>> GOOD AFTERNOON I'M SUSAN MOYER. WELCOME TO THE WEDNESDAY AFTERNOON LECTURE SERIES. AND TODAY WE HAVE ALSO A GUEST FOR THE INTRODUCTION. I'D LIKE TO INTRODUCE MY BOSS, DR. ANTHONY FAUCI, DIRECTOR OF NIAID, GIVING THE INTRODUCTORY REMARKS TODAY. THANK YOU AND ENJOY. >> THANK YOU VERY MUCH, SUSAN. IT'S REALLY A PLEASURE FOR ME TO INTRODUCE MAX. GOOD AFTERNOON TO ALL OF YOU. WELCOME TO THE NIH DIRECTOR'S WEDNESDAY AFTERNOON LECTURE SERIES. MAX COOPER WILL BE SPEAKING TO US ABOUT HIS INSIGHTS INTO THE EVOLUTION OF ADAPTIVE IMMUNITY, I WANT TO TELL YOU BRIEFLY ABOUT MAX AND A LITTLE BIT ABOUT HIS ACCOMPLISHMENTS. FIRST, A QUICK THUMB NAIL ABOUT HIS BACKGROUND. MANY OF YOU IN THE AUDIENCE KNOW HIM, BUT I THINK MORE PEOPLE HAVE JUST HEARD OF HIM. MAX WAS BORN IN RURAL MISSISSIPPI AND GRADUATED FROM THE UNIVERSITY OF MISSISSIPPI, AND EARNED HIS M.D. DEGREE AND COMPETED RESIDENCY IN PEDIATRICS AT TULANE, BECAME A TRAINEE AT UCSF AND ALONG THE WAY DEVELOPED A SPECIAL INTEREST IN CLINICAL IMMUNOLOGY FROM HIS EXPERIENCE CARING FOR CHILDREN WHO WERE UNUSUALLY VULNERABLE TO INFECTIONS. HE BECAME AN INSTRUCTOR IN PEDIATRICS AND TULANE AND MOVED TO THE UNIVERSITY OF MINNESOTA BEFORE SETTLING DOWN AT THE UNIVERSITY OF ALABAMA IN BIRMINGHAM BEGINNING IN 1967. AFTER SPENDING DECADES THERE, IN 2008 HE MOVED TO HIS CURRENT POSITION AS PROFESSOR IN THE DEPARTMENT OF PATHOLOGY AND LABORATORY MEDICINE AT EMORY UNIVERSITY, SCHOOL OF MEDICINE. NOW A BIT ABOUT HIS ACCOMPLISHMENTS. COULD I HAVE THE SLIDE PLEASE? BY THE ESTIMATION OF VIRTUALLY EVERY KNOWLEDGEABLE IMMUNOLOGIST WHO HAS A CORPORATE MEMORY, MAX COOPER IS CLEARLY ONE OF THE TRUE ICONIC GIANTS IN OUR FIELD. HIS EARLY WORK WAS CRISP, ELOQUENT AND TRANSFORMATIVE IN THE FIELD OF IMMUNOLOGY AND TO THIS DAY HE CONTINUES TO LEAD US. 50 YEARS AGO, AT THE UNIVERSITY OF MINNESOTA, BOB GOOD, RAY PETER AND AND MAX AND COLLEAGUE THE PUBLISHED A PAPER IN "NATURE" THAT BEGAN MAX'S REMARKABLE STRING OF RESEARCH ACCOMPLISHMENTS. HIS EXPERIMENTS WITH CHICKENS REPORTED ON IN THAT PAPER BEGAN THE ELUCIDATION OF SEPARATE T AND B-CELL LINEAGES AND IDENTIFICATION OF HEMATOPOIETIC AND THE ORIGIN OF B LINEAGE CELLS. BRIDGING THESE INSIGHTS TO THE CLINICAL ARENA, HE AND HIS COLLEAGUES HAVE FOR DECADES BEEN PERFORMING COMPARATIVE STUDIES OF LYMPHOCYTE DEVELOPMENT AND FUNCTION WITH AN AIM TO UNDERSTANDING DEFECTS IN MALIGNANCIES, IMMUNODEFICIENCY. HE WILL BE UPSET WITH ME FOR SAYING THAT BUT VIRTUALLY ANYTHING THAT ANY OF US DO IN THE FIELD OF IMMUNOLOGY AUTOIMMUNITY, HEMATOLOGIC MALIGNANCY, IMMUNODEFICIENCIES, HAS BEEN IMPACTED IN ONE WAY OR ANOTHER BY MAX'S SEMINAL WORK, ORIGINATING BACK DECADES AGO BUT CONTINUED TO THIS DAY. HOWEVER, SEMINAL RESEARCH ACCOMPLISHMENTS ARE NOT ALL THAT MAX IS ABOUT. HE HAS BEEN RECOGNIZED FOR HIS MENTORSHIP SKILLS, SERVING A A VALUED ADVISOR FOR STUDENTS, POSTDOCS, AND JUNIOR FACULTY, AND ONE OF THE REASONS THAT I'M INTRODUCING MAX IS A LITTLE BIT OF HISTORY. I FIRST MET MAX AROUND 1970, WHEN I WAS IN THE MIDDLE OF MY FELLOWSHIP HERE AT NIH. I WAS PRESENTING A POSTER AT AN ANNUAL MEETING OF THE AMERICAN ASSOCIATION OF IMMUNOLOGISTS, AND AT THAT TIME NO ONE BUT MY FAMILY KNEW WHO I WAS. MAX WAS ALREADY QUITE A FAMOUS INDIVIDUAL, AND I HAD ONLY HEARD ABOUT HIM. HE CAME OVER TO MY POSTER, AND SPENT A CONSIDERABLE AMOUNT OF TIME CHATTING WITH ME AND MAKING CONSTRUCTIVE SUGGESTIONS. I WAS AWE-STRUCK BUT HE TREATED ME LIKE A COLLEAGUE. I WAS TOTALLY ECSTATIC AND OVER THE YEARS TO MY GREAT BENEFIT WE'VE BECOME VERY CLOSE FRIENDS. I HAVE USED THAT EXPERIENCE AS MY STANDARD OF ALWAYS TREATING YOUNG SCIENTISTS AND TRAINEES AS COLLEAGUES. THEY WILL NEVER FORGET IT. I CERTAINLY DID NOT FORGET THAT FIRST DAY WITH MAX COOPER. MAX IS A MEMBER OF A NUMBER OF SOCIETIES INCLUDING AMONG OTHERS THE NATIONAL ACADEMY OF SCIENCES AND ITS INSTITUTE OF MEDICINE, AMERICAN ACADEMY OF ARTS AND SCIENCES, ASCI, FELLOW IN THE ROYAL