA IS REQUIRED FOR THIS PROCESS. IT'S A SUFFICIENT AND TO ADDRESS THIS QUESTION, WE ARE SWITCHED FROM TAX OWE PLASMA INFECTION TO VERY SIMPLE EXPERIMENT WHERE MICE WERE TREATED WITH INTER FEWER ON GAM APP AND WE SEE THAT IT'S SUFFICIENT FOR THE LOSS OF LIFE AND MACROPHAGES. SO, MANY CELLS CAN RESPOND TO INTERFERON GAMMA AND THE EXPERIMENTS WHICH I SHOWED YOU SO FAR DID NOT PROVE THAT INTERFERON GAMMA ACTS IN MACROPHAGES TO ADDRESS THIS POSSIBILITY WE GENERATED MICE WHICH LACK INTERFERON GAMMA RECEPTOR IN MACROPHAGES AND CROSS MICE TO INTERFERON KAMA RECEPTORS AND NOW INFECTED THOSE MICE. AS EXPECTED, THE CONTROL LOW ROW MARK BLEE REPRODUCING ABLE RESULTS AND IF THE TARGET NOW YOU CAN SEE THOSE CELLS ARE COMPLETELY PRESERVED AND CLEARLY INDICATING THAT THE DIRECT EFFECT OF INTRA FUR ON LIVES THROUGH THEIR DISAPPEARANCE AND WHEN WE TALK ABOUT THE LOSS, IT'S ACTUALLY CONVINCED A LOT OF THINGS, RIGHT. IT MEANS THAT THE RESPONSE TO INTERFERON GAMMA OR T TOXO GAMMA INFECTION SO CELLS CAN CHANGE AS SURFACE EXPRESSION THAT'S WHY WE DO NOT DETECT THEM BY FLOW SIM TOM TREE OR THEY CAN DIE. WE KNOW THAT IT'S NOT ULTRA-GENE EXPRESSION BECAUSE WE KNOW CELL TRACING EXPER EXPERIMENT AND WET THINK THIS MIGRATION, EVEN THOUGH IT CREATES MIGRATION T TO IT DOES NOT EXPLAIN THE MAJOR LOSS OF LIFE AND TO TEST A HYPOTHESIS, THAT INTER FUR AN GAMMA STIMULATION LEADS TO THE DEATHS OF PERITONEAL MACROPHAGES AND THIS IS SUPPORTED BY OUR PREVIOUSLY WORK ON CELLS AND WE'RE IN THE PAST, WE RECENTLY PUBLISHED THIS THAT'S WHY I AM NOT GOING OVER THE DETAILS, WE OBSERVED THAT IN RESPONSE TO TOXOPLASMA GONE DE. FOR THAT REASON, WE TESTED THIS IDEA THAT MAYBE MACROPHAGES ARE ALSO LOST IN RESPONSE TO INTERFERON GAMMA AND TO TEST THIS MODEL, WE SET UP A SIMPLE IN VITRO SYSTEM AND THE MACROPHAGES WAS STIMULATED WITH INTRA FUR ON GAMMA AND THIS IS, THIS IS AND WE DIDN'T SEE MANY DYING CELLS IF ANY. AND EVERYTHING CHANGED IF WE ADD INTERFERON GAMMA. WE CAN SEE THAT DIRECT STIMULATION OF MACROPHAGES IN VITRO RESULTING IN THE RAPID ACCUMULATION OF DYING MACROPHAGES IN VITRO AND CLEARLY INDICATING IN INTRA FUR ON GAMMA LEADS TO THE DEATH OF LARGE MACROPHAGES AND HOW IT'S HAPPENING, OBVIOUSLY WE'RE BIAS IN THE EXAMINING THE AND I WANT TO SHOW YOU A LOT OF NEGATIVE DATA BUT ALL THE CLASSICAL PATHWAY INCLUDING PLAY NO SIGNIFICANT ROLE IN DYING MACROPHAGES IN RESPONSE TO INTRA FUR ON GAMMA WE WITH YOU KNOW IN THE CONTEXT OF THE CELLS THAT IFN SUE PLUSES COMPLEX AND M.T.O. R AND IT'S A CENTRAL HAPPENS THE CELLS THAT CAN SEND NORMAL AVAILABILITY OF THE NUTRIENTS OR ENERGY AND ESSENTIAL FOR CELL AN BOLISM OR CAT A BOLISM AND I WANTED TO TEST THE IDEA THAT MAYBE MTOR DUE EXPRESSION IN THE DOWNSTREAM IS RESPONSIBLE FOR THE DEATH AND TO TEST THIS IDEA THIS MICE TREATED WITH SOME RISES OF DMSOE CAN SEE ALL THE MACROPHAGES. WE CAN DO THE TRACING EXPERIMENTS FOR THE CELLS AND YOU CAN SEE HERE THAT AGAIN IN THE CONTROL SYSTEM, WE CANNOT DETECT DYING CELLS BUT HERE IN RESPONSE TO WE CAN SEE DYING CELLS AMONG THOSE FEW REMAINING CELLS WHICH WE CAN STILL PICK UP BY FLOW CYTOMETRY AND IT'S INDICATING THAT MTOR1 AND LEADS TO THE DEATH OF LARGE PERITONEAL AND THIS IS THE QUANTIFICATION OF THE DYING CELLS. SO, BACK TO UPSTREAM EFFECT. WHAT HAPPENS? HOW MTOR IS SUPPRESSED IN MACROPHAGES OF INTERFERE AN GAMMA AND NOW A LITTLE BIT ABOUT PAN ETH CELLS SO IN THEIR DEPENDENCE ON MTOR SO SURPRI SURPRISINGLY, WE KNOW A LOT ABOUT MACROPHAGES BECAUSE THEIR EXISTENCE DEPENDS ON THE SIGNALING THROUGH MCF RECEPTORS AND CD115 AND IT LEADS TO THE SIGNALING PATHWAY REQUIRED FOR THEIR SURVIVAL AND PROLIFERATION AND WE FOCUS OUR ATTENTION ON CD115 AND SCHEER WHAT WE OBSERVED. IN MICE, MACROPHAGES EXPRESS LARGE AMOUNTS OF CD115 AND IF WE INFECT MICE WITH INTERFERON GAMMA AND THE PLASMA GONDII AND THE EXTRA CD115 AND CIRCULATING IN THE BLOOD IN PER TENILLE AND IT LEADS TO THE LEADING OF MCF ROW SCEPTER AND CD115 AND IT MAKES THE CELLS SUSCEPTIBLE TO INTERFERON GAMMA. IF WE INFECT THEM INJECT INTER FUR AN GAMMA NOTHING INTERESTING HAPPENS. IN TERMS OF CD115 LOSS AND SHEDDING MAKE ROW PSHEHAD MACROR THE CELL TIME YOU CAN SEE THE CELLS STILL LOSING CD115. AND AT THE SAME TIME, WHAT WE NEXT ASK WHETHER THIS LOSS OF LIFE MACROPHAGES PLAY A ROLE IN HOST RESISTANCE TO TAX OWE PLASMA GONDII AND HERE YOU CAN SEE YES WE PRE SERVED THE MACROPHAGES BUT THIS MICE ARE SUSCEPTIBLE TO THE INFECTION AND IN THEY'RE DYING BECAUSE OF THE UNCONTROLLED PARASITE BURDEN AND SO WHY IS THAT HAPPENING? LOOKS LIKE IF WE ANALYZE WHAT CELLS THAT TYPICALLY INFECT IT AND IN OUR HANDS WE'RE SEEING THE MACROPHAGES AND THE PATHOGENS TOXOPLASMA BONDI AND IT'S THE HOST DESK MECHANISM. EVEN WHEN WE DISAGREE OVER THE MULTIPLE YEARS I STILL LEARN, ALAN IS ALREADY RIGHT. ALAN SAID IT WAS A SMART PATHOGEN AND I'M NOT READY TO ACCEPT THAT BUT MAYBE HE IS RIGHT BECAUSE OF THE VIRTUE LENS FACTORS IN SUPPRESS INTRA FUR ON GAMMA AND ITSELF AND THE INFECTION AND I DON'T THINK THAT I NEED TO TRY TO SEE IT'S DIFFICULT TO CON FOR INSTANCE PEOPLE IN THIS ROOM BUT CLEARLY, REVIEWED THE KEY ASPECT OF RECOGNITION AND IMMUNO PATHOLOGY AND AS I CONTRIBUTED TO TLR INDEPENDENT AND INDEPENDENT MECHANISM AND CYTOKINE AND IMMUNE DEFENSE AND PATHO GOLLY AND WE DO BELIEVE FROM OUR RECENT WORK THAT METABOLIC AND. >> I DON'T THINK WE HAVE ESTABLISHED THE FORMAL BORDERS, MAYBE ALAN AND TOM KNEW WHERE THE LABS ENDS BUT FOR US IT DIDN'T ABSOLUTELY MATTER SO IT WAS A FANTASTIC ENVIRONMENT. I'M GRATEFUL TO MY OWN LAB. LABS ACTS DEPENDS NOT ONLY AND TRAIN THESE AND PHENOMENAL LAB MANAGER AND ALAN WAS ABSOLUTELY LIKELY TO HAVE SARAH HEINE AND HAVE SAM AND AND HE HAD PHENOMENAL PERSON AND ANDREW AND THE WORK ON MACROPHAGES AND OF HIS HELP OF ALL THE OTHER LAB MEMBERS AND ALAN IS SET UP INCREDIBLE STANDARDS FOR BEING A GOOD MENTOR SO I HAVE A LOT TO LEARN SO ON TRACK OR TENURE PROFESSOR SO A LOT TO BE ACCOMPLISHED AND THIS IS THE PEOPLE IN THE LAB ALYSE AND THANK YOU. [APPLAUSE] >> GREAT TALK, FELIX. AS ALWAYS. I WAS JUST CURIOUS IF LPS DOES THE SAME THING WHETHER OR NOT THAT'S INTERFERON DEPENDENT. SO WHAT IS CALLED MACROPHAGE DISAPPEARANCE REACTIONS SO MACROPHAGES AND SURPRISINGLY, THAT IS INTERFERON GAMMA INDEPENDENT MECHANISM AND ALSO INTER DEPENINTER DEPENDING MECH. >> THEY HAVE SHOWN A FASCINATING CAVITY WHERE MACROPHAGES CAN ACTUALLY CREATE THESE SHEET AND BASICALLY KIND OF BECOME A MEMBRANE AND IN THE CONTEXT OF INFECTION AND INFLAMMATION AND I KNOW THAT YOU HAVE SHOWN THAT SOME OF THOSE MACROPHAGES DIE. DO YOU KNOW THE PROCESS DURING THE CONTACT OF PLASMA AND CONTRIBUTE SOMEHOW TO MOR LIKE A SHIELD STRUCTURE AND GRANULAR. >> YEAH, THIS IS A BEAUTIFUL STORY FROM GWEN RANDOLPH. WE HAVEN'T SEEN IT TO THAT DEGREE SO IT'S TRAPPED IS ALMOST RIGHT AND COMPLIMENT THE LIST OF ACTIVATION AND ELIMINATION WITH FORMALLY HAVE NOT INVESTIGATED THE COMPLIMENT AND WE DIDN'T AND AS MUCH AND PREPARATION SO AGAIN, IT STUDLY AND NOT TO THIS BEAUTIFUL DEGREE AND NOT TO SAY AND AND. >> AND SO OUR NEXT SPEAKER IS TAKESHI KAWABE, A PROFESSOR AT THE UNIVERSITY OF TOHOKU WHERE HE HAS HIS OWN LAB. HE POSTDOC AND IS VISITING SCIENTIST HERE AT NIAID AND HE TALKS ABOUT THE ROLE OF INNATE ROLE OF T CELLS, I GUESS. >> THANK YOU. GOOD MORNING, EVERYONE. MY NAME IS TAKESHI KAWABE. FIRST OF ALL, THANK YOU VERY MUCH FOR IN INVITING ME TO SUCH A WONDERFUL SYMPOSIUM AND IT'S A GREAT HONOR I GREW UP WITH YOU IN THE LAST PART OF THE LONG HISTORY WHERE LPD AND TODAY, I'M GOING TO TALK ABOUT MEMORY PHENOTYPE C CELLS WHICH I STARTED IN THE LAB AND IN ALAN'S LAB FOR MANY YEARS AND MY OWN LAB IN JAPAN. T CELLS ARE GENERATED IN THE AND THEY ARE TESTED AND AND THOSE WITH ANTIGENS AND THEY DID IT BY AND THOSE WITH TWO CELLS ANTIGEN AND AND THE CONSEQUENCE ONLY SIGHTS CAN SURVIVE AND HIGH GRIT INTO THE PERIPHERY AS THE T CELLS AND IN THE PERIPHERY, THE INFECTION WHEN AND EFFECT US OUR CELLS TO CONTRIBUTE TO HOST PROTECTION AND AFTER MOST EFFECT OUR CELLS DYING WITH A POPULATION IN THE MEMORY CELLS WHICH PROVIDE MEMORY AND IT'S WELL-KNOWN THAT THIS MEMORY CELLS CAN BE DISTINGUISHIBLE USING MARKERS CD44 AND CD62L AND THIS IS AN EXAMPLE OF THE SPECIFIC CD14 EFFECTOR AND THEY'RE 44 LOW AND CD62 HIGH AND MEMORY CELLS AS CD44 HIGHS AND LOW. SO USING THESE MARKERS WE CAN DISTINGUISHED CELLS AND EFFECT OUR MEMORY CELLS AND BY THE WAY, ALL OF T CELLS SHOULD HAVE AFFINITY TO SELF-ANTIGENS THAT SELF-ANTIGENS BECAUSE OF THE POSTING SELECTION AND ACTUAL LOW THIS IS KNOWN TO PLAY ESSENTIAL ROLES FOR THE MAINTENANCE AND SURVIVAL OF NAIVE CD4 CELLS IN THE PERIPHERY HOWEVER IT'S NOT KNOWN WHETHER IT'S NAIVE T CELLS REGULATE SELF-ANTIGEN AND PROLIFERATE AND THE PHENOTYPE AND IF THIS IS THE CASE WHAT THESE FUNCTION. THE NEGATIVE CONVENTIONAL CD44 CELLS CONSIST OF TWO DIFFERENT CD44 HIGH NAIVE CELLS AND CD44 AND 62 LOW MEMORY CELLS AND PREVIOUSLY THIS CD4462 LOW THE MEMORY WERE OVERLY PRESENT SPECIFIC MEMORY CELLS AND THAT THE FOR COMMENCE OR ANTIGENS HOWEVER, THIS SIMPLE ASSUMPTION HAS BEEN ACCORDING TO QUESTION BAYS OF SEVERAL REASONS THAT I WILL TALK IN THIS TALK. AND SO, HERE ARE MY PRIMARY QUESTIONS. THE FIRST, HOW ARE THIS MEMORY-LIKE GENERATED IN CITY AND WHAT NEEDS THE FUNCTIONAL SIGNIFICANCE? AND THE SECOND, WHAT SAY MECHANISM OF THEIR DIFFERENTIATION AND THIRD, ARE THEY REALLY DIFFERENT FROM FOREIGN AG-SPECIFIC AND AUTHENTIC MEMORY CELLS? SO FIRST, THE FIRST QUESTION THAT GENERATION IN FUNCTION OF MEMORY-LIKE CD4T CELLS AND THE FIRST, TO START THIS PROJECT, FIRST, AS A POSTDOC, ACTUALLY IN POSTDOC I EXAMINED THAT THE DEVELOPMENT OF MEMORY-LIKE FOR T CELLS AND TO DO SO I FIRST COUNTED JUST COUNTED THE TOTAL NUMBER OF MEMORY-LIKE AND CD4T CELLS IN MICE OF DIFFERENT AGES AND AS YOU CAN SEE HERE, BOTH NAIVE AND MEMORY DEVELOPED WITH AGE AND TO EXAMINE IF THE DEVELOPMENT OF MEMORY-LIKE CELLS REQUIRED SOME 40 ANTIGEN RECOGNITION, NEXT I COMPARED THAT MEMORY NUMBER IN SPF AND AS YOU CAN SEE HERE, THERE'S NO DIFFERENCE IN THE NUMBER OF MEMORY-LIKE CELLS AND SHOWING THAT THE DEVELOPMENT OF MEMORY-LIKE CD4T CELLS DOES NOT REQUIRE 40 ANTIGEN RECOGNITION AND THIS COMMENCE ANTIGEN RECOGNITION. AND HOW ARE THESE MEMORY-LIKE CDF IN IT SHOULD BE NAIVE AND NEXT THE CELLS MAY CONVERT THE MEMORY CELLS IN THE PRO LIVE REIN AND THIS NEXT I TRANSFER ANY EVE CD4T CELLS AND JUST INTO THE KNEW OWE NIGHTAL AND COULD CON AGAIN I CAN ANIMALS THAT ARE NOT CHALLENGES WITH ANY ANTIGENS AND I USE A MICE ENVIRONMENT IS KNOWN TO BE WHICH IS A STRONGER GENERATES THE HOMO STATIC PROLIFERATION AND THIS GENERATES THE CELLS SO THIS IS JUST A CONTROL AND THIS IS THE GROUP OF INTEREST. AND SEVERAL WEEKS AFTER TRANS TRANSFER, I -- WHEN TRANSFERRED NEO NATURE, MANY MEMORY, WHAT IS INTERESTING, EVEN WHEN TRANSFERRED, SOME OF THE NAIVE CELLS CONVERTED TO MEMORY-LIKE CELLS AS YOU CAN SEE HERE. AND TO EXAMINE IF THIS MEMORY-LIKE CELL GENERATION REQUIRES ANTIGEN RECOGNITION OF THAT, NEXT TRANSFER CELLS INTO ONE TYPE AND MICE AND DID THE SAME EXPERIMENT. AND AS YOU CAN SEE HERE, ALL THE OF THE MEMORY-LIKE CD4 CELLS WERE GONE SHOWING THAT SOME OF THE CELLS SOME ANTIGENS AND CONVERT THE MEMORY-LIKE CELLS IN THE PERIPHERY AND EVEN IN THIS CITY STATE. HE REACH ANTIGENS CONTRIBUTE TO THE CELL GENERATION AND BECAUSE MEMORY LIKE CELLS WERE EQUALLY PRESENT IN SPF I THOUGHT SOME OF THE CELLS WERE DRIVEN BY SELF-ANTIGEN RECOGNITION AND I FOCUSED ON CD5 WHICH IS SAY GOOD MARKER OF SELF-ANTIGENS SO THAT MEANS THAT CD5 HIGH HAVE SELF-ANTIGENS AND CD5 HAVE LOW. SO USING THESE MARKERS, NEXT I TRANSFERRED CD5 HIGH AND CD4 CELLS TO MICE AND CD5 LOW CELLS FROM CD4.5 MICE MAKES THAT THE RATIO 1-1 AND TRANSFER INTO THE NEONATAL AND DID THE SAME EXPERIMENT. S YOU CAN SEE HERE IN NEONATE THE HIGH CELLS DID MORE MEMBER ROW-LIKE CELLS THAN THE LOW COUNTERPARTS. AND IT CAN GENERATE MORE MEMORY-LIKE CD4 CELLS AND THEY DO THE CELLS WITH LOW AFFINITY TO SELF-SO THIS FURTHER SUGGESTS THAT THE INVOLVEMENT OF SELF-ANTIGEN RECOGNITION IN THE MEMORY LIKE CD4T CELLS SO GOING BACK IT'S CLEAR THAT AT LEAST SOME OF THE CD4 CELLS RESPOND TO SELF-ANTIGENS AND THE MEMORY PHENOTYPE AND BECAUSE OF THIS REASON, WE HAVE BEEN DEFERRING TO THIS POPULATION AS MEMORY PHENOTYPE AND FARTHER AT RISK THE FUNCTION OF SIGNIFICANCE OF THESE MP CELLS. AND AS WE DISCUSS LATER, THE MANY MP CELLS BECAUSE OF THIS REASON FIRST NEXT MP CELLS HAVE TH1 FUNCTION TO TEST THIS NEXT I INFECTED THE ANIMALS AND CHECKED IN THE PRODUCTION BY CD44 LOW LEVELS AND SEVERAL DAYS POST INFECTION. AND NOW THE INFECTION CD44 HIGH MP CELLS DIDN'T PRODUCE GAMMA AS EXPECTED WHY DON'T DAY SEVEN CD44 HIGH CELLS AND THIS IS NOT SURPRISING BECAUSE ON DAY 7, NOT ALL OF THE HIGH SHOULD BE ANTIGEN DRIVEN BUT WHAT IS INTERESTING HERE IS ON DAY 2, AT AN VERY EARLY TIME POINT, POST TOXOPLASMA INFECTION ABOUT 20% OF MP CELLS PRODUCE GAM TO EXAMINE IF THIS GAMMA PRODUCTION ARE REQUIRES ANTIGEN RECOGNITION OR NOT, I TREATED THESE ANIMALS WITH ANTI-AND DID THE SAME EXPERIMENT AND SURPRISINGLY, THIS EARLY GAMMA PRODUCTION WAS NOT BROKE BY TREATMENT WHY DON'T DAY SEVEN THE GAMMA PRODUCTION WAS A SIGNIFICANT REDUCED. AND TO EXAMINE IF THIS GAMMA PRODUCTION REQUIRES IL12 AND NEXT I TREATED THE MICE WITH ANTI-IL12 AND DID THE SAME EXPERIMENT AND AS YOU CAN SEE, THE POST PRODUCTION WAS ALMOST COMPLETELY GONE BY THE IL12 SO THIS SUGGESTS THAT THEY CAN RESPOND TO IL12 AND PRO IN GAMMA EVEN IN THE ABSENCE OF THE ANTIGEN RECOGNITION. AND TO EXAMINE IF THIS GAMMA PRODUCTION BY CELLS HOST DEFENSE AGAINST TOXOPLASMA, I WANTED TO RECONSTITUTE THAT THEY WERE KNOCK OUT MICE WITH MP CELLS AND I TRANSFERRED CD4T CELLS INTO THE DOUBLE KNOCK OUT MICE AND WAITED FOR FOUR WEEKS SO ALL OF THE CD4 CELLS HAVE CD4 MEMBER ROW PHENOTYPE AND AFTER CONFIRMING THIS, THIS IS DRIVEN BY HOMEOSTATIC AND AFTER CONFIRMING THIS, NOW I INFECTED THESE ANIMALS WITH TOXO AND SURVIVAL AND THE DOUBLE KNOCK OUT MICE ARE SUN S SUSCEPTIBLE D IT'S SHOWING THE GREEN LIGHT AND PROLONG SURVIVAL AND WHEN THESE CELLS OBTAINED FROM KNOCK OUT AND RECEPTORS KNOCK OUT MICE THAT THESE SURVIVAL PROLONGATION EFFECT WAS BROKE AND SO THIS SUGGESTS THAT THE CELLS CAN PROLONG SURVIVAL IN GAMMA AND IL12 DEPENDENT MODEL AND I ALSO BROKE THE ANTIGEN RECOGNITION BY TREATMENT AND DID THE SAME EXPERIMENT AS CAN YOU SEE HERE, REGARDLESS OF THE ANTI-TREATMENT AND AND THE FOUR DAYS AND THE SURVIVAL PROLONG WAS ONLY THREE TO FOUR DAYS AND WE THOUGHT THIS MAY BE BECAUSE THE IL12 MAY BE DECREASED IN LAG GAMMA KNOCK OUT MICE AND ACTUALLY THIS WAS THE CASE AND WHEN I TREATED NISSAN MALLS AND DID THE SAME EXPERIMENT AND THE MP CELLS PROLONG SURVIVAL AND SO THIS MEANS THAT AT LEAST SOME OF MP CELLS RESPOND TO IL12 AND PRODUCE GAMMA CONTRIBUTING TO HOST DEFENSE IN THIS AND SO NOW, THE MP CELLS HAVE FUNCTIONED AND THEY CAN PRODUCE GAM IN THIS AND EVEN IN THE ANTIGEN RECOGNITION. AND SO NOW, LET'S MOVE INTO THE SECOND QUESTION, WHAT IS A MECHANISM OF THE DIFFERENTIATION? TO EXAMINE THE DIFFERENT STATE US OF CELLS WE ANALYZE THESE CELLS AND AS YOU CAN SEE HERE, THEY CONSISTED OF A DIFFERENCE OF CLUSTERS INCLUDING TH1 OR INDIFFERENTIATED ONES AND INCONSISTENT WHEN I USE THE TRIPLE MICE, AS YOU CAN SEE HERE ABOUT 50% OF CELLS EXPRESS WHILE VERY FEW PERCENT OF THE MP CELLS ARE POSTED AND ALSO INCONSISTENT WHEN WE LOOKED AT THE PROTEIN SPECIAL LEVELS OF T-BET AND P CELLS CONSISTED OF THROW DIFFERENCE POPULATIONS, THEY POSTED CELLS AND T-BET LOW POST CELLS AND T-BEST NEGATIVE SUCCESSFULLY NEGATIVE CELLS AND TO EXAMINE IF THESE T-BET HIGH CELLS HAVE FUNCTION NICE I GENERATED THE T-CELL TO DELETE TCL FOR MP CELLS AND CLOSE DEFINE WITH INTERFERON GAMMA TO REPORT THE MICE TO DETECT THESE AND FIRST I TREATED THESE ANIMALS WITH TAM OXY FEN AND AND THEY CONSISTED OF NEGATIVE AND LOW IN T CELLS, AFTER CONFIRMING H. NOW I INFECTED THESE ANIMALS WITH TOX AND THIS T-CELL AND NEGATIVE MP CELLS AS YOU CAN SEE HERE IN THIS RED LINE, HIGH MP CELLS ARE PROD THE HIGHEST LEVELS OF GAMMA EVEN IN THE ABSENCE OF THESE SIGNALING AND SO SUGGESTING THAT THE HIGH CELLS ARE THE MAJOR POPULATION AND POSSESSING INNATE FUNCTION AND ACTUALLY WHEN I TRANSFERRED HIGH AND LOW AND NEGATIVE MP CELLS INDIVIDUALLY INTO THE GAMMA THAT WERE KNOCK OUT NICE AND I DID THE SAME TOXOPLASMA SURVIVAL EXPERIMENT AS YOU CAN SEE HERE, THE HIGH M.P. CELLS AGAIN INSIGNIFICANT BUT AROUND SURVIVAL. AND HOW ARE THESE T-BET HIGH M.P. CELLS DIFFERENTIATED. TO ASK THIS QUESTION, NEXT, I CHECKED THE T-BET EXPRESSION AND T-BET HIGH CELLS AND IL-12P40 AND KNOCK OUT NICE AND FOUND THAT THE T-BET HIGH M.P. CELLS ARE SIGNIFICANT TO REDUCE IN THESE KNOCK OUT MICE SUGGESTING THE IL-12P740 PLAYS AN ESSENTIAL ROLE FOR THE ENHANCEMENT OF MP1 DIFFERENTIATION. AND THEN WHICH TYPES OF CELLS IN STEADY STATE AND WE EXAMINED DIFFERENT TYPES OF THIS SUBSET AND FOUND THAT CD8 ON DC1 BUT THERE'S NO SUBSET AND EXPRESSED IL12 AND ACTUALLY, WHEN WE LOOKED AT THE KNOCK OUT MICE WE SPECIFICALLY INDEBTED THE SUBSET THE HIGH MP CELLS WERE SHOWING THAT 50% REDUCTION. AND ALSO, WE TAKE THE ID12 SPECIFICALLY IN THE T-CELL ZONES OF THE SPLEEN BY IMAGING A YOU CAN SEE HERE. SO, NOW IT'S GREAT THAT MP CELLS CONSIST OF DIFFERENCE OF POPULATIONS INCLUDING THE HIGH AND LOW AND NEGATIVE CELLS AND HIGH CELLS HAVE THE INNATE DETECTOR FUNCTION AND THEY CAN PRODUCE GAMMA IN THIS AND HIGH LEVELS IN GAM OR I'M SORRY, POSITIVE IL12 AND CONTRIBUTE TO HOST PROTECTION AND THE NEGATIVE MP CELLS IS CREATE AND WE HAVE THIS ISSUE AND ALTHOUGH I DIDN'T HAVE TIME TO TALK ABOUT THIS BUT WE ALSO FOUND THAT THE T BET HIGH MP CELLS AND THEY WERE T BET NEGATIVE MP CELLS DO NOT AGAIN RIGHT THE HIGH MP CELLS SO WE THINK THAT THE HOMEOSTASIS IS MAINTAINED IN THIS MODEL AND LIKE I SAID, GIVEN THE HIGH MP CELLS IS PROMOTED BY IL12 WHICH IS PRODUCED BY THESE SUBSET AND THIS HOMEOSTATIC IL12 PROCESS BY DC1 IS BASICALLY PRIMED BY SOME SELF-AGO KNOW NIST AND THIS IS ACHIEVED BY THE MULTIPLE TRL AND MODEL SIGNALING DEPENDING MODEL AND OK, SO, THE FINAL QUESTION, SO IT WAS REALLY DIFFERENT FROM 40 ANTIGEN AND SO LIKE SAID IN THE BEGINNING OF THIS PRESENTATION, AT LEAST SPECIFIC OR MOST FOR ALL OF THE 40 ANTIGENS SPECIFIC MEMORY CELLS AS THEY ARE CD44 HIGH AND CD62 LOW SO THIS PHENOTYPE IS THE SAME AS A SELF-DRIVEN M.P. CELLS SO UNTIL RECENTLY WE DIDN'T HAVE EVIDENCE THESE DO CD44 ARE DIFFERENT SO OUR RECENT DATA SUGGESTED THAT THEY ARE DIFFERENT AND NOW I AM GOING TO TALK ABOUT THIS ISSUE. SO TO ADDRESS THIS QUESTION TO COMPARE THAT M.P. VERSUS MEMBER RE CELLS NEXT WE ISOLATED SPF M.P. CELLS AND SPECIFIC MEMBER ROW CELLS AND ANTIGEN SPECIFIC MEMORY CELLS AND I HAVE THESE AND COMPARED BY SINGLE CELL AND YOU CAN SEE HERE, THIS CD ARE DIFFERENT IN SOME GENES AND CAREFULLY, LOOKING AT THESE TWO POPULATIONS THE THROW MARKERS, ARE DIFFERENTIATABLE EXPRESSED BY THESE TWO CD40 CELL POPULATION AND THE MEMBER ROW CELLS ARE MARKERS ARE ALL HIGH AND THE M.P. CELLS HAD LOWER LEVELS OF THESE THROW MARKERS AND BY COMBINING THESE THREE MARKERS, ALL TOGETHER, WE