I WORK WITH ALAN FOR 12 YEARS. FIRST AS POSTDOC AND THEN STAFF SCIENTIST, SO, THIS MORNING'S SESSION IS REALLY DEDICATED, CELEBRATING ALAN'S LEADERSHIP IN THE COLLABORATION AND NIAID IMMUNOLOGY PROGRAM. AS SOME OF YOU KNOW, ALAN HAS LONGSTANDING INTEREST IN MICRO BACTERIA RESEARCH AND I THINK HE STARTED WITH MICRO BACTERIA AVIAN WITH MARK DOUGHERTY AND 2000, ALAN DECIDED TO REALLY MOVE INTO USE REAL BUGS AND MBT AND THEN HE HIRED MYSELF TO REALLY KICK-START THE PROGRAM AND ALAN USED TO CALL US THREE MUSKETEERS AND WE REALLY WORKED FROM SCRATCH AT THE TIME AND I REMEMBER THAT TIME WHEN YOU EVEN DON'T HAVE A FUNCTION OF PIECE OF THREAD SO WE HAVE TO BOR' SPACE. I HAD TO WORK WITH KAREN AND DAMIAN AND CHUCK HAS TO DRIVE TO TIM BROOKE TO USE TIMBER''S PC3 AND I REMEMBER WE ALWAYS TALK ABOUT HOW WE CAN MAKE A CONTRIBUTION TO THE TB RESEARCH AND HOW TO MARK OUR TERRITORY AS A NEW KID ON THE BLOCK. DON'T GET BULLIED BY OTHERS. SO, ANYWAY, SO THAT IS THE LONG TIME AGO SO I THINK UNDER ALAN'S LEADERSHIP, AS YOU CAN SEE NOW, NIAID PROVIDES THE BEST EQUIPPED AND LEADER IN THE TB IMMUNOLOGY IN THE WORLD. SO THAT IS REALLY REFLECTION OF ALAN'S LEADERSHIP AND THIS PROGRAM HAS TRAINED 15 FELLOWS AND POSTDOCS AND ALL OF US HAVE GONE TO ESTABLISH OUR OWN RESEARCH PROGRAMS. SO I THINK THAT IS TREMENDOUS CAREER AND GREAT CAREER FOR ALAN. WE ALL APPRECIATE IT. THANK YOU, VERY MUCH, ALAN. IT'S A GREAT HONOR TO BE PART OF THIS PROGRAM AND IN YOUR MENTORSHIP. IT'S GREAT FUN. I HAD A GOOD TIME AND WE HAD A FUN TIME. SO, BEFORE INTRODUCING AND INVITING OUR FELLOW FORMER TRAINEES TO PRESENT THEIR RECENT WORK, I JUST WANT TO INVITE BOB, BOB COFFMAN, TO HAVE SEVERAL WORDS AND OPENING REMARKS FOR ALAN. ROBERT COFFMAN HAS BEEN A STRONG SUPPORTER OF IMMUNO BIOLOGY SECTION. [APPLAUSE] ALL RIGHT. SO, IT SORT OF FOR THOUSANDS OF YEARS NOW, I'M SURE, WHEN THE ELDERS OF ANY TRIBE GOT TOGETHER AS WE'RE DOING HERE, THEY WOULD SIT AROUND THE CAMPFIRE OR WHAT HAVE YOU AND TELL ORIGIN STORIES TO PASS THEM ON ORALLY TO THE NEXT GENERATION, I SUPPOSE WAS THE PURPOSE OF THAT SO THAT'S WHAT I'LL TRY TO DO IN THE NEXT FEW MINUTES IS TELL A COUPLE OF ORIGIN STORIES. AND PROBABLY IT'S GOOD TO FALL TO ME SINCE I MAY HAVE KNOWN ALAN LONGER THAN ANYONE HERE, HAVING MET HIM ORIGINALLY HERE AT THE VERY BEGINNING OF 1970. SO PLAS TA STEEN SO STONE ARCHITECT TIRE AND IT'S A SOCK INSTITUTE. I WAS COMING IN AS A WET BEHIND THE EARS AND BEGINNING GRADUATE STUDENT TRYING TO LEARN WHAT ANTIBODIES WERE. ALAN WAS ON THE OTHER SIDE OF TBENCH. ALAN WAS TRYING TO CRAFT SOMETHING LIKE A PRESENTABLE THESIS AND GET OUT AT THE TIME. WE OVERLAPPED FOR ABOUT 18 MONTHS ACROSS THE BENCH. I CAN'T SAY I KNEW ALAN PARTICULARLY WELL. HE WORKED VERY HARD AND HE WORKED LATE AND HE WAS VERY SERIOUS. CHRIST, HE DIDN'T EVEN SURF SO WE HAD NO SHARED VALUES AT THAT POINT. BUT HE WAS A NICE GUY AND I ENJOYED TIME. HE IS LOOKING VERY DAPPER THIS IS FROM 1967. EVEN THEN TRYING TO SMILE. SO, OUR MENTOR WAS ON THE RIGHT, VERY PROMINENT IMMUNOLOGY AT THE TIME, VERY BRILLIANT MAN AND HAD A LOT OF INFLUENCE IN FORMULATING THE QUESTIONS AND CELLULAR AND MOLECULAR IMMUNOLOGY AND THERE IS JOHN WITH HIS WIFE SUZANNE AND A VERY YOUNG LOOKING GUS ON THE LEFT. MEL HAD GROWN UP AND DONE HIS FORMATIVE TRAINING FOR MANY YEARS IN PARIS AND STUDYING THE PIONEERS STUDIES OF BACTERIAL GENE REGULATION SO HE WAS, ALTHOUGH INTERESTED IN IMMUNOLOGY AT THIS POINT, A REALLY COMMITTED REDUCTIONIST IN TERMS OF HIS APPROACH TO SCIENCE AND A VERY MUCH COMMITTED TO USING SIMPLE SYSTEMS AND MODEL ORGANISMS AND THE IDEA OF ACTUALLY SIACTUALSTUDYING THE IE JUST SENT CHILLS DOWN HIS SPINE. AT THE SAME TIME, THERE WERE NEW WINDS BLOWING AROUND EVERYWHERE IN THE EARLY 70s AND THE ERA OF VIETNAM WAR PROTEST AND SO FOURTH AND SO ON AND ALONG WITH THAT A NEW SENSE OF SOCIAL RESPONSIBILITY THAT TOOK HOLD IN A LOT OF THE STUDENTS AND POSTDOCS AT SOCK INSTITUTE. I REMEMBER THIS VERY CLEARLY BECAUSE THIS IS WHAT WAS ALL FERMENTING OVER MY FIRST YEAR OR TWO THERE. I THINK IT INFLUENCED ALAN, BECAUSE HE MUCH TO THE DISGRACE OF THE LAB HE DECIDED TO BECOME A PARA SATOLOGIST WHEN HE LEFT THE LAB AND TRAINED THERE. AT THIS POINT, ALAN AND I, OUR PATHS DIVERGED A LOT. I WENT AHEAD AND FINISHED GRADUATE WORK AND DID POSTDOC AT STANFORD AND GOT INVOLVED IN STARTING A VERY INTERESTING RESEARCH INSTITUTE CALLED DEAN ACTS IN PALO ALTO WHICH SOME OF THE OLDTIMERS KNOW. AND I PROBABLY DIDN'T SEE ALAN FOR 12 OR 13 YEARS AT LEAST. IN THE MEANTIME, TIM AND I HAD DONE SOME OF THE WORK THAT WE'VE BEEN DOING IN OUR RESPECTIVE LABS ON TRYING TO DEFINE T-CELL CYTOKINES AND SOME OF THEIR FUNCTIONS. AND CAME OUT WITH SOMETHING THAT LOOKS FAMILIAR NOW AND I WON'T BELABOR HERE. WE'RE PROPOSING