SOCIETY OF MEDICINE, FORMER PRESIDENT OF THE AMERICAN SOCIETY OF IMMUNOLOGISTS, WON MANY AWARDS INCLUDING THE 3M LIFE SCIENCE AWARD, SANDALS PRIZE FOR IMMUNOLOGY, AND ON AND ON. HIS MOST RECENT STUDIES WHICH HE WILL DISCUSS TODAY HAVE FOCUSED ON A SEARCH FOR THE ORIGIN OF OUR ADOPTIVE IMMUNE SYSTEM AND REVEAL THE JAWED VERTEBRATES AND JAWLESS USED DIFFERENT STRATEGIES FOR GENERATING LYMPHOCYTE RECEPTORS. JOIN ME IN WELCOMING MAX COOPER TO DELIVER THE NIH DIRECTORS WALS LECTURE. [APPLAUSE] >> TONY, THANK YOU FOR THAT VERY WARM AND GENEROUS INTRODUCTION. AS YOU CAN SEE, I'VE BEEN AROUND FOR A LONG TIME, AND I'M A BIT OF A DILITANT IN RESEARCH IN THIS FIELD. I'D LIKE ALSO TO THANK SUSAN FOR HER KIND HOSPITALITY, AND FRANCIS, THANK YOU VERY MUCH, FOR THE HONOR OF PRESENTING THE NIH DIRECTORS LECTURE TODAY. NOW, AS ALL OF YOU KNOW, EVERY LIVING THING ON OUR PLANET IN THIS STRUGGLE FOR SURVIVAL HAS AN IMMUNE SYSTEM OF SOME SORT. EVEN AN ADAPTIVE IMMUNE SYSTEM AS WE KNOW FROM RECENT STUDIES OF THE CRISPR/Cas SYSTEM IN BACTERIA, AND EUKARIA AND THE RECEPTORS ELUCIDATED BEST FIRST IN INSECTS BUT IT'S ONLY IN OUR VERTEBRATE LINEAGE THAT WE HAVE A LYMPHOCYTE-BASED ADAPTIVE IMMUNE SYSTEM WITH A HUGE REPERTOIRE OF SPECIFIC RECEPTORS RECEPTORS THAT WILL ALLOW RECOGNITION OF CONFIGURATION, PATHOGEN, AND RESPOND WITH AND PRODUCE MEMORY. AND WE, BEING JAN KLEIN AND I, JUST AFTER THE TURN OF THE CENTURY, DECIDED TO JOIN FORCES TO GO BACK AND SEE IF WE COULD TRACE THE ROOTS OF OUR ADAPTIVE IMMUNE SYSTEM. WE KNEW ALL JAWED VERTEBRATES, SHARKS AND ONWARD, HAD THE SAME BASIC THYMUS, WHERE T-CELL RECEPTOR GENES WERE REARRANGED UNDER THE INFLUENCE OF RAG, CLASS 1, CLASS 2, TO PRESENT ANTIGENS, PEPTIDES TO T-CELLS, WE WOULD HAVE LIKED TO GO BACK TO SOME OF THE MORE PRIMITIVE OR EARLIER JAWED VERTEBRATES. BUT UNFORTUNATELY, THE ONLY TWO LIVING VERTEBRATE MEMBERS THAT ARE CONSIDERED TO BE MORE BASAL THAN JAWED VERTEBRATES ARE THESE TWO LAMPREY AND HAGFISH JAWLESS VERTEBRATES. NOW, WE CHOSE TO START WITH THE STUDIES OF LAMPREYS TO START WITH BUT AS I'LL MENTION WE ALSO HAVE STUDIED HAGFISH. AND WE'VE STUDIED THEIR MANY DIFFERENT KINDS OF LAMPREYS, AND THEY ARE WORLDWIDE IN DISTRIBUTION. ONE OF THE MORE THAN HALF OF WHICH ARE PREDATORY IS THE SEA LAMPREY AND WE STUDIED THEM MAINLY IN THE LARVAL STAGE WHERE THEY SPEND MOST OF THEIR LIFE BEFORE BECOMING PREDATOR, MATING AND DYING AND STARTING THE CYCLE OVER AGAIN. IF YOU CUT A LARVAE IN HALF AT 3 OR 4 YEARS OLD, ONE OF THE INITIAL FINDINGS THAT STARTED THIS ON THE STUDY WAS JAN KLEIN'S GROUP HAD FOUND A TRANSCRIPTION FACTOR THAT'S IMPORTANT IN HEMATOPOIESIS. A CELL IN THIS HEMATOPOIETIC TISSUE WAS EXPRESSING A MEMBER OF THE PU POINT ONE FAMILY, AND THE CELL WAS ROUND AND COULD HAVE BEEN A LYMPHOCYTE, SO JAN AND I DECIDED TO JOIN FORCES AND TO LOOK FOR THE ROOTS OF OUR ADAPTIVE IMMUNE SYSTEM. WE HAD SEVERAL QUESTIONS IN MIND, SOME SPECIFIC, SOME FAIRLY BROAD. FOR EXAMPLE, I WAS CURIOUS ABOUT WHICH CAME FIRST, B-CELLS OR T-CELLS? WE ALSO WANTED TO KNOW HOW AN ADAPTIVE IMMUNE SYSTEM BLENDED IN WITH THE PREEXISTING INNATE IMMUNE SYSTEM. AND WE THOUGHT IF WE COULD GET BACK CLOSER TO THE TIME OF THAT OVERLAYING, WE MIGHT GAIN MORE INSIGHT. OUR STRATEGY WAS TO JUST TAKE CELLS FROM THIS TYPE OF AREA AND BY SIMPLE 90 DEGREES, 180-DEGREE LIGHT SCATTER, PICK OUT CELLS THAT LOOKED LIKE LYMPHOCYTES WITH REGULAR BLOODSTAINS AND EVEN BY EM, HAD THIS VERY SIMPLE MORPHOLOGY OF A LYMPHOCYTE, NON-DESCRIPT, OF COURSE. WE TOOK THE CELLS AND MADE CDNA LIBRARIES AND GOT WHAT AT THAT TIME WAS HIGH THROUGHPUT SCREENING. WE FOUND MANY OF THE GENES OUR LYMPHOCYTES USE FOR ACTIVATION, MIGRATION AND SO FORTH BUT WE DIDN'T FIND ANY OF THE CARDINAL ELEMENTS OF OUR ADAPTIVE IMMUNE SYSTEM WHICH WAS DISAPPOINTING ESPECIALLY TO ME BUT WE WENT BACK AND STIMULATED THE LARVAE LAMPREY WITH A COCKTAIL AND MADE A SUBTRACTIVE LIBRARY AND PANCER MADE THIS AND WE FOUND IT