FOUND THAT THE 40 ANTIGEN SPECIFIC MEMORY CELLS AND M.P. CELLS ARE PHENOTYPICALLY DIFFERENT IN TERMS OF THESE THROW MARKERS AND ALSO IN THE STUDY, WE FOUND THAT M.P. CELLS THEMSELVES CAN BE DIVIDED INTO FOUR DIFFERENT SUB POPULATIONS USING CD1-27 AND WE FURTHER ADDRESSED THE FUNCTION OF THE FOUR M.P. SUBSETS. AND SO TO ADDRESS THIS QUESTION, I ROW HONORRIZED THE DATA OF M.P. CELLS SINGLE CELL AND SEQUENCE AND DATA AND WE DID THE ANALYSIS OF M.P. CELLS AND AS YOU CAN SEE HERE, AND ALSO IN BIAS MANNER AND M.P. CELLS CAN BE DIVIDED INTO FOUR DIFFERENT SUB POPULATIONS AND THE 1 SHOWING IN BLUE IS IL7 AND THE 2 IS DOUBLE HIGH AND 3 IS LOW AND HIGH AND THE 4 IS DOUBLE LOW SO IT'S REASONABLE TO THINK THAT THESE FOUR CRUSTERS RESPOND TO THE MP SUB POPULATIONS DEFINED BY CD127 AND FLOW SIGH TOM TREE AND MORE INTERESTING HERE, IS THE TWO CELLS WHICH ARE CD1 HIGH CELLS HAVE THE HIGHEST LEVELS OF TH1 ASSOCIATED GENES INCLUDING INTER FAN GAMMA AND CXR3 SO THEY HAVE THE MAJOR M.P. SUB POPULATION WITH EFFECT FUNCTION AND TO TEST THIS HYPOTHESIS, I THOUGHT IT FOR THESE FOUR SUB POPULATIONS AND STIMULATED THE CELLS AND OBVIOUS IL12 AND IL18 AND IN VITRO AND I DIDN'T STIMULATE THE T-CELL STIMULATION SO THIS CAN BE GOOD EVIDENCE THAT MP CELLS CAN WORK ON ALSO INVOTE ROW AND AS CAN YOU SEE HERE, THE CRUST 2 CELLS HAD THE HIGHEST LEVELS OF ININTEREST FUR ON GAMMA SHOWING IT'S BASICALLY TRUE. AND AT THE SAME TIME, THIS SUGGESTS THAT THESE CRUST 2 CELLS HAVE HIGH RESPONSIVENESS TO TH1 DIFFERENTIATING CYTOKINES SUGGESTING THEY MAY BE THE CLOSE FOR SOME INFLAMMATORY DISEASE AND TO TEST THIS HYPOTHESIS I THOUGHT THAT AGAIN FOR THESE FOUR SUB POPULATION AND TRANSFERRED INTO KNOCK OUT MICE TO INDUCE AND AS YOU CAN SHEEHY, THIS RED LINE ON THE CD1 27, THE CRUST 2 CELLS INDUCED IT AND THIS THE SCORE AND THIS IS A TOTAL NUMBER AND ON THE DATA SUGGESTS AND AND THE MOST SEVERE COR AND TRANSFER AND ALL TOGETHER, THIS IS A SUMMARY AND PROSPECTS AND I RESEARCH AND CD4 CELLS HAVE A AND IT'S AND NOW IT'S CLEAR THAT AND CELLS AND SELF-ANTIGEN AND I DIDN'T HAVE TIME TO TALK ABOUT THIS AND AND THEY STARTED DYING AND AND M.P. CELLS IS MAINTAINED AND BY THE SIGH IT HAS SEVERAL AND VI AND AND DEFENSE AND INFECTION AND WE ARE NOW GETTING EVIDENCE THAT AND ANTI-ACTIVITY AND WE ARE CURRENTLY ADDRESSING THIS ISSUE IN MY LAB AND ON THE OTHER HAND. AND IN THE LAB TRANSFER SYSTEM M.P. CELLS CAN INDUCE INFLAMMATORY DISEASE AND THIS IS REASONABLE AND M.P. CELLS SHOULD HAVE HIGHER SELF-ANTIGENS AND WE ARE ALSO ADDRESSING THIS ISSUE AND OF COURSE, M.P. CELLS DO NOT INDUCE DISEASE IN HEALTHY, NORMAL ANIMALS AND WE THINK THAT THIS IS BECAUSE OF THE PRESENCE OF REGULATORY T-CELLS AND AND IT PLAYS AN EX TENIAL ROLES OF THE HOMEOSTASIS OF M.P. CELLS SO WITH THAT I WILL RECOGNIZE THESE PEOPLE LISTED HERE AND FINALLY, THANK YOU VERY MUCH FOR EVERYTHING. IF YOU DIDN'T TAKE ME TO YOUR LAB, SEVEN YEARS AGO, WHEN WE WERE PASSED AWAY, MAY I WOULDN'T BE DOING SCIENCE NOW AND I WILL NEVER FORGET THAT WHEN WE HAD OUR FIRST PAPER IN SCIENCE TECHNOLOGY YOU SPENT ALL OF YOUR TIME WRITING WITH ME AND AT THE SAME TIME YOU DECLINED TO BE THE LAST AUTHOR AND INSISTED YOU COULD BE THE LAST. THANK YOU, VERY MUCH. [APPLAUSE] >> SO, IN THE FIRST PLACE, RIGHT, IN ORDER TO GET T BET YOU HAVE TO HAVE THE BETA CHAIN OF THE IL12 RECEPTORS OF REGULATED AND DIFFERENTIALLY IN THAT SUBSET. SO, DO YOU THINK THAT THERE'S A DISTINCT REP A TOUR OF SELF-ANTIGENS THAT THE T BET POSITIVE CELLS RECOGNIZE? THAT THEN DRIVES BETA TO RECEPTORS EXPRESSION? >> YOU ARE TALKING ABOUT THE T-CELL REPERTOIRE FOR THE DIFFERENTIATION? YEAH, SO WE THOUGHT SO AND IT'S A GOOD MARKER SO I ANALYZED THE RELATIONSHIP BETWEEN CD5 AND T BET BUT I DIDN'T FIND A GOOD CORRELATION SO AT THE MOMENT, I DON'T KNOW AND I THINK THIS IS STILL A T-CELL REPERTOIRE ISSUE AND I'M ANALYZING THE T-CELL REPERTOIRE IN LOW AND HIGH AND NEGATIVE CELLS. >> MANY YEARS AGO WE WERE DOING ADOPTIVE TRANSFER AND WE NOTICED THAT IL12 RECEPTOR AND TICK 2 KNOCK OUT T CELLS EXPANDED MUCH MORE THAN WILD TYPE T CELLS AND SO THEN IT'S PROBABLY SKEWING TOWARDS A NON T BED POSITIVE CELLS. >> THANK YOU SO MUCH, YEAH. >> SO, I WAS INTERESTED IN YOUR LAST POINT ABOUT WHETHER THESE M.P. CELLS ARE DIFFERENT FROM THE INFECTIOUS ANTIGENS STIMULATER CRIMINSTIMULATE CELLD THE EVIDENCE OF LVMV BUT THE LCMV WOULD HAVE RISEN IN STRONG ENVIRONMENT AND SO IT'S NOT AT ALL SURPRISING THE T CELLS WOULD BE DIFFERENT THAN AND ENVIRONMENT AND SO I'M STILL WONDERING THAT THE ALTERNATIVE POSSIBILITY AND CELLS YOU LOCK AT AND A LOT OF RESPONSES LIKE ENVIRONMENT ALAN TEE AGAINS FROM FOOD AND SELF-ANTIGEN RESPONSES AS WELL SO, CAN YOU DISTINGUISHED THAT KIND OF POPULATION FROM SOMETHING A LITTLE DIFFERENT WHICH I THINK IS WHAT YOU ARE SUGGESTING YOU ARE M.P. CELLS WERE? >> THAT'S A VERY INTERESTING POINT AND IT'S EXACTLY WHAT I'M DOING NOW BUT ONE THING IS THAT YEAH, SO, WE HAD MANY CANDIDATES AS A DIFFERENTIATE M.P. VERSUS MEMORY CELLS BUT AT THE MOMENT, THESE MARKERS WEREN'T THE ONLY ONES THAT CAN DISTINGUISHED IT BUT IT'S POSSIBLE THAT MEMORY CELLS ARE DRIVEN BY STRONG AND TRANSIENT ANTIGEN RECOGNITION AND I THINK THAT THIS IS WHY AS THEY EXPRESSED THE HIGH LEVELS OF CD127 AND ON THE OTHER HAND, YOU ARE RIGHT, THE M.P. CELLS ARE MAYBE CONTINUOUSLY AIRATED BY THE RECOGNITION OF CELL ANTIGENS AND YEAH, SO I THINK THAT MAYBE THAT IS THE REASON WHY THEY HAVE DIFFERENT LEVELS OF BCL1 AND CD127 AND WE THOUGHT SOME OF THESE CELLS MAY BE SPECIFIC FOR SOME COMMENCE OF ANTIGENS AND I ANALYZED THE SAME THING -- THE SAME MARKERS IN SPF AND ANTIGEN FREE MICE WHICH BOTH COMMENCE AND FOOD ANTIGENS BUT THERE ARE NO DIFFERENCE. SO M.P. CELLS STILL CONSIST OF FOUR DIP SUB APPROPRIATIONS AND EVERYTHING IS EASIER TO LOAD SO I THINK THAT SOME -- STILL, IT DOESN'T RULE OUT THE POSSIBILITY THAT'S THE ANTIGEN CONTRIBUTE TO THE GENERATIONS OF LOW AND M.P. LIKE CELLS BUT BASICALLY I THINK AT THE CONCEPT YOU ARE TALKING IS CONSISTENT WITH MY HYPOTHESIS SO I THINK THAT M.P. CELLS ARE CONTINUOUSLY DIFFERENTIATED BY RECOGNITION OF SOME WEAK SELF-OR POSSIBLY OTHER ENVIRONMENT ANTI-AND DIFFERENT MEMORY CELLS AND. >> IT WOULD BE VERY USEFUL FOR THAT. >> HELLO. >> AND YOU SPOKE OF IL18 SYNERGIES TO DRIVE THE GAMMA IN VITRO IN YOUR TOXO SYSTEM WITH THE M.P. WERE YOU BLOCKED WITH ANTI-IL12, HAVE YOU TRIED TO BLOCK WITH ANTI-IL18 AND BEFORE YOU ANSWER THAT I'LL JUST, THAT'S SOME WORK WE DID YEARS AGO THAT RESTING TH1 CELLS COULD RESPOND TO BOTH AND ANTIGEN AND KEN MURPHY FOLLOWED WHICH THAT PATHWAY OR NOT WHERE THE ANTIGEN DEPEND OR NOT AND THE OTHER MOL AMOLECULEWITH SOME VERY OLD WORE HAVE WITH GORGE YO WHO IS OVER THERE AND WHERE WE HAD SHOWN THAT B7, WHICH WOULD BE ON THE DC ALSO, COULD SYNERGIES AND L12 TO ACTIVATE AND INDEPENDENTLY OF ANTIGEN AND AND AT THAT TIME IT'S PEOPLE THAT DIDN'T LIKE THIS AND THEY REJECTED OUR PAPERS AND EVENTUALLY THEY GOT IT AND THEN PEOPLE HAVE BEEN TALKING ABOUT THESE BYSTANDER RESPONSES THAT ALWAYS HAPPEN IN VIRAL INFECTION SO YOUR M.P. CELLS SOUND LIKE THEY'RE ANSWERING IT ALL SELECTED WITH LOW AFFINITY ANTIGENS, NOT DOING MUCH AND YOU GET INFLAMMATION BUT THE QUESTION IS, IT COULD BE SEVEN AND L18 BE INVOLVED WHEN YOU ARE IN THE INFLAMMATORY SCENARIO. >> YEAH, WE TESTED INDIVIDUALLY STIM YOU LATE IL12, IL18 AND IL2 AND AS REPORTED AND IL18 ALONE CAN BE GENERATED GAMMA RESPONSE BUT WHEN SYNERGIESED WITH IL12, IT INDUCES THE HIGHER RESPONSE AND THIS IS TRUE IN VITRO BUT THIS WAS ALSO IN VIVO AND -- >> DO YOU BLOCK INVIVO? >> WHEN WE BLOCKED IL18, I DIDN'T SEE ANY DIFFERENCE IN PRODUCTION BUT WHEN WE STIMULATE IL12 AND 18 TOGETHER, AND M.P. CELLS PRODUCE HIGH LEVELS OF GAMMA AND FOR THE B7 PARTS YES, SO LIKE SAID, M.P. CELLS PROLIFERATING AND ACTUALLY ABOUT 30 TO 40% POSITIVE BUT IN STEADY STATE AND WHEN WE BLOCK THE B7 MOLECULES USING THE CTA THIS IS POSITIVE PRODUCTION IS SIGNIFICANT REDUCING SO THIS SUGGESTS THAT THE PROLIFERATION OF M.P. CELLS, IT'S DYNAMICALLY MAINTAINING BY THE ABOUT 7 MOLECULES SO YOUR POINT IS IMPORTANT SO I THINK IT'S POSSIBLE THAT IN THE PRESENCE OF SUFFICIENT LEVELS OF B7 SIGNALING, OF M.P. CELLS CAN PRODUCE MORE GAM WHEN STIMULATED WITH IL12. >> THAT'S FANTASTIC. THOSE STUDIES, WHAT WAS IT, MID 90 KMID'90s SO THEY'RE VERY OLD. >> IT'S GREAT. >> THANK YOU, VERY MUCH. >> THANK YOU. Dr. TAKESHI KAWABE. OUR NEXT SPEAKER IS GEORGE YAP FROM RUTGERS NEW JERSEY MEDICAL SCHOOL. IT WAS PART OF THE TOXO GROUP IN ALAN'S LAB AND HE WAS THE KING OF THE BONE MARROW TRANSFERS AND HE DID A LOT OF WORK ON IL12 AND TNF AND SO ON AND SO FOURTH. >> THANK YOU. >> THANK YOU, RICARDO. YES, SO I'M REALLY HAPPY TO BE HERE TODAY AND FIRST AND FOREMOST I WANT TO THANK ALAN FOR ACCEPTING ME AND ACCOMMODATING ME INTO HIS GROUP AND I STARTED ACTUALLY IN ALAN'S LAB AND TRANSFERRED TO ALAN'S LAB AND I STUDIED MOSTLY CHRONIC INFECTION IN I SAID, I REALIZED THAT IT WAS GOING TO BE HARD FOR ME TO MAKE A CAREER STUDYING HYSTE SO I SHIFTED TO STUDY TOXOPLASMA SIS TOGETHER WITH TANYA AND AT THAT TIME AND SO THIS IS JUST A PICTURE OF THOSE YEARS. WE HAD A LOT OF FUN. TANYA IS THERE AND MARK AND URG. I STILL HAD A LOT OF HAIR AT THE TIME. WE HAD AN ENTIRE MORNING DISCUSSING IMMUNE RESISTANCE, RIGHT, AND IMMUNE RESISTANCE WE KNOW IS IMPORTANT FOR PROTECTION AND WE'VE ALWAYS BEEN IN THIS MIND SET THAT THE PATHOGEN IS TRYING TO OUTSMART THE HOST IMMUNE RESPONSE BY PRODUCING VIRULENCE FACTOR THAT AN TAG A NICE THE IMMUNE RESPONSE FACTORS AND WE NEED TURN OUR VIEW DIFFERENTLY. IN THE END, WHAT NEEDS TO HAPPEN IS THE PATHOGEN AND THE HOST NEED TO MUTUAL LEISURE SERVE LY SURVIVE THISENCOUNTER AND THO REGAIN HOMEOSTASIS AND PERHAPS DIFFERENT SEC POINT AND TODAY WE GO BACK TO ARGUE IS THE AND STATE OF A VIRULENCE IS NOT A PASSIVE LACK OF FUNCTION BUT A ACTIVE FUNCTION OF THE PATH OWE AGAIN AND CAN HOST DISEASE AND MECHANISMS THAT WILL LEAD TO PROTECTION AND WE HAD THE BENEFIT OF THIS VIEW BECAUSE I STUDIED BOTH SISHOZOMES AND THE REASON THEY ARE SUCCESSFUL IS BECAUSE THEY BOTH DON'T KILL THEIR HOSTS, RIGHT. AND SO IN FACT, WHAT WE LEARNED AFTER, WELL BEFORE I LEFT THE LAB IT WAS ALSO VERY APPARENT THAT TOXOPLASMA IS A WONDERFUL EXAMPLE OF THE HOST RESPONSE FRAMEWORK PARABOLA AND SHOWING THAT IN FACT, THERE'S THIS OPTIMAL POST-RESPONSE AND BEYOND THAT A HIGHER IMMUNE RESPONSES ACTUALLY POST DETRIMENTAL. IN THE PRESENCE OF PATHOGENS THAT HAVE A OHIO POTENTIAL, YOU NEED TH1 RESPONSES IN ORDER TO RESTRAIN OR INHIBIT THIS INTRINSIC SIGINFORINS -- THE FIN RICARDO SHOWED THAT IL10 AND GAMMA WERE CRITICAL FOR THE SURVIVAL OF THE HOST FOR DIFFERENT REASONS. SO, WHAT IS INTERESTING IS THAT YOU KNOW, THERE IS INHERENT VARIABILITY ALSO IN THE PATHOGENS AND SO YOU HAVE DIFFERENT STRAINS OF A VIRULENT AND VIRULENT PARASITES AND ALL THOSE RULES WE LEARNED APPLIES ONLY TO THE A VIRULENT PARASITES BECAUSE THE HYPER VIRULENT TYPE 1 STRAIN RH WHICH WAS ACTUALLY ISOLATED BY 80 YEARS AGO, ACTUALLY DO NOT FOLLOW THOSE RULES AND THEY -- ONE INFECTIOUS PART KILL OF THE TYPE 1 PARASITE KILLS A PERFECTLY IMMUNO COMPETENT MOUSE SO WHAT'S WRONG WITH THAT? AND SO WHAT WE LEARNED SHORTLY AFTER LEADING THE LAB, WHAT WE LEARNED IS THAT THE TYPE 1 PARASITE ACTUALLY INTERFERES, USES VIRULENCE FACTORS WHICH ARE THOSE ROPE TREE PROTEINS THAT THE PARASITES SECRETS INTO THE MEMBRANE AND INTERACTS WITH HOST GTP ACE AND DISAB DISABLE THEM T WAS DISCOVERED IN THE LAB AND WT HAPPENS IS THE MOUSE GETS KILLED EVEN THOUGH THEY HAVE A PERFECTLY GOOD IMMUNE RESPONSE AND WORK FROM JOHNATHAN HOWARD'S LAB SHOWED THAT WE HAVE A VERY SHORT, NARROW SAM BELLING OF THE MOUSE GENOME AND IN WILD MICE THERE ARE ACTUALLY DIFFERENT TTPAs THAT COUNTERACT THE ACTIONS OF THESE ROPE TREE KINASES THAT PHOSPHORYLATED THE GDP AND YOU COULD NOW VIEW VIRULENCE AS A MISMATCH BETWEEN THE IMMUNE RESPONSE GENES AND THE VIRULENCE FACTOR REPERTOIRES OF THE DIFFERENT STRAINS OF MICE VERSUS PARASITES. SO IT BRINGS US BACK TO THE QUESTION OF MY TOPIC OF MY TALK TODAY, AND I CHOSE A TITLE THAT IS ABOUT CURIOSITY AND I THINK AGAIN I WANT TO THANK ALAN BECAUSE HE ACTUALLY GAVE ME A LOT OF FREEDOM EVEN WHEN I WAS IN HIS LAB TO PURSUE CURIOSITY-DRIVEN RESEARCH AND SO ONE OF THESE STORIES THAT WE'VE BEEN STUDYING A LONG TIME FOR A LONG TIME IS THIS FINDING THAT, YES, THE MOLECULAR BIOLOGIST, THE CELL BIOLOGIST STUDY THE VIRULENT STRAIN AND IMMUNOLOGISTS ALWAYS STUDY THE A VIRULENCE STRAIN BECAUSE THEY DON'T WANT THEIR MICE TO BE KILLED AND WHAT WE FOUND IS THAT ACTUALLY UNLIKE THE VIRULENCE STRAINS WHICH INRATE IS CELLS BY ACTIVE PENETRATION, THE STRAINS GO UP THE EVENT IN MACROPHAGES AND THEY INITIALLY ENTER A ENVIRONMENT AND THEN INITIATES THIS DIRECT INVASION PATHWAY WITHIN THE FAGOZOMW WHERE ONLY A VIRULENT DO IN THE CELLS SO THE AND WHAT SELECTIVELY ENGAGES AVIRULENT AND IT'S NOT THERE IN THE VIRULENT STREAMS AND I SAID OH, WHAT WOULD PEOPLE AT THE LPD DO? WELL, JUST THROW OUT ALL THE CELL LINES YOU HAVE IN THE FREEZER AND SEE WHICH ONES STICKS AND WHICH DOES NOT. AND RAW CELLS, THEY PHAGOCITE AND J774 CELLS DO NOT KNOW THIS PHONEFEENOPE TYPE SO WE SEE WHAS MISSING THAT THE RAW CELL HAS IN ABUNDANCE AND IT TURNS OUT TO BE CD36 WHICH WAS HIGHLY ABUNDANT IN THE RAW CELLS BUT ABSENT IN THE J774 CELLS. AND SO, WHEN WE LOOKED AT BONE MARROW FROM CD36 KNOCK OUT MICE, VERY QUICKLY, WE WERE ABLE TO CONFIRM IT WAS IN DEED CD37 THAT MEDIATES THIS PHAGOSOME AND GOING BACK TO MY NIH DAYS, I QUICKLY CALLED JOE SMITH, WHO WAS POSTDOC IN LOU MILLER'S LAB AND JOE SENT ME CHINESE HAMSTER OVARY CELLS THAT OVER PRESS CD36 AND IT'S SUFFICIENT TO CONFER ADHESIVENESS OF STRAINS TO THE CHINESE HAMSTER OVARY CELLS AND THIS CAN BE NEUTRALIZED BY SOME BUT NOT ALL ANTI-CD36 ANTIBODIES. SO, IT LOOKS LIKE THERE IS ANNAL CONTINUE TIVE PATHWAY THAT THEY USE TO ENGAGE WITH HOST CELLS AND VERY IMPORTANTLY, MONO SIGHT MACROPHAGES. ANTHE BONE MARROWMONOCYTE DERIVL MACROPHAGES AND WE SEE THAT THE SELECTIVE LOSS OF THE KNOCK OUT MICE OCCURS IN THE BONE MARROW DERIVED MONOCYTE DERIVED MACROPHAGES BUT NOT IN THE LARGE MACROPHAGES SO WE THINK THAT THIS IS THE INITIAL EVENT THAT CHANGES THE COURSE OF INFECTION AND WHAT HAPPENS IS THAT WE ACTUALLY DO NOT SEE ANY IMMUNODEFICIENCY IF ANYTHING, THERE IS AND THIS IS NOT ALWAYS CONSISTENT BUT THERE'S A GENERAL TREND FOR HIGHER INNATE AND ADAPTIVE IMMUNE RESPONSES SO WE'RE SHOWING THE MK CELLS HERE AND OVER ALL WHAT WE SEE IS PRESERVATION OF THE RESISTANCE PATHWAY SO THEY DON'T DIE LIKE GAMMA AND IL1 KNOCK OUT OR MYDA KNOCK OUT MICE AND MOST OF THEM ACTUALLY LIVE A LONG TIME AND THEY DIE OVER A PROTRACTED PERIOD AND THEY PRODUCE PERFECTLY GOOD IL12 GAM KA EVEN IL12 SO WHERE AT A LOSS BECAUSE THIS SEEMS LIKE IT'S I DON'T BEYONDIMMUNOLOGY NOW SO WE DID D PRO TEE OWE TICK ASSAY OF THE MICE AND WE WERE AND THEY WERE ELEVATED IN THE CD36 KNOCK OUT MICE AT THE TIME THEY WERE DYING AND WE REALIZED OH, WOW, THESE ARE REALLY INTERESTING MOLECULES THAT HAVE BEEN ASSOCIATED WITH SEVERITY OF DISEASE IN CANCER AND HART DISEASE AND KIDNEY DISEASE AND EVEN IN COVID. EL RATE ISEDSHOW THE MORTALITY T PAPERS SO WHAT WE SEE IS IN FACT BOTH WILD TYPE AND KNOCK OUT MICE ELEVATE GDF15 AND FG F-2 1 AND IN THE KNOCKOUS MICE THIS IS PROTRACTED AND IN THE WILD TYPE MICE, THIS IS RAPIDLY CONTROLLED. SO WE GO BACK AND JUST GUESS, OK, WHAT IF WE NEUTRALIZE INTRA FUR ON GAMMA IN DAY 12 AND WE WERE SURPRISED BOTH FG F21 AND GDF15 WERE INCREASED BY GIVING ONE DOSE ON DAY 12 AND THIS REVERSED THE WEIGHT LOSS THAT WAS OCCURRING IN THE CD36 KNOCK OUT MICE. SO IT'S FOREIGN TO ME AND IT TURNS OUT TO BE A VERY INTERESTING MEMBER OF THE TGF BETA FAMILY, VERY DISTANT MEMBER OF THE TGF BETA FAMILY AND WHAT IS INTERESTING IS THAT IT ENGAGES RECEPTORS WHICH ARE CALLED GFRAL RECEPTORS IN THE CONTROLLED APPETITE AND SO WE ACTUALLY LOOKED AT THIS EVEN MORE CAREFULLY AND I THINK I'M MISSING -- SO WE WENT BACK AND SAID OK, IN WILD TYPE MICE WHICH HAVE NOW DAY 12 LOW LEVELS OF GD IF, CAN WE INCREASE THAT BY GIVING THEM RECOM BIN ANT INTER FER AN GAMMA AND WE SEE IT'S CLEARED. EVEN THOUGH IT'S A OLD CYTOKINE IT SEEMS LIKE IT CONTROLS THINGS LIKE THE METABOLIC TISSUE RESPONSE CYTOKINES THAT WE HAVE YET TO LEARN MORE ABOUT AND SO THIS IS WORK THAT ONE OF MY GRADUATE STUDENTS IS DOING CURRENTLY AND SO YEAH, YOU CAN STILL EVEN OLD CYTOKINES STILL HAVE NEW TRICKS UP THEIR SLEEVES. AND SO YEAH, SO, ESSENTIALLY, WHAT WE FOUND IS THAT THIS, WE THINK THIS TROPPISM TUNES IN IMMUNE RESPONSE THAT LEADS TO A DIFFERENCE IN THE TISSUE STRESS AND SICKNESS RESPONSE IN THE APP SENSE OF CD36 SO TWO AVIRULENT TRAINS AND VER LINS STRAINS WE SEE NIGHT AND DAY SO IT BINDS TO THE AVIRULENT STRAINS BUT NOT TO THE RH STRAIN. WHEN WE LOOKED AT THE ROPE KNOCK OUT THAT PHOSPHORYLATED THE GTP ACES WE FOUND OH, THEY DO NOW BIND TO CD36 WHICH WAS COMPLETELY UNEXPECTED. AND SO IT SEEMS LIKE -- IT SEEMS LIKE RH STRAINS HAVE THE CAPACITY TO BIND TO CD36 AND THE VIRULENCE FACTOR IS SUPPRESSING THE BINDING OF CD36 TO AN UNKNOWN LIGAMENT SO WE TESTED THAT BY LOOKING AT A SCREEN, A TYPE OF THREE STRAIN AND THEN EXPRESSED EITHER THE WILD TYPE OR THE KINASE DEAD ROPE 18 VERSIONS DERIVED FROM A TYPE 1 STRAIN AND IT'S EVEN THAT KEEN THE KINASE ROPE 18, WHICH IS INACTIVE IN INHIBITING THE IS STILL ABLE TO SUPPRESS BINDING OF CD36 AND SO WE STILL DON'T KNOW EXACTLY WHAT CD36 IS BINDING TO AND BUT ONE OF THE SUSPECTS THAT WE HAVE IS THIS MOLECULE WHICH IS A GPI ANCHOR PROTEIN CALLED SRC29C IS WE THINK THAT BECAUSE WE DID A PROTEOMIC SCREEN AND WE DID FIND 29C BINDING TO A PULL DOWN OF CD-36 AND ALSO WORKED FROM MIKE GREGRIG'S LAB SHOWED THE TOPIC F SRS29C CAN MAKE A ENTEROVIRUS D68 LENS STRAIN AND WE'VE GONE BACK AND LOOKED AT THE ROPE 18 KNOCK OUT AND YOU CAN SEE THAT WHEN WE PURIFY ALL THE GPI ANCHORED PROTEINS, AND OF THE ENTEROVIRUS D68 LENS FACTOR SO OUR CURRENT WORKING MODEL, WE THINK THERE'S AN ACTIVE SYSTEM THAT PO MOTEL PATHWAYS IN THE HOST AND CRITICALLY THESE ARE BOTH INTERACTING WITH AVIRULENCE FACTORS BUT IN DIFFERENT WAYS. SO, THE EFFECT ON THE HOST IS KINASE DEPENDENT AND THE EFFECT WITHIN THE PARASITE IS COMPLETELY KINASE INDEPENDENT. AND SO I'LL END HERE BY SHOWING NOW THE PICTURES FROM THE LAB AND BEFORE COVID AND ALAN ACTUALLY CAME TO VISIT FOR A TBRU PROGRAM AT RUTGERS AND HE DEVOTED I THINK A MORNING AND WE HAD A LAB MEETING WITH ALAN SO MY STUDENTS PRESENTED TO ALLEN AND WE WANT OUT FOR A NICE LUNCH IN NEWARK AND THESE ARE SOME MORE PICTURES OF MY CURRENT STUDENTS WHO SOME OF THEM HAVE JUST GRADUATED AND WE'RE HAVING SOME CELEBRATIONS. THANK YOU. [APPLAUSE] >> I HAVE A QUESTION. >> I'M VERY INTERESTING THE CD36 STORY AND LOOKING AT THE PHENOTYPE OF THE MOUSE IN VIVO IT SEEMS THAT MAYBE ACTIVE INVASION DO YOU HAVE AN IDEA WHAT CD36 IS INVOLVED IN PASSIVE OR ACTIVE UPTAKE OF THE PARAS PARASITE? >> YOU NEED CD36 FOR THE INVASION PATHWAY SO WE LOSE IT IN TROPISM IN THE ABSENCE OF CD36. THERE'S STILL A LOT OF THINGS THAT WE NEED -- YOU