THAT BASICALLY THE CD4 T-CELL RESPONSE TO ANTIGENS COULD REALLY SORT ITSELF OUT AS TWO MAJOR T-CELL SUBSETS AND NAMED TH1 AND TH2. THESE DELIVERED BY THEIR PATTERNS OF CYTOKINE EXPRESSION BUT ALSO BY THE DIFFERENCES IN FUNCTIONS THAT ARE MEDIATED BY THOSE DIFFERENT EVERY SHALL CYTOKINE PATTERNS AND IN ADDITION IT'S SHOWN IN THE BLUE ARROWS, THERE WERE A NUMBER OF RECIPROCAL INHIBITORY INTERACTION THAT'S ALLOWED FOR INTERESTING POSSIBILITIES FOR THEIR UNDERSTANDING IMMUNO REGULATION. DURING THE BEGINNINGS OF THIS WE WERE CONVINCED THIS WAS RELEVANT TO ALLERGIC DISEASES BUT WE'RE JUST, TIM AND I WERE TRYING TO FIGURE OUT DOES IT HAVE RELEVANCE BEYOND THAT? WHAT ABOUT REAL INFECTIONS AND CONTROLLING INFECTIONS? SO BY CHANCE, I WAS ON THE NIH CAMPUS AND I THINK WE WERE REALLY JUST RANDOMLY BUMPED INTO ALLEN WE GOT TO TALKING AND I TOLD HIM ABOUT THAT AND HE GOT EXCITED AND SAID THIS MAY BE THE ANSWER TO ALL OF THE UNANSWERED PHENOMENON IN SOME OF THE PARASITE IMMUNOLOGY SYSTEMS I STUDIFUL LET'S TRY TO DO SOME THINGS TOGETHER AND WE DID AND A NUMBER OF YEARS OF LOVELY AND VERY FRUITFUL COLLABORATIONS AND THE FIRST OF THEM I THINK IS VERY IMPORTANT. PHIL SCOTT ALLUDED TO IT YESTERDAY BUT LET ME SHOW YOU THE BOTTOM LINE. THIS IS THE FIRST OUTCOME OF THE WORK WETOGETHER AND THIS IS WITH PHIL AND PEARCE AND AT THE TIME, THAT ALAN AND I MET UP THEY WERE DEVELOPING A T-CELL LINES AND WE'RE NOT FORMALLY CLONES BUT CLOSE, T-CELL LINES TO ANTIGENS THAT HAD INTERESTING PROPERTIES WHEN YOU TRANSFERRED THEM IN NORMAL MICE AND INFECTED AND THIS IS A GOOD EXAMPLE. THIS IS LYNN ONE ACTUALLY DID SOMETHING QUITE REMARKABLE WHICH IT MADE THE FOOT PADS SWELLING AND THE DISEASE WORSE THAN IF YOU JUST PUT IN NORMAL T CELLS OR NO T CELLS AT ALL WHERE OTHERS SUCHS A THE LINE NINE GAVE COMPLETE PROTECTION WHEN YOU CHALLENGED. THE DIFFERENCES CORRELATED PRETTY ACCURATELY WITH THE DISTINCTIONS THAT TIM AND I HAD PROPOSED AND NOT SHOWN HERE IS INTERFERON GAMMA BUT ALIGN NINE AND IN ADDITION TO MAKING IL2 BUT NOT IL4 AND 5 MADE GAMMA AND LINE ONE DID NOT. THESE WERE CLASSIC PHENOTYPES BUT THIS WAS THE FIRST PAPER TO REALLY SHOW THAT THEY WERE LEADING TO REALLY DRAMATICALLY DIFFERENT OUTCOMES IN AN INFECTIOUS DISEASE SYSTEM. EVERYTHING ELSE BEING COST IN THIS FIELD. IT WAS A LANDMARK TIME. SO THIS WAS A REAL I AM FLECKION POINT FOR ALAN AND I. ALAN BECAME A T-CELL AND IMMUNOLOGIST HERE AND I BECAME A LEASH MANIAC AND STARTED FROM ALAN AND THEY LET ME SET UP A LEISHMANIA SYSTEM IN MY LAB AND WE DID STUDIES USE PARTICULAR AS A COOL FOR STUDYING IMMUNE REGULATION AND I'LL JUST SHOW YOU ONE OF THE FAVORITE PAPER BECAUSE IT'S A GREAT EXAMPLE OF A YOU DON'T ALWAYS GET WHAT YOU WANT BUT SOMETIMES YOU GET WHAT YOU NEED PHENOMENON TO PHOTO MICK JAGGER AND SO A PAPER WITH TWO VERY RIGHT PEOPLE, RODRIGO FROM ALAN'S LAB AND FIONA AND BASICALLY THEY WERE TRYING TO CHASE UP WHAT WE AND OTHERS WAS THE FACT THAT IN MICE THAT DON'T HEAL AND DEVELOP A THC RESPONSE WHEN THEY'RE INFECTED WITH LEASH MANIA THAT THERE'S ALWAYS EVIDENCE OF SOME LEASH MANIA SPECIFIC INTERFERON GAM AND WHETHER IT'S TH1 CELLS AND WHY IN THEY THERE AND IT WAS REVERSD BY ANTI-IL4 BUT AN INTERESTING THING IS THE INTERFERON RESPONSE WAS AMONG THE PHENOTYPE WHICH WAS ASSOCIATED WITH NAIVE T CELLS AND THE RBIT CELLS AND THEY WERE TRANSFERRED HEALING TH1 RESPONSE TO THIS ANIMAL AND THAT WAS ALL VERY INTERESTING EXCEPT THE MICE THAT HEALED WEREN'T REALLY OUT OF T WOODS BE IN SIX OR EIGHT WEEKS THEY STARTED SUFFERING FROM A SEVERE INFLAMMATION OF THE COLON, SO FIONA DID THE OBVIOUS CELL AND SASS IF THIS HAPPENS IF WE TAKE LEISHMANIA OUT OF THE EQUATION, WE DO ISOLATE THE CELLS FROM LEISHMANIA INFECTED MICE AND WE JUST DID NORMAL CELLS AND FRACTION EIGHT THEM AND TRANSFER THEM INTO SKID MICE NO LEISHMANIA INFECTION TO THAT AND YOU GET THE SAME PHENOMENON MANY OF NORMAL MOUSE COLONS AND IT'S TYPICAL SECTIONS AND TRANSFERRING THE CELLS LED TO GREAT ENLARGEMENT OF THE COLON, SEVERE DISRUPTION OF THE ARCHITECTURE AND CELL TYPES AND LOSS OF T CELLS IN THERE BUT INTERESTINGLY ENOUGH, IF YOU MIX BACK IN AND CO TRANSFERRED AN PRE PRE AT NUMBER OF THE CD45RB LOW CELLS WITH THOSE, NONE OF THIS HAPPENS AND THEY WERE NORMAL AND YOU CAN LET THEM DIE OF OLD AGE AND SEE ANYTHING. SO, THIS TURNED OUT AFTER ANOTHER SERIES OF VERY ELEGANT EXPERIMENTS FROM GERMAN BUY FIONA TO BE ONE OF THE THREE OR OUR DIFFERENT LINES THAT LED TO OUR CURRENT SYNTHESIS OF REGULATORY T CELLS THE STORY BEING THEY'RE LOW POPULATION CONTAINED A POPULATION OF T CELLS AND LEISHMANIA THAT WERE REGULATORY. NOW, SO, THE LAST BIT OF ORIGIN AND SET US OFF TO SOME INTERESTING THING PEOPLE LIKE ALAN AND STEPHANIE IN PARTICULAR AND KEEP LOOKING AT OUR DIE A TRAMS AND LOOKED AT