WAS RICH WITH LEUCINE RICH REPEATS. A STUDENT SEQUENCED THE OPEN READING FRAMES TO SHOW SOME STRIKING FEATURES THAT MADE US RECOGNIZE THAT THIS MIGHT BE WHAT WE WERE LOOKING FOR ALTHOUGH WE HADN'T REALIZED THAT. THESE WERE THE FEATURE THAT -- THE FEATURE THAT STRUCK ONE IMMEDIATELY IS THAT EVERY ONE OF THEM HAD A DIFFERENT SEQUENCE. THEY DIFFERED IN A NUMBER OF THESE LEUCINE RICH REPEAT MODULAR UNITS AND EVEN THE ONES THEY SHARED ARE DIFFERENT SEQUENCES, ONE FROM THE OTHER. NOW, THE LONGEST ONE, HERE A STICK MODEL OF IT, IT SHOWS THAT LIKE MANY LEUCINE-RICH REPEAT PROTEINS, AND EVERY LIVING THING ON THE PROTEIN AS FAR AS I CAN RECOGNIZE USES LEUCINE-RICH REPEATS FOR VARIOUS PURPOSES, AND THESE HAD -- ALL HAD AN N TERMINAL AND C TERMINAL CAPPING VLR THAT PROTECTED THESE SOLENOID STRUCTURES, CORE HYDROPHOBIC SEQUENCES, AND THEY ALL HAD 18 RESIDUES AND CONNECTING PEPTIDE OF 13, IN BETWEEN THESE GREEN BLOCKS WERE THE ONES THAT WERE VARIABLE IN NUMBER, THEY WERE INVARIABLY 24 RESIDUES. THE STALK REGION WAS RICH AND HAD EIGHT CYSTEINES IN THIS TERMINAL, CARBOXY TERMINAL REGION, THE HYDROPHOBIC SEQUENCE HERE. IN ADDITION THOUGH, THERE WAS A PREDICTED GPI CLEAVAGE SITE, AND WHEN WE -- WHEN EARHEART EXPRESSED A CONSTRUCT FOR THIS, WITH A TAG IN MOUSE, YOU FOUND IT WAS ON THE SURFACE AND IT COULD BE CLEAVED FROM THE SURFACE BY THE ENZYMEATIC ACTION OF LIPO ENZYME, THAT WOULD CLEAVE GPI-LINKED PROTEINS OF BACTERIAL ORIGIN. NOW, WHEN WE SEQUENCE MORE OF THESE FROM MANY DIFFERENT LARVAE, AND BOTH ADULTS AS WELL AS THE LARVAE, THE SEQUENCE IS DEPICTED ON THIS CARTWHEEL PHYLOGENETIC MAP AND YOU CAN SEE FOR EACH ONE IS PAIRED WITH ITS CLOSEST RELATIVE IN TERMS OF SEQUENCE SIMILARITY, SO THIS ONE IS ABOUT 10, 20-SOMETHING RESIDUES DIFFERENT FROM THE OTHER ONE WHICH IT'S CLOSELY RELATED TO AND ALL WERE DIFFERENT EXCEPT FOR THREE PAIRS, WHICH CAME FROM THESE IMMUNOSTIMULATED LARVAE THAT I MENTIONED EARLIER. THERE'S ANOTHER FEATURE SHOWN HERE ON THIS SLIDE. THE ASTERISK INDICATES SINGLE-CELL PCRs, AND IN EACH INSTANCE ONLY ONE SEQUENCE WAS FOUND, SUGGESTING MONO ALLELIC ASSEMBLY, OR EXPRESSION. NOW, WHEN WE LOOKED WITH CHRIS IN WASHINGTON, WHO HAD A LIBRARY, GENOMIC LIBRARY MADE FROM THE SPERM OF A SINGLE LAMPREY, AND USED OUR PRIMERS TO SCREEN THIS LIBRARY, CDNA LIBRARY, WE COULD ONLY FIND ONE VLR GENE, NOT MULTIPLE, THAT WE COULD ACCOUNT FOR THE PRODUCTS WE WERE SEEING AND IT WAS VERY INCOMPLETE. IT ONLY ENCODED FOR PART OF THE N TERMINAL FOR THE LRR AND PARTS OF THE C TERMINAL, STALK REGION AND SIGNAL PEPTIDE. IN BETWEEN THERE WERE LARGE STRETCHES OF NON-CODING INTERVENING SEQUENCE, 6KB. WHEN WE GOT SEQUENCES, GENOMIC SEQUENCES OF FLANKING REGIONS WE FOUND THERE WERE LRR CODING CASSETTES, IF YOU WILL, THAT IF SOMEHOW STITCHED IN THIS GERMLINE VLR GENE WITH DELETION OF INTERVENING SEQUENCES COULD ACCOUNT ARE TO THE MATURE VLR SEQUENCES WE WERE SEEING INDEED WHEN WE LOOKED, ERYTHROCYTEV MADE THIS PRIMER, VARIABLE LYMPHOCYTE RECEPTOR GENE, WHEREAS IN LYMPHOCYTES WE ALSO FOUND SMALLER ONES, AND WHEN THESE WERE CLONED AND SEQUENCED, ALL WERE DIFFERENT JUST AS THE ONES WE HAD BEEN SEEING. NOW, WE DON'T UNDERSTAND ALL OF THE BIOCHEMICAL BASIS OF THIS ASSEMBLY PROCESS, BUT THIS SLIDE ILLUSTRATES SOME OF THE THINGS WE KNOW ABOUT IT PRESENTLY. THESE FLANKING SEQUENCES ARE BROUGHT IN AND BASED ON SHORT STRETCHES OF SEQUENCE HOMOLOGY BETWEEN DONOR AND RECIPIENT PORTION OF INCOMPLETE GERMLINE GENE, THESE SEQUENCES GET COPIED INTO THE GENE. AND REPLACING THE INTERVENING NON-CODING SEQUENCES AT THE SAME TIME. THAT PROCESS OCCURS OVER AND OVER IN A STEP-WISE AND PIECE-WISE FASHION UNTIL THERE'S COMPLETE ASSEMBLY INTO A MATURE VLR GENE. THE PROCESS CAN START ON EITHER END, BUT NOT ON BOTH, AND GO THROUGH TO COMPLETION. NOW, THE -- ALL OF THESE PIECES, AND WE DIDN'T RECOGNIZE IT SO WELL TO BEGIN WITH, ONLY BITS OF THEM ARE USED BECAUSE OF STOP CODONS AND SO FORTH. THIS PROCESS, HOW