KNOW, THERE ARE HUGE GAPING GAPS AND SO WE NEED FUNDING FROM THE NIH. [LAUGHTER] AND LEE IS HERE TODAY. >> THE QUESTION IS, BECAUSE CD36 IS A LIPID RECEPTORS, RIGHT? AND WHY DO YOU THINK IT'S SELECTIVE FOR ONE OF THE PROTEINS? >> I THINK -- WHAT IS INTERESTING ABOUT IT IS IT'S REALLY INTERESTING BECAUSE FOR ME, WHEN I FOUND CD36, I WAS LIKE OH MY GOD, NO. YOU CAN IMAGINE ALAN SHAKING HIS HEAD NO. FOR ME IT'S A D A GENTLEMAN VIEA AND HOW THAT TURNED OUT I WAS LIKE WAIT I DON'T WANT TO REPEAT THAT STORY. NO, BUT RICARDO, I THINK CD36 IS SUCH A MULTI FUNCTIONAL MOLECULE THAT INTERACTS WITH A MYELOID AND A TLR2 OR TL6 AND IT CAN SENSE LIPIDS AND IT'S A CHANNEL ACTUALLY FOR LIPIDS. THAT'S WHY YOU FEEL HAPPY EATING BUTTER AND FAT FRENCH CHEESE AND IT'S ALSO ACTUALLY ALSO, FOR EXAMPLE, INVOLVED IN ESTABLISHMT OF TUMOR MICRO ENVIRONMENT SO THERE WAS A SLEW OF PAPERS THESE PAST TWO YEARS SHOWING THAT CD36 ACTUALLY KILLS, RIGHT. IT PROMOTES MAYBE M2 MACROPHAGES BUT CELLS THE CD8 THEMSELVES THROUGH THERIP TOE SIS AND OTHER MECHANISMS. >> HELLO, GEORGE, THAT WAS FANTASTIC. REALLY FASCINATING THIS VIRULENT VERSUS AVIRULENT AND HOW THEY BIND OR DON'T BIND TO CD36 AND IMMEDIATELY WHAT CAME TO MY MIND, BECAUSE, I USED TO BE SURROUNDED BY MALARIA PEOPLE, EVEN INTEREST DNAC WHEN RUSSELL HOWARD WAS THERE, AND IT MADE ME THINK, CD36 AND MALARIA AND MAYBE JEAN CAN COMMENT WHETHER THERE ARE ANY PLASMA STRAINS THAT DIFFER ACCORDING TO THIS BINDING OR DIFFERENTIAL EFFECTS ON CD36, DO WE KNOW? >> I DIDN'T WANT TO TREAD IN THE MALARIA ZONE. HAVING BEEN A MALARIA PERSON MYSELF AND NO, WE ACTUALLY TRIED TO SEE BECAUSE ALL OF THESE SRS, ALL OF THESE ANCHOR PROTEINS ARE ALSO EXPRESSED IN JASPER A AND SO HOW -- I COULDN'T SLEEP JUST THINKING ABOUT THAT BECAUSE YOU KNOW, ALL THESE GPI ANCHOR PROTEINS AND THEN YOU CAN IMAGINE, RIGHT, LIKE, IN TOXOPLASMA IT'S A MOUSE TO CAT PREDATION PATH WAY AND IT'S SHEEP TO FOX PRETATION PATHWAY BUT WE DIDN'T SEE A LOT OF CD36 BINDING. >> IT MIGHT BE DIFFERENT? THE RECEPTORS THERE THAT ARE KEY ARE ON THE CELLS AND HERE IT SEEMS TO BE MONOCYTE MACROPHAGES THAT ARE KEY. >> THANK YOU. >> WHO WANTS TO GO FIRST? >> BEAUTY BEFORE WISDOM OR WISDOM BEFORE BEAUTY? >> [LAUGHTER] >> IN WHICH CASE IF WE WANT TO GO FOR EITHER IT'S JASMINE. >> SO, WHO IS DOUBLE POSITIVE AND WHO IS DOUBLE NEGATIVE? [LAUGHTER] >> I WILL SHUT UP BEFORE I GET COMPLETELY -- >> FINE, WHATEVER. SO, FASCINATING TALK. I HAVE TO Z. I MISS YOU SO MUCH. YOU ACTUALLY SUCH A WONDERFUL FEATURE IN THE LABORATORY AND YOU ARE ALWAYS ASKING THE BEST QUESTIONS SO WE HAVE MISSED YOU FOR MANY YEARS. >> THANK YOU. >> THE QUESTION I HAVE IS ABOUT THIS REALLY INTRIGUING OBSERVATION ABOUT OF COURSE, THIS MOLECULE THAT MAY REGULATE FOOD INTAKE AND ENERGY EXPENDITURE AND SO MY QUESTION IS WHAT IS ACTUALLY UNDERSTOOD ABOUT A SICKNESS BEHAVIOR HA HAPPENED IN THE CONTEXT OF INFECTION WHEN THE ANIMALS STOP EATING AND IT COULD IN SOME CASES PROMOTE RESISTANCE SO MY QUESTION WHAT'S IS ACTUALLY KNOWN ABOUT TOXO AND ACTUALLY HOW FORCE FEEDING OR ACTUALLY PREVENTING THE SICKNESS BEHAVIOR AND MORE IMPORTANTLY, BASED ON YOUR BEAUTIFUL OBSERVATION, HOW MUCH HIGH LIPID CONTENT IN THE CONTEXT OF THIS INFECTION COULD CHANGE THE PATTERN OF INFECTION? >> WE HAVEN'T DONE ALL THE FEEDING EXPERIMENTS THAT THE INVESTIGATORS DO, BECAUSE THEY HAVE A TON OF MONEY. THERE WERE PAPERS FROM SWITZERLAND, MANY, MANY YEARS AGO SHOWING THAT THERE IS LOSS OF FOOD INTAKE BUT THERE'S ALSO HYPER METABOLISM THAT IS DRIVEN BY INFLAMMATION AND NEUTRALIZATION OF INTERFERON GAMMA IS ABLE TO BLOCK ALL THAT HAVE AND SO IT GOES BACK TO OUR OBSERVATIONS NOW THAT WHEN WE NEUTRALIZE INTERFERON GAMMA, BOTH SYSTEMIC LEVELS OF FG F21 ARE SHUT DOWN AND GDF. WHEN WE LOOK AT THE GDF LEVELS, I DIDN'T HAVE TIME TO SHOW THEM, IN THE TISSUES, THERE IS ABSOLUTELY NO CHANGE WHEN YOU NEUTRAL LIES GAMMA AND GDF IS NOT A GAMMA RESPONSIVE GENE SO THERE'S NO REASON WHY GAMMA SHOULD BE EFFECTED, GDF15 OR FG F21 SO THAT'S JOJO'S PROJECT. HOW DOES GAMMA ACT ANOTHER A DISTANCE TO REGULATE SYSTEMIC, RIGHT, BECAUSE ESSENTIALLY FGD AND I DIDN'T KNOW THIS, UNTIL YOU GOT THE PROTEOMIC DATA. SO TIM MITCHENSON HAD A MOL HE CAN EYE LAR SYSTEMS BIOLOGY SYSTEM WHERE THEY AGO AGREE GATED USING ARTIFICIAL INTELLIGENCE AND ALL THE DATA OF THE NIEHS. TOXICOLOGY DATA AND SO USING ARTIFICIAL INTELLIGENCE THEY CAN BOIL DOWN ALL OF THAT LIKE WHATEVER TOX CAN'T YOU USE, KIDNEY, LIVER, ET CETERA, ET CETERA, IT ALL SPOILED DOWN TO FG F21 AND GDF15 SO IT'S REALLY NOT SURPRISING THAT IT DOESN'T MATTER, PEOPLE WHO DO BIOMARKER RESEARCH, RIGHT, IT DOESN'T MATTER, CANCER, HEART AND KIDNEY DISEASE, GDF15 POPS UP AS . MARKER OF SEVERE OR MORTAL DISEASE. >> FASCINATING, THANK YOU. >> THAT WAS GREAT. I'M REALLY GLAD YOU MENTIONED THE IRGs BECAUSE YOU KNOW, THOSE DATASETS FROM ALAN'S LAB WITH GREG TAYLOR REALLY KNEW CREATED THE WHOLE FIELD AND IT KILLS PATHOGENS AND MY QUESTION FOR YOU IS, I DIDN'T QUITE UNDERSTAND THE RON 18 EFFECT ON CD36. >> IT'S GOING TO BE RE COMPLICATED RIGHT. HOW DOES IT ACT WITHIN THE PARASITE NOT OUTSIDE -- >> WHAT DO YOU THINK THAT MEANS? DO YOU THINK THAT'S AN INFECTED CELL PROTECTING ITSELF FROM OTHER PARASITES? >> RIGHT NOW, I'M MORE, I WANT TO UNDERSTAND HOW, WHAT'S THE INTRA CELLULAR MECHANISM AND SHOWING THAT IN FACT THERE IS RSR29CMRNA IN THE VIRULENT STRAIN BUT HOW SOME PROTEIN SO SOMEHOW SOMEWHERE THE PROTEINS GETS LOST IN THE RH AND SO, ROP 18 MIGHT BE WHAT IS CAUSING THAT LOSS RIGHT NOW SO WE HAVE TO FIND OUT WHEN IS SRS29C AND OTHERS LOST AND WE TRIED TO KNOCK OUT SR29C BUT THE PROBLEM IS STILL BINDS CD36 BECAUSE WHEN YOU DO THE PROTEOMICS, OTHER PROTEINS ARE NOW SUDDENLY EXPRESSED. IT'S SO FASCINATING BECAUSE IT'S A VERY DIFFICULT PROBLEM. >> THANK YOU. >> SO WHAT ABOUT TH FAG A SEEM M THE CELL SERVICE IT'S A COMPLETELY DIFFERENT ENVIRONMENT? DO YOU THINK THERE'S A COMPLETELY NEW MECHANISM OF INVASION OF PEOPLE HAVING PREVIOUSLY APPRECIATED? >> I WOULD RATHER THINK THAT IT'S THE OPPOSITE, RIGHT. THAT PERHAPS, PERHAPS THE WAY EVOLUTION OCCURS IS THERE WAS A UPTICK FIRST AND SO THEN THE PARASITE LEARNED TO INCOVID-19 FROM WITHIN AND THE HR AND THEY WERE SO PROFESSIONAL AND SO GOOD THAT THEY'RE SO FAST AT INVADING ON THE CELL SURFACE THEY DON'T NEED TO GO IN. IF YOU LOOK AT MALARIA IN THE LIVER RIGHT AND EVEN IN AVIANS AND REPTILES, MALARIA ELEVATES MACROPHAGES FIRST AND EVEN IN THE EXTENT OF VERTEBRAE MALARIA THEY INTERACT EXTENSIVELY WITH MACROPHAGES FIRST IT'S LIKE THEY'RE TESTING THEIR METAL AND THEY FINALLY INVADE HIGH PATH OWE SIGHT. >> DO YOU THINK THE MECHANISM OF INVASION ITSELF THE MECHANICS ARE THE SAME? >> WE THINK THE SECOND HALF WHEN IT'S ALREADY IN THE PHAGO SEEM, AND IT FORMS ITS OWN VACUUM FROM WHAT WE CAN TELL MICROSCOPICALLY THERE'S A ROP TREE NECK MOVING FUNCTION AND THAT IS FORMED AND SO YEAH. AND ACTUALLY I DIDN'T HAVE TIME TO TALK ABOUT IT BUT OF COURSE WE, YOU KNOW, BEFORE WE TRIED THE LOOM YOU LAR APPROACH OF THAWING OUT ALL THE CELL LINES IN THE FREEZER, WE OF COURSE DID PROTEOMIC SCREENS TO LOOK FOR WHAT IS -- WHAT ON THE HOST IS BINDING TO PTG PARASITES BUT NOT RH PARASITES AND WE DID NOT FIND ANYTHING THERE. I THINK WHAT WE FOUND WAS 245 THERE WERE ALL THESE OTHER PROTEINS THAT WERE BINDING TO BOTH RH AND PTG SO WE PERSUADE THOSE AND UNLIKE WHAT OUR FIELD IS SAYING THAT THE TOXOPLASMA DOES NOT USE HOST RECEPTORS WE HAVE RECEPTORS, RIGHT, THAT ARE MEDIATING, EVEN THE CLASSICAL ACTIVATE PENETRATION PATHWAY AND SO WE'RE NOT THAT DIFFERENT FROY HOST RECEPTORS TO INVADE RED CELLS SO WHY COULDN'T TAX OWE PLASMA USE HOST RECEPTORS. THANK YOU. [APPLAUSE] >> SO I THINK WE'RE HAVING GLOBAL TRANSMISSION OF THIS EVENT AND SOME OF MY COLLEAGUES IN BRAZIL THAT COULDN'T BE HERE ARE SENDING WARM REGARDS TO ALAN, RECOVERING WELL AND ANDY SIMPSON AND RODRIGO THAT WAS SUPPOSED TO BE HERE, ANTI-AGO SO OWE AND OTHERS. SINCE NO ONE MENTIONED ANOTHER CHARACTERISTIC OF ALAN, HE HAS A VERY GOOD NOSE FOR RESTAURANTS AND WHEN I LEFT NIH I HAD 50 EXTRA POUNDS AND EXTRA PAPERS. SO ANYWAY, I WOULD LIKE TO THANK YOU ALAN, FOR EVERYTHING AND ALL THE SUPPORT OF THESE YEARS AND I'M SURE WE'RE GOING TO CONTINUE HARASSING HIM FOR LETTERS OF SUPPORT AND ADVICE AND SO ON. AND NOW I GUESS DAVID SACKS FROM NIAID I THINK HE MIGHT HAVE BEEN THE FIRST ALAN POSTDOC AND CAME FROM HARVARD TO NIH AND AND CHRIS KARP. AND THAT I POSTDOC AND AROUND AROUND TODAY AND WAIT AND TODAY'S AND IMMUNOLOGY PROGRAM OF GATES FOUNDATION AND THEY'RE GOING TO GIVE US SOME REFLECTIONS AND HOPEFULLY AND ENTERTAINING STORIES. >> SO FIRST OF ALL, THANK YOU, THIS IS WONDERFUL. SO DON'T LOOK AT THAT TREE YET AND JUST A FEW COMMENTS. I'VE KNOWN ALAN FOR A LONG TIME AND I'VE KNOWN HIM SINCE HIS TIME AT HARVARD AND WHEN HE WAS A SLENDER MAN AND SMOKED A LOT OF CIGARETTES. AND ALAN WAS ON MY THESIS COMMITTEE THERE AND HE ENCOURAGED ME TO TO GO TO MILL HILL TO DO A POSTDOC WHERE HE DID HIS POSTDOC. HE RECRUITED ME BACK FROM THERE TO JOIN HIS GROUP AT THE NIH IN THE LPD WHERE FRANK RECRUITED HIM FROM HARVARD TO START A IMMUNOLOGY PROGRAM IN THE LPD AND SO OUR GROUP AT THAT TIME WAS JUST ALAN AND STEPHANIE AND SARAH HEINIE AND MYSELF AND SO IT WAS A SMALL GROUP AND I THINK WE JUST HAD ONE OR TWO AND IT WAS A CERTAIN INTIMACY AND GETTING TO KNOW ONE ANOTHER. THEN, SO I WAS PRIVILEGED TO BE PRESENT AT THE GENESIS OF THE IMMUNO BIOLOGY PROGRAM AND WAS ABLE TO EXPERIENCE CLOSED HAND THE FLOURISHING OF THIS PROGRAM SO THIS TREE, THIS IS A GNARLY OLD TREE THAT -- IT IS STRONG LIKE AN OAK BECAUSE THAT'S WHAT IT IS AND IT HAS SUPPORTED OVER THESE DECADES THESE VERY STRONG BRANCHES OF DIFFERENT KINDS OF RESEARCH AND NURTURED THE SCIENTIFIC CAREERS OF THIS REALLY BOUNTY OF WONDERFUL INVESTIGATORS. SO, TONY MENTIONED WITHIN ALAN'S OWN FAMILY THERE WERE SCIENTISTS WHO INFLUENCED HIS FUTURE CAREER SO HIS PARENTS WERE BOTH SCIENTISTS AND HIS UNCLE AND AUNT AND THIS PERSON HERE I THINK WAS THE UNCLE'S WIFE'S THIRD COUSIN OR SOMETHING WHO WON A NOBEL SURPRISE SO THAT'S VERY COOL ALTHOUGH ALAN WE'RE ALL WITHIN SIX DEGREES OF SEPARATION OF A NOBEL SURPRISE E WINNER BUT I THROUGH IT IN THERE HE WENT TO THE SULK INSTITUTE AND TO STUDY IMMUNOLOGY WITH MEL CONE AND THEN HE WENT AND DID HIS FIRST POSTDOC WITH RON SMITHERS AT MILL HILL AND THIS HAD SUCH A POWERFUL INFLUENCE ON HIS FUTURE AND THE DIRECTION OF HIS FUTURE RESEARCH AND NOT JUST THE OTHER FOLKS WITHIN THE LAB AND WITHIN MILLHILL ITSELF AND ALL OVER THE U.K. THAT WAS THE REALLY EARLY BEGINNINGS OF IMMUNE LOGICAL STUDIES AND IN PARA SAID I CAN INFECTION. HE BROUGHT HIS OWN RESEARCH GROUP TO HARVARD AND LEE HALL WHO WAS HERE IS ALAN'S FIRST PH.D STUDENT. AND STEPHANIE JAMES WAS HIS POSTDOC AT THE LAB IN HARVARD AND THEN JUST TWO OR THREE YEARS AFTER SETTING UP THE PROGRAM AT HARVARD, HE WAS RECRUITED AS I SAID BY FRANK TO THE LPD AND STEPHANIE CAME WITH HIM AND CARRIED ON THE SIS TOE IMMUNOLOGY STUDIES AND THE LPD AND WE BROUGHT ME ON TO START SOME RESEARCH ON ANALYSIS AND PHIL FOLLOWED SHORTLY AFTER ME AND THIS BRANCH OF RESEARCH AND ALAN'S LAB SAID SOMEHOW WITHERED AFTER THAT AND I DON'T KNOW WHAT HAPPENED. [LAUGHTER] BUT CERTAINLY THE SHISTO PROGRAM FLOURISHED AND THERE WAS ALAN DABBLED IN SOME RESEARCH WITH SOME INVESTIGATORS LOOKING AT IMMUNE RESPONSES. THROUGHOUT THE YEARS THERE WERE A NUMBER OF SENIOR SCIENTISTS ON SA BAD CAL LEAVE WHO WERE IN THE LAB AND REALLY OFFERED THE WONDERFUL COMPONENT TO THE -- IT ADDED TO THE RESEARCH ENVIRONMENT OF HIS GROUPS OVER THOSE YEARS AND SO IMPORTANTLY, HE ALSO INITIATED A PROGRAM AND RESEARCH ON THE IMMUNE RESPONSES TO TOXO AND I THINK THAT THIS WAS REALLY INSIGHTFUL AND FORESIGHTFUL TO HAVE THESE TWO COMPLIMENTARY PROGRAMS OF A RESEARCH OF SHISTO AND TOXO VERY DIFFERENT CLASSES OF PARASITES AND I THINK IT WAS DURING THIS TIME THAT THEY PRODUCED ONE PAPER AFTER ANOTHER WITH THE PATHWAYS INVOLVED IN THE EVOLUTION AND CONTROL OF TYPE ONE AND TYPE TWO RESPONSES. AND THEN OF COURSE HE SHIFTED AGAIN AND BROUGHT ON SOME INVESTIGATORS TO LOOK AT IMMUNE RESPONSES TO TTB AND IT SPEAKS TO THE RESOURCES AND THE LATITUDE THAT THE NIH OFFERS THAT ALLOWS THEE SORTS OF PRAB PROGRAMMATIC CHANGES AND IT TAKES COR AGE TO MAKE THOSE WHOLE SCALE CHANGES IN THE LAB AND EVERY TIME ALAN DID THIS, THE WORK THAT CAME OUT OF THOSE PROGRAMS WERE WORLD-CLASS AND IMPACTFUL. AND NIH I THINK ALAN AND THE NIH ARE SO WELL SUITED TO ONE ANOTHER, THE NIH ENCOURAGES LOTS OF MULTI-DISCIPLINARY APPROACHES AND A LOT OF COLLABORATION AND TO ALAN COLLABORATION IS JUST ANOTHER FORM OF SCHMOOZING. [LAUGHTER] AND ALAN IS A NATURAL-BORN SHMOOZERS. SO LET ME JUST END BY SAYING THAT TO HAVE -- I HAVE JUST BEEN SO PRIVILEGED TO HAVE HAD ALAN AS A MENTOR, COLLEAGUES AND A FRIEND FOR 40 PLUS YEARS NOW. THANK YOU. THANK YOU, ALAN. AND YOU H THIS IS FOR YOU. TO HANG IN A BATHROOM OR WHERE YOU LIKE. [APPLAUSE] [OFF MIC] >> I THINK WE ALL WANT A COPY OF THIS. ALAN, WHAT A PLEASURE IT IS TO BE HERE TODAY. AND THANK YOU DRAGANA FOR SETTING UP THIS. THERE ARE MANY SIDES TO ALAN I CAN TALK ABOUT AND MANY OF THEM TALKED ABOUT. WE CAN TALK ABOUT ALAN AS A PROLIFIC AUTHOR OF SEMINAL PAPERS AND IMMUNO BIOLOGY. AS A SCIENTIST WHO FOCUSED THE BENEFITS OF HIS PRIVILEGED ON FUNDAMENTAL WORK AND NEGLECTIVE DISEASES OF THE POOR, AS A TRULY GENEROUS, GRACIOUS COLLABORATOR, AND A SEEDER AND BUILDER OF SCIENTIFIC COMMUNITIES, OFTEN SEEDING THEM SEED AND THEN CEDEING THEM TO HISS TRAININGS, AS A DYED IN THE WOOL MOUSE MODELER WHOSE MID LIFE CRISIS INVOLVED MOVING INTO STUDYING THOSE MORE COMPLICATED ABSTRACT BY PEDESTRIANPEDS OR PIPE 2 ABIE AND BY THE WAY, MEMORY AS WELL AS SOME PHOTOGRAPHIC EVIDENCE INDICATES THAT THE LAST TWO ARE SOMEHOW RELATED AND I COULD CELEBRATE ANY OF THESE BUT I REALLY WANT TO FOCUS ON A SPECIAL QUALITY OF ALAN THAT IS BOTH REFLECTED IN THE BRED OF THOSE WHO ARE HERE TODAY AND WATCHING ON-LINE AND THEN I THINK ACTUALLY REPRESENTS WHAT I THINK AN OUT SIZED PART OF HIS LEGACY WHICH IS VERY ACTIVE ROLE AS A MENTOR AND SPONSOR OF SCIENTISTS. A ROLE PLAYED AS MUCH OUTSIDE THE GENERATIONS OF SCIENTISTS THAT TRAINED IN HIS LAB AS WITHIN IT AND ALAN, YOU HAVE A WONDERFUL, BRED GENEROUS RECORD OF SUPPORTING THE SCIENCE AND CAREERS OF OTHERS AND SOMETHING THAT'S EVIDENCE ON MANY, MANY LEVELS. AS RICARDO MENTIONED, I POSTDOCKED WITH DAVID SACKS SO I'M A SCIENTIFIC GRAND CHERNOFF AND I CAME BACK HERE IN THE LATE PALEO SEW I CAN IN 1989 AND EVER SINCE THEN, AT EVERY STAGE OF MY CAREER, ALAN HAS BEEN THERE, PROVIDING ADVICE, THAT I SOMETIMES EVEN THOUGHT AS WELL AS SPONSORSHIP AND IT'S SOMETHING THAT I'M DEEPLY APPRECIATIVE OF AND I TRIED TO PASS OUT THROUGH MY CAREER. IT'S THE SAME WITH MY WIFE, SO, I HAD A SECOND CLASS MD DEGREE BEFORE COMING TO THE LPD AND I WENT TO A COURSE AND TO GO TO THE BENCH FOR THE FIRST TIME AND MARSHA CAME TO VISIT ONE WEEKEND AND HEARD ALAN LECTURE ON THIS NOVEL PH1 AND 2 DARA TIME AND SHE WORKED TON AND TRANS DUCK AND ASTHMA AT THE TIME AND IT WAS A LIGHT BULB AND CRIMINOLOGY AND AND THROUGHOUT, ALAN WAS A GRACIOUS SPONSOR AND INTRODUCING THE POTENTIAL COULD BE AT RATERS ADVICE AND HELPS POD LIED AND -D IN THE ROOM AND OF COURSE BY MANY MORE AROUND THE WORLD SO THANK YOU, ALAN AND YOU'VE HAD A HUGE I AM IMPACT ON PEOPLE AND PROVIDE A ROLE MODEL FOR OTHERS. I ALSO WANT TO THANK YOU FOR YOUR TOLERANCE FOR PUNS. THAT'S FOR ANOTHER TIME. THANK YOU. >>GOOD "AFTERNOONSKI" I GUESS I'M PARTLY GUILTY FOR TEACHING ALONG THE SWEDISH ME, LENA AND TONY AND SO THAT WAS THE LEVEL OF SWEDISH WE HAD IN THE LAB. AND SO THIS IS REALLY GREAT TO BE HERE AND ALAN HAS BEEN SUPPORTIVE TO ME AS MY TIME AS A SCIENTIST. I CAME ALREADY AND I WAS ACTUALLY A PH.D STUDENT IN THE LAB AND STARTED COLLABORATION WITH ALAN'S LAB AND FIRST, AS A PH.D STUDENT, I MEAN, AND THEN AS A POSTDOC IN THE LAB AND LOTS OF GREAT MEMORIES OF THAT. SO, TODAY, THIS AFTERNOON, WE HAVE IN THIS SESSION WE HAVE FIVE FORMER POSTDOCS AT THE NIH HERE IN THE 1980s AND 1990s AND BECAUSE I WAS HERE A LONG TIME AGO, I'VE OVERLAPPED WITH SEVERAL OF THEM. THREE OF THESE POSTDOCS ON THE SLIDES WERE IN ALAN'S LAB AND IN ADDITION WE HAVE THREE LONG-TERM COLLEAGUES AND COLLABORATORS AND FRIENDS OF ALAN WHO WILL TALK IN THIS SESSION. WE'LL START WITH REMARKS WHO IS ONE OF THE LONG-TERM COLLABORATORS AND GEORGE YO JOINED THE NIH IN 2006 AND HE WAS NOT AND HE IS NOW SENIOR INVESTIGATOR AND CHIEF OF THE LABORATORY OF CANCER IMMUNOLOGY CANCER RESEARCH AT THE N. >> Catherine: SO OVER TO YOU. >> SO, IT'S WONDERFUL TO BE HERE AND I WILL DISCUSS WHY I FEEL THE FRIENDSHIP WITH ALLEN MORE THE COP AGO RACE HAS BEEN IMPORTANT FOR ME FOR MANY YEARS BUT FIRST, I WANT TO JUST SEND THIS MESSAGE SO BOTH ALAN AND I FOR A YEAR A FEW YEARS AGO AND IT'S THIS MESSAGE AND THEY WANT TO BE SURE THAT EVERYBODY SEES THE MESSAGE THAT HE REGRET NOT TO BE ABLE TO BE HERE AND YOU SAY WHAT WE CAN ALL SAY, YOU HAVE BEEN A INSPIRATION FOR AND YOU MAY BE STEPPING DOWN FROM SOME TASKS BUT NEVER RETIRE FROM YOUR DEEP INSIGHTS INTO INFECTION AND IMMUNITY. I MET ALAN IN THE LATE 80s AND THE WAY I MET HIM IS HE CALLED AND THERE WAS AN ORGANIZE AND NIAI MEETING AND HE WANT TODAY GET A TALK THAT WAS IL12 AND YOU HAVE TO ASK GEORGE TO SHARE IN THE SESSION AND ALAN SAYS I'M A GOOD FRIEND OF GEORGE'S NO PROBLEM. I NEVER METAL AN BEFORE THAT WAS THE PROBLEM. [LAUGHTER] BUT THAT WAS ACTUALLY THE START OF A REALLY CLO COLLABORATION AD FRIENDSHIP THAT'S BEEN LASTING AT THIS POINT ALMOST 35 YEARS. WE STARTED WORKING TOGETHER WHEN TNF WAS REPLACED BY GAMMA INDUCER SO AT THE TIME, ACTUALLY, WE WERE HAVING A MEETING AND THEY WERE ALL COMING UP AND I ALWAYS WONDER IF THEY'RE COMING TO TO PHILADELPHIA BECAUSE OF MY SCIENCE OR THEY WERE COMING BECAUSE OF MY WIFE'S FOOD AND ALAN CAN SAY WHAT THE REASON IS. [LAUGHTER] AND THEN I THINK THERE'S BEEN A COMPLICITY BETWEEN ALAN AND PHIL AND THEY TRIED TO MAKE ME A (INAUDIBLE) AND THEY ALMOST SUCCEEDED AND I REMEMBER BEING INVITED TO A MEETING, IT WAS REALLY SURPRISING FOR ME AND I ALWAYS WORKED A LITTLE BIT ON TOXOPLASMA BUT ANYWAY, IT'S -- I THINK THAT'S TOO GOOD TO RESOLVE. ONE IS THAT I START TO BE INVOLVED WITH BRAZILIAN SCIENTISTS AND I START TEACHING IN BRAZIL AND NOW INTERACTING WITH CANCER BIOLOGISTS THERE, NOT MUCH WITH PARA SITTOLOGISTS BUT TWO-THIRDS OF MY LAB IS BRAZILIAN SO THAT WAS A GOOD RESULT. AND THE OTHER THING WAS THE COMPARATIVE IMMUNOLOGY AND I THINK BOTH I AND A LOT OF RUM EVEN A IS REALLY BUILDING IN CANCER IMMUNOLOGY IN WHAT ARE THE COMMONALITY AND DIFFERENCE BETWEEN ACUTE INFECTION AND CHRONIC INFECTION AND CHRONIC INFECTION IN TUMORS AND THE ACTIVATION OF INFLAMMATION IN IMMUNITY IN CHEMOTHERAPY SO THAT WAS REALLY SOMETHING THAT INFLUENCED LIKE INFLUENCED MANY AND MY WORK IN CANCER AGAIN STARTS FROM THE -- THE ONE THING THAT REALLY I AM GRATEFUL TO ALAN IS THAT MANY OF YOU MAY KNOW THAT A OPPORTUNITY POINT I WAAT ONE POINT I WORKED FORINDU. TO GO FROM