IL5 WHICH WE KNEW WAS STRONG STIMULATING FACTS OR AND BUT IT WAS NOT UNTIL WITH THE ASSISTANCE FROM TIM'S LAB WE MADE THE FIRST MONO CLONE ANTIBODY AND ASKED THE QUESTION, INITIALLY IN A SIMPLE SYSTEM WHICH WE HAD RUNNING IN OUR LAB AND SUBSEQUENTLY WITH ALAN AND IN A MUCH MORE COMPLICATED, RELEVANT SYSTEM WHICH IS HERE. AND WHAT ROLE WAS ID5 AND THE WISDOM OF THE TIME SMITH IT WOULD BE A MULTI FACTORIAL RESPONSE TO REALLY GET THE DRAMATIC YOU SEE IN A COMPLEX SIS SEES SUCH AS INFECTION. AND ANSWER TRYING TO BE SIMPLE AND ANTI-IL5 BLOCKS AN EFFECTER AND REDUCED THE BLOOD LEVELS TO ALMOST ZERO AS YOU SEE HERE. ANTI-IL4 AS NO EFFECT ON EITHER OF THESE MODELS AND I'M SHOWING YOU THE DATA AND RESIP I CANNILY IT HAS NO EFFECT ON IT BLOCKS IGE PRODUCTION SO REALLY NAILED DOWN THE DIFFERENCE BETWEEN THESE DIFFERENT FUNCTIONS AND LED TO THE DEVELOPMENT OF SEVERAL ANTI-IL5 TREATMENTED WHICH ARE NOW COMMONLY USED IN A NUMBER OF ALLERGIC DISEASES. SO ABOUT THIS TIME, 1990s, ALAN SAID A LOT OF PEOPLE ARE LOOKING AT THE INFLUENCE OF CYTOKINES AND T-CELL CYTOKINES AND T-CELL SUBSETS IN A WIDE RANGE OF DISEASES. A LOT OF PEOPLE WORKED A WORORGANIZED AWORKSHOP SO WITH Y TAPPING ALL THE PEOPLE, PRESSING STEPHEN COFFMAN AND I COULD RAISE MONEY, HE PUT TOGETHER A WONDERFUL CONFERENCE AND IT WAS 1992, HERE ON THE NIH CAMPUS THAT I THINK WAS REALLY A SEMINOLE JUNCTURE OF T-CELL IMMUNOLOGY BECAUSE IT BROUGHT TOGETHER WHOLE GROUPS OF PEOPLE FROM DIFFERENT FIELDS BASIC IMMUNOLOGY, AUTOIMMUNE DISEASE O FOURTH THAT REALLY DIDN'T TALK TO EACH OTHER BUT NEVER GO TO THE SAME MEETINGS AND WE GOT TOGETHER FOR SEVERAL DAYS AND IT WAS REALLY A FORMATIVE TIME. THERE'S ACTUALLY THE CONFERENCE PICTURE THERE. YOU SEE OUR FEAR LESS LEADER HERE LOCKING VERY GO AHEAD. YES, THAT'S ME WITH A WOMAN ON MY LAP. HERE IN THAT AND I REALIZED THAT WOULD NEVER FLY THESE DAYS. I GUARANTEE YOU. IT WAS POLY PATSINGER. IT'S A LOT OF THINGS HAVE CHANGED AROUND HERE AND NOT THE LEAST OF WHICH THAT WOULD NEVER BE ALLOWED ON THE NIH CAMPUS. I'LL FINISH BY SAYING ONE THING AROUND NIH THAT HASN'T CHANGED AT ALL SINCE THAT TIME, IS ALAN AND ALAN'S LOVE OF SCIENTISTS AND HIS ENORMOUS AFFECTION FOR THE PEOPLE THAT DO SCIENCE, WHICH COMES OUT AND IT'S COMING OUT OBVIOUSLY IN SO MANY TALKS WE'VE HEARD TODAY. ALAN'S INFECTIOUS ENTHUSIASM AND HIS LEGENDARY GENEROSITY NOT MUCH MORE I CAN SAY ON THAT. THAT HASN'T ALREADY BEEN SAID. I WANT TO THANK ALAN FOR HAVING SUCH A STRONG INFLUENCE ON THE DIRECTION OF MY CAREER TOOK EVER SINCE AND MANY OF THE PEOPLE IN MY LAB. THANK YOU. [APPLAUSE] >> THANK YOU, BOB. SO OUR NEXT SPEAKER IS ALSO A VERY STRONG SUPPORTER OF IMMUNO BIOLOGY SECTION AND ANNE O'GARRA FROM THE CRICK AND ANNE, PLEASE. ANNE REALLY INTRODUCED SYSTEM BIOLOGY'S INTO TB FIELD AND MAKE A -- >> HOW DO I DO THIS? >> TECHNOLOGY IS THE MOST CHALLENGING BUT IT ALL WORKS HERE, WHICH IS GREAT. >> SO, IT'S MY HONOR AND MY PLEASURE TO REALLY BE HERE TO CELEBRATE ALAN'S CAREER. WHICH THINK IS GOING TO CONTINUE FOR A LOT LONGER ANYWAY. AND THAT I'VE BEEN SELECTED TO ALSO GIVE A SCIENCE TALK AS WELL AS MY MEMORIES OF ALL THE INTERACTIONS WE'VE HAD. WE KEEP HAVING AND WE'RE GOING TO KEEP HAVING. SO, BEFORE I GO ONTO THE WORK THAT WE'VE BEEN DOING IN TUBERCULOSIS, I JUST WANT TO SAY A FEW WORDS ABOUT ALAN SHER. AND LOOKING AT IT FROM THE POINT OF VIEW OF ALAN, A PEOPLE PERSON, COMPLETELY CURIOUS ABOUT SCIENCE, EXCITED, FINDING NEW AREAS TO WORK ON AS HIS YOUNG POSTDOCS GO ON TAKING PROJECTS AND DEVELOPING THEIR OWN LABS AND FULLY SUPPORTIVE AND MEETING WHEN PROJECTS AND DIFFERENT LABS COME TOGETHER COLLABORATING, HAVING FUN, AND REALLY BEING INSPIRED. SO WITH A SMILE, I'VE BEEN INSPIRED THROUGH ALL MY INTERACTIONS WITH ALAN. I'M GOING TO RECAP JUST A FEW THINGS THAT'S BEEN SAID BUT JUST TO SAY HOW WHEN I FIRST METAL AN, SO, YOU'VE HEARD FROM RON AND OTHERS THAT ALAN DID A POSTDOC AT THE NIMR AT MEL HILL THAT LOVELY OLD BUILDING WITH A GUY CALLED RON SMITHERS. I KNEW RON SMITHERS BUT I WASN'T THERE BETWEEN 70 AND '74 BUT ALSO HEARD FROM DAVID SACKS WITH HIS FAMILY TREE THAT HE WAS A POSTDOC AFTER ALAN IN '77 TO '7S BECAUSE THEY CAPTURE THE ERA. SO LOOK AT DAVID SACKS AND THIS GUY JERRY WAS ACTUALLY THEN BECAME MY SUPERVISOR SO ED PEARCE WHO THEN DID HIS PH.D ALSO AT NIMR IN RON SMITHERS DIVISION ON SCHISTO SO I WAS ACQUAINTED WITH PATHOLOGY AND ALAN SHER AND DAVID SACKS, THE WORK THERE ALTHOUGH I WAS A MICROBIOLOGIST LOOKING AT BACTERIAL ADHESION AND I DID A POSTDOC ON IMMUNOLOGY AND I HAD THE LUCK TO BE INTRODUCED TO BOB COFFMAN AND I CAME AS A POSTDOC TO DEAN X AND S DNAX. ALAN IS NOT THERE YET BUT YOU WILL SEE SOME PICTURES. SO, IT WAS THAT PAPER THAT BOB HAS JUST TOLD YOU ABOUT THAT REALLY STARTED THAT COLLABORATION