THE DNA GETS CUT, AND THE DONOR SEQUENCES ARE USED AS TEMPLATES TO BE COPIED IN BIT BY BIT AND STEP BY STEP, IS THOUGHT TO BE CATALYZED BY TWO AID-LIKE GENE PRODUCTS, THAT WERE IDENTIFIED BY ROGOZIN AND PANCER AND COLLEAGUES. ONE LOOKS VERY MUCH LIKE OUR AID GENE, THAT AS YOU KNOW IS INVOLVED IN CLASS SWITCHING, SYMPTOMATIC HYPERMUTATION AND ALSO IN GENE CONVERSION, IT'S USED BY SOME SPECIES, CHICKENS AND RABBITS, FOR EXAMPLE. THE OTHER ONE IS MORE LIKE THIS, ANOTHER ONE IS ENTRON AND COULD HAVE GOTTEN INTO THE GENOME POTENTIALLY BY RETRO TRANSCRIPTION. I'LL COME BACK TO THOSE TWO CDA 1 AND 2 AGAIN IN A MOMENT. BUT AT THAT TIME, THE MODELS SUGGESTED THAT THESE MUST BE ASSEMBLED AND EXPRESSED ON LYMPHOCYTE-LIKE CELLS, AND THEY ESTIMATED FROM SEQUENCES THAT WE PROVIDED THEM THAT THE REPERTOIRE OF THESE WOULD BE GREATER THAN 10 TO THE 14th, AT LEAST THE SAME GENERAL REPERTOIRE THAT OUR B-CELLS GENERATED BY LIGHT CHAIN, VDGA REARRANGEMENT. AND JUST TO SEE IF THOSE WERE ACTUALLY CAPABLE OF BEING MADE INTO ANTIBODIES, WE REPEATED AN EXPERIMENT THAT HAD BEEN DONE BACK IN THE '60s, BY A COUPLE GROUPS, WHEREBY THESE LAMPREY LARVAE, AND YOU CAN SEE THEY ARE SMALL, THERE ARE THE FINGERS OF THE PERSON INJECTING WITH THE INSULIN SYRINGE, YOU INJECT A SMALL AMOUNT, THE ANTIGEN IN THIS CASE WAS ERYTHROCYTE, HUMAN, O-POSITIVE ERYTHROCYTE, RESULTING IN ANIMALS MAKING GLUTENS OF A FAIRLY LOW TITER, BUT IF WE BOOSTED THEM TITER WAS INCREASED 20-FOLD. THESE AGLUTININ, BIOCHEMISTRY EVADED INVESTIGATORS WE MADE AN ANTIBODY TO THE STALK REGION OF OUR VLR AND USED IT TO ABSORB AND ALL OF THIS ANTIBODY ACTIVITY WAS REMOVED. NOW, THE CELLS THAT WERE PRODUCING THESE AGLUTININS, RECEPTORS THAT WERE SECRETED ON SHOWN HERE IN THE STAINED GREEN, IN THE SECTIONS OF GUILD FOLD, THESE BEING FILAMENTS, AND BEFORE AND AFTER IMMUNIZATION WITH ANTHRAX EXOSPORIUM, INJECTED WITH BRD U, SUGGESTING THESE RESPOND TO ANTIGEN STIMULATION WITH PROLIFERATION. THE CELLS THAT ARE ACTUALLY SECRETING THE AGLUTININS, VLR ANTIBODIES AS WE CALL THEM, BEING MADE BY CELLS LIKE PLASMA BLASTS AS SHOWN HERE, EVEN MATURE PLASMA CELLS. BRANT HERON, WHO DID WORK TO DEFINE THESE LAMPREY ANTIBODIES, USED THIS ANTI-STALK REGION ANTIBODY TO SHOW A WESTERN BLOT OF THE SERUM IS -- THESE ARE VERY LARGE MOLECULES, VARIABLE IN SIZE AS YOU MIGHT EXPECT BECAUSE THEY HAVE DIFFERENT NUMBERS OF THESE BLOCKS, BUT TREATMENT TO REDUCE THE DISULFIDE, BRANT DEVISED A WAY TO MAKE RECOMBINANT MONOCLONAL ANTIBODIES BY IMMUNIZING, MAKING CDNA LIBRARIES, EXPRESSING THEM CLONE BY CLONE, HUMAN EMBRYONIC KIDNEY CELLS AND PICKING OUT ONES THAT SECRETED THE VRL THAT COULD BIND ANTIGEN. LOOKING AT A PURE VLRB ANTIBODY WE COULD SHOW BY NEGATIVE EM THAT THEY LOOK MUCH LIKE AN IGN ANTIBODY MOLECULE EXCEPT THEY ARE ONLY MADE OF ONE KIND OF CHAIN. THEY ARE BONDED AT THEIR BASE, THE STALK REGION, BY CYSTEINES I MENTIONED EARLIER, AT LEAST FOUR OF THE EIGHT ARE REQUIRED FOR THIS BINDING AND CONFIGURATION. SO THESE ARE MULTIMERIC UNITS WITH HIGH AVIDITY EVEN WHEN THE INDIVIDUALS REACT ACTIVITIES HAVE A LOW AFFINITY FOR THE ANTIGEN. THE ONE THAT WAS SPECIFIC FOR THIS H TRISACCHARIDE, HUMAN O-POSITIVE THE DETERMINANT, HAN AND WILSON WERE ABLE TO OBTAIN CRYSTALS AND SHOW THE STRUCTURE. HERE IS THE H-TRISACCHARIDE ANTIGENS. THESE ARE HYDROGEN BONDS, TWO ON THE END, AND I HAVEN'T MENTIONED THAT THERE'S A HIGHLY RARABLE VARIABLE INVERT IN THE CARBOXY THAT FORMS A LOOP AND FIGURES STRONGLY IN THE HYDROPHOBIC BONDS TO THE H-TRISACCHARIDE. NOW, THAT LOOKS MORE LIKE THIS IN A SPACE-FILLING MOLECULE, AND ROY AND HIS COLLEAGUES WORKING WITH ZEB PANCER IS SHOWN FOR PROTEIN ANTIGENS, SO HAVE WE, TO A PREDOMINANT COAT PROTEIN OF ANTHRAX EXOSPORIN. THE SPECIFICITY IN THE SLIDE, RBC 36 RECOMBINANT MONOCLONAL, IT REACTS WITH EVERYTHING THAT HAS THE TWO TERMINAL SUGARS OF THIS TRISACCHARIDE IN THE PROPER CONFIGURATION OF WITH A ONE-TWO LINKAGE TO THE GLUCOSE WITH