UNIVERSITY TO INDUSTRIES AND INDUSTRY TO BACK TO ACADEMY IS NOT EASY SO ALAN SAID TO ME AND IT WAS SUPPOSED TO BE A SA BAD CAL FOR SIX MONTHS AND I STAYED FOR TWO YEARS. IN THE MEANTIME, THEY OFFERED ME A POSITION AT NCI AND IT TOOK ABOUT COUPLE OF YEARS FOR NCI TO CLEAR ME AND SO I STAYED IN HIS LAB AND THE FUNNY THING IS IS THAT THE PEOPLE AT NCI TALK WITH ALAN AND SAY WOULD YOU BE INTERESTED IN A POSITION IN NCI AND I SAID NO WAY AND A WEEK LATER WHEN WE'RE CHATTING, I SAID THEY TAUGHT ME ABOUT THAT AND YOU KNOW, WE BETTER LESSON AND IT'S SOMETHING GOOD SO THAT WAS 18 YEARS AGO AND OBVIOUSLY I'M STILL HERE SO IT'S BEEN GOOD. AS I SAY, AS MUCH AS I APPRECIATED THE MENTORSHIP AND THE SCIENTIFIC COLLABORATION WITH ALAN I REALLY APPRECIATE THE FRIENDSHIP OF STEPHANIE AND ALAN AND FOR ALL THESE YEARS. I REALLY RATHER THAT REMEMBER THE MEMORY AND I THINK THIS IS WHERE ALAN WE HAVE THIS WONDERFUL IMMUNOLOGY WAS THERE AND MY WIFE WAS VERY SCARED BECAUSE WE'RE ALL IMMUNOLOGISTS BUT WE WERE THERE FOR ONE WEEK AND NO ONE TALKED ABOUT IMMUNOLOGY. WE WERE SO IMPRESSED BY THE PLACE WE WENT BACK AND COVID CAME AROUND AND WE HAVE TO DO THE 51st A ANNIVERSARY BUT EVENTUALLY I GOT THERE WITH MY FAMILY. SO AGAIN, ALAN, THANK YOU VERY MUCH. AS I SAY, FIRST FOR YOUR FRIENDSHIP AND ALSO FOR SCIENTIFIC COLLABORATION AND BEING YOUR FELLOW HAS BEEN A LOT OF MENTORSHIP OR PSYCHOLOGICAL HELP, WHATEVER YOU WANT TO CALL IT. THERE'S ONE REGULAR THAT YOU HAVE NOT PROMISED FOR MANY YEARS TO VISIT UP IN ITALY SO THE PLACE IS THERE. SO THE OPPORTUNITY WILL COME SOON. THANK YOU, VERY MUCH. [APPLAUSE] >> SO, OUR NEXT SPEAKER IS GOING TO BE YASMIN BELKAID. SHE DID A TESTIFIED WITH MILL ON AND POSTDOC HERE AT THE NIH WITH DAVID SACKS AND AFTER THAT, SHE MOVED TO CINCINNATI TO TAKE UP A POSITION AT THE CHILDREN'S HOSPITAL RESEARCH FOUNDATION AND AS AN ASSISTANT PROFESSOR AND IN 2005 SHE JOINED NIH AGAIN AND I GUESS IT WAS PARTLY ALAN WHO BROUGHT HER BACK TO THAT, YES. AND AT THE MOMENT, YASMIN IS IS NOW CHIEF OF THE LABORATORY OF HOST IMMUNITY AND MICROBIOME AT THE NIH-NIAID. OVER TO YOU. >> I HAVE THE GREAT PRIVILEGE TO GIVE A TALK TO HONOR ALAN AND I HAVE TO SAY IT'S VERY INTIMIDATING AT MANY LEVELS BECAUSE I'M GIVING A TALK IN FRONT OF EVERY BOSS I EVER HAD. THIS IS TRUE. DAVID SACKS, ALAN SHER AND RON GERMANY IS AROUND SO IT'S TERRIFYING. I JUST WANTED DISCUSS WITH YOU, FIRST, YOU THINK IT'S AN EXTRAORDINARY MOMENT. I WAS SAYING TO ALAN I FEEL MISTY. THE MEMORIES WITH ALL THE PEOPLE AROUND AND ALL THE JOURNEYS WE HAVE MADE TOGETHER AND I WANTED TO SHARE WITH YOU SOME WORD OF WISDOM THAT I WAS GIVEN BY ALAN ALONG THE WAY, IT'S VERY TAME, DON'T WORRY, IT'S REALLY FOR GENERAL PUBLIC. [LAUGHTER] I'M GOING TO JUST GIVE YOU A LITTLE BIT OF IN SIGHT ON SOME OF THE WORK WE HAVE BEEN DOING WHICH VERY MUCH HAS BEEN INSPIRED BY THE JOURNEY OF ALAN ON LOOKING AT HOW MIKE ROBES AND BEHAVING AS A WAY TO TEACH US ABOUT THE IMMUNE SYSTEM. >> SO, THAT WAS DARK AGES AND IMMUNOLOGY WAS COMPLY ASIDE AND I REMEMBER SEEING IMMUNOLOGY AS THOSE DEMI GOD, THE STRANGE CREATURES WORKING IN BACK SOME HAD BEARDS AND THEY WERE TALKING A STRANGE LANGUAGE AND ARGUING ALL OF THE TIME. IT WAS INCOMPREHENSIBLE. BEFORE THEN WAS ADD AND WE HAD TONY FAUCI AS A LEADER WHICH OF COURSE BECAUSE IS STILL AND AFTER THAT, IT'S VERY IMPORTANT DISEASE, MALARIA AND I WAS IN THE LPD SO THAT WAS DISEASE THAT WAS THE CENTER -- N THE BOTTOM IT WAS US. ALL THE OTHER PARASITES. THAT'S ACTUALLY HOW IT WAS. THAT'S WHERE I LANDED BECAUSE YOU CAN SEE I WAS VERY STRATEGY IN MY CAREER BUT IT WAS ALSO WHERE I THINK ALAN MADE A TRUE TRANSFORMATION AND REALLY HAD ENORMOUS IMPACT. SO, OF COURSE, THIS MICROBES WHICH ARE OF COURSE A VERY IMPORTANT PUBLIC-HEALTH PROBLEM AND THEY STILL ARE. THIS IS REALLY SOMETHING THAT REMAINS TO BE ADDRESSED AT A HIGH LEVEL BUT THEY'RE INCREDIBLE TOOLS AND INCREDIBLE MASTER TO TEACH US ABOUT THE IMMUNE SYSTEM THAT WAS THE GENIUS OF ALAN TO CREATE AN ENTIRE FIELD BY SAYING, YOU KNOW WHAT, WE'RE NOT BRIGHT ENOUGH WE HAVE TO LOOK AT THEM AND OBSERVE. THIS IS EXACTLY WHAT THIS FIELD WAS. IT WAS TRANSFORMATIVE. IT WAS NOT THIS KIND OF LIKE TOP DOWN UNDERSTANDING OF THE SYSTEM IT WAS FOLLOWING THE BUGS AND FOLLOWING WHAT THEY WERE TEACHING US AND I REALLY THINK THE IN SIGHT WE HAD ABOUT THE IMMUNE SYSTEM, BASED ON THIS FIELD THAT WAS VERY MUCH CREATED BY ALAN, WE CAN ALL RECOGNIZE THAT AND HAS REALLY COMPLETELY TRANCE FIELD THE FIELD OF IMMUNOLOGY AND PROVIDING IN SIGHT INTO THE INFECTION. AND YOU KNOW, IN A WAY THIS IS A UNIQUE FIELD. THE PATHOLOGY WITH ALAN AS A LEADER, WE WERE THE FIRST WAN THAT SOUGHT ABOUT PHYSIOLOGY IN THE CONTEXT OF IMMUNOLOGY AND THEY HAVE TO BECAUSE THE MICROBES GO TO TISSUE AND THEY HAVE TO THINK ABOUT TISSUE INVENTED BY THIS FIELD AND WE CAN REMEMBER THE BEAUTIFUL WORK OF TOM WIN FOR EXAMPLE AND THE CONTEXT OTHER THAN MANY OF THE PEOPLE THAT HAVE WORKED WITH ALAN AND THIS CONCEPT AND YOU HAVE TISSUE SPACE IN THE COMMUNITY CAME FROM THE FIELD OF IMMUNO PAR SITOLOGY. IT WAS NOT A VERY NICE TERM FOR IMMUNOLOGY. CONTEXTALITY BUT IT WAS VERY COMFORTABLE WITH IT AND THE FACT THAT YOU CAN HAVE DIFFERENT GENOTYPE AND CONTEXT AND THAT DROVE PATHO BEGIN WAS IT WAS VERY ACCEPTED BYPATHOLOGISTS AND IT WAS BY A SINCE A MULTI-DISCIPLINARY SCIENCE AND ALAN DEVELOPED MULTI DISCIPLINARY SCIENCE AND YES, YOU DID. I CAN GO OVERBOARD. IT WAS REALLY BRIDGING THE TWO EXTRAORDINARY FIELD THAT MADE SOMETHING UNIQUE AND TODAY THIS IS REALLY THE NORM. WE ARE REINVENTING THE WHEEL BUT THAT HAPPENED THERE AND THE CONCEPT OF IMMUNO REGULATION AND IT WAS TRULY VERY MUCH DEVELOPED IN THE CONTEXT OF THE EXPLORATION OF THE WORK AND WE KNOW SOME. WORK DONE IN THE CONTEXT OF IL10 AND MANY OTHERS. ALL OF THESE FINDINGS HAD ENORMOUS CONSEQUENCES OF UNDERSTANDING OF OTHER FIELDS AND I THINK THAT OF COURSE I'M GOING TO JUST BE VERY BIASED THERE BUT THERE'S A LOT OF TRANSFORMATION ABOUT HIGH IMMUNATION IS A DRUG AND SOMETHING WE CAN TARGET FOR THERAPY AND STEMS FROM THE WORK THAT'S BEEN DONE IN THE CONTEXT SO COMPLETELY BIAS BUT I'M RIGHT. IT SAVED THE WORLD. AND SO WHERE ARE WE TODAY? EXACTLY WHERE WE WERE BY THE WAY. THERE'S THIS ONE THAT HAS TAKEN OVER BUT IT'S EXACTLY THE SAME THING BUT THERE'S A HUGE, HUGE DIFFERENCE IS THAT WE ALL KNOW. WE IN THIS ROOM WE KNOW WHERE IT FRAME AND MAYBE THIS IS NOT ACKNOWLEDGE ENOUGH BUT WE KNOW THE IMPORTANCE OF THE WORK THAT HAS BEEN DONE HERE AND THE WORK THAT'S BEEN DONE BY ALAN IN SHAPING THE WAY UNDERSTANDING OF THE HUMAN SYSTEM. WHAT I'LL GOING TO DO NOW IS GIVE YOU A COUPLE INSIGHTS INTO MY OWN JOURNEY AND MY RELATIONSHIP WITH ALAN AND HOW HE HAS INFLUENCED ME AND IF THERE'S A TRAINING WATCHING US REMOTELY MAYBE THEY CAN LEARN SOMETHING THERE, OR MAYBE NOT. THE FIRST THING THAT I HAVE LEARNED TO ALAN, IS THE IMPORTANCE OF BEING ADOPTED BY THE RIGHT MENTOR. THIS TRANSFORMED BY LIFE AND MY CAREER. WHEN YOU ARE ADOPTED SOMEONE LIKE ALAN, THIS IS FOR LIFE. IT'S NOT A LITTLE. IT'S REALLY HISS FAMILY. AND WE ALL JOKE HERE, YOU KNOW, WE ARE THE GRAND CHILDREN, THE CHILDREN BUT THIS IS REAL. THE FACT THAT I'M NOT ONLY I'VE BEEN BENEFIT INTERESTED THIS EX DIN TARRY MAN TO BE SHIP BECAUSE I WAS A GRANDCHILD AND TRAINEE OF DAVID AND EVEN MY TRAINING, THE GREAT GRANDCHILD OF ALAN BENEFITS FROM THIS INTERACTION AND IT'S UNIQUE. I DON'T THINK ANYONE IN THE FIELD OF SCIENCE HAS DONE AS MUCH AS ALAN TO CREATE A SENSE OF COMMUNITY AND FAMILY. SO OF COURSE NOW THERE'S A PROBLEM BECAUSE HE HAS HIS OWN GRANDCHILDREN SO THERE'S A COMPETITION THERE AND I DON'T KNOW HOW TO FEEL ABOUT THAT BUT YOU KNOW, I WANTED TO SHARE A SHORT STORY THAT ILLUSTRATES THE BEGINNING OF OUR RELATIONSHIP AND I THINK THE WHOLE BEGINNING OF MY RELATIONSHIP WITH ALAN IS BASED ON THE COMPLEX UNDERSTANDING. I THINK SOME OF YOU IN THIS ROOM REMEMBER THIS TALK I GAVE WHEN I HAD ARRIVED MOVING THE STATES NOT SPEAKING ENGLISH I THOUGHT IT WAS A GREAT IDEA TO MOVE TO THE STATES. I WAS ASKED TO GIVE A TALK EIGHT MONTHS AFTER I JOINED THE LABORATORY OF DAVID. DAVID DIDN'T MIND AT ALL BY THE WAY. I DON'T KNOW HOW EXACTLY AT ONE POINT WE SHOULD TALK ABOUT THIS YEAR I SPENT NOT TALKING ENGLISH. I LEARNED EVERYTHING BY HEART. I WAS VERY SERIOUS AND I LEARNED EVERYTHING AND I GAVE A TALK. OF COURSE, WHEN QUESTION CAME, I HAD ABSOLUTELY NO IDEA WHAT PEOPLE WERE ASKING AND I REMEMBER I INSULTED CHRIS HUNTER THAT HE IS HERE AND OTHERS. THE FIRST THAT CAME OUT OF MY MOUTH WAS ABSOLUTELY NUTS. NEEDLESS TO SAY, EVERYBODY WAS PISSED OFF AND THOUGHT I WAS A HORRIBLE PERSON AND NOT ALAN, HE WAS VERY AMUSED. I THINK AT THE END OF ALL THESE STORIES, WHAT HE GOT FROM THAT SHE'S A TOUGH WOMAN. [LAUGHTER] SO, THIS FACE TAS TICK MISUNDERSTOODING THAT I WAS ILLITERATE WENT WELL. IT WAS THE START OF A LONG RELATIONSHIP. THE SECOND LESSEN LEARNED FROM ALAN IS ALAN FOR ME IS A HUMAN FACE OF SCIENCE AND I KNOW THAT MAY SOUND STRANGE, IT WAS A STRANGE IMMUNOLOGY AND ALL THESE PEOPLE AND IT WAS VERY MALE-DOMINATED, LET'S FACE IT AND ALAN WAS SO DOWN TO EARTH, SO EASY TO TALK TO, SO VERY EX CENTER VERT AND FRIENDLY AND FOR ME IT WAS VERY MUCH THE HUMAN FACE OF SCIENCE. THIS INSPIRED ME ENORMOUSLY. I WANT TO SHARE ANOTHER STORY IF AWE LAW ME. I GAVE MY FIRST DOCK, I HAD MOVED NOW AND CHRIS KARP DEPARTMENT IN CINCINNATI AND GIVING MY FIRST TALK AS AN INDEPENDENT INVESTIGATOR AND I'M TERRIFIED. MANY OF YOU REMEMBER THIS MOMENT AND WHEN YOU GIVE YOUR FIRST TALK AS A P.I. I WAS SITTING IN THE FRONT ROW AND I'M GOING TO PASS OUT. THIS IS THE WORST THING AND ALAN WAS SITTING CLOSE TO ME. HE WAS GIVING A TALK AND WE HADN'T SEEN EACH OTHER FOR A FEW MONTHS AND HE TURNS AND SAYS, YOU KNOW, I'M SO SCARED AND I HAVE NOTHING NEW TO SAY AND I'M LOOKING AT HIM AND TWO THINGS CAME TO MY MIND. WE'RE ALL CRAZY AND THIS IS GOING TO BE FOR LIFE AND THE SECOND ONE THAT I REALIZED IS, HOW GENEROUS THIS WAS OF HIM. I DON'T KNOW IF HE REALLY REALIZED HOW SCARED I WAS BUT HE SHOWED VULNERABILITY AND SOME HUMANITY AT A MOMENT WHERE I NEEDED IT AND I REALLY THINK THIS IS SOMETHING THAT TINY STORY IS A SYMBOL OF HOW ALAN HAS BEHAVED TO BE THIS HUMAN, KIND AND ALWAYS SAYS THE RIGHT THING AT THE RIGHT TIME. THE OTHER THING THAT IS IMPORTANT AND MANY OF YOU ARE TORTURED WHEN YOU WRITE A PAPER OR PRESENT A TALK, I WAS NOT A TRAINEE OF ALAN'S AND I AVOIDED THE PAPER PART BUT I HAD TO DEAL WITH TALKS. WHEN I WAS GIVING MY BSE PRESENTATION AND SO FOURTH. ALAN IS INCREDIBLY PECULIAR WILL PRESENTATION. THE FONT, THE SAYING, IT'S VERY DIFFICULT SO YOU SPEND A LOT OF TIME WORKING WITH HIM AND I LEARNED THROUGH HIM THAT IT'S TRUE, EVERY DETAIL COUNTS AND YOU NEED TO PAY EXTREME ATTENTION HOW YOU PRESENT YOURSELF AND I LEARNED HOW TO PRESENT MYSELF IN THE LAB. EVERYTHING THAT CAME ABOUT FASHION AND ABOUT HOW YOU ACTUALLY GIVE A TALK CAME FROM ACTUALLY THIS REMARKABLE INTERACTION WITH THIS MASTER. [LAUGHTER] BUT MOST SERIOUSLY I THINK WHAT I LEARNED ONE THING ABOUT ALAN AND SOMETIMES IT'S IMPORTANT TO REMEMBER AND THIS IS ONE THAT HE IS A VERY IMPORTANT LESSON, DON'T TAKE YOURSELF TOO SERIOUSLY AND THIS IS REALLY WHAT ALAN TAUGHT US. YOU CAN FIGHT. YOU CAN DO WHATEVER YOU WANT. AT THE END OF THE DAY, REMEMBER WHERE YOU COME FROM AND TRY TO REMAIN HUMBLE AND BE THE PERSON YOU ARE WHEN YOU STARTED. ALAN IS PROBABLY EXACTLY THE SAME PERSON HE IS NOW THAN HE WAS WHEN HE STARTED HIS CAREER AND I THINK IT'S ANNA ACHIEVEMENT. SO THE LAST LEGACY AFTER THAT OVER A PROMISES, I THINK MOST IMPORTANT ONE I HAVE LEARNED FROM ALAN IS HE IS VISION OF LEGACY. THE VISION OF ALAN LEGACY IS NOT TO SELF-CENTER LEGACY WHAT HE IS ABOUT ME AND HOW AIM GOING TO BE IN MY OWN -- IT'S ALL ABOUT EMPOWERING OTHERS AND NEVER EVER GIVING UP ON THEM AND THIS IS VERY MUCH THE KIND OF MENTOR AND THE KIND OF PERSONAL AN IS AND NOT ONLY ABOUT THE SCIENCE HE HAS BROUGHT FORWARD BUT THE PEOPLE HE HAS BROUGHT TOGETHER AND THIS INCREDIBLE LOYALTY HE HAD WITH PEOPLE THAT HE HAD ACTUALLY THAT HAD PASSED THROUGH HIS LIFE. AND SO FOR ALL THESE REASONS, I REALLY WOULD LIKE TO SAY A HUGE HOMAGE TO ALAN FOR THE MENTOR HE HAS BEEN TO ME AND THIS IS NOT ACTUALLY JUST BECAUSE IT'S YOUR RETIREMENT THING, WHATEVER, I REALLY NO THAT MY CAREER WOULD NOT BE WHERE IT IS IF IT WAS NOT BECAUSE OF YOU. THANK YOU, VERY MUCH, ALAN. [APPLAUSE] I'M NOT DONE. [LAUGHTER] NOW I'M GOING TO GIVE 15 MINUTES OF SCIENTIFIC TALK. THAT IS AS VERY MUCH INSPIRED BY THE JOURNEY OF ALAN AND I DON'T KNOW WHAT HE DID. I LOGGED AT THE BOOK AND FOLLOWED THE PATH AND OBSERVED FROM THEM AND TRIED TO LEARN THROUGH THE LENS OF HOST MICROBE. THIS IS WHAT I'M DOING. WHEN I MOVED BACK AT NIH 15 YEARS AGO SOMETHING LIKE THAT, I DECIDED TO EXPLORE COMPLETELY EMPTY FIELD OF HOST MICROBIOME AND IT WAS NOT VERY STRATEGICALLY SMART BECAUSE CLEARLY THIS WAS NOT AN EMPTY FIELD TWO YEARS LATER BUT AT THE TIME IT WAS EMPTY AND IT WAS REALLY REMARKABLY FUN TO WORK AT THE NIH AND REINVENT MYSELF AFTER HAVING LEFT PATHOLOGY AND I AM STILL A PATHOLOGIST AT HEART AND I HAD FANTASTIC PEOPLE IN THE LAB AND THEY WERE ADOPTED BY ALAN WHO THEY ALSO BECAME THE MENTOR AND WHAT HELPED ME PAVE THE WAY OF UNDERSTANDING THE BEGINNING OF THIS RELATIONSHIP AND YOU KNOW, EVEN TO THIS DAY WE SCRATCH THE SURFACE AS A COMMUNITY AND THE COLLECTIVE WORK THAT HAS BEEN DONE HERE IN PART BY MANY PEOPLE INCLUDING IN THIS ROOM, GEORGE YO WAS A INSTEALEADER IN TUMOR IS THE UNDERSTANDING THAT THE IMMUNE SYSTEM IS SHAPED BY THE MICROBE AND THIS LEADS TO THE ABILITY OF THE MICRO BUY OT TA AND ALSO AS AN AGENT OF INFLAMMATORY PROCESSES. >> WE KNOW A LOT WHAT WAS LEARNED WAS GENERATED BY THE PEOPLE AT THE NIH OF TRYING TO UNDERSTAND THE SIGNAL THAT DICTATES THE ABILITY OF THE MAIN SYSTEM TO BE ENGAGE IN THE CONTEXT OF INFLAMMATION AND TISSUE DAMAGE AND MANY REMARKABLE WORK DONE BY MANY OTHERS HIGHLIGHTED THE FACT THAT TO INITIATE HUMAN RESPONSES YOU NEEDED SOME DEGREE OF TISSUE DAMAGE AND INFLAMMATION BUT THIS LED US WITH THESE PARADOX AND SOMETHING THAT WAS NOT VERY CLEAR FROM THE WORK OF ANDREW McPHERSON, A GROUP OF MANY OTHERS, WHICH IS THE MICROBE LEAVING OTHER SURFACE THE SKIN, THE G.I. TRACK AND LUNG AND SO FOURTH WAS ABLE TO INDUCE IMMUNE ARY RESPONSES WITH T CELLS AND ANTIBODY RESPONSES IN THE ABSENCE OF INFLAMMATION. SO THIS RAISED THE PARADOX THAT WE'RE NOT CLEAR AND WE DECIDED OVER THE LAST FEW YEARS TO EXPLORE WHEN WHAT THIS MEANT. TRYING TO UNDERSTAND WHAT PROPERTY AND FUNCTION THAT IS NOT HERE TO ELIMINATE THE INVADING AGENT AND AND SO I'M GOING TO DISCUSS BUT AND ALSO SOME ON GOING WORK AND ANOTHER PARTIES, WHAT IS THE ORIGIN OF THIS RELATIONSHIP. THAT ALLOWED THE HUMAN SYSTEM TO RECOGNIZE I THIS SEARCH OF THE WAY. THIS IS TO SUMMARIZE THE WORK OF ALAN. OVER THE LAST FEW YEARS, IF IT HAS ITS OWN MICROBE AND THEY WILL BE A NEURO RESPONSE THAT CAN BE GENERATED AND TO ENHANCE THAT BY AND ENHANCING BROADLY DEFENSE IN THE TISSUE AND THEY ALSO VERY PLASTIC AND THESE CELLS CAN CHANGE FUNCTION BASED ON CIRCUMSTANCES AND CAN BE REPURPOSED IN THE CONTEXT OF TISSUE DAMAGE AND ALSO PROMOTE TISSUE REPAIR AND THIS OF COURSE LED US TO MANY TO TRY TO UNDERSTAND THE SIGNAL THAT ALLOW THE MICRO TO ENGAGE THE SYSTEM AND THE FIRST HINT OF THE MECHANISM OR POTENTIALLY THE REASON FOR THIS RELATIONSHIP CAME FROM THE WORK THAT WAS DONE BY THE LABORATORY AND SHE LIKED IN A SIMPLE PAY POST ASSOCIATION WITH A MICROBE AND WHAT SHE WAS ABLE TO SHOW IS WHEN SHE LOOKED AT THE SITE THIS HAD VERY STRONG ANTI-VALVE SIGNATURE SO IT LOOKED LIKE THE TISSUE WAS INFECTED BY A VIRUS WHICH OF COURSE WAS SURPRISING. THERE WAS NO INFLAMMATION AND IT WAS A ASSOCIATION AND EVERYTHING AND WAS REMINDED BY THE WORK AND I REMEMBER YEARS AGO HIM TALKING ABOUT THIS IDEA OF CROSS TALK BETWEEN MICRO AND THE VIRUSES AND THIS ALWAYS STAYED IN THE BACK OF MY MIND AND I'M SURE I'M CONTINUING TO BE INSPIRED BY HIS WORK FOR YEARS TO COME. WHAT WE ALL KNOW IS THE FACT THAT THE HUMAN GENOME, 10% IS INDIGENOUS RETRO VIRUSES THAT ARE REMNANT OF INFECTION THAT HAVE EMBEDDED THEMSELVES IN THE GENOME AND MECHANISMS AND SOME OF THEM ALTHOUGH FRAGMENTED HAVE MEAN TRANSCRIPTIONAL ACTIVITY AND INFERENCE HOST PHYSIOLOGY SO FOR EXAMPLE THE PLACENTAL DEVELOPMENT THE FACT THAT WE HAVE A PLACENTA WAS ACQUIRED FROM THE ACQUISITION WHICH I FOUND INCREDIBLE SO THE FACT WE HAVE THESE ANTI-VALVE SIGNATURE IN THE CONTEXT OF THE EXPOSURE TO THE MICROBE POINTED IT COULD BE AN INTERACTION BETWEEN THE VIRUS AND THE MACRO BUY THAT ISEN DODGE US IN AND THIS WAS A WORK THAT WAS DONE IN THE LABORATORY AND TEAM UP TO ISEN DODGE US IN AND THIS WAS A WORK THAT WAS DONE IN TH >> IF YOU PULL THE CONTROL MICE THEY HAVE FEW TEE CELLS AND THEY ACCUMULATE IN THE SKIN AND THE TREATMENT WAS HAS A SIGNIFICANT IMPACT ON THE ABILITY OF THE T-CELL TO BE GENERATED. AND THIS CAN BE VISUALIZED BETTER AND BY IMAGING WHICH I THINK IS SOMETHING WE HAVE LEARNED FROM RON AND IF YOU LOOKED IN THE MICE THAT IS THERE'S AN ASSOCIATION AND ACCUMULATION OF THE T CELLS IN THE EPI DERMIS AND INTRODUCED BY TREATMENTS SO THIS REALLY SURPRISINGLY POINTED TO THE IDEA THAT TRANSCRIPT IS ACTIVITY THE HOST AND IT WAS PLAYING A FUNDAMENTAL ROLE IN THE ABILITY OF THE SYSTEM TO DEVELOP T-CELL RESPONSE TO A NEW MICROBE SO TO MAKE A LONG STORY SHORT. THEY CROSSED DIFFERENT CELL LINEAGE TO SEE WHICH CELL WILL BE REALLY THE IMPORTANT ONE FOR THIS RELATIONSHIP AND I'M SHOWING YOU THE ONE THAT OF COURSE WORKED WHICH IS. HE WAS ABLE TO SHOW IS THAT IF YOU ELIMINATE EXPRESSION THAT IS OF COURSE THE MISSION AR SEE REQUIRED FOR DNA SENSING IN THE SITE OWE PLASMA, IN THIS CONTEXT IS YOU SEE THE ACCUMULATION OF T CELLS REDUCED WITH A STRONG REDUCTION IF YOU HAVE ASSOCIATION OF IN ABSENCE OF THE SITE AND VERY SIMILAR WE HAVE A SHUT DOWN OF MATE CELL RESPONSE WE HAVE SHOWN FOR INDUCED BY THE MACRO BIO TIN AND THE ABOLISH THAT IS RESPONSE TO THE MICRO SO THE CARA TIN IS REALLY ARE THE ONES ABLE TO SENSE THOSE ELEMENTS AND WE ENGAGE IN THE HUMAN SYSTEM. WE HAVE RECEPTORS FOR THIS RESPONSE AND DOWNSTREAM TO MAKE TO BE FULLY UNDERSTOOD AND THERE'S A CLASS O ERV AND EXPRES IT IN CLASS AND I THINK THIS IS POORLY UNDERSTOOD. THESE ELEMENTS CANNOT BE REVERS TRANCE DESCRIBED AND CREATE A MINI HOTSPOT IN THE TISSUE THAT ARE NOT STRONG ENOUGH TO CREATE INFLAMMATION BUT STRONG ENOUGH TO ACTUALLY VERY DISCRETE WAY ENGAGE THE LOCAL CELLS RESPONSE AND OTHER CELLS ALLOWING THE INITIATION OF THE IMMUNE RESPONSE IN A PHYSIOLOGICAL WAY SO WE THINK THAT THAT PROCESS CAN PLAY AN IMPORTANT ROLE AND WE HAVE EVIDENCE THAT THIS MAY NOT BE RESTRICTED TO THE SKIN BUT PLAY A IMPORTANT ROLE IN OTHER TISSUE SUCH AS THE LONG AND THE G.I. TRACK. SO, WHAT THEY STUMBLED INTO AND IT WAS REALLY A FUNDAMENTAL AND THE COMMUNITY AND YOU CAN ABSOLUTELY FASCINATED BY THE RELATION BETWEEN AND THE REASON HE IS INTERESTED BY THESE AND IS WE THINK ABOUT IT AS A PATHOGEN AND IT'S A COMMON SOUL THAT LIVE AT A SURFACE WITH THE SKIN WITHOUT CAUSING