WITH BOB AND DNAX AND I WAS A POSTDOC AT XNAX AND I WAS LUCKY TO PROGRESS AND GET A JOB THERE AND INSTEAD OF SAYING TWO YEARS I STAYED 15 YEARS AND BOB I HAD JOINT LAB MEETINGS TOGETHER AND WE SAW ALAN AND ALL HIS LAB MEMBERS, PHIL SCOTT, EDMONTON PEARCE AND THROUGH THE YEARS, WE BECAME FRIENDS. I DIDN'T COLLABORATE SO MUCH, ALTHOUGH ON SOME PAPERS I DID ON THIS ERA. WE HAVE A COLLABORATION, ALAN BUT FROM THE BOB COFFMAN DNAX TO TIM MONTHSMANNIL-10 THIS IS THE FIRST THINGTHEFIRST PAPER YOU PH ED PEARCE AND THIS WAS IN SCHISTO AND SO THAT IS WHEN I WAS LUCKY TO REALLY NOT HAVE JUST JOINED DNAX AND INTERACT WITH BOB AND TIM AND MEET ALAN, PHIL, DAVID, ED AND I ALREADY KNEW EACH OTHER. SO, SPIN FORWARD A LONG WAY BECAUSE I'LL GET TO THE SCIENCE BUT I REALLY WANT TO REALLY EMPHASIS THAT ALAN, YOU KNOW, I'VE HAD REASONS TO LOOK AT ALAN, I WON'T GO INTO THEM. IT'S EASIER THAN LOOKING AT PUBMED BY I NOTICED LOTS OF PEOPLE LEAVING HIS LAB TAKE THE PROGRAMS AND THEN PHIL LEFT TOOK THE PROGRAM AND THEN TOXO AND THEN ALL THESE MODELS WE'VE HEARD OF AND ALAN'S GENEROSITY AND ALSO CREATIVITY AND IMAGINATION IN LOOKING FOR OTHER MODELS EXPERIMENTAL MODELS TO TELL US ABOUT IMMUNO REGULATION AND HE HAS BEEN THE KING ON THIS. NOW, TB IS WHAT, AS CARL THANKS FOR THAT INTRODUCTION BECAUSE I COULDN'T INTRODUCE ALL THE SEMINOLE PAPERS THAT HAVE BEEN PUBLISHED FROM ALAN'S LAB I THINK IT WAS SINCE 2000 THAT YOU GUYS STARTED WORKING ON TB. I SUFFER WITH YOU BECAUSE WE STARTED IN 2003 AND IT WAS HARD AND IT WAS HARD FOR MANY YEARS THAT WE'LL DISCUSS IN A BAR OVER WINE, NOT JUST CONTAINMENT 3 BUT LOTS OF DIFFERENCE. SO ALAN'S CONTRIBUTIONS ARE RECOGNIZED ALL OVER THE WORLD IN PATHOLOGY AND ALSO IN TB AND BASICALLY THIS IS EX EMPTIED OF THIS TWITTER OF FEED WHICH IS A RESEARCH REBOOT OF TUBERCULOSIS AT THE SYMPOSIUM 50th ANNIVERSARY WHICH ALAN IS LED ORGANIZERS AN AND IT WILL HAPPEN FROM JANUARY AND THE FACT THAT ALAN IS LEADING THIS MEETING IS PRETTY IMPORTANT BECAUSE YOU KNOW, THE TB FIELD HAS BEEN KIND OF TRACTION AND HARD AND HERE WE HAVE CAT AND DAN WHO ARE CONTINUING THE LEGACY OF WORKING IN TB AT THE NIH. SO, WITHOUT FURTHER ADIEU, I WANT TO SHOW YOU A BIT MORE OF THE HUMAN ASPECT OF ALAN THAT LOOK AT HIM SMILING. AND BOB AND THIS IS A KEYSTONE WHICH ALAN WAS INSTRUMENTAL ACTUALLY IN MY BECOMING INVOLVED AND BOB AND ALAN THERE AND I HAD SUCH WONDERFUL TIMES AND YOU SEE US WITH DRINKS AND THIS IS THE LAKE BY KEYSTONE AND THAT IS THE PATH MUCH WE WOULD GO FOR A WALK AND YOU KNOW, ALAN MENTORS IN A WAY IT DOESN'T COME ACROSS AS MENTORING BECAUSE HE IS CHATTING BUT HE IS GIVING YOU WISDOM AND GEMS OF HOW TO THINK ABOUT THE WAY FORWARD IN RESEARCH PROGRAMS AND RUNNING A LAB, THE PEOPLE, THE DIFFICULTIES, THE HARDSHIPS, THE JOYS AND SO THAT GIVES ME SUCH PLEASURE WHEN I LOOK AT THOSE PHOTOS AND I'LL BE HAPPY TO SEND YOU SOME OF THESE AS OUR MEMORIES. THANK YOU FOR YOUR GENEROSITY AND HELPING ME COME INTO THE TB FIELD BECAUSE IT WASN'T EASY. I STARTED BECAUSE IT SEEMED IMPORTANT THAT I WORK ON A DISEASE THAT ACTUALLY KILLED PEOPLE AND HAVING COME FROM DNAX AND I WAS SUPPOSED TO INTERFACE IMMUNE OLE GOUGH AND INFECTION SO I'LL COME AND IT WILL INTERFACE WITH ALAN AND CAT MAINLY THROUGH THE WORK AND SO, I'LL TALK TO YOU ABOUT HOW THEY HELPED US AND TO GIVE YOU BACKGROUND OF WHAT WE KNEW IN 2003 ABOUT THE IMMUNE RESPONSE AND MTB AND MTB AND IT WOULDN'T GO INTO BUT COULD INFECT KNEW TRA FILLS AND WE FOUND MACROPHAGES AND THEY MIGRATE TO THE LYNCH NODE AND THE RIGHT T CELLS WERE ACTIVATED AND THEY WOULD ACTIVATE AND THAT COULD PROTECT AND IT WAS FUNDAMENTAL IN EXPERIMENTAL MODEL BY ANDREW COOPER AND JOANNE FLYNN AND ALAN'S LAB ADVANCED A LOT OF THAT AND TNF WAS FUNDAMENTAL AND WHAT WAS REALLY AMAZING IS THESE FACTORS WERE ALSO SHOWN TO BE IMPORTANT IN HUMAN DISEASE AND THE TH1 ACCESS AND SUSCEPTIBLE DISEASES AND ANTI-TNF HAVE BEEN SHOWN TO ACTIVATE TD IN INDIVIDUALS WHO HAD BEEN EXPOSED BUT THAT WAS IT. THAT WAS THE TIP OF THE ICEBERG. I CAME FROM DNAX AND WORKED COLLABORATE BEING WITH TIM AND BOB FOR MANY YEARS AND SO, I THOUGHT OK, LET'S HAVE A LOOK TO SEE WHAT IL-10 DOES AND STOPS DEVELOPMENT MUST HAVE A ROLE AND I'M NOT GOING TO SHOW YOU THE DATA THIS WAS ALREADY PUBLISHED AND IN DEED IT DID HAVE A ROLE AND IN THESE RESIST APARTMENT MICE THERE WAS A SLIGHT ROLE BUT IT WASN'T PROFOUND SO THIS WAS A TIP OF THE ICEBERG. SO HOW ELSE TO GO AND THIS BRINGS INTO MIND THE COLLABORATIVE NATURE OF THIS FAMILY FOR PEOPLE LIKE ALAN, BOB, AND JACK WHO RAN THE INSTITUTE THAT WAS PARALLEL BUT MOVED TO DAMN AS AND WHERE I THEN WAS THERE FOR MEETING AND I SAID TO ALAN, TO TUBERCULOSIS IS A PROBLEM AND I'M STARTING TO WORK ON IT AND IT'S DIFFICULT. THOSE WHO DON'T WORK ON TB 1.5 MILLION