ONE-FOUR LINKAGE TO THE NEXT CARBOHYDRATE. THIS IS A GLYCAN GLYCOMIC CENTER THAT RICK CUMMINGS AND COLLEAGUES AT EMORY DESIGNED, IT HAS 611 CARBOHYDRATE MOIETIES ON HERE, IT REACTS WITH NOTHING ELSE AND HAS THE LAST TWO TERMINAL TWO TO SHOW THAT SPECIFICITY. NOW, SO AT THAT POINT, IT LOOKED AS IF THESE INCOMPLETE GENES WERE PUT TOGETHER IN THE FASHION THAT I'VE JUST ALLUDED TO, AND THE CELLS RESPONDED TO ANTIGEN, TO PROLIFERATE, DIFFERENTIATE AND SECRETE THESE ANTIBODIES. SO I THOUGHT THAT PROVED MY BIAS THAT B-CELLS CAME FIRST. HOWEVER, WHEN WE HAD LOOKED IN HAGFISH WE HAD FOUND TWO VLR GENES, THEN PANCER AND COLLEAGUES FOUND A SECOND LAMPREY GENE AND MOST RECENTLY KASAHAR AND COLLEAGUES FOUND A THIRD VLR GENE, ALL INCOMPLETE, EVEN SOME MORE INCOMPLETE THAN THE ORIGINAL VLRB GENE. AND THEY ALSO, NOT SHOWN HERE, ARE FLANKED BY DONOR LRR SEQUENCES. WHEN PANGOA AND MASA MADE THE STALK REGIONS INVARIANT FOR EACH ONE OF THESE, AND THEY DIFFER FROM EACH OTHER, SO MONOCLONAL ANTIBODIES WERE MADE THAT DISCRIMINATED BETWEEN VLRA, B AND C. AND THOSE WERE USED TO SHOW THAT EACH ONE OF THEM IS EXPRESSED BY AN INDIVIDUAL POPULATION OF LYMPHOCYTE. SO VLRBs ARE DIFFERENT FROM A, C AND B ALSO DIFFERENT, ALLOWING US TO ENUMERATE, ISOLATE AND LOOK AT FUNCTIONAL PROPERTIES. ONE OF THE STRIKING FEATURES IS THAT THE VLRAs IN RED AND Cs IN BLUE PREFERENTIALLY RESPOND TO INJECTION OF A CLASSIC T-CELL MITOGEN IN JAW VERTEBRATES WHEREAS B RESPOND VERY LITTLE. MOREOVER, WHEN WE LOOKED AT A LIMITED GENE EXPRESSION PROFILE, FOR THE As, Bs AND Cs, AS YOU CAN SEE, MANY GENES WERE SELECTIVELY OR PREFERENTIALLY EXPRESSED BY THE VLRAs AND Cs TOGETHER. NOTE TO START WITH THAT THE CDA 1 IS PREFERENTIALLY EXPRESSED BY A AND C, NOT BY B, AND CDA 2 IS PREFERENTIALLY EXPRESSED BY THE VLRB IMPLYING PERHAPS THEY ARE DIFFERENTIALLY INVOLVED IN THE ASSEMBLIES OF THESE DIFFERENT TYPES OF VARIABLE LYMPHOCYTE RECEPTORS. THE VLRAs HAVE A NUMBER OF GENES THAT ARE TRANSCRIPTION FACTORS, CHEMOTACTIC FACTORS, AND SIGNALING ELEMENTS THAT ARE USED BUT FOR T-CELL DEVELOPMENT AND FUNCTION, ON THIS SCALE, AND I WON'T READ ALL OF THEM OFF. IN ADDITION TO SOME CYTOKINES AND MIGRATION INHIBITORY FACTOR, WHEREAS THE VLRBs EXPRESSED TRANSCRIPTION FACTORS IN OTHER SIGNALING COMPONENTS AND ELEMENTS THAT ARE CHARACTERISTIC FOR MAMMALIAN B-CELLS. I'M COME BACK TO THE DIFFERENCES IN A MOMENT. SO ONE OF THE QUESTIONS THAT IMMEDIATELY WE WISHED TO ANSWER IS WHERE ARE THESE GENES ASSEMBLED? AND IN OTHER WORDS, WHERE IS A THYMUS EQUIVALENT-LIKE TISSUE FOR THESE T-CELL-LIKE CELLS THAT RESPOND PREFERENTIALLY TO PHA, THEY ALSO DON'T SECRETE THEIR PRODUCTS, THEY ALSO, AND I WON'T SHOW THE DATA, PREFERENTIALLY RESPOND WITH PROLIFERATION TO ALLO STIMULATION. AND SO WE LOOKED AT THE MOST LIKELY PLACE, TURNED OUT TO BE IN THE GILL REGION, AND THERE WERE SEVERAL CLUES THIS MIGHT BE THE CASE. WITH THOMAS BOONE, AND HIS COLLEAGUES, USING IN SITU, WE COULD SHOW FOXN1, A GENE ESSENTIAL FOR STROMAL CELL OF THE THYMUS AND MUTATED GENE IN NUDE MICE AND HUMANS AS WELL AS EXPRESSED AS A LEVEL ONLY DETECT IN THE THE TIPS OF THE GILLS AND ENDS OF THE FIRST FILAMENT. THIS WAS ALSO ONLY DETECTABLE IN THAT SITE, AND EVEN IF YOU STIMULATED WITH PHA OR IMMUNIZED IN OTHER WAYS, THIS IS THE ONLY PLACE THIS ENZYME THOUGHT PERHAPS, OR IMPLIED IN IN ASSEMBLY OF VLRA IS EXPRESSED IN THIS LOCATION. VLRA IS ALSO EXPRESSED HERE, VLRB IS NOT. IT'S NOT A VERY IMPRESSIVE TISSUE BUT THERE ARE LYMPHOCYTES IN HERE, IN CLOSE JUXTAPOSITION WITH EPITHELIAL CELLS, IN THIS ENVIRONMENT, AND WE CALL IT THE THYMOID. EACH ONE OF THE TIPS OF THE GILL FILAMENTS COMPRESSED CDA1 AND A FEW OF THE CELLS IN THIS THYMOID REGION EXPRESSED A BUT NOT B. THEY ALSO CAN EXPRESS THE C PROTEIN. WHEN THOMAS USED MICRO DISSECTION TO GET SAMPLES OF DNA FROM THIS THYMOID REGION HE FOUND THERE WERE LOTS OF NON-PRODUCTIVE ASSEMBLIES OF VLRA AND THESE WERE NOT FOUND IN BLOOD, WHICH WAS