INFLAMMATION OR TISSUE DAMAGE SO IT WAS PRESENT IN MICE AND IT'S A CORRELATION AND WHAT THEY POSTURE LATED BECAUSE OF THE ABILITY OF THIS TO IT PROMOTES TISSUE FOR ACCIDENT DAMAGE AND THEY GENERATE TOGETHER WITH THE TOOLS TO BE ABLE TO TRACK AND WHEN YOU TRANSFER AND ASSOCIATED THERE'S A STRONG RESPONSE THAT IS USE AND NOTHING SURPRISING AND IT'S CLASS FOR RESPONSE AND WHAT WAS SURPRISING IS TO SHOW TO STAFF TEND ARE CLOSE IT TEE TO SENSORY NEURON THAT CAN BE IN THE SKIM VISUALIZED BY THE EXPRESSION AND THIS IS WAY CLOSER FROM THE SENSORY NEURON AND THAT THEY WERE ABLE TO SHOW AND IF YOU HAVE HAVE STUFF AND AFTER INJURY AND THAT CAN BE ACTUALLY SEEN HERE AND YOU CAN SEE THAT IF YOU HAVE A CONTROL THEY ARE THE EDGE OF THE WOUND AND YOU HAVE MORE OF THOSE AND THIS IS HIGHLY DENSE FEED IF YOU HAVE PRIOR TO THE WIND INDEX AND CAN BE QUANTIFIED ON THE RIGHT SO HAVING THE HIGH GROBE PROMOTES THE NEURON AND IT COULD BE THE HIGH CROW SO WHAT MICHELLE DID IS UTILIZE A APPROACH WHERE HE DELETED AND FROM THE T CELLS THAT THE CELLS AFTER THEY HAVE ENTERED IN THE SKIN AND YOU CAN SEE HERE THAT IF YOU HAVE AN ANIMAL THAT IS NAIVE THEY HAVE A GENERATION RING IF YOU HAVE ANIMAL THAT IS CONTROL ANIMAL THAT EXPRESS OUR ON THE T CELLS THEY HAVE A MASSIVE ACCELERATION BUT YOU CAN SEE THE ANIMALS ARE TH17 LOOK OUT THIS IS REDUCE SOD DESPITE THE FACT THAT THERE'S 17 COMES FOR MANY DIFFERENT PLACES, THE ONLY RELEVANT SOURCE APPEAR TO BE THE T CELLS GENERATED IN THE CONTEXT OF THE ASSOCIATION. THE NEXT SPOT TO UNDERSTAND WHERE THIS WAS WORKING AND WHAT HE WAS ABLE TO SHOW IS THAT ACTUALLY NEURON THEMSELF NEEDED TO SENSE IL17 AND IF BOTH OF THE MICE CONTROL AND THERE'S A MASSIVE REAGAIN RACE IN CONTROL AND THESE ARE SENSORY NEURON AND YOU CAN SEE THE EXPRESS AND YOU SEE APPLY THE NEURON CAN NO LONGER RESPOND TO AND THIS IS SEVERELY DECREASED AND THIS IS VISUALIZED AND THIS IS QUANTIFIED ON THE RIGHT SO THEY PROPOSE THAT AND RESPONSE TO A SEEMS TO ALSO HAVE THE ABILITY TO PROMOTE TO THE DIRECTING INTERACTION AND WITH IL17 SO THE MODEL THAT MICHELLE IS PROPOSING AT THE MOMENT IS A FOLLOWING ONE. AND IT'S REALLY DEFAULT RESPONSIVE MOST AND THESE CALLS ACCUMULATE IN THE TISSUE AND THEY ARE IN CLOSE PROXIMITY AND WORK IN THE LABORATORY FROM LOOKING AT THIS OTHER PART OF THE EQUATION AND IN THE CONTEXT OF A BREACH WHAT YOU HAVE NOW IS ENHANCED ACCESSIBILITY OF ANTIGEN BECAUSE THEY LEAVE AT THE SURFACE OF THE SKIN AND THIS INDUCED A LOCAL RESPONSE AND AND I HAVEN'T HAD TIME TO SHOW YOU BUT THE DRG UP REGULATE THE 17 RECEPTORS SO THEY HAVE THE RESPONSE AS A RESPONSE FOR TISSUE DAMAGE SO THIS RELATIONSHIP ALLOWED TO ENGAGE IN THIS COMPLEX REPAIR WHICH A DEEPER BUT IMPORTANTLY THE REGENERATION OF NEURO SAY FUNDAMENTAL ASPECT OF TISSUE BIOLOGY AND THEY COORDINATE THE REPAIR OF THE TISSUE FROM THE CLOSURE OF THE WOUNDS SO THE SENSORY CONNECTING WITH THE VASCULAR SYSTEM AND IT'S A FUNDAMENTAL IMPORTANCE AND SO THE LAST BASICALLY WHAT I HAVE SHOWN YOU TODAY AND FOLLOWING AND JUST OBSERVING WHAT THE MICRO IS TEACHING US AND WITH THE VIRUSES AND IT'S A REALLY AND BROAD SENSE AND RANGING FROM DEFENSE, REPAIR AND REGENERATION AND AS I MENTIONED BEFORE I DO THINK THAT WIRE ALL AS A COMMUNITY SCRATCHING THE SURFACE OF THE EXTRAORDINARY POWER OF THE SYSTEM TO THE MICRO IN THE CONTROL TISSUE AND SO IN THE CON AND REGENERATION OF IN THAT IS A CLINICAL ISSUE AN AND IT COULD PREVENT THIS REGENERATION OF NEURO THAT MAY CONTRIBUTE TO NEURO PATHING BUT MORE IMPORTANTLY AND LIKE ALAN, I DO BELIEVE THAT BY OBSERVING MICROBES, YOU CAN GET EX STORY DANE' INSIGHTS INTO THE LOGIC OF TISSUE AND THAT'S WHY I LOVE THIS FIELD AND WHY AT HEART, I REMAIN HERE AND THE LAST SLIDE, I HAVE TO THANK ALL THE FANTASTIC PEOPLE THAT HAVE DONE THE WORK AND I MENTIONED THEM ALONG THE WAY. NICOLE A HAS BEEN IN MY LAB FOR YEARS. A LOT OF THE TRAINING I GOT WHEN I STARTED AT NIH THEY APPLIED TO ALAN SO THERE ARE SO MANY LAYERS OF HELP ALAN HAS DONE AND NICOLA HAS BEEN ONE OF THEM AND ALL THE PEOPLE HAVE PRESENTED THE WORK AND WE HAVE FANTASTIC COLLABORATION WITH GEORGE, EYE SACK AND MANY PEOPLE WHO HAVE HELPED US ALONG THE WAY. AND I WANT TO FINISH ON THESE SLIDES. AND I AM WITH CHRIS HUNTER AND MANY PEOPLE ARE HERE AND SOME AS TRAINING AND I DIDN'T REALIZE THEM, CHRIS KARP WAS A TRAINING THERE AND ALAN AND I WERE TEACHING TOGETHER. THANK YOU VERY MUCH FOR YOUR ATTENTION AND ONCE AGAIN, THANK YOU, VERY MUCH, ALAN. [APPLAUSE] >> YASMIN, BOTH HALVES OF YOUR TALK WERE FANTASTIC. I HAVE A QUESTION AND A COMMENT. IS THERE SOMETHING ABOUT THE PATHOGENIC STAFF ARIUS IMPORTANT FOYER NEURO REGENERATION THAT STAFF EP DETERMINE ADVERTISE A . >> AS I MENTIONED BEFORE, FIRST OF ALL IT'S NOT A PATHOGEN PER SE IT'S A CONTEXTUAL PATHOGEN AND THERE'S STRAINS AND FOR THE MOST PART IT'S A COMMENCE SUL AND SO WE HAVE A NEUTRALIZED SOME THAT EXPRESS HIGH LEVELS OF TOX THAT COULD INTERFERE WITH THIS PROCESS AND THE STAFFERS WE UTILIZE IT DOESN'T CAUSE INFLAMMATION BUT YOUR POINT IS CORRECT, MICROBE THAT COULD CAUSE INFLAMMATION COULD INTERFERE WITH THIS PROCESS AND INFECTION WITH A VERY SAME MICROBE MAY NOT PREVENT THE REGENERATION PROCESS AND IT'S SOMETHING THAT MICHEL IS EXPLORING AT THE MOMENT BUT THEY'RE DIVERSE AND THEY CAN HANG WITH US WITHOUT CAUSING INFLAMMATION AND THE VAST MAJORITY OF MICROBE THAT CAN CAN DISEASE START IN THE G.I. TRACK, LUNG AND SKIN. THEY HAVE TO PREPARE THE HOST FOR SUBSEQUENT DAMAGE. >> IT SEEMS EVOLUTIONARILY SURPRISING THAT YOU WOULD NEED A PATHOGENIC MICROBE TO PROMOTE SENSORY NERVE REGENERATION. >> I DON AGAIN I DON'T AGREE, TE ARE FEW MICROBE THAT ARE REALLY PATH OWE GOWNS MOST OF THE MICROBES ARE COMMON SILLS. I JUST FEEL LIKE IT'S A RARE OCCURRENCE IN BIOLOGY. OF COURSE, THE LAST COUPLE OF YEARS MAY HAVE PROVEN US WRONG. [LAUGHTER] THEY USED TO SAY THAT. >> THE COMMENT IS, I WONDER IF THIS HAS ANY BEARING ON WHICH PEOPLE INFECTED WITH SARS-CoV-2 DEVELOP NERVE DAMAGE AND THE SPEED OF REGENERATION BECAUSE OBVIOUSLY, THE ORO IS MICROBES AND -- THAT COULD EFFECT MAYBE NOT THE DAMAGE DONE BY THE VIRUS ITSELF BUT THE RATE OF REGENERATION. >> YOU ARE ABSOLUTELY CORRECT. I'M NOT GOING TO SPEAK ABOUT THE HUMAN BUT IF YOU THINK ABOUT WILD ANIMAL AND WILD MICE FOR EXAMPLE WILL HAVE BY DEFAULT WAY MORE IL17 IN THEIR TISSUE. WE HAVE STERILIZED OUR ENVIRONMENT AND WE HAVE DEVELOPED COEXISTENCE WITH HIGH ESCROWS THAT ARE DOING THE JOB BUT DIFFERENTLY NOT AND YOU ARE ABSOLUTELY CORRECT AND WE PROBLEM IN A SITUATION OF FAILURE TO REPAIR FOR NUMEROUS REASONS, HAVING THE WRONG MICROBE AND PUT US IN A SITUATION WHEN THE ABILITY TO REPAIR IN A PHYSIOLOGICAL WAY IS NOT OPTIMAL AND IT'S FOR THE LUNG AND NASAL CAVITY. AND THIS MICROBE HELPED TO REGENERATE AND IF WE DON'T HAVE THE TONE OF THE IMMUNE SYSTEM IT'S GOING TO HAVE CONSEQUENCES. [APPLAUSE] >> THANK YOU. >> SO, OUR NEXT SPEAKER IS ISABEL OSWALD AND ISABEL WORKS WITH BOTH ALAN AND STEFANI BACK IN 1990 TO 1994. AFTER HER STAY AT NIH, ISABEL TURNED EVER RETURNED TO FRANCE AND CLIMBED THE RANKS IS NOW A RESEARCH DIRECTOR AND HEAD OF THE JOINT RESEARCH UNIT. SHE'S SINCE LEAVING THE NIH ISABEL WORKED ON TOXINS PRODUCED BY -- THE TITLE OF HER TALK IS "THE FOOD EXPOSOME, A NEW FRONTIER IN RESEARCH." >> THANK YOU, A LOT. I'M HAPPY TO BE HERE WITH ALL PEOPLE THAT I KNOW AND IT WAS 30 YEARS AGO. I WORK TOX A LIMB I WAS RECRUITD FOR INRAE AND TO DEVELOP RESEARCH ON MY CO TOXINS SO I TRIED TO GET AS MUCH AS POSSIBLE FROM WHAT I LEARNED WITH ALAN AND STEPHANIE AND WHAT I'M GOINO PRESENT MORE TODAY IS WHAT THE EFFECT AND DESTRUCTION AS MIXTURE AND IN THE CONTEXT OF THE EXPOSOME IN 2019, I P.M. BECOME THE LEAD OF THE LAB AND DOING MORE AND MORE PAPERWORK THAN SCIENCE UNFORTUNATELY BUT THE WORK THAT I'VE BEEN DOING WAS FAIRLY SUCCESSFUL AND I GET IT'S BASICALLY AS INDICATED BY THE NAME COX INPRODUCED BY FUNGI AND THERE ARE A LOT OF DIFFERENT TOXINS AND SO WITH DIFFERENT CHEMICAL STRUCTURE AND WHAT IS ALSO VERY WORRYING IS THAT THERE CHEMICALS STRUCTURE AND THEIR PROPERTY ARE CONSERVED DURING BOTH STORAGE AND PROCESSING COOKING THUS ISSUE UNFORTUNATELY I'VE CONTAMINATED THE FINER MATERIAL WILL BE ALSO CONTAMINATED WITH THOSE TOXINS. AND THIS IS A VERY OLD PROBLEM AND IT HAS BEEN SAID THAT EVEN THE DECLINE OF THE BUE TO THIS COX TIN BUTOXIN BUT NO ONE WILL DEMONSTRATE THAT. MORE SERIOUSLY, IT'S BEEN DETECTED AND IT'S KNOWN IN THE MIDDLE AGE AND SENT UNDER FIRES IS KNOWN TO BE DUE TO THIS REICH ROW TOXIN AND HERE IS A STABLE OF THE PAINTING, THE BEGGAR AND IT'S DO YOU TO MICRO TOXIN PRODUCED BY CLAVICEPS. IT HAS A CURE AND WE HAD SOME TROUBLE IN THE IT'S NOT SURE ANYMORE AND THERE HAVE RECENTLY IN THE 50s IN FRANCE IT HAS BEEN A PROBLEM OF STORIES CALLED THE IT CAME FROM BREAD AND IT'S KNOWN TO BE DUE TO THIS KIND OF TOXIN. SO, TODAY'S THIS IS A GLOBAL THREAT BECAUSE WE KNOW THAT EVERYWHERE IN THE GLOBE, UNFORTUNATELY, FUNGI BE ABLE TO GROW AND ABLE TO PRODUCE TOXINS BUT DEPENDING ON THE PLACE ON THE EARTH IT WON'T BE THE SAME FUNGI AND NOT BE THE SAME TOXIN. SO 60% TO THE 70% OF THE RAW MATERIAL ARE CONTAMINATED, BECAUSE I'M WORKING IN A AGRICULTURE INSTITUTE, MANY ARE WORKING ON PIGS BECAUSE IT'S EXPOSED TO THIS TOXIN BECAUSE OF ITS CEREAL RICH DIET BECAUSE IT'S SIMILAR TO HUMAN. IT HAS EFFECT ON RIVER, KIDNEY AND I'M MAINLY INTERESTED IN INTESTINAL EFFECT ON THE IMMUNE SYSTEM AND ESPECIALLY INTERESTED BY ONE MICRO TOXIN. IT'S INTRODUCE BY GROWING AND IT CONTAMINATE HUMAN AND ANIMAL FOOD FOR BOTH WHEAT AND IT'S STABLE DURING STORAGE AND COMMON TOXIC TECHNOLOGY TREATMENTS OF SUCH AS EATING, MILLING AND EXPRESSING USING COMMONLY TO MAKE FOOD. AND IT'S RAPIDLY METABOLIZED AND IT'S TOX SIT TEE IS DO YOU TO REDUCED INTAKE AND VOMITING AND AN IMMUNE RESPONSE AND DECREASE BARRIER FUNCTION OF THE INTESTINE AND THE MECHANISM ACTION IS NOT THAT IT INTER FIERCE WITH THE RYE BOW SOME AND PROTEIN SEN THAT SIS AND ACT ON THE NERVOUS SYSTEM. SO, I'VE BEEN WORKING ON THIS TOXIN FOR 20 YEARS BUT THEN, IT IS -- WE REALIZE THAT WE NEEDED TO GO NOT JUST TO WITHIN MYCO TOXIN AND THE MOTION OF EXPOSOME IT WAS INTRODUCED BY CHRIS WHITE WAS AT THIS TIME DIRECTOR. REALLY A LOT TO STUDY THE SURROUND THE IMPACT THE EARTH AND HAVE BEEN INTERESTED ESPECIALLY FOR HUMAN AND ANIMAL. AND ALSO WITH PATHOLOGY AND I'M JUST GOING TO GIVE YOU SOME EXAMPLES. WE KNOW IT'S A REALITY BECAUSE UNFORTUNATELY IT CAN BE CONTAMINATED AND OUR THIS IS ON A SURVEY IT SHOWS THAT 71% GET MORE THAN ONE TOXIN SO THIS IS WHEN YOU DO A MIXTURE AND IF YOU WANT TO DETERMINE THE IT'S TO HAVE A TWO-STEP APPROACH. THE FIRST TO DO IS A DOSE-EFFECT RELATIONSHIP WITH BOTH TOXIN AND THIS WILL ALLOW YOU TO PREDICT THE COMBINATION AND THIS IS JUST AS AN EXAMPLE THAT DEPENDING ON THE DOSE RESPONSE, YOU MIGHT NOT HAVE THE SAME RESPONSE AND YOU CAN ONLY QUALIFY THE INTERACTION AND IT'S ADDRESS BEING MIKE OWE TOXIN INTERACTION AND I WOULD SAY DIRECTION ARE DIFFICULT TO INTERPRET AND YOU CANNOT QUALIFY THE INTERACTION BECAUSE YOU DON'T HAVE A DOSE RESPONSE. AND ONE WAY TO DO THAT IS TO USE SOME THINGS THAT ARE USED FOR DRUGS AND BASICALLY THE ICE OWE BOLOGRAM OR THE APPROACH CALLED COMBINATION INDEX, THAT INDICATES WHAT IS A TYPEIN OFN T IS BASICALLY WHAT YOU ARE DOING IS AND IT'S YOUR TWO TOXIN AND AND YOU PLOT ON ONE GRAPH WHAT DOSE YOU HAVE FOR ONE TOXIN AND THE OTHER TOXIN AND THE MIXTURE AND IT'S BECAUSE YOU HAVE A SYNERGY AND IT'S ABOVE THE LINE IS BECAUSE YOU HAVE ANNAN TAG NISM AND THAT CAN YOU DO FOR TWO AND FOR THREE IF YOU HAVE A THREE DIMENSIONAL AND WITH A MATHEMATICAL APPROACH IS A COMPLEX CALCULATION AND THE RESPONSE IS QUITE EASY AND IF YOU HAVE A RESPONSE THAT IS ONE, IT'S AGITATED AND IT'S BELOW ONE IT ABOVE ONE IT'S ANTAGONISTIC. WHAT IS MORE WORRYING IS IT THAT AT LOW DOSES, WHAT YOU ARE OBSERVE IS A SYNERGISTIC EFFORT AND LOW DOSES OF ANY CONTAMINATE IS WHAT YOU ARE TRYING TO REACH. SO, THIS IS EXPERIMENT THAT WE DID WITH DIFFERENT TOXIN WITH HUMAN INTESTINAL CELL AND BASICALLY WHAT WE FIND THAT AT LOW CONCENTRATION IT'S A MAIN TYPE OF INTERACTION OBSERVED BETWEEN THIS TOXIN AND THAT IS IS WHAT IS DONE ALSO WHEN LOOKING AT THE INFLAMMATORY RESPONSE AND HERE, WE LOOK A LITTLE BIT MORE AND IT MEANS THAT WE HAVE TWO AND THEY NEED TO HAVE 22 TIMELESS AND KITTING YOU CAN HAVE SYNERGY WITH A HIGH IAMPATTITUDE SO THERE'S NOT MUCS YOU KNOW, THERE'S NOT JUST MICRO TOXIN IN THE FOOD, WE ALSO LOOK AT THE INTERACTION BETWEEN MYCOTOXIN AND HEAVY METAL AND AS EXAMPLES, WE TAKE THE INTERACTION BETWEEN MYCOTOXINS AND HEAVY METALS AND ESPECIALLY CAN BE WIDESPREAD IN THE AND WE LOOK AT THE INTERACTION IN DIFFERENT ORGANS. THE FIRST THING WE LOOK AT WAS THE DIFFERENT RATIO AND BETWEEN THE DIFFERENT RATIO OF THESE TWO CONTAMINATES WE BASICALLY HAVE THE SAME RESPONSE WHICH WAS ENCOURAGING BUT WITH WE LOOK AT THE DIFFERENT CELL TYPES, HERE WE HAVE MAINLY INTERESTING LINE AND WE HAVE ALSO ANNAN TAG NISM BUT NOT IN THE SAME RANGE. BUT WHEN WE LOOK AT BLOOD OR LIVER CELL, WE HAD SINNER SO MAKING THE STORY EVEN MORE CAMP INDICATED THAT THE INTERACTION CAN ALSO DEPEND ON THE ORGAN AND THE LAST EXAMPLE IS SOME EXPERIMENT THAT I DID WITH HIGH ROMYCOTOXIN AND ERIC MY HUSBANDS WORK ON E. COLI SO WE WORK TOGETHER ON THIS TOXIN. SOME E. COLI ARE ABLE TO PRODUCE "TOXIN CALLED COLLIE BACK TIN AND THIS IS CAN CARRY THAT IN TS INTESTINE SO WHAT WE DID WAS JUST EXPOSING CELL TO BOTH MIKE CO TOXIN, PRESENT AND THIS SEE COL EYE PRODUCING COLIBACTIN AND WE LOOK AT THE GENERAL TOXICITY AND BASICALLY WHAT YOU SEE IS THAT WE DIDN'T HAVE ANY TOXICITY WITH JUST' COL EYE AND MYCOTOXINS AND WE HAD THEM TOGETHER WE HAVE AN IMPORTANT GINO TOX SIT TEE AND THAT WAS NOT SEEN ONLY IN VOTE ROW AND ALSO IN VIVO. THEY WERE WITH CONTAMINATED THERE'S A CLEAR EXACERBATION SO WE CONCLUDE THAT DIET, CONTAMINATED WITH THIS MICRO TOXIN AND EXACERBATED TOX SIS TIGHT AND AND THIS IS JUST A CONCLUSION SO BASICALLY, WHEN WE HAVE THIS MYCOTOXIN IN OUR FOOD AND WE HAVE THIS TOXIN IN OUR INTESTINE WE INCREASE DNA DAMAGE SO THE HIGH CROW BIO TA EXPOSED TO CONTAMINATED DIET. CLIMATE CHANGE THAT HAVE THE FUNGAL FLORA AND HERE IS A SCALE OF TOXIN RISK ESPECIALLY NOW PLUS 2¡ AND PLUS 5¡ AND YOU SEE THAT IT WILL SPREAD OVER EUROPE. THE INTERACTION REQUIRES THOSE AFFECT RESPONSE FOR INDIVIDUAL COMPOUND AND MIXTURE AND FOR MYCOTOXIN IT DEPENDS ON MANY FACTORS INCLUDING RATIO AND CONCENTRATION. AT LOW CONCENTRATION, THE MAIN TYPE OF INTERACTION OBSERVED IS A SYNERGY WHICH MAY POSE A THREAT TO PUBLIC-HEALTH AS LOW CONCENTRATION WANTS TO ASSESS AND T AT RANGE REALISTIC. MYCOTOXIN INTERACT WITH OTHER CONTAMINATES. I GIVE YOU THE EXAMPLE HERE AND ARE WITH THE MICRO BIOTA SO THE TOXICITY OF FOOD CONTAMINANTS SHOULD BE CONSIDERED IN THE GLOBAL CONTEXT OF EXPOSOME. THIS IS MY TEAM AND THIS IS A POOR QUOTE BUT THIS WAS AT A BABY SHOWER. THREE WEEKS LATER, WE HAD A SMALL ALEX COMING AND THROW YEARS LATER, HIS SISTER WAS THERE AND WHAT IS NOW ALEX IS BIG, MARRIED LAST YEAR AND THIS IS HIS SISTER WITH HIS BOYFRIEND AND LAST MONTH WE ARE TWO HAPPY GRANDPARENTS. THANK YOU FOR THAT. >> IT'S A VERY LOVELY DEMONSTRATION OF THE IMPORTANCE OF THE DETAILS, THE TYPE OF CELLS AND THE CON SENSATION AND SO ON. SEEING ALL OF THOSE COMMON INTERACTIONS IT MAKE ME WONDER ABOUT THE INTERACTION AND MUCH MORE COMPLEX POSSIBILITIES WHICH I'M SURE EXIST NORMALLY. AND I WAS WONDERING IF SOME OF THE NEW DATA MINING METHODS THAT ARE ABLE TO SEE PATTENS IN LARGE BUT RELATIVELY SPARSE DATA IF YOU CAN USE THOSE TO GET AT SOME OF THE INTERACTIONS BEYOND THE BINARY AND (INAUDIBLE). >> YES. WHAT YOU SAY TO REAL QUESTION BECAUSE REALLY HERE IT WAS, I WOULD SAY EXAMPLE WITH A MINIMUM INTERACTION AND WE REALLY NEED TO DEVELOPED AND I TOTALLY AGREE WITH YOU THAT ALLOW US TO DO DO METHODS. TODAY WHAT IS MORE DONE IS THAT PEOPLE TRY TO LOOK AT EXACTLY WHAT WE ARE EXPOSED TO AND THEN LOOK WHETHER THE MIXTURE WE'RE EXPOSED TO HAVE A CONSEQUENCE BUT THEN IT HASN'T BEEN SO YET METHOD TO REALLY GO TO A ONE BY ONE COMBINATION BECAUSE THE REAL PROBLEM IS THAT WE CANNOT WORK ON AVERAGE COMBINATION. BECAUSE, EVERYONE IS INDIVIDUAL AND THE COMBINATION THAT YOU MAY EXPOS TO MIGHT NOT BE THE SAME. >> YES. >> VERY QUICKLY IT GETS OUT OF HAND WITH THE NUMBER OF POSSIBILITIES AND ALL THE CONCENTRATION EFFECT THAT'S YOU SHOWED WERE SO IMPORTANT AND THAT'S WHY I WAS WONDERING WHETHER SOME OF THE COMBINATIONS IN A MUCH MORE SPARSE WAY INSTEAD OF THE BEAUTIFUL SYSTEMATIC WAY THAT CAN BE EFFECTIVE ON SMALLER NUMBERS, IF YOU DO SOMETHING MUCH MORE SPARSE ON A LARGE-SCALE, YOU MAY BE ABLE TO MINE THAT DATA FOR SOME. >> TODAY WAS WHAT PEOPLE ARE MAINLY DOING IS TRYING TO REALLY EXACTLY KNOW TO WHAT PEOPLE ARE EXPOSED BUT IN A LEVEL TO SEE WHETHER THEY HAVE EFFECT ON THAT BUT THEN T. WITH THAT YOU ARE NOT ABLE TO CHARACTERIZE THE REAL INTERACTION BETWEEN THE DIFFERENT COMPOUND BUT I GROW. GROW.