DEATHS DIAGNOSIS IS DIFFICULT AND NOT GOING TO GO INTO THIS FOR THE SCOPE OF THIS TALK AND THE FASCINATING POINT OF IMMUNOLOGIST IS THAT A THIRD OF THE WORLD ESTIMATED A QUARTER OF WORLD TO BE LATENT AND THOSE WILL PROGRESS TO TB AND YOU CAN SAY BY DEFINED BY IMMUNE REACTIVITY BUT YOU CAN'T SAY WHO WILL GO. I'LL COME BACK TO THAT AT THE END OF THE MY TALK. TO GET TO THE QUESTION WE ASKED AND HOW THE HELP FROM OUR FRIENDS AND THIS WAS DAMIAN WHO IS HERE WHO REALLY HELPED THIS SO THE LINE LINE WAS I WILL HELP YOU CONQUER TB AND THOSE WHO KNOW YOU AND OUR QUESTION WAS MOUSE MODELS AND THEY WEREN'T AND THEY TOLD US ABOUT RESISTANT BUT WHY DO INDIVIDUALS REMAIN LATE ANT AND HEALTHY AND OTHERS GO ON TO GET TB WHAT FACTORS OF THE IMMUNE RESPONSE CONTRIBUTE TO TB AND THAT WAS THE QUESTION THAT CAME AFTER I LOOKED AT VIRGINIA PASQUAL'S COMPUTER AND SEEING THE IMMUNE SIGNATURES SHE DEFINED IN AUTOIMMUNITY SO TOGETHER WE ASKED THE QUESTION, CAN HUMAN BLOOD TRANSCRIPTIONAL SIGNATURES DISTINGUISH INDIVIDUALS WITH LATENT AND ACTIVE TB AND THIS IS PUBLISHED SO I'M GOING TO GO THROUGH IT QUICKLY, BUT TO SAY WE DEFINED AN INTERFERON BLOOD TRANSCRIPTIONAL SIGNATURES THAT WAS MISSING IN LAT ENTER AND HEALTHY INDIVIDUALS AND I'M GOING TO SHOW YOU A LITTLE BIT OF THESE DATA AND SHOW THE CONTRIBUTION THAT THEY HELPED US TO DECIPHER THIS TOGETHER WITH VIRGINIA. SO THIS SIGNATURE OF ACTIVE TB WAS DOMINATED BY AUGUST REGULATED GOWNS IN DOWN REGULATED SCENES IN ACTIVE TB BUT NOT IN LATENT AND THIS WAS IN LONDON AND WE REKA PIT RATED THAT IN SOUTH AFRICA AND THERE EVERYBODY WAS POSITIVE AND THE INTERFERON GAMMA AND ANTIGENS BUT STILL, THEY DIDN'T HAVE THE SIGNATURE BUT THE ACTIVE TB PATIENTS DID AND I'LL COME BACK TO THE HETEROGENEITY LATER. NOW, HOW DO YOU GET ALL THESE GENES AND DAMIAN HAD COME UP WITH A MODULAR APPROACH AND HE SHOWED IT COULD PUT INTO MODEL AND THEY COULD BE ANNOTATED LOOKING THEM AT THEM SO THEY HAD A APPROACH THAT WE USED IN TESTING AND OUR TB TRAINING SET IN LONDON AND OUR SOUTH AFRICA VALIDATION ZENDAYA SET AND YOU CAN SEE THIS IS SUPER IMPOSE ABLE THREE DIFFERENT YEARS AND THIS WAS GENES AND I TOLD YOU GAMMA INTER FEWER ON CAME UP IN TB PROTECTIVE SO COULD BE SEEN SO I THOUGHT IT'S GAMMA INTERFERON BUT THIS IS WHY TOOKING TO COLLABORATORS SAID NO, NO, IT'S NOT GAMMA INTERFERON BUT WAS IMPORTANT THAT S THESE CONFOUNDING DISEASE OF GROUP A STREP AND STAFF, DID NOT HAVE THE SIGNATURE BUT THIS OTHER BLOB WHICH IS A PLASMA GENE SIGNATURE SO YOU CAN DISTINGUISH TB FROM OTHER DISEASES AND WE'VE DONE A LOT MORE ON OUR WORK SHOW BUT VIRGINIA'S POINT AND THAT HAS TO BE TYPE ONE BECAUSE IT'S TYPE ONE INTERFERON AND I SAID NO, NO, IT HAS TO BE GAMMA, AND WHEN THE PHYSICIAN LOOKED AT THIS 393 SIGNATURE IN DETAIL HE FOUND THAT IT WAS GENESM D AND INTERFERON ALPHABET TA UP REGULATED YOU COULDN'T SEE CYTOKINES THEMSELVES SO THEY HAVE TO INDUCE THE GOWNS THAT GIVE THEM THE SIGNATURE AND ON PURCHASEIFICATION OF CELLS FROM THE BLOOD WHICH HAD A SIGNATURE, CHRIS GRAHAM IN THE LAB SHOWED THAT IT WAS NEWT TRA FILL AND MONOCYTES THAT EXPRESS THIS INTERFERON INDUCE ABLE SIGNATURE. NOW WHY WERE WE SURPRISED ABOUT TYPE ONE INTERFERON BECAUSE NOTHING WAS REALLY TALKED ABOUT IN TB IN TYPE ONE INTERFERON AND TB AS YOU KNOW TYPE ONE WAS FAMOUS FOR PROTECTING AGAINST VIRUSES AND CANCER AND IT'S CERTAINLY MODELS LIKE LISTERIA IT COULD EXACERBATE INFECTION SO WHAT WAS GOING ON SO THIS MADE US DIG DOPE INTO THE LITERATURE AND THERE WERE TWO PAPERS BY KAPLAN THAT SHOWED THAT TYPE ONE INTERFERON CORRELATED WITH SHORTER SURVIVAL TIMES IN MICE AND THE BLOCKADE COULD GIVE LONGER SURVIVAL TIMES OF THE TYPE 1 INTERFERON IF NOT SIGNALING. WHAT WAS DIFFERENT? RE RACKED OUR BRAINS SO WE WEREN'T -- SHE WASN'T USING THE H37 LAB STRAIN OF TB SHE WAS USING A CLINICAL ISOLATE AND SHE WAS USING F-1 INTRA BRED MICE OF GOD KNOWS WHAT GENETIC BACKGROUND THEY WERE AND SO PROBABLY GENETIC BACKGROUND STARTED TO BE IMPORTANT. WHAT STARTED TO COME OUT AS WELL IN PARALLEL, IS THE INCREASING LEVELS OF TYPE 1 INTERFERON DURING TB INFECTION BY OTHER MEANS COULD EXACERBATE TB SO ALAN, DOING THE TREATMENT TO PROTECT AGAINST TB AND LOW AND BOW HOLD IT LED TO TWO LOG FOLD INCREASE IN THE LUNG TB LOAD BUT NOT IN THE RECEPTOR KNOCK OUT MICE SO TYPE 1 INTERFERON IS BAD. IN OUR LAB WE HAD TO EXACERBATE AND THAT WAS A COLLABORATION WITH ALAN AND ANDRE AS WHERE WE FOUND THE INFLUENZA VIRUS A WILL IMPAIR THE PROTECTION AGAINST MTB AGAIN THROUGH A TYPE ONE INTERFERON PATHWAY AND A GENETIC APPROACH GETTING RID OF THIS TPL2 PATHWAY IN MICE AND WE FOUND IT WAS ELEVATED AND FIN MACK NAB SHOW THIS COINCIDED AND DEPENDED ON TYPE ONE INTERFERON SIGNALING SO THIS WAS MEANS THAT YOU HAVE TO USE TO PUT THIS TYPE ONE INTERFERON