CONSISTENT WITH THE POSSIBILITY THAT THE ASSEMBLY WAS BEING SELECTED FOR -- IN THIS LOCATION, FOR THE Cs AS WELL, THE SAME STORY. EXCEPT THAT THERE WERE ALSO NON-FUNCTIONAL ASSEMBLIES IN THE BLOOD, AS WELL AS IN THE THYMOID THOUGH THEY WERE MORE COMMON IN THE THYMOID REGION, MANY WERE INCOMPLETE. YOU CAN SEE HOW THE VLRC IN THIS INSTANCE IS BRINGING IN THESE DONOR LRR SEQUENCES. FIRST THIS, THEN ADDING MORE FOR THE LRR1 TO COMPLETE IT, AND LRRV AND CAN BEGIN INCOMPLETE ASSEMBLIES INVOLVING FROM FIVE PRIME OR THREE PRIME LOCATIONS. I SHOW THIS VLRC BECAUSE IT'S A SIMPLER DONOR SEQUENCE, FIVE POTENTIAL DONORS, THEY WOULD HAVE TO COME IN IN A SEQUENTIAL MANNER FOR EXPRESSION. THERE ARE OVER 180 OF THEM, AND I WON'T FOR TIME REASONS GO INTO MORE. WHEN HIRANO PULLED TOGETHER CELLS FROM ALL OF THE DIFFERENT TISSUES, AND DISCRIMINATED BETWEEN THE GERMLINE AND MATURE VLRA, C AND B GENES, HE OBTAINED SEVERAL BITS OF INFORMATION, I'LL POINT OUT JUST A COUPLE OF POINTS. ONE IS THAT THE VLRA-B-POSITIVE CELLS ASSEMBLE AND EXPRESS VLRB TRANSCRIPT, THE A AND C LOCI ARE SILENT. SIMILARLY A-POSITIVE CELLS, OR THAT -- THE B, VLRB IS TOTALLY SILENT, TRUE FOR C, BUT SHARED ASSEMBLY OF A AND C THAT ARE BOTH, AS I'VE SHOWN YOU EVIDENCE SUGGESTING THAT THEY ARE ASSEMBLED IN THIS THYMOID REGION. DATA SUGGESTS A WORKING MODEL THAT A CELL OF A COMMON PROGENITOR AGAINST ASSEMBLY ON ONE ALLELE OR THE OTHER. IF PRODUCTIVE, CELL CAN SURVIVE. IF IT'S NOT, IT CAN TRY ON THE OTHER ALLELE. IF IT MISSES ON GETTING PRODUCTIVE ASSEMBLY ON BOTH ALLELES IT CAN ASSEMBLE VLRA. THEY BEGIN TO BEGIN ASSEMBLY LATER AND GO THROUGH THE SAME ALTERNATIVE POSSIBILITIES. NOW, THE VLRAs AND Cs, WHICH ARE BEGINNING TO LOOK MORE AND MORE LIKE OUR T-CELLS, THEY MIGRATE PREFERENTIALLY INTO ENDOTHELIAL SURFACES. THIS IS THE TYPE OF CELL, THIS IS THE INTESTINE, YOU CAN SEE THESE GREEN AND RED ONES HERE, IN THE INTESTINE MIXED TOGETHER, THERE ARE NO Bs. IN THE SKIN, VLRCs ARE PREFERENTIALLY EXPRESSED BY THEMSELVES, AND MOREOVER, IN THE SKIN THE VLRCs ARE OLIGO CLONAL, NOT VERY OLIGO CLONAL IN OTHER TISSUES, AND VLRAs ARE NOT OLIGO CLONAL IN ANY OF THESE SITES. SO IT'S BEGINNING TO LOOK MORE LIKE ALPHA, BETA AND GAMMADELTA T-CELLS, NOW LET ME COME BACK TO THE DIFFERENTIALLY EXPRESSED GENES, SOX 13, WHICH IS AN INHIBITOR TRANSCRIPTION FACTOR IS ESSENTIAL FOR T-CELL DIFFERENTIAL IN MICE. GENES ARE HOMING MOLECULES FOR MIGRATION TO EPITHELIUM. SO IT LOOKS AS IF THERE'S A STRIKING RESEMBLANCE OF VLRCs TO GAMMADELTAS, NOT EXACTLY THE SAME, YOU WOULD NOT EXPECT THEM TO BE FOR TIME OF DIFFERENCE SINCE EMERGENCE IN EVOLUTION, AND THE Bs, OF COURSE, MUCH LIKE THE Ds. B-CELLS THAT WE HAVE. SO WHAT ABOUT THE HAGFISH? AND HAGFISH TURN OUT TO HAVE ALSO THREE VLRs THAT WE'VE NAMED VLRA, C AND B, AND THESE ARE EXPRESSED PREFERENTIALLY BY THREE DIFFERENT LYMPHOCYTE LINEAGES. THEY ARE PRODUCTS, LAMPREY AND HAGFISH, VLA A,C AND B A SIMILAR, PREDICTED STRUCTURE AND ACTIVE STRUCTURE IS SIMILAR, EVEN WITH THE SAME PREFERENTIAL SELECTION OF THE NUMBER OF THESE INSERT LINKS WHICH IS SHORT FOR THE VLRC SO IT DOESN'T HAVE THE LOOP THAT I MENTIONED EARLIER. PERHAPS A CLUE TO WHAT IT'S ACTUALLY SEEING. SO I WON'T GO INTO THAT PART OF OUR STUDY AND WE'VE CONDUCTED OTHER STUDIES WITH ROY AND HIS COLLEAGUES TO GAIN EVIDENCE SUGGESTING SELECTION IN THE THYMOID REGION AND SAID THREE LYMPHOCYTE LINEAGES, THE PROGRAM MUST HAVE BEEN PRESENT IN THE ANCESTORS 500 MILLION YEARS AGO, JAWED AND JAWLESS, BUT THE VLLRA AND B-CELL AND T-CELL MUST HAVE BEEN CONVERGENT SOLUTIONS TO A MECHANISM FOR GENERATING A HIGHLY DIVERSE REPERTOIRE. WHAT THE RECEPTORS MIGHT HAVE BEEN SEEING BEFORE CONVERSION EVOLUTION OF THESE TWO TYPES WE CAN ONLY GUESS, BUT ONE OF THE QUESTIONS THAT I'D LIKE TO ASK QUICKLY IN THE LAST FEW MINUTES IS WHEN DID THE INNATE LYMPHOID CELLS, NATURAL KILLER CELLS EVOLVE? IN THE LAST FIVE YEARS THE NON-CYTOTOXIC LYMPHOID CELLS OF DIFFERENT