>> FASCINATING. I WAS JUST WONDERING, IF THERE ANY EVIDENCE THAT YOU CAN PRIME PROTECTION OR IS RESISTANCE OR TOLERANCE TO THE TAX INWITH A LOW DOSE, RIGHT, AND INVOKE A RESISTANCE OR A CLEARANCE OR TOX TYING MECHANISM ENDOGENOUSLY. >> THAT'S NEVER BEEN DEMONSTRATED AND I DOUBT THAT IT WILL EXIST BECAUSE REALLY, USUALLY WE ARE REALLY EXPOSED TO LOW DOSE AND EVEN (INAUDIBLE) SO, THAT HAS NEVER BEEN DEMONSTRATED. >> I MEAN LIKE, IF YOU SAY EXPOSED ANIMALS OR CELLS TO A LOW DOSE, WOULD THEY BE RESISTANT TO THE HIGHER TOXIC DOSE? >> THAT IS NOT KNOWN. CHRIS, YOU WANT TO ADD SOMETHING? >> I WAS GOING TO FOLLOW THAT BY ASKING, ONE WOULD EXPECT AN UP REGULATION OF THE RELEVANT SEEN OWE METABOLISM PATHWAYS SO THAT YOU CAN IMAGINE A BETTER WAY OF BETTER CLEARANCE AFTER A LOW DOSE EXPOSURE. >> IT'S NOT THAT POSSIBLE. [APPLAUSE] >> SO, OUR NEXT SPEAKER WILL BE SPEAK WHO ARE WILL COME WITH SOME PERSONAL REMARKS AND THAT'S Dr. RON GERMAIN BUT HE WILL TALK TO US BY ZOOM. HE JOINED IN 1998 AS HAD MANY DIFFERENT ROLES AT THE NIH. CURRENTLY CHIEF OF THE LABORATORY IMMUNE SYSTEMS BIOLOGY AND DIRECTOR OF LABORATORY SYSTEMS BIOLOGY AND OF THE CENTER FOR HUMAN IMMUNOLOGY INFLAMMATION AND IMMUNITY AND VERY SIMILAR TO ALAN, RON HAS ALSO BEEN A MENTOR FOR A LOT OF PEOPLE AND A LOT OF POSTDOCS AT THE NIH WHO HAVE GONE ON TO HAVE DISTINGUISHED POST OF OTHER UNIVERSITIES AND MEDICAL SCHOOLS SO, ARE WE READY TO GO? EXCELLENT. >> HELLO TO ALL MY FRIENDS AND COLLEAGUES IN THE AUDIENCE AND SEE I'M NOT PHYSICALLY THERE BUT MAYBE I'LL SEE YOU LATER TODAY. I WANTED TO START WITH DRAGANA'S SORT OF SCENARIO FOR ALAN'S PAST BUT A LITTLE EARLIER THAN SHE MENTIONED, SO HE GOT HIS DEGREE AT UCS D AND THERE WORKED WITH NELL COHEN AND MANY OF YOU KNOW THAT MEL WAS A FAMOUS AND INFAMOUS INDIVIDUAL IN IMMUNOLOGY AND HE HAD HAD TENDENCY TO DECIDE THE THEORY IS MORE VALUABLE IN MANY CASES THAN ACTUAL DATA. SOMEHOW ALAN ESCAPED THAT PHENOTYPE, AS WE'VE ALL HEARD IN EARLIER DISCUSSIONS TODAY AND KNOW FROM HIS WORK OVER THE YEARS. HE WENT TO MILL HILL. I'M SURE YOU REMEMBER THIS SCENEMENT TO HARVARD WHICH IS WHERE WE FIRST OVERLAPPED. AS DRAGANA SAID HE LEFT A YEAR EARLIER THAN I DID AND I WAS SMART ENOUGH TO REALIZE THAT ALAN HAD MADE A GOOD CHOICE COMING TO NIH AND DECIDED TO FOLLOW ON AND JOIN HERE AND HE WAS IN BUILDING 5 AND BUILDING 4 AND BUILDING 50 AND NOW IN BUILDING 33 AND I DON'T KNOW WHERE HE IS GOING TO SHOW UP NEXT, MAYBE HE IS SETTLING THERE, NOT ABSOLUTELY CLEAR. AND ALAN AND STEPHANIE SHOWN HERE IN THIS PICTURE HAVE BEEN NOT ONLY COLLEAGUES BUT FRIENDS OF MY WIFE GALE AND I FOR MANY, MANY YEARS BECAUSE OUR CHILDREN WERE THE SAME AGE AND I'M SPARING SU ALL THE PHOTO AT PARTIES WITH CLOWNS AND SO ON THAT CHARACTERIZED OUR EARLY INTERACTIONS BUT THIS IS ONE OF THE EARLIER PHOTOS OF ALAN AND I HAVE BEEN ABLE TO BE AT SOME OF THE PREVIOUS CELEBRATIONS AND SO HERE WAS HIS 60th BIRTHDAY. AND PEOPLE SAID HE WAS STILL GOING STRONG AND I GATHER WE ALL AGREE THAT'S STILL TRUE AND SO HERE ARE SOME OF TODAY'S SPEAKERS BACK AT THAT TIME AND I'LL LEAVE IT TO YOU IDENTIFY WHO IS WHO AND WHAT HAPPENED OVER THE YEARS AND SOME MORE OF TODAY'S SPEAKERS. ALAN HAS ANOTHER FAMILY AND I DON'T KNOW IF IT'S BEEN MENTIONED BUT YOU CAN SEE HIM HERE WITH HIS TWO DAUGHTERS AND AGAIN, HE TENDS TO HAVE THOSE PICTURES WITH THE TWO OF THEM BUT OF COURSE, THE REAL PARTNER OVER THE LONGEST PERIOD OF TIME IS STEPHANIE. AND IF YOU PAID ATTENTION THERE'S SOMETHING CHARACTERISTIC OF ALL OF THEM. WHICH IS ALAN IS ALWAYS SMILING. THIS IS PERHAPS THE MOST CHARTER TICK FEATURE I CAN POINT OUT SO I TRIED TO FIGURE OUT EXACTLY WHY THIS IS THE CASE. AND I HAVE ONE GOOD PIECE OF EVIDENCE HERE OF WHAT ACTUALLY HAPPENED. THE OTHER IS, FOR THOSE OF YOU WHO DON'T KNOW WHAT THIS IS, BRAZILIAN BARBECUE WHICH IS ANOTHER THING THAT HE IS COMPLETELY ENAMORED BY. THE THIRD IS DRIVING FANCY CARS. AS YOU CAN TELL IT'S PROBABLY NOT ALAN IN THIS PARTICULAR FANCY CAR IT'S ACTUALLY ME. AND SO ALAN IN THIS CASE, I THINK I HAVE GONE ONE STEP FURTHER. AND AS I SAY, I DID GOOGLE SEARCHES TO TRY TO SEE IF I MISSED SOMETHING, JUST BY KNOWING ALAN WELL FOR A LONG PERIOD OF TIME AND YOU HAVE TO BE CAREFUL. IF YOU LOOK DOWN IN THE HIGHLIGHTED YELLOW, THIS IS ALAN AND SHERRY'S 2019 PHOTO IN FRONT OF THE TAJ MAHAL AND THAT'S NOT THAL AND CHER AND THIS IS CLOSER AND 75th BIRTHDAY AND AL AND SHER BUT IT'S NOT THE ALAN SHER. SERIOUSLY AND I HOPE THAT I HAVEN'T GONE TOO FAR OVERBOARD IN MY PRIOR COMMENTS AND THIS WAS A REALLY FANTASTIC INTERACTION THAT LEADS ME TO THE INTRODUCTION OF THE NEXT SPEAKER, CAETANO STRUCK UP A FRIENDSHIP THAT LASTED 25 YEARS AND ALSO SCIENTIFIC COLLABORATION AND I DID REALLY SEMINAL STUDY ABOUT THE ROLE OF CERTAIN DENDRID CELLS AND ALTHOUGH CAETANO WAS IN THE LAB, IT WAS THE PRODUCT OF ALAN AND CAETANO AND BUILDING ON THE THINGS THAT WE HEARD ABOUT THAT WENT ON IN ALAN'S LAB SO TO MAKE THE SEG WAY FROM ALAN TO CAETANO THIS IS HIM AT THE TIME OF THE PAPER AND AS A POSTDOC, AT MANY MEETINGS, I'VE ALWAYS WIND UP BEHIND THE CAMERAS, MANY OF YOU KNOW, SO I HAVE PICTURE OF ALAN AND CAETANO BUT I'M NEVER IN THE PICTURES. THIS IS PARALYZED MEETINGS AT THE MEETINGS. MOVING AWAY FROM ALAN TO OTHER THINGS THIS IS AN EARLY HOUSE MEETING WITH CAETANO DANCING WITH HIS WIFE TO BE, AND THEY MANAGED TO GENERATE A REALLY TERRIFIC FAMILY AND UNFORTUNATELY BECAUSE OF THE PANDEMIC I ONLY HAVE AN OLDER PHOTO OF AWFUL THEM TOGETHER AND SEVILLEIA AND ALEX, CAETANO AND THEY'VE GONE UP A BIT SINCE I DID THIS PHOTOGRAPH. HE ALWAYS HAS HIS OWN PEACEFUL SMILE, ESPECIALLY IN THE HOME ENVIRONMENT BUT EVEN AT THE OFFICE AND SO LET ME SEG WAY TO THE SCIENCE PART AND I'M REALLY ESPECIALLY PLEASED THAT HE IS MANAGED TO KEEP ON THE WEBSITE BLURB THAT HE WAS A POSTDOC IN THE LAB BUT OVER THESE 25 YEARS OF INDEPENDENT ANT WORK, HE HAS PROGRESSED TO BE THE SENIOR GROUP LEADER AND ASSISTANT RESEARCH DIRECTOR AT THE NEW FRANCE CRICK INSTITUTE AND UNIVERSITY COLLEGE, LONDON AND KINGS COLLEGE AND HIS TERRIFIC CONTRIBUTIONS HAVE BEEN RECOGNIZED BY AWARDS AND I JUST HIGHLIGHT TWO OF THEM HERE AND PERHAPS MOST HIS ELECTION TO THE ROYAL SOCIETY IN 2019. TODAY HE IS GOING TO TALK TO US ABOUT HIS FAVORITE CELL FOR SURE. SO CAETANO, THE FLOOR IS YOURS. [APPLAUSE] >> THANK YOU. VERY MUCH FOR THAT. THANK YOU RON IFOR THAT INTRODUI HONORRAL IN THIS WAY. IT HAS TO BE SAID THAT ALAN'S INFLUENCES ARE EVERYWHERE AS YOU HAVE HEARD AND I LANDED YESTERDAY IN DALLAS AND THE FIRST THING I DID WHEN I CLEARED PASSPORT CONTROL WAS TO CALL FOR AN UBER. THIS IS THE UBER THAT SHOWED UP. [LAUGHTER] AND I THOUGHT, THIS REALLY HAS TO BE A GOOD OMEN AND IT JUST SHOWS THAT HIS INFLUENCE EXTENDS FAR BEYOND SCIENCE. IT ALSO TELLS US I SUPPOSE OF THE ANSWER TO THAT ONE THAT ALWAYS ASKS, WHICH IS, IF YOU WERE A CAR WHICH ONE WOULD HE BE. WINE WAS VERY KIND TO PRODUCE ME AND POINT OUT THAT HE IS ALWAYS BEHIND AND NEVER IN ANY PICTURE BUT HERE HE IS ALAN AND RON AND ONE OF THE PICTURES IT HAS CULMINATED THE BEGINNING OF A LONG AND MEANINGFUL RELATIONSHIP. I'M VERY FORTUNATE TO BE FRIENDS WITH ALAN AND TO HAVE KNOWN HIM FOR MANY YEARS AND AS ALREADY BEEN MENTION HERE A FEW TIMES, ALAN SORT OF ALWAYS REMAINS YOUR FRIEND AND ALWAYS WILL PROMOTE YOU AND SO HE DID. OVER MANY YEARS HE HAS BEEN A STRONG ADVOCATE FOR ME AND MY RESEARCH AND IN PARTICULAR AT THE GUNG OF MY CAREER AND HE WAS THANKS TO ALAN AND OTHER FRIENDS THAT I WAS INVITED TO GIVE SEMINARS AND TO PROMOTE ME AND ALSO, HE OF COURSE BECAME A FRIEND OF THE FAMILY SO HE IS A LITTLE BIT OF A THIS IS A VIDEO AND YOUR MICROPHONE MIGHT PICK IT UP. YOU CAN HEAR, I HOPE. YOU CAN'T HEAR I'M AFRAID BUT YOU WOULD HAVE HEARD ALAN PRACTICING GRANDFATHERHOOD BY READING TO MY KIDS AND A BOOK ABOUT THE GRINCH AND IN FACT WHEN MY DAUGHTER HAS SLIGHTLY FEARFUL EXPRESSION MOVING AWAY FROM HIM AND ALAN IS GOING RAAAAWWW! >> HOW DID WE ACTUALLY STRIKE UP OUR FRIENDSHIP AND SCIENTIFIC COLLABORATION? AND IN FACT, ALAN MAY NOT REMEMBER THIS, IT ALL STARTED WITH A CONVERSATION ABOUT IL12 AND A LONG TIME AGO AT THE KEYSTONE MEETING, AND ANNE HAD GIVEN A TALK WHICH SHE DESCRIBED HER WORK SHOWING IN RESPONSE TO LISTERIA MACROPHAGES WILL PRODUCE IL12 DRIVING TH ONE RESPONSES AND I REMEMBER AFTER THAT, I WAS STANDING BEHIND ALAN AT THE LUNCH QUEUE AND WE STARTED TO TALK AND I WAS SAYING, PERSPECTIVE THAT IT WOULD MAKE SENSE FOR THE DENDRIDIC CELLS TO AND NOT JUST MACROPHAGES IN ORDER TO DRIVE TH1 RESPONSES AND ALAN SAID YES, AND YOU SHOULD TROY IT WITH TOXOPLASMA AND THERE'S A LOT OF IL12 AND NOTHING MUCH CAME OUT OF THAT PARTICULAR CONVERSATION ALTHOUGH IT WAS IN THE BACK OF MY MIND AND A FEW YEARS LATER, BACK HERE AT NIH, WE HAD SIMILAR CONVERSATIONS ABOUT THE NOTION THAT DENDRIDIC CELLS MIGHT RESPONSE IL12 IN RESPONSE TO MICROBIAL CONTACT AND IT WAS ALREADY KNOWN FROM THE WORK OF ANNE AND OTHERS THE CELLS DID PRODUCE IL-12 BUT THERE WAS CONCEPT A LOT WAS DRIVEN BY T CELLS AND THINGS LIKE CD40 AND THE CONNECTION WITH MICROBES WAS NOT EXPLICITLY MADE NECESSARILY AND SO ALAN SAYS WE CAN USE STAG AND SO WE DESIGNED SOME EXPERIMENTS AND WHICH THEY WERE ACTUALLY COLLABORATIVE NOT ONLY INTELLECTUALLY BUT PRACTICALLY. ALAN WOULD COME IN THE MORNING AND DO INTRAVENOUS INJECTIONS OF MASS WITH STAG AND I WOULD COME IN HIS LAB LATER AT DIFFERENT TIME POINTS AND HARVEST THE SPLEANDIT LED TO A PAPER THAT RN MENTIONED WHICH MADE A FEW IMPORTANT POINTS IN RETROSPECT. THE FIRST ONE IT SHOWED THAT YOU COULD JUST STAIN TISSUE SECTION WHICH WAS A CRAZY PROPOSAL AT THE TIME BUT IT DID¡ AND YOU CAN SEE THAT THIS IS THE AREA OF THE SLOAN. THE OTHER THING THAT IS THIS IS IN VIVO AND THIS IS THE DENDRID I CAN CELLS AND I WILL MENTION IT IN A MINUTE. IT LED TO A CONCEPT THAT DENDRIDIC CELLS CAN INTERACTION WITH MICROBES AND DECODE SIGNALS FROM MICROBES IN ORDER TO BE ACTIVATED AND OBVIOUSLY CYTOKINE PRODUCTION IS ONE OF THE FEATURES OF THAT ACTIVATION PROGRAM AND REMEMBER, THIS IS BEFORE THE TIME WHEN WE ACTUALLY KNEW ABOUT THE MOLECULAR DETAILS OF PAT END RECOGNITION BUT IT DID CRATE THESE WERE INTERESTING CELLS OF THE IMMUNE SYSTEM IN WHICH TO GO AND LOOK FOR PATENT COG I GUESS AND AND IF HAS TO DO WITH DENDRITIC CELLS AND INFECTION AND FOLLOWING THE THEME OF THIS SYMPOSIUM. SO JUST TO REMIND YOU THAT DENDRITIC CELLS ARE THE PRODUCT OF THIS DIFFERENTIATION OF PATHWAY AND THAT STARTS IN THE BONE MARROW AND GIVES RISE TO THESE TWO MAJOR FLAVORS OF DENDRITIC CELLS AND I'M GOING TO CALL CONVENTION CELLS TYPE ONE AND TYPE TWO AND ALL OF THESE EFFECTIVELY START BY BEING BORN IN THE BONE MARROW AND THEN THE PRE CURSERS OF THESE CELLS TRAVEL THROUGH THE BLOOD TO GIVE RISE TO THIS NETWORK AND WHETHER YOU ARE DEAL WITH CDC 1 OR 2s THESE ARE THE CELLS THAT WILL END UP IN THE TISSUE TO FORM THIS SURVEILLANCE NETWORK THAT WE'RE ALL SO FAMILIAR WITH AND SO I GUESS, THE POINT THAT I WOULD LIKE TO DISCUSS TODAY, IS HOW IT IS THAT YOU CREATE AND MAINTAIN THESE HOW DO YOU KNOW HOW MANY CELLS TO SEED AND MAINTAIN AND THIS REALLY I THINK BOILS DOWN TO UNDERSTANDING THAT POINT AT WHICH THE SO-CALLED PRE DENDRITIC CELL FROM THE BONE MARROW HOW THESE CELLS ARE MOBILIZED AND THEY SEED THOSE TISSUES AND OUR APPROACH TO THIS PROBLEM AND TO USE A GENETIC LYNN YANG TRACING AND METHOD AND THAT WAS AND AND PRE CURSERS HELL AND? PARTICULAR AT THE PRE DC LEVEL BUT ALSO, AT THE ONE JUST PRECEDING THAT CALL AND PRECURSOR OR CDP AND THEY EXPRESS THESE RECEPTOR CALLED THE NGL1 AND I DON'T UNDERSTAND WHY WE HAVE LOST THE SLIDES SUDDENLY. YES, I CAN SEE THEM HERE SO I KNOW MY DEVICE TOGETHER AND THERE WE GO. AND SO THE AT THIS POINT WHICH WE START TO IN WHICH THEY WANT TO PROMOTE THE COLLECT NINE DAY DRIVES THE CONVERSATION DURING AND YOU CROSS AND REPORT AND THIS EXAMPLE AND AT THE POINT WHICH THESE PRE CURSERS STARTED AND THEN YOU HAVE EXPRESSION OF CREE AND THE STOP CASSETTE AND INDELL ABLE MARKING WITH DRIVEN BY THE PROMOTER WHICH MEANS THAT THESE ARE NOW EXPRESSING TOMATO AND AS THEY LEAVE THE BONE NARROW AND THEY MOVE INTO THE BLOOD INTO TISSUES, WE CAN EFFECTIVELY DETECT THEM AND WE CAN DO THIS WITH A SINGLE FLUORESCENT PROTEIN BUT WE CAN ALSO DO THIS WITH MULTIPLE FLUORESCENT PROTEINS USING THE MILES AND REPORT OF MILES GENERATE IN WHICH YOU HAVE NOT WON BUT FOUR DIFFERENT PROTEINS AND WHY WOULD YOU WANT TO DO THAT WELL BECAUSE YOU CAN START TO ASK QUESTIONS ABOUT WHAT HAPPENS TO THESE CELLS AS THEY ENTER THE TISSUE SO HERE IS A SIMPLE QUESTION, YOU CAN HAVE A SYSTEM WHERE THESE PRE CURSERS SEED THE TISSUE AND INTER MINGLE IN WHICH CASE ALL OF THESE WOULD BE DISBURSING AND YOU CAN SEE ON THE LEFT AND I DON'T KNOW HOW TO POINT THIS AND I CAN'T POINT TO IT AND YOU COULD HAVE LOCAL EXPANSION AND OF THE PRE CURSERS AND TO GENERATE MINI CLONES OF A SINGLE COLOR. AND SO THIS WAS A QUESTION THAT WAS ADDRESSED BY MARK AND YANIC A SO I'M JUST GOING TO SHOW YOU SOME EXAMPLES OF THESE CELLS IN THE LUNGS YOU CAN SEE THEM HERE THE AUTO FLORES ENTERS TAKING AWAY HERE AND YOU CAN APPRECIATE THAT WE CAN SEE ALL OF THESE DIFFERENT INDIVIDUAL FLUORESCENT PROTEINS BUT ALSO IT LOOKS TO YOU, I HOPE, THAT THESE ARE FOUND IN CLUSTERS OFTEN OF ONE COLOR AND THIS IS PRECISELY WHAT WE CAN OBSERVE WHEN AND WE CAN LOOK AT OBSERVED SCENARIO AND VERSUS A SIMULATED SCENARIO IN WHICH WE RANDOMLY DISBURSE THE CELLS A CROSS THE TISSUE THAT WE ARE IMAGING AND IN ALL CASES THE OBSERVED SCENARIO IS SIGNIFICANTLY DIFFERENT FROM THE SIMULATE ORSEN AR YO AND INDICATING OUR EYES ARE NOT LYING AND THERE TENDS TO BE SIGNAL COLOR CLUSTERING OF THESE CELLS AND THEY ARE QUIET AND COMPARED TO WHAT YOU EXPECT DISBURSED AND THAT IS BECAUSE IT TURNS OUT THERE ARE PRE DENDRITIC CELLS THEY RETAIN CAPACITY WHICH ALLOWS THEM TO GIVE RISE TO LOCAL CELL AND THAT IS ALSO TRUE TO SOME DEGREE TO THE FULLY DIFFERENTIATED DENDRITIC CELLS AND THIS IS BEEN NOTED BY OTHERS AND WHAT HAPPENS DURING INFECTION THIS IS ALL IN THE STEADY STATE AND THE MODEL AND I'M GOING TO DISCUSS WITH YOU TODAY IS NOT A MODEL OF PARASITE INFECTIONS IT'S VIRUS INFECTION WHICH INFLUENZA A VIRUS AND SO THE FIRST THING THAT YOU SEE WHEN YOU INFECT MICE WITH THIS VIRUS, OVER TIME THE NET NUMBERS OF DENDRITIC CELLS IN LUNGS IS INCREASING BOTH CDC 1s AND CDC 2s AND REMEMBER, THERE'S A LOT OF DYNAMIC GOING ON HERE BECAUSE THERE ARE CELLS EXITING THE BUNS BUT THERE'S AFTER OVER ALL INCREASE SO I TOLD YOU ALREADY THAT YOU HAVE GOT LIMIT ABILITY OF CELLS IN THE LUNGS AND YOU CAN VISIT THAT THEY ARE SIMPLY INCREASING THEIR PROLIFERATION TO MAKE UP NUMBERS BUT IT'S NOT THE CASE. IF ANYTHING, THERE ARE PROLIFERATION THAT GOES DOWN NOT UP SO THIS NET INCREASE IN THE NUMBER OF DENDRITIC CELLS AND INFECT OF OF THE PRE CURSERS IS NOT DUE TO INCREASED PROLIFERATION AND WHAT IT IS DUE TO IS INCREASED INFLUX OF THE PRE DCs COMING FROM THE BONE MARROW AND MOOING INTO THE INFECTED LUNGS AND IN FACT, UNINFECTED LUNGS, THERE ARE VERY FEW PRE DCs AND IT'S A TRANSIENT STAGE AND THEY AND THEY FULLY DIFFERENTIATE THE CELLS AND BUT YOU CAN SEE THAT THESE NUMBERS NOW INCREASE AND IF YOU LOOK AT WHAT IS GOING ON YOU CAN SEE A TRANSIENT DIP OF THE CELLS IN THE BONE MARROW AND YOU CAN ALSO SEE THE APPEARANCE OF THESE CELLS IN THE BLOOD AND THEY BASICALLY IS CONSISTENT WITH A NOTION THAT WHAT YOU ARE DOING IS ACTUAL LOW MOBILIZING THESE CELLS FROM THE BONE MARROW WHERE THEY NEED TO BE AND SO WHAT I THINK THESE DATA SUGGESTS IS IN THE STEADY STATE MUCH OF THE DENDRITIC NETWORK IN TISSUE AND I SHOWED YOU EXAMPLES FOR THE LUNG BUT THE SAME THING IS TRUE IN THE GUT AND WE'RE THINKING OTHER TISSUES BUT MUCH OF THIS NETWORK IS MAINTAINED AS MINI CLONES OF SISTER CELLS AND THAT DIVIDE THE LOCALLY AND I DIDN'T SHOW YOU THIS DATA BUT EACH ONE OF THESE CLONES IS EITHER A CBC ONE OR IMPLIES THAT THESE PRE CURSERS THAT ARE COMING IN ARE ALREADY COMMITTED THEY'RE NOT POTENTIAL THEY HAVE DECIDED WHETHER THEY'RE GOING TO BE DC1s OR DC2s. THIS IS ALSO SHOWING US THAT THERE IS AN INCREASE THAT CAN TAK PLACE WHEN THERE'S A CHALLENGE AND INSULT AND THERE'S THE DEMAND FOR THESE DENDRITIC CELLS AND IT'S NOT MET BY LOCAL PROLIFERATION BEING INCREASED BUT EFFECTIVELY SENDING SIGNALS ALL THE WAY BACK TO THE BONE MARROW AND CAUSING A INFLUX INTO TISSUE AND THESE ARE NOT MONOCYTES SO WE ARE VERY CARRYINGFULLY MY TOMORROW INOLE GOUGH AND FROM MONO SID I CAN THERE'S SUCH A TERM TO DECREE CELL THAT'S NOT WHAT WE'RE TALKING ABOUT AND HERE AND THESE ARE BONA FIDE DENDRITIC CELLS THEY ARE COMMITTED TO THOSE LINEAGES SO THIS IS VERY INTERESTING BUT DOES IT ACTUAL LOW MAKE ANY DIFFERENCE AND DOES IT ACTUAL LOW MATTER AND SO, THE MODEL THEY SATURDAYED IT IT AND HE WAS CONTINUED BY CARLOS AND MAIRIANNA ACOSTA. SO THIS IS JUST A RECAP OF WHAT I TOLD YOU WITH SOME SLIGHTLY MORE DETAILED CONNETICS SHOWING YOU IN AN INFECTED LUNG AND CAREFULLY THE PRE CDCs COMMITTED TO THE CDC 1 OR CDC 2 LYNN YANG AND YOU CAN SEE ALL OF THEM GO UP AND YOU CAN APPRECIATE AND PERHAPS YOU ALREADY APPRECIATE THAT THERE IS MOUSE TO MOUSE VARIATION AND YOU CAN GET THESE DIFFERENCE AND SEE THEY BECOME STATISTICALLY SIGNIFICANT OVER TIME. NOW THIS IS LOOKING AT THE ENTIRE LUNG AND OF COURSE, THE INFECTION IS PATCHY AND IF YOU INFECT WITH A VIRUS YOU HAVE LOCALIZED IF YOU LOOK AND HERE WE ARE USING A GROWN DYE AND I'LL TELL YOU IN A MINUTE HOW WE DID THAT TO IDENTIFY THOSE CELLS AND IN ORANGE AND WE HAVE A KWAN TA TATING VIRAL PROTEIN AND WE SEE A REMARKABLE A LOCALIZATION OF THESE RECRUITED AND THESE INFECTION AND SO NOT ONLY THESE CELLS ARE GOING TO THE LUNG AND BY THE WAY THAT OR NOT GOING ELSEWHERE SO IF WE LOOK IN INFECT OF MICE WE DO SEE AN INCREASE OF DENDRITIC CELLS IN THE SPLEEN OR OTHER ORGANS THEY'RE GOING TO THE LUNG BUT WITHIN THE LUNG THEY'RE GOING VERY SPECIFICALLY TO THE SIDE OF INFECTION AND THIS IS CONSISTENT IN REMINISCENT OF A CHEMOKINES DRIVEN PROCESS AND SO YOU CAN START TO LOOK AT WHAT THE RECEPTORS ARE EXPRESSED IN PRE BCs AND THIS IS NOW UNINFECTED IN BON MARROW LOOK INTERESTING RAC DATA AT ALL OTHER RECEPTORS AND IN THE ONE THAT IMMEDIATE PLEA AND CAN YOU SEE IT'S HIGHLY EXPRESSED IN NON MONO SIT I CAN CELLS AND NOW THERE'S NOT MUCH IN TERMS OF LIGAMENT THIS IS THE LUNG AND THEY'RE VERY RAPIDLY GOING UP AFTER AND YOU CAN SEE THAT HERE THIS IS KWAN TATING THEM BY RTPCR AND HERE WE'RE ALSO KWAN TATING TWO OF THE CCL TO SO, CONDITIONALLY ABLATING IN THIS RECURSOR WAS ACHIEVED BY MAKING USE OF THIS MILES SO IF E HAVE THE GENOTYPES RIGHT THERE, IF YOU HAVE ONE ALLELE DELETED YOU CAN HAVE GFP EXPRESSION BUT THIS IS SUFFICIENT SO YOU DON'T ACTUALLY LOSE CCR2 AND THEY WILL REPORT NOW ON PRE DCs AND WITH GFP AND IT'S IN FACT THE REPORT WE USED IN THE IMAGES I USED A LITTLE WHILE AGO IN LUNG BUT IF WE HAVE NOW TWO ALLELES WE GENERATE A NULL MOUSE AND THIS IS USING OUR CROW MOUSE TO SPECIFICALLY TARGET THESE RECURSEERS AND EVEN THOUGH YOU HAVE INCOMPLETE PEN A TANTS OF THIS EVENT, YOU CAN SEE SOMETHING INTERESTING. IF YOU LOOK IN UNINFECTED MICE, AND THE ONES ON THE LEFT, IT MAKES NO DIFFERENCE. HAVING CCR2 OR NOT DOES NOT CHANGE THE LOW NUMBER OF PRE CDCs THAT YOU HAVE AND THE OVER ALL NUMBER WHEN YOU LOOK AT PRE CDC 1s OR 2s BUT IF YOU NOW LOOK SPECIFICALLY AT THE INCREASE THAT IS BROUGHT ABOUT BY THE INFECTION, THEN YOU CAN SEE THAT IN THE MICE IN THE PRE CDCs AND YOU'VE GOT FEWER OF THESE CELLS COMING INTO THE LUNGS AND IF YOU LOOK AT THAT LOCALIZATION AT THE FULL SITE OF INFECTION AND HERE WE SORT OF ARE DOING THIS BY A STATISTICAL DISTANCE METRICS AND WHAT YOU CAN BASICALLY SEE IS THAT YOU LOSE SOME OF THAT CO LOCALIZATION SO IT'S LESS CO LOCALIZATION AND IT'S WHAT IS QUANTITATIVE ON THE RIGHT HYSTE GRAMS AND OF COURSE, IF YOU LOOK AT THAT THE CDCs IN THE LUNG, THEY ARE NOT ELECTRICKED IN THE MICE SO IN THE STUDY STATE NUMBERS OF CDC IN THE LUNG AND ARE EXACTLY THE SAME AND HOWEVER, THE INCREASE IN CDCs THAT YOU SEE AS A RESULT OF INFECTION NET INCREASE THAT IS REDUCED. NOW, THE IDEA OF GETTING AND IN THE RUNNING IT'S NOT NECESSARY THEY'RE GOING TO DO WHAT THEY NEED TO DO IN AND YOU ALL KNOW IT'S TO DO THEIR JOB OF LOCALLY ACQUIRING INFORMATION SO THEY CAN TRANSPORT THAT VIA THE LIMP INTO THE TRAINING NODES WHERE THEY CAN NOW PRIME YOUR T CELLS AND WHAT YOU SEE AND YOU LOCK AT THE HIGH GRA TORI POLL THAT IS ARRIVING IN THE CLUB AND INFECTION AND THERE'S NO DIFFERENCE IN THE OVER ALL NUMBERS IN UNINFECTED MICE AND THERE'S A REDUCE IN THESE HIGH GRA TORI DENDRITIC CELLS WHICH ARE ANTIGEN LADEN AND HAVE SOME FROM THE LUNGS AND SO IT'S NOT ALL OR NONE EFFECT AND THERE'S A QUANTITIVE EFFECT HERE AND DOES IT HAVE FUNCTIONAL CONSEQUENCES SO IF YOU LOOK AT THE ANTIBODY RESPONSE IT MAKES NO DIFFERENCE AND IT'S EXACTLY THE SAME AND AND TOTAL TIGHTERS OR YOU LOOK AT NEUTRALIZING ANTIBODY TIGHTERS YOU DON'T SEE A DIFFERENCE AND THERE'S A SLIGHT INCREASE IN WHICH THEY HAVE BEEN ABLATED FROM THESE PRE CURSERS AND IF YOU LOCK AT THE C CELLPHONES, WHAT YOU SEE SAY QUANTITIVE DECREASE IN THIS RESPONSE AND YOU CAN SEE THAT AT THE LEVEL OF T CELLS AND ON THE LEFT AND IN THE MIDDLE