UP AND THE MICE SUCCUMB SO TO SUMMARIZE A LOT OF WORK PUBLISHED BY A LOT OF PEOPLE, TYPE 1 INTERFERON COULD AT EARLY STAGES, MAYBE BE PROTECTIVE AND THERE'S STILL A LOT TO LEARN BUT WHEN IT WAS LATE SUSTAINED HIGH DOSES IN THE SYSTEMS IT COULD BLOCK IL-12 AND TNF AND INDUCE IL10 AND MY PROGRAM HAS A WHOLE PROGRAM ON IL-10 AND TYPE ONE INTERFERON COULD ALSO BLOCK ACTIVATION OF MACROPHAGE BY FAMILIAR A INTERFERON TO KILL BACTERIA AND ALSO PRO THESE PROTECTIVE CYTOKINES. SO, THAT WAS USING THESE METHODS TO TURN IT ON. I WANT TO REMIND YOU THOUGH OF OF SOME DATA THAT THEN CAT AND ALAN REALLY LED TO THEIR NEXT SEM SEMINOLE FINDINGS IS ALAN HAD FINDINGS IN HIS LAB THAT MY D88 MICE DEFICIENT MICE DISPLAY A PROFOUND LOSS IN RESISTANCE TO MTA AND CAT CAME IN VERY TENACIOUS AND LIKE HANGING ON AND I'VE GOT TO WORK THIS OUT AND I'LL NEVER FORGET SAYING ANNE, WHAT DO YOU THINK IS GOING ON? IT WAS TOUGH TO FIGURE OUT WHY IL-1 PROTECTION WAS REQUIRED DOWN TREATMENT MYD88 AND I'LL TOUCH ON THIS SEMINOLE PAPER THAT FOLLOWED AFTER A LOT OF THINKING AND DISCUSSION OF ALAN AND KAT AND KAT GOING TO CHINA AND REALLY PUSHING UNTIL THIS STAGE WHERE SHE'S ABOUT TO HAVE A BABY AND SHE'S FOUND OUT THE Y AND THE HOW AND LOOK THEY'RE SMILING AND CAT, THIS IS A MIRACLE, SHE WAS SUBMITTING THIS PAPER AND GOING FOR AN AWARD AND JUST REALLY PREGNANT. SO, THIS IS FOR ALL YOUNG PEOPLE TO KNOW ABOUT. KAT AND ALAN AND COLLABORATE HOST DIRECTIVE OF TUBERCULOSIS WAS BASED ON R1 AND TYPE 1 INTERFERON CROSS TALK AND SO THE THE KNOCK OUTS DIED YET WHEN YOU CROSS THEM TO THE KNOCK OUT THEY GOT BETTER SO THIS CROSS REGULATION THAT BOB HAD BEEN TALKING ABOUT WITH GAMMA AND L4 THEN THERE'S BEEN IL-10 AND GAMMA NOW IT WAS TYPE 1 AND L1 AND THERE'S A HUGE AMOUNT TO BE DONE AND A LOT OF THINKING HAS BEEN GOING ON DURING THESE TYPES AND KAT AND ALAN TOGETHER WITH ANDRE HE IS, MARK AND MY LAB MEMBERS WE'VE WRITTEN REVIEWS ON TYPE 1 FOR INFECTIONION DISEASE AND TB AND OCCASIONALLY FRIEND THAT WAS LUCIA WITH KAT AND ALAN AND THIS IS BEEN SO MANY GREAT DISCUSSIONS OF WHERE THIS IS GOING AND TYPE 1 INTERFERON IS TOUGH AND TB IS TOUGH AND WE KNOW THAT KAT AND DAN FOLLOWING ON FROM ALAN WHO WILL BE THERE TO GIVE TIPS AND WHATEVER, WE WERE TRYING TO FIND A MODEL WHERE WE DIDN'T HAVE TO KNOCK OUT A GENE SO WE LOOKED FOR MOLDS AND THERE WAS A MOUSE WHICH HAD BEEN HAVE LUNG RESEMBLING HUMAN CASES GRANULOM A AND COULD IT BE THE STRAIN BASED ON THIS PAST THINKING AND ALL OUR DISCUSSION. WHAT WE DID WAS WE FOLLOWED ON THE BLOOD SIGNATURES OF HUMAN TB AND I'M TAKING YOU THROUGH A LOT OF DATA. THIS IS WHOLE BLOOD OF HUMAN FROM LONDON, SOUTH AFRICA LESTER AND I SHOWED YOU BEFORE WITH A DIFFERENT APPROACH WHERE WE SEE A NEW TRA FILL SIGNATURE AND ANOTHER INDUCE ABLE SIGNATURE IN THE HUMAN. WE USED LOW AND HIGH DOSE OF THE BUG AND EACH BUG TOGETHER WITH EACH STRAIN AND YOU HAVE TO LOOK ON THE RIGHT-HAND SIDE AND SEE WHEN IT'S HIT WITH THIS CLINICAL ISOLATE REALLY STARTS TO LOOK LIKE HUMAN DISEASE. AND SO, ALL OF A SUDDEN WE HAD A MODEL AND SO GOING IN AND LOOKING AT THIS MODEL, IN FACT YOU COULD SEE THAT THE B6 WHEN YOU GAVE IT A HIGH DOSE OF H78 HIGH DOSE, ALSO COULD RESEMBLE A HUMAN DISEASE SO HIGH DOSE, DIFFERENT STRAINS, ALL OF THIS MAKES A DIFFERENCE AND MAKES A DIFFERENCE IN HUMANS AND SURE ENOUGH, LOW DOSE IN THE MICE IF NO KNOCK OUTS HAVE NO EFFECT HIGH DOSE, YOU CAN SEE THE BACTERIAL LOAD BUT YOU CAN SEE THAT IN THE MOUSE ACTUAL LOW THIS IS EVEN A LOW DOSE YOU SEE THE ANTI-IF NOT ANTIBODIES SINCE IT'S THE WRONG BACKGROUND FOR THE KNOCK OUT CAN HAVE A AN AMELIORATING EFFECT. >> THIS WAS AN INTERACTION WITH CAT AND ALAN HELPING US AND CONTRIBUTING, PARTICULARLY WHEN OUR MOUSE CL3 WAS DOWN AND ALSO ON THE THINKING SO LUCIA SHOWED THE MICE REDUCTION OF LESION BURDEN AND CFU AND PRODUCTION OF KNEW TRA FILL TRAP FORMATION AND SHOWN BY H3 AND YOU SEE THAT THE MPO STAINING IS REDUCED BUT STILL THERE AND IN THAT NOT ALL THE KNEW TRA FILLS WERE GONE BUT THE NETS WERE GONE. THE GREN IS FOR THE B CELLS AND THE RED YOU SEE THE T CELLS AND JUST GOING RESISTANCE AND SUSCEPTIBLE LAB STRAIN CLINICAL ISOLATE AND YOU CAN SEE AS SOON AS YOU START GETTING THE PATHOGENESIS, THE B AND THE T CELLS DIMINISH AND WE NEED TO KNOW CHICKEN AND EGG AND WHAT'S GOING ON SO THERE'S A LOT MORE TO EXPLORE THAT WE'LL BRANCH IT AND CONTINUING THE LAB BUT THIS IS FROM ACTUALLY, THIS IS THE EARLY FIGURE REMEMBER ALAN IN THE REVIEW WE'VE JUST WRITTEN WITH ALAN AND ANDRES AND IT'S MUCH CHANGED THAT IT ACTIVATES NEUTROPHIL FOR MICE AND WE STILL ARE NOT SURE WHERE THE TYPE ONE INTER FOUR ON DRIVES RECRUITMENT MONOCYTES AND NEUTROPHIL FROM THE BONE MARROW TO THE LUNG. KAT WILL FOLLOW ON THE IL1 AND HOW THE TYPE ONE INTERFERON CONTINUES AND