TYPES THAT SOME CALL HELPER LYMPHOID CELLS, NONE OF WHICH EXPRESSED THE T OR B-CELL RECEPTORS, HAVE BECOME MORE AND MORE INTERESTING AND SEEM TO BE INVOLVED IN DEFENSE, PARTICULARLY ALONG MUCOUS SURFACES, INITIATION OF THE ADAPTIVE RESPONSES OF T AND B-CELLS AND THEY HAVE BEEN CATEGORIZED INTO SEVERAL DIFFERENT LINEAGES, THESE ILC LINEAGES, FOR NATURAL KILLER CELLS AND THREE DIFFERENT TYPES OF ILCs, ONE, TWO AND THREE. AND SO WE WONDERED IF WE MIGHT BE ABLE TO FIND ANY EVIDENCE FOR WHEN THESE EVOLVED. SO WE WENT BACK AND LOOKED AT THE CELLS IN IT THE LYMPHOCYTE LIGHT SCATTER GATE THAT WERE NEGATIVE FOR ALL THREE OF THESE DIFFERENT SPECIFIC VLR TYPES OF RECEPTORS. AND AS YOU CAN SEE FROM THE RED, ALL THE TISSUES THAT WE LOOKED IN, BLOOD, KIDNEY, ESPECIALLY THE GILL REGION THERE WAS ABUNDANCE OF TRIPLE NEGATIVE CELLS. YOU MIGHT HAVE NOTICED TRIPLE NEGATIVES ARE JUST A BLANK FOR ANY OF THE ORTHOLOGS OR LOOK ALIKE GENES THAT I TALKED ABOUT SELECTIVELY EXPRESSED BY A AND C AND B LINEAGES OF CELLS. WHEN WE LOOKED AT MARKING GENES SHOWN TO BE IMPORTANT IN DIFFERENTIATION OF ILCs, AND NATURAL KILLER CELLS, ELG, KIT AND SO FORTH, YOU CAN SEE THESE ARE PREFERENTIALLY EXPRESSED BY THESE TRIPLE NEGATIVE CELLS, ALSO A CELL SURFACE MOLECULE THAT CAN BE FOUND ON NK CELLS, AND A COUPLE OF OTHER GENES THAT WE DON'T KNOW THE SIGNIFICANCE OF. SO IF YOU LOOK ON THIS MAP THAT DAVID ARTIS AND JURGEN SPITS PRODUCED FOR THE FACTORS IMPORTANT FOR DIFFERENTIATION OF IL-1, 2, 3, LYMPHOID TISSUE INDUCERS, WHICH ARE ESSENTIAL FOR DEVELOPING OF LYMPH NODES, NONE OF WHICH THE LAMPREYS HAVE, BY THE WAY, AND NK CELLS THE GENE ORTHOLOGS ARE SUGGESTIVE THAT INDEED THIS AT LEAST THIS ILC 2-LIKE LINEAGE, THE TRANSCRIPTIONAL PROGRAM, IS AWFULLY REMINISCENT OF THAT SHOWN TO BE REQUIRED FOR ITS DEVELOPMENT OR THEIR DEVELOPMENT IN MICE. AS ALSO THE NK CELLS WITH THE CD 56, AND THESE ARE LARGE GRANULAR LYMPHOCYTES, THE FUNCTION OF WHICH WE'VE NOT YET SHOWN, SO FOR AN NK-LIKE, IN AT LEAST THIS ILC-LIKE 2, IT'S SUGGESTIVE, SO THE WAY WE WOULD TRY TO PUT THESE FINDINGS TOGETHER IS THAT TWO MAIN POINTS, THAT T AND B-CELLS HAVE BEEN INSEPARABLE COMPANIONS FOR OVER 500 MILLION YEARS. THE QUESTION IS WHY, AND WHY HAVE THEY BEEN SELECTED FOR FURTHER EVOLUTION, OVER THIS LONG, LONG PERIOD? I WOULD SUBMIT THAT IT'S PROBABLY BASED ON THEIR VALUE IN PROTECTION AGAINST INFECTIONS. ONE COULD ALSO USE PARTICULARLY RECENT DATA TO SUGGEST THAT THE OLD IDEA THAT THE T-CELLS WERE SELECTED AND THE IMMUNE SYSTEM EVOLVED FOR PROTECTION AGAINST MALIGNANCIES, THAT THAT IS ALSO A POTENTIAL TENABLE REASON FOR THE CONSTANT EVOLUTION OF T AND B-CELLS, FOR THIS VERY LONG PERIOD OF TIME. AND THE SLIM DATA THAT I SHOWED YOU AT THE END MAY SUGGEST THAT BASIC GENETIC PROGRAMS EVOLVED EVEN EARLIER THAN THE T AND B CELLS FOR THIS TWO CONVERGENT SOLUTIONS WERE OBTAINED TO GIVE THEM SPECIFIC RECEPTORS. THESE ARE PEOPLE WHOM I MENTIONED TWO MENTIONED WHO DID THE WORK I WAS TALKING ABOUT. BRANT HERRIN, MASA HIRANO, THE SEPARATION OF THE DIFFERENT POPULATIONS AND GENETIC PROGRAMS, SABY DAS IS A COMPUTATIONAL BIOLOGIST WHO DID MUCH OF THE PHYLOGENETIC COMPARISONS, JIANXU LU AND CUILING YU, AND OTHERS WHOSE WORK I DIDN'T MENTION SO MUCH. I WOULD LIKE TO CLOSE WITH THANKS TO THE NATIONAL INSTITUTES OF HEALTH, WHO SUPPORTED MY WORK OVER THE MANY YEARS THAT I'VE BEEN WORKING, AND NOT MENTIONED HERE, HOWARD HUGHES MEDICAL INSTITUTE, WITHOUT WHOSE SUPPORT WE NEVER WOULD HAVE STARTED THESE FISHING EXPETITIONS, AND ALSO EMORY VACCINE CENTER AND GEORGIA RESEARCH ALLIANCE FOR HELPING SET UP THE LABORATORY AND THANKS TO YOU ALL FOR LISTENING TO THIS LONG FISHY TALE. THANK YOU. [APPLAUSE] >> ALL RIGHT. I WOULD LIKE TO THANK DR. COOPER FOR SUCH A GREAT AND ENLIGHTENING TALK THIS AFTERNOON, AND OPEN UP THE DISCUSSION TO QUESTIONS PEOPLE MIGHT HAVE ON THIS PRESENTATION. ALL RIGHT. MAYBE I'LL ASK ONE. I WAS STRUCK BY SOME OF THE CYTOKINES