PANELS AT PEP SIDES ON THE RIGHT AND CD4 TEE SHELLS AND IT'S NOT THAT THERE'S NO RESPONSE, BUT THERE IS AN OVER ALL DIMINISHMENT AND IT'S WHAT IS AND OR IN THE LUNG AND YOU CAN ALSO LOOK MORE LONG-TERM AT THE ESTABLISHMENT OF CELLS AFTER THE INFECTION HAS CLEARED SO THIS IS MEMORY CELLS AND THESE ARE TISSUE RESIDENT MEMBER ROW CELLS IN THE LUNG AND AGAIN YOU CAN SEE THAT THERE'S NOT A LOT IN THIS RESPONSE AND IN FACT THIS QUANTITATIVE DECREASING THE RESPONSE HAS IMPACT AND WE CAN SHOW THAT AND EXPERIMENT AND WE ACTUALLY NEED TO SWITCH STRAINS SO DON'T HAVE A PROBLEM WITH NEUTRALIZING AND THAT'S WHAT WE'VE DONE HERE AND SO WE HAVE CROSS PROTECTION THAT CAN ONLY BE MEDIATED BY T CELLS AND NOT ANTIBOD'S AND IF YOU JUST LOCK AT WEIGHT LOSS, WHAT YOU FIND IS THE MICE THAT HAVE LET CCR2 IN THE PRE CDCs AND TEND TO LOSE MORE WEIGHT AND THEN HAVE EFFECTIVELY HIGHER IN THEIR LUNGS SO THIS A LOUSE US TO SAY IS SOMETHING THAT IS QUITE IF RETRO TEXT BOOK SAYS. THEY'RE IN TISSUE AND THEY HAVE IMMUNE RESPONSES AND WHAT THEY'RE TELLING US IS THAT THEY ARE IN TISSUES AND THEY'RE SENT INALS BUT THE NUMBER OF CELLS AND TISSUES IS NOT SUFFICIENT TO ACTUALLY SUSTAIN IMMUNITY TO INFECTION AND YOU NEED TO GET THIS BACK UP AND YOU GET THIS BACK UP BY CALLING IT ALL THE WAY FROM THE BONE MARROW BY ENSURING THAT YOU'VE GOT AN APPROPRIATE CHEMO KEEN AGREED CN INGREDIENT AND EVEN IF YOU HAVE ALL THE CAN CELLS YOU NEEDED BUT THEY WERE DISBURSED THROUGH OUT THE LUNG AND YOU HAVE A FOCUS OF INFECTION HERE YOU CAN'T EXPECT THEM TO ALL CROSS AND ALL OF THE AN ATOMICAL STRUCTURES OF THE LUNG TOE GET HERE THE ONLY WAY THEY CAN GET THERE COMING IN VIA THE BLOOD AT THE BLOOD SO THIS IS PRESUMABLY WHY THIS SYSTEM IS WORKING AND THAT I HAVE DESCRIBED AND NOW IN THE CASE OF FLU, IT DEPENDS ON AND I DON'T WANT TO GIVE YOU THE IMPRESSION THIS IS ALWAYS THE CASE AND I'M NOT SHOWING YOU DATA BUT WE'VE DONE IT WITH A PARA DIET AND WE HAVE A STRONG LUNG DAMAGE FACE AND WE CAN SEE THERE'S AND THE DENDRITIC CELL AND AS A RESULT OF NIP OWE INFECTION AND THERE IS COMPLETELY INDEPENDENT OF CRR2 SO WE PRESUMED THERE MUST BE OTHER CHEMOKINE RECEPTORS AND I THINK THAT YOU MAY HAVE SEEN IN THAT RNA SEEK ANALYSIS THAT THERE WERE OTHER RECEPTORS THAT CAN BE EXPRESSED BY PRE DCs AND THE FINAL POINT IS, THAT THIS IS A MECHANISM WE COULD THINK ABOUT HARNESSING AND FOR BOOSTING IN PARTICULAR T-CELL RESPONSES BUT PERHAPS FOR T-CELL RESPONSES YOU COULD THINK ABOUT THE DELIVERY SYSTEMS THAT MIGHT NOT JUST RELY ON THE CELLS LOCALLY BUT ACTIVE WILL HE BRING IN CELLS TO THE SITE OF ANTIGEN DEPOSITION SO THAT'S WHAT I WANTED TO SHARE WITH YOU TODAY AND THESE ARE THE MEMBERS OF MY LAB CURRENTLY AND I MENTION THE PEOPLE WHO CONTRIBUTE THIS SPECIFIC WORK AND I PRESENTED AS A WENT ALONG AND WE HAVE QUITE A NUMBER OF COLLABORATORS AND THEY HAVE HELPED US INCLUDING RON HERE AT THE NIH AND I WANT TO LEAVE YOU WITH THIS FINAL SLIDEF A SMILING ALAN SHER AND THANK HIM FOR ALL HIS FRIENDSHIP OVER THE YEARS. [APPLAUSE] >> SIMPLE QUESTION, TO WHAT EXTENT DO YOU THINK WHAT YOU ALL EMERGENCY IS REALLY DRIVEN BY WHAT YOU ARE PROBABLY DOING AT MOST EXPERIMENTALIST GIVING A VERY LARGE DOSE OF FLU AS OPPOSED TO SORT OF THE NORM AND IT'S EXPOSURE ONE THAT AN ORGANISM MIGHT EXPECT TO A RESPIRATORY VIRUS? >> THERE'S ALWAYS A QUANTITATIVE COMPONENT TO THIS AND THE EXTENT OF THE CHALLENGE WILL BE THE EXTENT OF WHAT THIS BACK UP IS NEEDED SO FOR US TO DETECT IT, WE NEED TO TAKE IT TO THE POINT WHERE WE CAN SEE IT BUT I DON'T THINK IT WILL BE A DIFFERENCE AND IT WILL BE A QUANTITIVE DIFFERENCE AND I THINK WE ARE GOING TO SEE THAT IN A VARIETY OF MODELS AND WE DO SEE IT IN MODELS, WITH TUMOR CHALLENGES AND DIFFERENT INFECTIONS SO I THINK IT'S GOING TO TURNOUT TO BE SOMETHING THAT IS DEPENDING ON THE EXTENT OF THE NEED. >> SO, I WAS FASCINATED BY YOUR VERY FIRST STARTING POINT YOU SHOWED A MOVIE OF THE DENDRITIC CELLS IN TISSUE AND I THOUGHT YOU WERE GOING TO TALK ABOUT THE CONTROL OF THE INITIAL RESTING NUMBERS OF THE CELLS IN THE TISSUE AND IT LOOKS LIKE YOU HAD A UNIFORM SPACING WHICH WOULD MAKE SENSE AND I WONDERED IF YOU HAVE ANY THOUGHTS IS THAT CONTROLLED BY THE DENDRITES TOUCHING EACH OTHER AND REPEL OR IS THAT A SOL YOU ABLE MEDIATOR GIVING YOU A NUMBER HOMEOSTASIS AND. >> AND THE ANSWER IS WE HAVEN'T LOOKED AT IT AND BUT YOU ARE QUITE RIGHT THERE ARE DATA FOR EXAMPLE WHERE THE HEMO SIGHT NETWORK THAT FORMS IS CONTROLLED PRO SIGHSLY BY SOME REPULSIVE MECHANISM THAT ALLOWS ALL OF THE CELLS TO OCCUPY A CERTAIN TERRITORY AND I BELIEVE THAT THERE ARE SIMILAR PRINCIPLES HAPPENING AND I DON'T THINK HE HAS BEEN LOOKED AT NOT BY US AND THE CONTEXT OF HOW IT FORMS AND HOW THOSE HIGH ROW ISSUES ARE POCK I'D. THERE ARE STRATEGIES IF YOU ARE REALLY CAREFUL YOU CAN DISTINGUISH THEM AND YOU CAN EVEN FIND A TYPE OF AND AND HE ARGUES IS NOT MONOCYTE AND DERIVED AND WE FIND EXACTLY WHAT HE HAS FOUND AND HE ALSO MONITOR THE APPEARANCE OF THE CELLS IN THE LUNG SO AND WE USED A PLUS LOTS OF GENETIC MODELS INCLUDING LOOKING AND THROUGH TO ASCERTAIN THE MARKERS WE'RE USING HERE ARE DEFINING SOMETHING AND AND BY THE WAY, MONOCYTE AND THEY'RE NOT EFFECTED IN THE DELTA CCR2 AND AND I CAN TALK TO YOU ABOUT SEPARATELY ABOUT IT. >> GO AHEAD. >> ALL RIGHT. I WAS WONDERING, DO YOU KNOW WHETHER THIS CCR2 ARE DEPENDENT STEP IS IN THE EGRESS OUT OF THE BONE MARROW OR IN THE ENTRY TO THE LOUNGE. IT'S A POINT IN THE LUNG BECAUSE OF THE LOCALIZATION SO INFECT IN THE MIO SIN WHICH WE DON'T HAVE CCR2. YOU STILL HAVE SOME ENTRY ALTHOUGH NUMBERS ARE DECREASING BUT THE CELLS ARE MORE DISBURSED SO CLEARLY, IT'S PLAYING A ROLE THERE AND WE THINK IT'S PLAYING A ROLE IN BECOMING EXTRA. WE THOUGHT IT DIDN'T PLAY A ROLE IN THE EXIT FROM BONE MARROW AND WE ARE REVISITING THAT AT THE MOMENT AND WE NOW THINK THAT IT MIGHT IN THE CASE SPECIFICALLY OF THE FLEW FLU. THE MAIN ROLE IS THE ENTRY AND LOCALIZATION WITHIN THE LUNG. >> I WOULD LIKE TO GO BACK TO YOUR REGIONAL AND VERY INSPIRING AND IN USING MODERN TERMINOLOGY AND AND WE DO KNOW THAT NOT DCA2 AND THE STIMULATION AND IN THIS REGARDS AND TO ASK WHAT CELL FEATURES OF ACTIVATION IN VIVO ARE TRULY DEPEND AN ON DIRECT RECOGNITION VERSUS INDIRECTION RECOGNITION WITH TLR9 AND TLR9 DEFICIENT AND GIVING THINGS LIKE A LOT OF MIGRATION IS NOT NECESSARILY THE DEPENDENT ON DIRECT ACTIVATION AND EVEN THINGS LIKE TNF CAN DO THAT IN A MANNER AND MANY GO UP ON THE CELLS UPON ACTIVATION AND B7, ET CETERA, ALL OF THESE CAN BE DRIVEN BY THE INFLAMMATORY AND DON'T REQUIRE DIRECT STIMULATION. WHEREAS THINGS LIKE CYTOKINE PRODUCTION INCLUDING ALCOHOL DEPEND ON THE DIRECTION SIGNAL. SORRY? >> WHETHER MIGRATION OF DENDRITIC CELLS ALONE WOULD BE SUFFICIENT FOR -- ALONE? >> SO, ALONE IT'S NOT STUFF AND IN FACT, WE SHOWED MANY YEARS AGO WITH THIS MIXED RESISTANT THAT ONLY DIRECTLY ACTIVATED CELLS WERE FULLY COMPETENT TO PRIME. >> THANK YOU. >> VERY NICE. WHEN YOU LOOK AT THE NUMBER OF PRE DCs, IT'S VERY LOW AND YOU SEE A HUGE INFLUX COMING FROM THE LUNG. IS THIS CHANGING THE MDP SO THE MULTIPLE WITHIN THE BONE MARROW AND IS THIS A TRAINED IMMUNITY TYPE OF THINGS OR ARE THEY EXPANDING? >> THAT'S A GREAT QUESTION. WHAT EXACTLY IS HAPPENING UPSTREAM AND SO WHAT WE -- THIS IS INFERENCE FROM LOOKING AT NUMBERS AND CONNETICS AND OUR INFERENCE IS THERE'S A TRANSIENT DIP IN THE BONE NARROW IN THE POOL OF PVC AND A VERY RAPID REFLYNNISHMENT CALLING UP FROM UP TREATMENT AND THEY PRESUMABLY NOW FILL UP THE NICHE AND NOW THE EXACT DYNAMICS OF THAT AND WHAT IS THE EXACT UPSTREAM THAT IS FEEDING THE NICHE IN THE AND YOU KNOW ABOUT INJECTING AND SEEING A -- I DON'T REMEMBER WHAT HE CALLED IT. THE WAVE OR WHATEVER IT IS,. >> DO YOU KNOW HOW MUCH GM DEPEND ANT WHICH IS HIGHLY ABUNDANT IN THE LUNGS, SORRY? >> WE DON'T THINK IT'S DEPENDENT HOWEVER WE HAVEN'T FORMALLY SHOWN THAT AND IT MIGHT BE DEPENDENT BUT WE'RE NOT SURE. >> THIS IS JUST FOLLOWING UP ON THAT QUESTION. SO I THINK MAYBE YOU WERE ASKING OVER HERE BUT GMCSF AND YOU THINK IT'S LIE BEGAN AND THEY GET THE BONE NARROW AND MOBILIZE IT. >> SO IN THE CASE OF THE FLU THERE'S ANOTHER SIGNAL THAT IS IS DRIVING THE SYSTEM HERE AND BECAUSE ALSO THESE CCR2 SIGNIFICANTMENTS AND THE UP REGULATIONS CCR2 IS PART TYPE 1 DEPENDENT IT MAY NOT BE THE CASE FOR OTHER SYSTEMS WHICH WE STARTED. >> YOU THINK THIS IS INTERFERON LEADS TO THIS MOBILE SAYS IN THE BONE NARROW. >> IT WOULDN'T BE THE CASE IN NIPA WHERE IT'S COMPLETELY IN TYPE ONE INDEPENDENT SO THERE ARE MULTIPLE WAYS TO GET THERE AND IT'S GOING TO BE DEPENDENT ON PARTICULARLY INFECTION MODEL THAT YOU ARE USING? >> THANK YOU. >> VERY NICE TALK. THE LAB HAS SEEN THAT CCR2 IS INVOLVED IN MIGRATION OF MYELOID DERIVED SUPPRESSER CELLS INTO TISSUES SO WITH RECRUITMENT OF DENDRITIC CELLS OR PRE DENDRITIC CELLS, DO YOU SEE MYELOID DRIVE SUPPRESSER CELLS BEING ATTRACTED AS WELL? >> SO WE HAVEN'T SPECIFICALLY STARTED. >> THIS WAS IN THE CONTEXT OF INFECTION IN MONKEYS BUT I'M JUST WONDERING ABOUT THE BALANCE BETWEEN POSITIVE AND NEGATIVE DECISIONS IN TERMS OF REGULATING THE IMMUNE RESPONSE? >> >> THANK YOU. >> [APPLAUSE] >> THANK YOU. I THINK YOU OUGHT TO GET A SPECIAL PRIZE FOR THE MOST CLEVER TITLE AS WELL. BEING ABLE TO MENTION ALAN FOUR TIMES. THAT WAS VERY GOOD. SO, OVER TO OUR NEXT SPEAKER WHO IS TANYA SCHARTON-KERSTEN AND WORKING ON IT AND AFTER LEAVING THE NIH AND TANYA WENT INTO INDUSTRY AND BEEN IN DIFFERENT PLACES BOTH IN THE U.S. AS WELL AS IN EUROPE AND IN 2018 SHE STARTED HER OWN COMPANY AND WORKING ON RECOGNIZE LATORRE AFFAIRS SO OVER TO YOU, TANYA. HERE I AM, AND YEAH, I WAS PHIL SCOTT'S FIRST GRADUATES STUDENTS AND AFTER I GRADUATED FROM HIS LABORATORY, HE HANDED ME OVER TO ALAN AND I AM AN INDEPENDENT NOW AND I WORK IN REGULATORY AFFAIRS AND SO THIS WILL BE A VERY DIFFERENT TALK THAN THE ONES YOU'VE SEEN SO FAR AND I'M GOING TO SPEND SOME TIME EXPLAINING TO YOU SORT OF WHAT PEOPLE IN REGULATORY AFFAIRS DO AND WHAT ALAN AND PHIL GAVE ME IS A TREMENDOUS ABILITY TO WRITE AND I REMEMBER SO MANY TIMES SITTING IN ALAN'S OFFICE AND THIS WAS SOMETHING THAT PEOPLE ROTATED THROUGH AND IF YOU HAVE A PAPER DUE, YOU HAD TO SIT IN HIS OFFICE UNTIL IT WAS FINISHED. AND WHEN HE WAS DONE WITH ONE PERSON'S PAPER AND THE NEXT PERSON CAME IN AND SAT THERE AND EVERYONE WAS LIKE QUICK, FINISH YOUR WRITING. IT'S A SKILL I'VE USED FOR THE REST OF MY LIFE SO I'M SO GRATEFUL TO PHIL AND ALAN AND I REALLY TURNED TO DO SOMETHING THAT I'M PASSIONATE ABOUT AND I'VE CONTINUED IN THE SCIENCE AREA AS WELL AND SO, I'M GOING TO GIVE YOU MY REFLECTIONS. RIGHT NOW IN HEALTHCARE, GENE THERAPY IS A HUGE TOPIC AND USING A VIRUS INJECTING 10 TO THE 13th VIRUS PARTICLES INTO VARIABLE NERVAL PATIENTS IN ORDER TO CORRECT GENETIC DISEASES AND IT'S FASCINATING AND FROM MY PERSPECTIVE AND THIS HAS REALLY PAVED THE WAY FOR WHAT IS NOW BEING DONE AND GENE THERAPY. SO, A DISCLAIMER. I REMEMBER 30 YEARS AGO, AND PHIL'S LAB AND WE HAD THESE HUGE CARO SELLS AND YOU HAD TO GET YOUR SLIDES DONE WAY IN ADVANCE AND THEN THERE WAS THE WEEK BEFORE YOU GOT YOUR SLIDES DOWN, THEN THERE WAS THE EMERGENCY FIX. >> WE HAVE IDEAS AND THEN TAKE THOSE IDEAS AND GET THEM INTO THE CLINICS SO THAT THEY CAN TEST TO SEE WHETHER THOSE ARE ACTUALLY GOING TO WORK. AND WHAT DOES THIS MEAN CMC, CHEMISTRY MANUFACTUR MANUFACTURD CONTROL IT MIGHT SOUND BORING BUT IT'S VERY INTERESTING TO TRY TO UNDERSTAND WHAT IS THE PRODUCT AND WE HAVE A VERY SPECIFIC WAY THAT WE WRITE THESE THINGS UP AND SO WHAT IS THE PRODUCT AND HOW IS IT MANUFACTURED AND WHAT IS THE SCIENCE BEHIND IT AND WHAT ARE THE METHODS TO PROVE THE PRODUCT IS WHAT IT IS AND WHAT IT SAYS AND SO, FOR ME IT'S REALLY INTERESTING BECAUSE WE HAVE LIKE LITTLE BOXES THAT YOU FILL IN AND THERE'S THE INTERNATIONAL CONFERENCE OF HARMONIZATION AND WHAT IS HAPPENING IS THERE'S A COMMON APPLICATION SO YOU CAN DO A TRIAL IN GERMANY, YOU CAN DO A TRIAL IN JAPAN AND IN EUROPE AND IN U.S. AND EVERYBODY HAS A VERY SIMILAR APPLICATION SO THAT YOU ARE ALL SPEAKING THE SAME THING AND IT'S REALLYBALIZED THE HEALTHCARE INTERVENTIONS FIELDS. AND SO, I'M GOING TO GIVE YOU INFORMATION FROM MY VANTAGE POINT WHICH IS NOT AS SORT OF LIKE DEEPLY SCIENTIFIC AS WHAT WE'VE HEARD THIS MORNING BUT I THINK THAT EQUALLY IT'S IMPORTANT WHEN IT COMES TO DEVELOPING VACCINES AND TO THERAPIES SO IF WE LOCK AT ELEPHANT, I THINK OF MYSELF WHEN I GO INTO WORK WITH SCIENTISTS AND I'VE WORKED WITH PEOPLE ALL OVER THE WORLD AND AT HARVARD AND SCRIPPS AND HIV AND EBOLA AND COVID AND EVERY TIME I GO INTO WORK WITH THE SCIENTISTS THEY TELL ME HOW NOVEL AND INCREDIBLY EXCITING AND COMPLETELY DIFFERENT THEIR APPROACHES SO IT'S LIKE THE FRONT OF THE ELEPHANT AND THEN MY JOB IS TO SAY, NO, NO, IT'S NOT THAT DIFFERENT TO TELL THE REGULAR, THIS IS EX LOW WHAT HAS BEEN DONE BEFORE IT'S NOT DIFFERENT AT ALL AND SO LIKE NEGOTIATING >> EVERYBODY HAS A BUT. IT'S LIKE TRYING TO FIGURE OUT WHAT IS COMMON AND SORT OF LIKE IT'S A VERY INTERESTING FIELD WORKING WITH THE SCIENTISTS AND ENCOURAGING THEM AND HELPING THEM TO REALIZE OF COURSE THEIR IDEA IS NOVEL AND TELL ME ALL THE PARTS THAT AREN'T NOVEL SO I CAN RELATE IT TO SOMEONE ELSE ALREADY DONE SO WE DON'T GET HUNG UP WITH THE REGULATORS AND NOT ABLE TO MOVE FORWARD. IT'S AN INTERESTING POSITION. AND WHEN I DO MY JOB WELL, NOBODY KNOWS WHO I AM. WHOLE LIKE THIS INVISIBLE PERSON BEHIND THE SCENES AND THE CLINICAL TRIALS GET STARTED AND IF THE ONLY TIME PEOPLE SORT OF KNOW WHO I AM IS IF THINGS GO WRONG AND WE HAVE DELAYS AND WE CAN'T START THE TRIALS OR IF THEY GO ON CLINICAL HOLD. SO THE LEGACY OF VACCINES PRECEDED THE GENE THERAPY SPACE AND A WANTED TO EMPHASIZE A LITTLE BIT ABOUT YOU ALL KNOW THIS AND IF WE LOOK AT VACCINES AND INTERVENTIONS. THE VACCINES STARTED IN 1700, 1800s A VERY LONG TIME AGO AND THERE'S NOW DOZENS OF VACCINES ON THE MARKET AVAILABLE. WE ALL KNOW ABOUT THE DIFFERENT COVID VACCINES THAT CAME OUT, HOW QUICKLY THEY CAN DEVELOP AND THE DIFFERENT TECHNOLOGIES. VACCINES ARE REALLY ADVANCED. THEY'RE VERY MATURE IN TERMS OF WHAT WE CAN DO AND THE TECHNOLOGY. ON THE OTHER HAND GENE THERAPIES THERE TO CORRECT ENZYMATIC AND OTHER DEFICIENCIES AND ADENOVIRUS IS A NEW AREA OF SCIENCE AND I THINK A LOT OF THE WORK THAT'S BEEN DONE IN INFECTIOUS DISEASES HAS PAVED THE WAY AND MADE IT EASIER FOR THE GENE THERAPIES TO MOVE FORWARD. THERE ARE HUGE DIFFERENCES. WHEN YOU GO TO VACCINES GOING TO MILLIONS OF PEOPLE WITH LIKE A VAT OF YOUR VACCINE. WHEN WE TALK ABOUT GENE THERAPIES YOU ARE POSSIBLY GOING INTO THE 12 COUNTRIES WITH ONLY 50 PATIENTS. SO EACH ONE OF YOUR VACCINES IS GOING TO A FEW PEOPLE. IT'S A LITTLE BIT OF A COMPLETELY DIFFERENT MIND SET. SO THERE'S ONLY TWO GENE THERAPIES LICENSED. ONE IS LEXTERNA FOR VISION AND THE OTHER IS FOR SPINAL MUSCULOATROPHY SO A DIFFERENT FIELD THAN THE INFECTIOUS DISEASE SPACE AND ALL THE WORK COMING BEFORE IS HELPING. AGAIN, THIS INJECTING VIRUSES INTO PEOPLE AND MAKING SURE THEY HAVE THE RIGHT RESPONSE. SO IF WE MOVE FORWARD HOW DO WE SEE THIS IS PAVING THE WAY FOR GENE THERAPY INNOVATION AND WE'RE LOOKING AT HOW TO INDUCE IMMUNE RESPONSES AND HOW TO MODULATE UP AND DOWN. IN GENE THERAPY IT CAN INHIBIT LOCALIZATION OF YOUR VIRUS AND DECREASE GENE FUNCTION IF YOU HAVE AN IMMUNE RESPONSE AND KILLS THE VIRUS BEFORE IT GETS TO WHERE IT NEEDS TO BE AND CAN BE UNSAFE. CYTOKINE STORMS ARE UNSAFE IN PERSONS RECEIVING THE GENE THERAPIES AND VIRUSES. SO INTERESTINGLY THE MANUFACTURING PRACTICES BETWEEN THERAPIES ARE COMMON AND USUALLY THERE'S A HUMAN CELL LINE YOU'RE ABLE TO REPLICATE THE VIRUS IN AND GROW IT UP AND HARVEST IT AND YOU GET RID OF SOME OF THE EXTRA DNA THERE THAT'S COMING FROM YOUR HOST CELLS SO YOU DON'T CAUSE ANY KIND OF POTENTIAL CANCER EVENTS. YOU CONCENTRATE YOUR VAT OF MATERIAL THEN YOU PURIFY AND CREATE THE FINAL PRODUCT THAT'S SAFE. ALMOST THE SAME WITH A VACCINE AS GENE THERAPY. I THINK WITH ALL THE DIFFERENT MANUFACTURERS THEY HAVE TAKEN THE INFRASTRUCTURE FOR VACCINES AND ABLE TO CAPITALIZE ON THAT. THE LANDSCAPE IS THE REGULATORY ENVIRONMENT. THE REGULATORS ARE VERY FAMILIAR WITH A LOT OF THE TECHNOLOGIES USED FOR GENE THERAPIES. IT'S MAKING IT MUCH EASIER FOR THE GENE THERAPY ORGANIZATION TO MOVE FORWARD WITH CLINICAL TRIALS. THERE'S BEEN A GREAT EXAMPLE OF COLLABORATION AND INFORMATION SHARING IN THE VACCINE SPACE. THE BILL AND MELINDA GATES AND NIH FUNDED THINGS IN THE NIH SPACE AND IT'S A BIT COMPETITIVE OPPOSED TO COLLABORATIVE BUT HOPEFULLY THEY CAN LEARN FROM THE VACCINE SPACE AND THE CLINICAL INFRASTRUCTURE MADE IT EASIER TO INTRODUCE THE GENE THERAPIES. ONE OF THE MANUFACTURING PROCESSES INFLUENCED BY VACCINE DEVELOPMENT. AS I WAS SAYING, DURING VACCINE DEVELOPMENT, MANUFACTURIE IING FACILITIES ARE ABLE TO HANDLE BSR2 MATERIALS AND ABLE TO SEGREGATE THINGS AND USE SINGLE-USE TECHNOLOGIES, BIO REACTORS THAT CAN ALLOW DIFFERENT PRODUCTS TO BE HANDLE IN THE SAME FACILITY AND DISPOSED OF TO MANUFACTURE FASTER. AND THERE IS THERE'S NO CROSS-CONTAMINATION BETWEEN THE DIFFERENT PRODUCTS. SO IT'S ESTABLISHED A STRONG MANUFACTURING DEVELOPMENT TALENT POOL OF PEOPLE. SO IN THE VACCINE SPACE THERE'S BEEN A LOT OF DEVELOPMENT OF YOUNGER TALENT. WE WERE TALKING ABOUT THIS IN THIS GROUP. PEOPLE ARE COMING RIGHT OUT OF THEIR GRADUATE SCHOOL EDUCATIONS AND GOING INTO INDUSTRY. THEY HAVE A STRONG SCIENCE BACKGROUND AND THAT'S BEEN REALLY IMPORTANT FOR THE GENE THERAPY SPACE TO THRIVE AS WELL PARTICULARLY BECAUSE THERE'S SO MANY GENE THERAPY COMPANIES NOW AND THEY ALL NEED A SMALL FLEET OF PEOPLE TO GET THESE PRODUCTS INTO THE PEOPLE. AND THEN THERE'S BEEN THE VACCINES HAVE ALSO PREPARED RAW MATERIAL SUPPLIERS FOR THE INFLUX OF HIGH QUALITY MEDIAS AND BUFFERS AND GETTING RID OF ANIMAL-DERIVED PRODUCTS IN THE MEDIA BECAUSE YOU CAN'T USE THAT FOR VACCINES OR GENE THERAPIES AND THE PROTEIN AND VIRUS PURIFICATION SYSTEMS. THE ANALYTICAL METHODOLOGIES IS ALSO HELPING WITH GENE THERAPIES. HERE WE'RE DESCRIBING DIFFERENT METHODOLOGIES BEING APPLIED FOR DISCOVERY BUT THE SAME TOOLS ARE USED IN MANY WAYS FOR ALLOWING YOU TO RELEASE YOUR VACCINE PRODUCT OR YOUR GENE THERAPY. SO YOU'RE LOOKING AT PRODUCT RELATED IMPURITIES LIKE INCAPSIDS AND GLYCOSYLATED FORMS. ALL THE FORMS YOU MAY NOT WANT IN YOUR PRODUCT AND PROCESS-RELATED IMPURITIES LIKE HOST CELL DNA AND EACH ONE OF THESE REQUIRES A DIFFERENT METHOD. FOR GENE THERAPIES, OFTEN TIMES WHEN WE'RE WRITING UP OUR DOSSIERS THERE'LL BE UP TO 30 METHODS THAT HAVE TO BE DEFINED AND VALIDATED. IT'S INTENSIVE AND HEAVY. AND ADVANTVANTAGEOUS EVENTS AND THERE'S DOCUMENTS WHICH ALLOW GLOBALIZATION AND ALLOWS TO YOU HAVE 50 PATIENTS DISTRIBUTED OVER 12 COUNTRIES AND HAVE THE CLINICAL TRIAL FINISHED IN SEVERAL YEARS. ACCEPTANCE OF THE USE OF IMMORTALIZED HUMAN CELL LINES FOR MANUFACTURING PROCESSES. WE TAKE THAT FOR GRANTED BUT IT'S REALLY NEW. I WAS ABLE TO WORK WITH KYLON INTRODUCING THE FIRST FLU CELL CULTURE VACCINE AND I WAS PART OF THE GROUP THAT