WHEN YOU GET CONTROL VERSUS DEATH AND SUSCEPTIBLE AND WORE STILL ASKING THIS QUESTION DOES TYPE ONE INTERFERON AT A LOW LEVEL EARLY HELP ON THESE BNT RESISTANT MICE. SO THERE'S STILL A LOT TO ANSWER IN THE EXPERIMENTAL MODELS BUT I STARTED WORK ONG THE HUMAN TB TO TRY AND UNDERSTAND WHAT WAS GOING ON. AS AN IMMUNOLOGIST, I SAID, WHAT REALLY WAS FASCINATING IS THE PROBLEM OF LATENT SEE, A QUARTER OF THE WORLD ESTIMATED TO BE INFECTED BUT ALL YOU CAN SAY IS THAT THEY'RE POSITIVE FOR THIS INTERFERON GAMMA RELEASE ASSAY AND YOU JUST DON'T KNOW WHO IS GOING TO GO AND IN ALL THE TEXTBOOKS IT SAID, THESE LATER INDIVIDUALS CAN PROGRESS TO ACTIVE DUTY AT ANY TIME DURING THEIR LIFETIME AND THIS SOMETHING THAT I'D LUKE TO REVISIT HERE AND SUGGEST THAT IT'S A SHORTER WINDOW WHEN PEOPLE CAN PROGRESS TO TB AND AFTER THAT, IT'S POTENTIALLY REINFECTION AND SO HOW DID WE GET AT THIS? SO THE QUESTION WAS, OF THESE LATENT INDIVIDUALS, THEY COULD BE FIVE TO 10% PROGRESSIVE AND CLEARED THE INFECTION AND IT COULD BE CONTROLLED OR PERSISTENT AND SUB CLINICAL DISEASE AND ALL YOU CAN SAY IS THAT THEY'RE POSITIVE FOR THE TEST WHICH SHOWS THAT THEY REACTED AND LOOKING BACK AT OUR OLD DATA, WHAT I DIDN'T POINT OUT IS THAT THESE LATENT INDIVIDUALS WHERE HETERO GONE YUS THAT THEY LOOKED LIKE THE ACTIVE TBN AND IF YOU JUST LOOK, THESE ARE LATENT BUT THEY HAVE A SIGNATURE OF ACTIVE TB SO THIS IS STATIC. IF YOU LOCK AT A MODULAR LEVEL YOU CAN SEE THAT YOU TAKEAWAY THE ACTIVE TB OUT LIES, THESE THAT WERE THE HETEROGENEOUS ONES AND LOOK AT THE ACTIVE TB BLOOD SIGNATURE AND YOU SEE THE INTERFERON AND THE OUTLIERS AND THE OTHER LATE ENTERS, NO SIGNATURE. THIS IS STATIC. CAN TRANSCRIPTOMIC TELL US ABOUT PROGRESSION TO TB SO TO DO THAT IS QUITE TOUGH AND WE'RE VERY LUCKY TO HAVE COLLABORATORS IN LESTER WHO ARE THIS MINIMIZES REINFECTION OF MTB WITH ANOTHER STRAIN AND SO WE DID A LONGITUDINAL STUDY WITH THEM THAT WE PUBLISHED IN 2018 SO I'M SUMMARIZING IN A CARTOON HERE AND LED IN LESTER AND IN MY LAB LOOKING AT THE ANALYSIS AND THE BOTTOM LINE OF THESE 53 ACTIVE TB PATIENTS, THESE CONTEXT, 108 CONTEXT WERE FOLLOWED SOME WERE NEGATIVE AND 49 WERE POSITIVE BUT STAYED HEALTHY AND THEY'RE STILL HEALTHY FIVE YEARS LATER. THE NEGATIVE WENT POSITIVE WENT ON TO DEVELOP TB AND THIS WAS 10% AND THEY WERE CHARACTERIZED BY A PERSISTENT HIGH BLOOD SIGNATURE IN MOST OF THEM. SO WE WANTED TO LOOK FORWARD AND WE INCREASED THE NUMBERS AND DID A QUITE A DETAIL STUDY WHERE THEY WERE FUNDAMENTAL AND WE LEARNED A LOT AND LINKING CLINICAL PHENOTYPES WITH GENOME DATA SO THE BUG WAS SEQUENCED IN THE ACTIVE TB PATIENTS AND THEN IN THE CONTEXT WHEN THEY PROGRESS TO TB SO WE GOT THE INDEX CASE AND THEY HAD THE SAME SEQUENCE WHICH IS NOT BEEN DONE IN OTHER STUDIES, WE COULD DO IT. SO, THIS IS THE DESIGN OF THE STUDY THAT WE'VE JUST PUBLISHED IN THE JOURNAL OF EXPERIMENTAL MEDICINE AND IT WAS HARD AND IT CAN TAKE MONTHS TO DETERMINE THAT SOMEONE HAS TB BUT, THE ONES WHO HAVE TB ARE RECRUITED TO THE STUDY AND THEN OF 365 IF I CAN FIND MY POINTER. THESE WERE MONITORED OVER THREE YEARS AND IT'S NOW FIVE YEARS AND MEETING WITH HE KNOWS ME NONE OF THE OTHER DEVELOP MOST OF THEM ONE TIGER WOODS AND TWO AND A FEW OF THREE YEARS AND THOSE THAT THEY WERE DEFINITELY CONTEXT OF THOSE TB AND NOW, FROM THE CLINIC, THE PROGRESSIVE TL IT COULD BE PEOPLE WHO HAVE NO SYMPTOMS AND THEY'RE POSITIVE AND NOW CLINICAL AND NO RADIOLOGICAL ABNORMALITIES AND NO EVIDENCE OF ACTIVITY TB AND WE FOUND THAT THESE TOOK BETWEEN 41, 125 AND SOME 200 TO 300 DAYS TO PROGRESS TO T AND THE SUB CLINICAL ARE ONES THAT EITHER HAVE NO CLINICAL SYMPTOMS. THEY REALLY ARE FINE AND THEY'VE BEEN TAKEN BECAUSE THEY WERE CONTACTS OR FOR SOME OF THE REASONS THEY LOOKED AT THEIR LUNG AND THESE MAY HAVE A CHANGE IN THEIR LUNG X-RAY BUT THIS COULD BE ANOTHER LUNG DISEASE, IT COULD BE CANCER, PNEUMONIA OR THEY COULD HAVE GROWN A BUG FROM THESE PATIENTS BUT THEY HAVE NO SYMPTOMS AND NO LUNG SYMPTOMS AND THAT COULD BE CROSS CONTAMINATION OR AN OLD SCARRING. AND THESE ONES WE FOUND DEVELOPED TB WITHIN 20 TO 40 DAYS AND THEN THEY HAD RADIOLOGICAL FEATURES AND EVIDENCE OF ACTIVE AND PARTICULARLY FOLLOWING THESE CONTEXT THE NEW SCENARIO AND SO, WE LOGGED AT THE GONE SIGNATURE AND I'M ONLY SHOWING YOU PART OF WHAT IS IN THE PAPER AND YOU CAN SEE CERTAIN GENES ARE STARTING TO GO UP AND THEN BY SUB CLINICAL, THEY'RE GETTING HIGHER AND THEN AT CLINICAL TB THEY'RE HIGHER AND THESE ARE THE CONTEXT BUT THEY'RE NOT EVEN NEARLY AS HIGH OF WHEN YOU HAVE HIGH FULLY BLOWN ACTIVE TB AND WE HAD A LOT OF THINKING ON THIS AND ANALYZED ALL THE DATA AND THERE ARE A COMMON PATHWAYS ENRICHED IN UP REGULATED GOWNS IN ALL THROW OF THE CLINICAL GROUPS THAT ARE TYPE ONE INTERFERON TIM YOU