WE SEE IN THE LAMPREY, WHY ARE SOME KEPT, ARE MAYBE SOME NOT KEPT, OR SOME THAT ORIGINATE SUCH AS IL-17 AND Bcl6, WHY NOT IN THE T-CELLS? >> I WISH I COULD ANSWER ALL OF THOSE QUESTIONS. INTERLEUKIN 17, AS YOU MENTION, HAS BEEN RELATIVELY LATE IN DISCOVERY BY IMMUNOLOGISTS, WHAT IS THOUGHT TO BE AN ORTHOLOGUE OF INTERLEUKIN 17. WHAT'S SURPRISING TO US IS THAT THE ONE THAT WE'VE NOT BEEN ABLE TO FIND, AND IN PARTICULAR, INTERFERONS, THEY PLAY SUCH AN IMPORTANT ROLE IN OUR DEFENSE SYSTEMS, WE WERE SURE WE WOULD SEE THOSE, AND THEIR RECEPTORS. THERE ARE MANY OF THE GENES THAT ARE INTERFERON-RESPONSIVE GENES AND SOMETHING MUST BE ACTIVATING THEM. WE HAVE FOUND ONE INTERFERON RECEPTOR GENE, INTERFERON-1 RECEPTOR GENE LOOK-ALIKE SO IT'S POSSIBLE WE JUST MISSED IT SO FAR. THE GENOME SEQUENCING IS NOT PERFECT. IT'S ALSO POSSIBLE THAT SOMETHING THAT'S STRUCTURALLY SERVES THE SAME PURPOSE THAT WOULDN'T BE IDENTIFIED BY NUCLEOTIDE SEQUENCING IS ALSO PRESENT. I WON'T GO ON. >> IN THE SEA-LIVING ANIMALS ARE THERE LOTS OF VIRUSES AND BACTERIA LIKE IN THE ATMOSPHERIC SYSTEM WE FACE EVERY DAY? WHY THEY ARE MAKING THESE DIFFERENT ADAPTIVE IMMUNE SYSTEMS? >> I CAN'T ANSWER THAT QUESTION, BUT IT SEEMS STRIKING TO US THAT EVERY JAWED VERTEBRATE THAT HAS SURVIVED, HAS ONE OR THE OTHER OF THESE SYSTEMS. IT COULD BE THAT THEY ARE -- HAVE BEEN KEY TO SELECTIVE SURVIVAL OF LAMPREYS AND HAGFISH, AND THE JAWED VERTEBRATES THAT WE KNOW ABOUT. WE DON'T KNOW A LOT ABOUT THEIR PATHOGENS, FOR LAMPREYS THERE HAVE BEEN OUTBREAKS THAT HAVE CAUSED WIDESPREAD DEATH AMONG LAMPREYS IN CAPTIVITY, BACTERIAL IN ORIGIN. WE DON'T KNOW MUCH ABOUT VIRUSES AT ALL. BUT ALL OF THEM, BOTH OF THEM, ARE SWIMMING IN A SEA OF BACTERIA, OF COURSE, AND THERE ARE ALSO VIRUSES IN THEIR ENVIRONMENT. AND AS I -- THE CRISPR/Cas SYSTEM, FOR EXAMPLE, IS A WAY THAT BACTERIA DEFEND THEMSELVES FROM PLASMIDS AND VIRUSES, SO ONE WOULD EXPECT THAT THESE JAW LESS VERTEBRATES WOULD HAVE ENCOUNTERED VIRAL -- VIRUSES, POTENTIAL PATHOGENS AS WELL. BUT WE'RE MAINLY JUST, AS FOR MANY OF THE FEATURES OF THIS IMMUNE SYSTEM AND ITS PROTECTIVE VALUE, HAS SO LITTLE INFORMATION THAT IT'S MOSTLY HAND WAVING. >> THE MYLAR SYSTEMS, MAMMALIAN SYSTEM, YOU MENTIONED EXTENSIVELY THE LYMPHOID SYSTEMS. >> OH, THEY DO HAVE -- THEY HAVE ALL OF THE DIFFERENT TYPES OF BLOOD CELLS THAT WE HAVE ESSENTIALLY. THEY HAVE GRANULAR CYTEV, LOOK ESINOPHILS, THEY HAVE MONOCYTES, AND NUCLEATED THROMBOCYTES. A WHOLE REPERTOIRE. A WHOLE PUBLICATION PROVIDES EVIDENCE ENDOTHELIAL CELLS IN THE BLOOD VESSELS FOR A VERTEBRATE ARE MENTIONED, AND NOT SEEN IN THE SISTER GROUP, NOR IN THE ANTHYOCIX, THE HEAD OF THE CHORDATES. MOREOVER, IN THE ANTHYOCIX, THEY HAVE A HEART AND VASCULAR SYSTEM, THE REPORTS SAY THAT THE BLOOD IS ACELLULAR, THERE ARE NO CELLS FLOATING AROUND BUT WHEN THEY EVOLVED, THE HEMATOPOIETIC SYSTEM, WE'RE STILL LEFT GUESSING AT THIS POINT. >> THANK YOU FOR A BEAUTIFUL TALK. IS THERE ANY RELATION BETWEEN THE GENE PRODUCTS AND COMPLEMENT SYSTEM, SOME INTERACTION, MAYBE SOME MECHANISM OR SOMETHING LIKE THAT? >> PROFESSOR LEE AND HIS COLLEAGUES HAVE PUBLISHED STUDIES SHOWS VLRB ANTIBODIES COUPLED WITH C-3 AND C1Q ORTHOLOGUE FROM LAMPREYS CAN TOGETHER INDUCE BACTERIA IN TUMOR CELLS, SO AT LEAST IN VITRO, THESE CAN ENGAGE, COMPLEMENT STUDIES SUGGEST FOR ALYTIC -- POTENTIALLY PROTECTIVE ACTIVITY. >> A SHORT QUESTION, DO YOU THINK THAT SIMILAR TO EGN, THERE'S A CHANGE OF CONFIRMATION, YOU KNOW, BECAUSE IT'S VERY SIMILAR TO THIS ANTIBODIES IN THE HUMAN. >> WE ASSUME THAT THE CONSTANT REGION, LIKE THE STALK REGION, LIKE OUR REGION, MAY ALSO HAVE A RECEPTOR TO ALLOW THEM TO FACILITATE VAGOCYTSIS ACTIVITY, BUT SO FAR IT'S AN ASSUMPTION WITHOUT ANY SUPPORTIVE DATA. >> THANK YOU VERY MUCH. >> ALL RIGHT. THANK YOU, DR. COOPER, THANK YOU, EVERYONE, FOR YOUR ATTENTION. [APPLAUSE] THERE'S A RECEPTION IN THE LIBRARY, NIH LIBRARY, IF PEOPLE WANT TO JOIN. THANK YOU.