WENT TO TALK ABOUT WHY IT WAS OKAY TO USE THESE CELLS AND WAS INTERESTING TO HEAR THE CONCERNS OF THE REGULATORS AND NOW THAT IT'S BEEN ACCEPTED THERE'S BEEN AN EXPLOSION AND THERE'S MODELS AND THEY'RE FAMILIAR WITH THOSE. AND DEMONSTRATING THE PRODUCT AND VIRUSES WILL INDUCE THE BIOLOGICAL AFFECT THAT'S NECESSARY AND AGAIN THE COMPLEX MULTI TRIAL CLINICAL TRIAL OPERATIONS. IN THE AREA OF REGULATORY AFFAIRS THERE'S APPLICATIONS. PEOPLE ARE DEEPLY INVOLVED IN THIS AND THERE'S CELLS AND HEX CELLS USED FOR THE COVID VACCINES. LOTS OF DIFFERENT CELL SUBSTRATES THAT CAN BE USED. INTERESTINGLY IS A LOT OF THESE ARE FIBROBLASTS THAT NORMALLY GROW AND A PLATE AND IN THIS PARTICULAR SPACE FOR GENE THERAPY AND VACCINES, THE CELL LINES HAVE BEEN ADAPTED TO GROW IN SUSPENSION BECAUSE GROWING IN SUSPENSION IS MUCH EASIER TO HANDLE IN A MANUFACTURING FACILITY. WHAT IS ON THE HORIZON FOR GREEN THERAPY? THERE NEEDS TO BE AN ACCESS TO THERAPY ACROSS ALL REGIONS. AT THIS POINT MOST THE TRIALS AND GENE THERAPIES AVAILABLE ARE IN THE UNITED STATES OR EUROPE, JAPAN THERE'S VERY FEW GOING FORWARD AN AFRICA OR INDIA OR THE OTHER COUNTRIES THAT CAN DEFINITELY USE THESE TYPES OF INTERVENTIONS BUT THEY'RE NOT AVAILABLE AND TOO EXPENSIVE. THAT'S GOING TO BE NEED TO BE ADDRESSED IN THE FUTURE FOR EQUAL ACCESS. THERE NEEDS TO BE DECREASE OF THE COST OF MANUFACTURING. IF YOU THINK OF AN ENTIRE MANUFACTURING CAMPAIGN FOR 50 PATIENTS ACROSS 12 COUNTRIES IT'S EXPENSIVE TO HAVE AN INFRASTRUCTURE TO SUSTAIN THAT AND THAT ENDS UP COMING FORWARD WITH THE CHARGES TO THE HEALTH CARE PROVIDERS AND THE PATIENTS THAT RECEIVE IT AT THE END. AND DEVELOP COMMON PLATFORMS TO MAXIMIZE RESOURCE. RIGHT NOW EVERY SINGLE GENE THERAPY COMPANY HAS ITS OWN TECHNOLOGY THOUGH THEY'RE ALL WORKING INDEPENDENTLY AND CONTRIBUTES TO THE EXPENSE AND RESOURCES THAT NEED TO BE USED TO DEVELOP THE PRODUCTS. >> COVID HAS HELPED BUT THERE'S RELUCTANCE OF PEOPLE TO RECEIVE VACCINES AND I THINK WE'VE LOST SOME OF THE HUMAN AND IN THE GENE THERAPIES IT'S GONE AFTER THE HUMAN PERSONS THAT ARE DEEPLY AFFECTED BY THEIR DISEASE THERE'S A LOT OF PATIENT ADVOCACY GROUP AND THE ADVOCACY GROUPS WORK WITH THE PUBLIC AND I THINK IT WILL HAVE A KNOCK-ON EFFECT ON VACCINES. I THINK THE WAY THE GENE THERAPY CLINICAL TRIALS ARE RUN IS MUCH MORE EFFICIENT THAN WHAT'S BEING DONE IN THE VACCINES FOR NEW CLINICAL TRIALS. I THINK THAT WILL HAVE A KNOCK-ON EFFECT. AND THAT'S CHANGED WITH THE ADVENT OF GENE THERAPY AND WILL HAVE A POSITIVE EFFECT ON VACCINES. AND THERE'S MORE AND YOU'LL DEVELOP A TECHNOLOGY AND YOU'LL DO THE MINIMUM YOU NEED TO DO TO START THE CLINICAL TRIAL AND THEN ONCE YOU SEE YOUR INTERVENTION WILL BE SUCCESSFUL YOU SCALE UP OR MOVE TO A DIFFERENT FACILITY AND THERE'S A HUGE NEED TO WRITE SCIENTIFIC DOCUMENT TO DEMONSTRATE THOUGH THE PROCESS HAS CHANGED OR THE CONSTRUCT HAS CHANGED FOR YOUR TRANS GENE THAT HASN'T AFFECTED WHAT YOU PUT IN THE PHASE IS AREA. -- PHASE 1 AREA. I THINK IT WILL HELP BECAUSE IN VACCINES IT'S'S SLOW-MOVING PROFIT AND PEOPLE ARE AFRAID OF MAKING THE CHANGES AND HAVING TOO MANY REGULATORY HURDLES. I HOPE THAT WILL AFFECT THE GENE THERAPY AS WELL. OR VACCINES AS WELL. AND AFTER I LEFT THE NIH WITH PHIL OR WITH ALAN I WORKED WITH A COMPANY TO HAVE BAND-AIDS TO PUT ON THE SKIN AND GET IMMUNE RESPONSES TO TETANUS AND THE SAME COMPANY TURN INTO NOVA VAX. WHEN I HAD MY FIRST CHILD I HAD AN OFFICE THAT WAS A CONVERTED MEN'S BATHROOM AND LIKE NOVAVAX IS A MEGA CONGLOMERATE. I'VE GONE TO GLOBAL CLINICAL TRIAL SITES AND TRAVELED TO AFRICA TO LOOK AT THEIR CLINICAL TRIAL SITES AND BEEN TO INDIA AND TRAVELED ACROSS EUROPE. IT'S PROVIDED ME AN OPPORTUNITY TO SEE DIFFERENT WAYS OF DOING THINGS THAT I WOULD HAVE NEVER SEEN AND FOR THREE YEARS A RAN A REGULATORY AFFAIRS TEAM AND THAT WAS FASCINATING AND WORKED WITH THE BILL AND MELINDA GATES FOUNDATION AND I SAW HIV VACCINES AND THEN I JOINED ROCKEFELLER UNIVERSITY THE POPULATION COUNCIL FOR A SHORT STINT TO DO WOMEN'S HEALTH. DEVELOPMENT OF IUDs AND DIFFERENT TYPES OF POPULATION OR BIRTH CONTROL METHODS. THAT WAS FASCINATING. I WAS WITH NOVARTIS VACCINES AND GLAXOSMITHKLINE AND NOW WHOLE INDEPENDENT WORKING FOR MYSELF. I HAVE FOUR BOYS. THIS IS A RECENT PICTURE OF ALL OF THEM. THAT FEELS LIKE A RESEARCH EXPEDITION. THE ONE ON THE RIGHT SAY FASHION DESIGNER AND THE NEXT ONE IS STUDYING TO BE A PARAMEDIC AND THE NEXT ONE IS DEAF AND STILL TRYING TO FIGURE OUT WHAT HE'S DOING BUT AMAZING KID. I FEEL LIKE IT'S A RESEARCH ENDEA ENDEAVOR. I CAME FROM A FAMILY OF FIVE DAUGHTERS SO HAVING FOUR SONS IS JUST -- WHEN I THINK OF ALAN I THINK OF BEING IN BUILDING FOUR AND HIM COMING IN AND TALKING TO ME ABOUT LAUREN AND ALISON AND HOW THEY WOULD GET UP TO MISCHIEF AND I REMEMBER ONE PARTICULAR STORY HE SAID THAT THEY'RE WATER BILL HAD BEEN GOING UP AND REALIZED THE TWO WERE IN THERE DOING DISHES AND RUNNING THE WATER FOR HOURS WHILE ALAN AND ANY STEPHANIE WERE DOING SCIENCE AND A REMEMBER THINKING THIS COULD NEVER HAPPEN TO ME. BOY, HAS IT EVER HAPPENED TO ME IN SO MANY DIFFERENT WAYS. I THINK OF THAT STORY OFTEN ABOUT ALAN. THEN DURING COVID WE HAVE A VERY SMALL HOUSE AND WITH THOSE FOUR BOYS IT GOT TO BE QUITE UNMANAGEABLE AND I NEEDED AN OFFICE TO WORK FROM AND SO IN FINDING AN OFFICE IN OUR TOWN I LIVE IN NEW YORK NOW I GOT A STOREFRONT AT THE SAME TIME SO YOU CAN'T SEE IT BUT I HAVE A NICE OFFICE RIGHT IN THE MIDDLE AND WE MOVED MY SON DOWN STAIRS WITH FOUR SEWING MACHINES SO HE SEWS AROUND THE CLOCK AND AN ART GALLERY AND IT'S A WEIRD TIME TO BE SURROUNDED BY ART AND MY DESK IN THE MIDDLE AND MY SON DOWNSTAIR SEWING. ALAN INSPIRED ME TO DO GREAT THINGS WITH MY CAREER AND SCIENCE AND OFF THE BEATEN TRACK WITH WHAT YOU ALL ARE DOING BUT I FEEL WHOLE CONTRIBUTING TO SCIENCE AND MENTORING YOUNG STUDENTS TO GO ON TO VACCINES AS WELL. THAT'S IT. THANK YOU. >> THE LAST 15 MINUTES WE'LL ARE REFLECTIONS FROM SPEAKERS. OUR SECOND POST-DOC AND ALAN'S FIRST Ph.D. STUDENT. HE'S NOW PROFESSOR OF MICRO BIOLOGY AND IMMUNOLOGY AT THE UNIVERSITY OF PENNSYLVANIA AND VICE DEAN FOR ACADEMIC RESOURCES AND LEE HAS 2005 IN THE INTERNATIONAL PROGRAMS BRANCH AT NIAID. WHOLE NOT SURE EVERYBODY BOUGHT INTO IT BUT I DEFINITELY BOUGHT INTO IT. THAT FAMILY DID A LOT OF DIFFERENT THINGS TOGETHER. WE HAD PARTIES. VERY FAMOUS PARTIES RUNNING ALL OVER A MAILBOX LEAVING ALAN'S PARTY ONCE. ALAN WAS PRETTY FORGIVING ABOUT THINGS. HE PROBABLY SHOULDN'T HAVE LEFT ME DRIVING AT THAT POINT BUT IT WAS A DIFFERENT TIME. I WANTED TO MAKE SURE PEOPLE GOT THE THINGS THAT WERE UNIQUE TO ALAN. I WROTE DOWN SOME BUT MOST HAVE ALREADY BEEN MENTIONED. A COUPLE HAVEN'T YET AND I'LL MENTION A FEW WHO HAVE NOT. EVERY KNEW LAB MEMBER WAS BRILLIANT. IT WAS REALLY -- EVERYBODY WAS BRILLIANT. SO ED AND I WOULD LOOK AT EACH OTHER AND SAY NOT AGAIN. DAVE LINAR CAME AND WAS GOING TO DO WESTERN BLOTS. OOPH. YEAH. AND THE OTHER THING IS NO COOL RESULT NOT EVEN NECESSARILY A COOL RESULT COULD BE TOLD ALAN WHEN HE MEANDERED THROUGH THE LAP DROPPING PIPETTE TIPS BECAUSE AS SOON AS YOU TOLD HIM HE WAS IN THE HALL TELLING EVERYBODY THE RESULT WHICH YOU WEREN'T EVEN SURE WAS REPRODUC REPRODUCIBLE. WE MENTIONED ALAN IS MULTI-LINGUAL SO I'LL LEAVE THAT AND HOW WRITING PAPERS WITH ALAN WAS PAINFUL. I'LL LEAVE THAT. WE HAVEN'T MENTIONED GOING TO THE RESTAURANT WITH ALAN WAS EASY BECAUSE HE'LL ORDER FROM YOU. I CAME FROM UPSTATE PENNSYLVANIA AND THEY DON'T HAVE SUSHI IN UPSTATE PENNSYLVANIA AND HE DID TEACHING ME ABOUT CULINARY THINGS AND THAT'S ACTUALLY ONE OF THE IMPORTANT THINGS ALAN TAUGHT ME. OFFERING ADVICE. ALAN'S VERY GOOD AT OFFERING ADVICE. AND SOME UNSOLICITED. TELLING MY WIFE AFTER A CRAB PLACE THAT SHE MIGHT WANT TO GO IN THE LADIES ROOM AND CLEAN HER FINGER NAILS IS SOMETHING SHE NEVER FORGOT. I THINK WE TALKED ABOUT IT A MONTH AGO. HALLOWEEN PARTIES, ETCETERA. THOSE ARE THE KINDS OF THINGS WE THINK ABOUT WITH ALAN BUT THIS IS SUPPOSED TO BE A SYMPOSIUM CELEBRATING ALAN AND I THINK YASMINE SAID IT'S BEEN WISTFUL FOR SOME OF US IN THE SENSE THAT IT WAS SUCH A GOOD TIME WHEN WE WERE HERE. AND MANY OF THE PEOPLE THAT HAVE WORKED WITH ALAN, IT'S NOT UNIQUE ALAN HAS BEEN CRITICAL FOR THEIR CAREERS AND THAT'S CERTAINLY TRUE FOR ME. I JUST WANTED TO TALK ABOUT THREE EXAMPLES WHERE ALAN WAS CRITICAL FOR MY CAREER AND MADE ME THINK ABOUT THE SYMPOSIUM AS NOT SO MUCH ABOUT ALAN AS MAYBE ABOUT US. IT'S WHERE WOULD WE BE IF ALAN HAD NOT BEEN PART OF OUR LIVES. I CAN GET WISTFUL ABOUT THAT BUT I'LL TRY NOT TO. SO THREE THINGS I'LL MENTION SOMEWHAT BRIEFLY. I'LL MENTION BRAZIL. WHOLE GOING MENTION BIOLOGY PARASITISM AND CELLS. SO BRAZIL. IN 1983 ALAN WAS INVITED TO ST. PAOLO FOR A MEETING AND HE COULDN'T GO. THANK GOODNESS BECAUSE HE SENT ME. I'VE BEEN IN LAB TWO YEARS AND I DON'T KNOW IF I HAD ANYTHING INTELLIGENT TO SAY AT THAT MEETING. I LEARNED NOT TO GO LIKE THIS BECAUSE THAT WAS INAPPROPRIATE AND I COULDN'T HELP DOING IT EVERY TIME I WAS TALKING BUT THAT'S ANOTHER STORY. AND SO I WENT TO BRAZIL AND STARTED A RELATIONSHIP WITH COLLEAGUES IN BRAZIL AND OTHERS WHICH CONTINUED TO THIS DAY. ED GAUR AND I COLLABORATE AND HIS SON AND I COLLABORATE AND IT'S BEEN SCIENTIFICALLY ONE OF THE GREAT EXPERIENCES OF MY LIFE. ACTUALLY BOTH SCIENTIFICALLY AND PERSONALLY BUT IT WOULD NOT HAVE HAPPENED WITHOUT THE CONNECTION. AND ALAN WAS CRITICAL FOR THAT. WE'VE ALL HEARD HOW ALAN HAS A SPECIAL LOVE OF BRAZIL AND LUCKILY ENOUGH HE PASSED THAT ON TO ME. BIOLOGY OF PARASITISMS. SO ED AND MYSELF, DAVID, WE WERE LUCKY WE WERE IN ALAN'S LAB WHEN HE WAS INVOLVED IN THE IMMUNOLOGY OF THE BIOLOGY OF PARASITISMS AND WE WENT THERE AND WE WERE ASSISTANTS IN THE COURSE. IT WAS PROBABLY FIVE YEARS WE DID THAT. IT WAS QUITE A PERIOD OF TIME. THAT LED US TO MEET A LOT OF PARASITOLOGISTS AND OUT OF THAT GREW THE RELATIONSHIP OF WHAT THE PARASITOLOGY AND WHO DID WE GO TO FOR ADVICE? ALAN. DID ALAN TAKE OVER? NO. HE WAS BEHIND THE SCENES AND GAVE ED AND I THE ADVICE TO START THE MEETING. AND THE LAST THING I'LL MENTION TH1TH2 CELLS. IN MAYBE 1987 OR SO, ALAN BEFORE THE PUBLISHED PAPER FROM KIM AND BOB ON TH1 AND TH2 CELLS. HE VISITED AND WE HAD BEEN LOOKING AT T CELL CLONES AND LINES AND NOT SURE WHAT WE KNEW WHAT TO DO WITH THEM AND HE SAID THERE'S A TH AND TH2 STORIES AND LET'S LOOK AT THE CELL LINES AND THE CONVERSATION ABOUT DOING THAT WAS THE GENESIS OF THEN STUDIES WE DID WHERE WE SHOWED THAT THE TH CELLS HAD A PHENOTYPE A NOT ONLY THAT BUT IMPORTANTLY THEY COULD CONTROVERSY DETECTION AND 5:AND I LEFT IN 1989 AND TH1 AND TH2 CELLS WAS HOT AT THAT PARTICULAR TIME. ALAN LET ME TAKE THE WHOLE PROJECT WITH ME TO PENN. I CONTINUE TO WORK ON TH1 AND TH2 CELLSALAN LET ME TAKE THE W PROJECT WITH ME TO PENN. I CONTINUE TO WORK ON TH1 AND TH2 CELLS AND IT SPEAKS TO HIS GENEROSITY IN SO MANY WAYS. I WANT TO PUBLICLY THANK YOU. I WOULD NOT HAVE HAD THE LIFE, NOT TALKING CAREERS, THE LIFE I'VE HAD WITHOUT THE EXPERIENCE OF BEING ABLE TO BE YOUR POST-DOC AND I WANT TO THANK YOU PUBLICLY FOR THAT. IT'S BEEN A WONDERFUL TIME. WITH THAT I'LL STOP. THANK YOU VERY MUCH. >> SO THIS HAS BEEN A MARVELOUS DAY. WHOLE LEE HALL. AS DAVID SAX TOLD YOU I WAS ALAN'S FIRST STUDENT. IT'S ALMOST EXACTLY 45 YEARS AGO TODAY THAT I TOOK A BUS OVER TO HARVARD MEDICAL SCHOOL AND MET ALAN IN VANDERBILT HALL AND HE INTERVIEWED ME FOR A JOB IN HIS LAB. IT WAS A STUDENT JOB AND IF IT WORKED OUTSIDE GO ON AND STAY. I HAD JUST GRADUATED FROM COLLEGE. I HAD A DEGREE IN ECONOMICS. I HAD ALMOST NO SCIENCE EXPERIENCE AND DECIDED TO GO TO MEDICAL SCHOOL AND THE ONE COURSE I HAD WAS AN IMMUNOLOGY COURSE AND SAT DOWN WITH ALAN AND TOLD ME A LITTLE BIT ABOUT WHAT THEY WERE DOING AND WHEN HE SAID ONE THING MY EARS PERKED UP. I HAD AN INTEREST IN TROPICAL MEDICINE AND KNEW HOW TO SPELL IT. I THOUGHT THAT WAS A MAIN THING. SOMEWHAT TO MY SURPRISE I GOT A CALL A DAY OR TWO LATER AND ALAN SAID COME ON OVER. YOU'RE IT. I SAID WHAT AM I DOING TO DO? HE SAID TO START I THINK YOU'LL BE A SNAIL TECHNICIAN AND A THOUGHT TO MYSELF I WANT TO GO TO MEDICAL SCHOOL AND HOW IS THIS GOING TO LOOK SNAIL TECHNICIAN? WORKS AT THE PACE OF A SNAIL? IT TURNED OUT THAT WAS NOT THE CASE AND ENDED UP NOT BEING THE SNAIL POSITION BECAUSE SOMEBODY ELSE CAME TO TAKE OVER THAT RESPONSIBILITY. I GOT IN THE LAB AND ALAN AND I WERE TALKING ABOUT DIFFERENT PROJECTS. HE HAD A NUMBER OF IDEAS AND HAD JUST COME FROM RON SMITHERS LAB AND MADE THE OBSERVATION AND WE'LL DO THIS AND THAT AND DO AN IN VIVO ACQUISITION AND EX VIVO ACQUISITION AND I LEARNED FROM ALAN EVERYBODY SHOULD HAVE TWO PROJECTS. ONE SHOULD BE A SURE THING AND THE SECOND SHOULD BE A LONG SHOT. OH, OKAY. THIS IS GREAT. APPEALS TO MY ECONOMIC BACKGROUND, PORTFOLIO DIVERSE FICTION. I UNDERSTAND WHAT HE'S TALKING ABOUT HERE. WE START OFF ON THESE THINGS AND I WOULD MEET PERIODICALLY WITH ALAN AND I LEARNED ABOUT EXPERIMENTAL DESIGN AND SCIENCE FROM THAT. AND THE AMAZING THING WAS THE PROJECTS WERE MOVING ALONG. BUT NOT ALWAYS PERFECT. THE SECOND LESSON I LEARN FROM ALAN WAS THAT WELL, SOMETIMES THOSE SURE THINGS BOMB AND SOMETIMES THOSE LONG SHOTS PAYOFF? AND RELATED TO THAT, YOU SHOULD ALWAYS CELEBRATE YOUR SUCCESSES. MAKE SURE YOU HAVE THE BOTTLE OF CHAMPAGNE IN THE COLD ROOM. SOMETHING ALAN HAD LEARNED AND SO WHEN WE FOUND MHC ANTIGENS ON CHISTOSOMES WE CRACKED OVER THE CHAMPAGNE BOTTLE AND FOR YEARS I KEPT IT WITH THE LABEL WHICH READ NOT JUST H2, H2NA AND MY WIFE FINALLY GOT ME TO THROW OUT THE BOTTLE BUT I CAN'T TAKE A PICTURE OF THE LABEL TO PHOTOGRAPH SO I APOLOGIZE FOR THAT AND HAVING THAT ONE-ON-ONE TIME FROM ALAN TO LEARN HOW TO THINK ABOUT SCIENCE WAS AN IMPORTANT ASPECT. AT THE TIME THERE WAS A QUOTE FROM THOMAS KUEHNE GOING AROUND AND I WITH REFLECTED ON THIS REY FOR OTHER REASONS BUT THE QUOTE WAS THE ANSWERS YOU GET ARE DETERMINED BY THE QUESTIONS YOU ASK. AND ALAN COULD HAVE ADDED A COROLLARY TO THAT WHICH WOULD HAVE BEEN ALONG THE LINES AND THE WAY YOU ASK THEM. BECAUSE HE ALWAYS HAD CLEVER AND INSIGHTFUL WAYS OF DESIGNING EXPERIMENTS TO ANSWER QUESTIONS. BUT THERE WERE SOME OTHER ASPECTS WHICH WERE IMPORTANT AND THE FIRST WAS ASK INTERESTING QUESTIONS THAT MATTERED. AND ALAN WAS ALWAYS GOOD AT IDENTIFYING THOSE INTERESTING QUESTIONS THAT MATTERED. THE OTHER THING WAS THAT IT WAS REALLY IMPORTANT TO MAINTAIN ALTERNATIVE VIEWS. AND LOOK AT THE DATA IN DIFFERENT WAYS. AND I THINK THIS IS ACTUALLY ONE OF THE THINGS THAT HAS LED ALAN TO MULTIPLE COLLABORATIONS WITH PEOPLE. AND I THINK IT'S ALSO SOMETHING ABOUT THE WAY HE BRINGS IN PEOPLE IN TO THE LAB BECAUSE HE'S REALLY INTERESTED. HE HAS HIS VIEW BUT HE'S INTERESTED IN THE VIEW AND DISAGREEMENT IS A SIGN OF RESPECT. HE VALUES WHAT YOU THINK AND WANTS TO HEAR WHAT YOU HAVE TO SAY AND ALSO WANTS YOU TO THINK ABOUT WHAT YOU'VE DONE AND WHETHER OR NOT THAT'S THE RIGHT PERSPECTIVE. AND ALAN IS THE SORT OF CONSTANT TEACHER. I DECIDED GROWING UP IN AN AREA WHERE PUBLIC TRANSPORT WAS READILY AVAILABLE, I DIDN'T KNOW HOW TO DRIVE. I ASKED ALAN IF I CAN TAKE DRIVING LESSONS DURING LAB HOURS. ALAN SAID, SURE. AS LONG AS WORK GETS DONE TAKE DRIVING LESSONS. I THOUGHT THAT'S GREAT. THREE DAYS A WEEK I LEARNED HOW TO DRIVE AND LEARNING HOW TO DRIVE IN BOSTON IS A LIFE-THREATENING EXERCISE BUT IT WAS ACTUALLY GREAT. THEN ONE DAY WE WERE WORKING IN LATE AND ALAN GAVE ME A LIFT HOME BECAUSE I LIVED IN CAMBRIDGE NEARBY AND WE'RE DRIVING ALONG AND SUDDENLY ALAN GOES, DAMN, FLAT TIRE. LEE, YOU'RE LEARNING HOW TO DRIVE AND I'M GOING SHOW YOU HOW TO CHANGE A TIRE. SURE ENOUGH, HE GETS DOWN AND RUNNING ME THROUGH WHAT I NEED TO DO TO CHANGE A TIRE AND I'M NOT SURE I'M GOING GET A LICENSE BUT I DO KNOW HOW TO CHANGE A TIRE NOW. TO PHIL'S POINT, A LOT OF WHAT I COULD POINT AND I COULD GO ON, I HAVE 45 YEARS WORTH OF MEMORIES, A LOT OF IT HAS ALREADY BEEN SAID. I DO WANT TO REITERATE THAT MY LABORATORY EXPERIENCE IN HARVARD CHANGED MY LIFE. I DECIDED NOT TO PURSUE ECONOMICS AND INSTEAD WENT INTO BIOMEDICAL RESEARCH AND INSTEAD WENT TO MEDICAL SCHOOL AND A YEAR AFTER MEDICAL SCHOOL I GOT A CALL FROM THE DEAN'S OFFICE AND I THOUGHT OH, MY GOD, THEY'RE GOING TO THROW ME OUT. BUT THEY CALLED ME DOWN AND SAY WE UNDERSTAND YOU'VE GIVEN INTERESTING TALKS. THE TALKS I HAD GIVEN IN THE RESEARCH JOURNAL CLUB WAS LARGELY ABOUT THE WORK I HAD DONE IN ALAN AS LAB. THEY SAID IF YOU'RE INTERESTED IN RESEARCH WE HAVE AN MD Ph.D. PROGRAM AND THINK YOU SHOULD JOIN IT SO I SAID, OKAY. I HAVE TO WRAP UP HERE, GUYS. SORRY ABOUT THAT. WE'RE UNDER A THUNDERSTORM WATCH. SO ANYWAY, I CALLED ALAN AND SAID HE WAS HAPPY FOR ME AND WAS THINKING WHO TO WORK WITH AND YOU ALL THE THINGS THAT ARE IMPORTANT. I ENDED UP HAVING A GREAT RESEARCH EXPERIENCE AND WENT IN INTERN AND RESIDENCY AND CAME TO THE NIH AS A CLINICAL FELLOW AND ABLE TO RENEW MY COLLABORATION WINS ALAN AND HE INTERN SET UP COLLABORATIONS WITH ED AND LOOKED AT HISTOSOMES AND WORKED ON COMPLIMENT RECEPTOR MEDIATED UPTAKE AND MADE MANY FRIENDS SITTING IN THE AUDIENCE AND I'M GLAD TO SEE AND THEN THE OTHER THING THAT IS A TESTIMONY TO ALAN THERE WAS AN INITIATIVE AT THE TIME AND A WAS WORKING IN SOMEBODY ELSE'S LAB BUT WHEN THE TIME CAME UP, I WAS A FELLOW AND AN ENDED UP BEING A CO-P.I. ON THIS INITIATIVE WHICH WAS REALLY KIND OF EXTRAORDINARY. IT WAS GREAT BECAUSE I GOT TO GO TO BRAZIL ONE OF ALAN'S FAVORITE PLACES AND MET PEOPLE HERE AND ENDED UP COLLABORATING WITH THEM SUBSEQUENTLY. AND ALAN MENTIONED TO THE PARASITOLOGY AND TROPICAL DISEASES BRANCH I WAS LOOK FOR A JOB SO I GET A CALL FROM STEPHANIE AND SAID I HEAR YOU'RE LOOKING FOR A JOB. LET ME TELL YOU ABOUT THIS. I HAD LOOKED AT A NUMBER OF JOBS IN INDUSTRY AND LOOKED AT TROPICAL MEDICINE AND PARASITIC DISEASES AND I'VE BEEN THERE EVERY SINCE. -- EVER SINCE. THAT WAS ANOTHER TIME AS NOW ON THE EXTRAMURAL SIDE KNOWING ALAN WE WERE ABLE TO CONVERSE AND ENGAGE ON DIFFERENT TOPICS OF INTEREST. IN FACT, ALAN WAS INSTRUMENTAL IN ORGANIZE THE IL12 MEETING THAT BECAME A LARGE EFFORT BUT GEORGIO REFERRED TO IT AND THERE WERE A NUMBER OF INVESTIGATORS THAT REFER TO HOW SEMINOLE IT WAS IN WHAT THEY WERE THINKING ABOUT. I THINK WHAT WE HEARD TODAY IS ALAN IS INCREDIBLY INSIGHTFUL AND PRODUCTIVE AND HAS BEEN A PROLIFIC SCIENTIST. HE'S BEEN AN INCREDIBLE MENTOR TO MANY OF US. HE HAS BEEN A TRUSTED ADVISER SOMETIMES ON ISSUES THAT CAN BE CHALLENGING. AND HE'S A LONG TIME FRIEND AND HOPE HE CONTINUES TO STAY THAT WAY AND HOPE WE'LL CONTINUE TO BENEFIT FROM YOUR MENTORSHIP AND GUIDANCE AND WISDOM. AND NO MATTER WHAT HAPPENS I'LL STILL BE UNCLE LEE TO YOUR GIRLS. BUT I DON'T THINK THEY NEED ME TO BABYSIT ANYMORE. ANYWAY, THANK YOU VERY MUCH, ALAN. IT'S BEEN A REAL PLEASURE AND COMPLETELY CHANGED THE TRAJECTORY OF MY CAREER AND WHERE I WAS GOING. I APPRECIATE THAT. >> SO THAT CONCLUDES THIS AFTERNOON'S SESSION AND TODAY'S SESSION. I GUESS THERE IS TOMORROW AT 9:00 SAME PLACE. LOOK FORWARD TO SEEING EVERYBODY THEN.