LATED GOWNS AND HIGHLY ENRICHED SO EARLY ON BEFORE ANYBODY HAS ANY SYMPTOMS, THESE TYPE 1 ARE COMING UP AND THE GLAD SIGNATURE IS MORE AND MORE PRONOUNCED AS THEY PROGRESS. YOU CAN'T -- IT ALSO DEALS US A LOT ABOUT THE POTENTIAL PATHOGENESIS AND WITH THE EXPERIMENTAL MODELS. USING THE MODELS AGAIN WE COULD TEASE OUT A LITTLE BIT MORE AND ACTUALLY WHAT WE CAN SEE HERE IS THAT IN DEED IN THE SUB CLINICAL YOU CAN SEE THE INTERFERON SIGNATURE AND THE SECOND INTERFERON SIGNATURE WHICH USING WGCNA WE TEASE OUT MORE. WE SEE THOSE T-CELL GOWNS ARE STARTING TO GO DOWN IN THIS ACTIVE, IN THIS PROGRESSION FROM NO SYMPTOMS TO THEN HIGH SYMPTOMS AND THIS IS WHAT NEEDS TO BE TEASED OUT MORE BUT OF COURSE, THE BLOOD WE WERE LUCKY TO SEE A SIGNATURE AT ALL IN THE BLOOD OF TB PATIENTS NEVER MIND PROGRESSES SO OUR FUTURE STUDIES IS WHAT I'M GOING TO LEAVE YOU WITH, I'M NOT SHOWING ANY DATA. THIS IS WHAT WE'RE IN THE MIDDLE OF. CONTINUING OUR STUDIES WITH POUR COLLABORATORS ON PATIENTS WITH ACTIVE TB WHERE SAMPLING 30 EXPOSURE GET THE RECENT CONTEXT IN AND WHEN THEY'RE POSITIVE WE WILL BASICALLY TAKE AN AIRWAY SAMPLE AT ONE VISIT THAT WE CAN DISCUSS SO DEEPLY WITH AND REALLY GET TO THE BOTTOM OF IT ALL AND THIS IS JUST TAKING US THROUGH THE JOURNEY WITH MATTHEW BERRY THE FIRST CLINICIANS AND LOTS OF CLINICIANS FOLLOWING. WE STARTED IN LONDON AND WE HAD TO LEAVE THERE BECAUSE THE CLINICIAN, THE TB PROFESSOR DIDN'T WANT TO SHARE SAMPLES SO THEN WE WANT TO LESTER AND IN LESTER THEY HAVE SUCH A FANTASTIC SET UP FOR GETTING TB PATIENTS CONTACTS IN EARLY TO THE CLINIC AND OF COURSE NO RISK OF REINFECTION AS YOU GET IN AFRICA WHEN PEOPLE ARE SUGGEST THAT YOU ARE REACTIVATING TB AND ROB WILKINSON IS A LONG-TERM COLLABORATOR AND THE BEGINNING WAS IN THE HUMAN STUDY WAS REALLY JAQUES AND VIRGINIA WHO GAVE US THE STRENGTH AND THE KNOW HOW TO GO FORWARD AND ALL MY LAB MEMBERS USED TO GO THERE AND GET TRAINED NOW WE'RE VERY HAPPY WE DO THE BIO FORMAT TICKS OURSELVES WITH OUR OWN LAB AND THESE ARE ALL THE PEOPLE CONTRIBUTING TO THE MOUSE WORK AND THROUGH THE YEARS I HIGHLIGHTED FIN SAY LONG-TERM MEMBER OF MY LAB WHO IS STILL WITH US DURING THE MTB MOUSE STUDIES WHICH ARE HARD. WE HAD TO GO TO LONDON SCHOOL AND TAKE A TAXI AT THE BEGINNING OF OUR TB STUDY SO WE EMPATHIZE AND WE'RE LUCKY WITH OUR SCIENCE TECHNOLOGY PLATFORMS AND ANDRES IS A FRIEND AND KAT AND ALAN, I HAVE TO SINGLE OUT THIS IS BEEN SUCH A REWARDING INTERACTION ON WHAT WE ALL AGREE IS A DAMN DIFFICULT DISEASE TO WORK ON AT ALL LEVELS. WE CAN DRINK TO THAT FOR THE FUTURE. GOOD LUCK TO ALL OF YOU ALUMNI OF ALAN'S AND ALL THE TRAINEES WHO HAVE NOW GONE OUT TO THE WORLD TO CURE TB. [APPLAUSE] >> DO I HAVE ANY QUESTIONS? IT'S TOO ARTICLE IN THE MORNING I THINK. >> I WOULDN'T TO CONTRIBUTION [OFF MIC] >> ALAN SAID HE IS GOOD I THINK HE WOULD BE PERFECT BUT HE SITS AT HIS DESK A LOT. [LAUGHTER] YOU LED THE WAY. AND YOU KNOW WHAT, I THINK IT'S WORTH, THANKS FOR HIGHLIGHTING THAT BECAUSE DAMIAN IS BACK IN NEW YORK SINCE FEBRUARY AND WE'RE ALL DELIGHTED THAT WE CAN INTERACT. NEW YORK IS A HOP FROM LONDON AND IT WAS GREAT TO SEE HIS KID AND SEE THAT IT'S ALL SETTLING AND ANOTHER ALUMNI OF ALAN'S. HIM SITTING AT HIS DESK WAS USEFUL. [LAUGHTER] >> THANK YOU FOR THAT FASCINATING TALK. HAVE YOU LOCKED AT EXTRA PULL PULMONARY TB VERSUS DISSEMINATED OR. >> WE PUBLISHED A PAPER THAT WAS SIMON BLACKLY'S WORK WHICH I COULDN'T GO INTO EVERYTHING AND EXTRA PULMONARY TB HAS AN IDENTICAL SIGNATURE AND WE THOUGHT IT WAS WEAKER AND ACTUALLY WHEN WE STUDIED IT, IT WAS MORE SIM POM RELATED. SO, WE PUBLISHED IN OUR FIRST PAPER AND THEN SUBSEQUENTLY THAT THE EXTENT OF LUNG RADIO GRAPHIC DISEASE IN PULMONARY TB CORRELATES WITH THE SIGNATURE AND EXTRA PULMONARY TB WE THOUGHT IT WAS HAVING A WEAKER SIGNATURE BUT WHEN WE SEPARATED THE SYMPTOMS, WE FOUND AGAIN STRONG SIGNATURES WHEN THEY HAVE BAD DISEASE AND WEAKER. IT'S VERY, VERY SIMILAR AND SO THIS IS A QUESTION WE'VE ASKED FOR MANY YEARS AND THE DISSEMINATION AND WHY DOES IT HAPPEN AND IT'S SOMETHING WE'RE NOT FOLLOWING AND IT'S OBVIOUS LOW A HARD ONE BECAUSE IT'S DIFFICULT TO DIAGNOSE. THERE HERE ARE FOLLOWING TO TRY AND GET A SUPPORT FOR DIAGNOSIS OF TB IN THE SIGNATURE TO SUPPORT OTHER ASPECTS LIKE EXTRA PULL MINPULMONARY. >> THANK YOU. ALL RIGHT. THANK YOU. [APPLAUSE] OUR NEXT SPEAKER IS TRAINEE FROM ALAN'S LAB AND ANTONIO ROTHCHS. >> THESE PODIUMS IN THE U.S. MADE FOR MUCH SHORTER PEOPLE THAN THE SWEDISH ONES. I REALLY APPRECIATE THAT. SO, WHERE IS DRAGANA BY THE WAY. I HAVE TO START BY SAYING DRAGANA FOR ORGANIZING THIS FAMILY ROW UNION WHICH HAS BEEN AMAZING SO THANK YOU FOR ORGANIZING THIS AND IT'S A TREMENDOUS HONOR FOR ME TO BE HERE AND TO PRESENT. WE'VE SEEN A LOT OF PICTURES OF ALAN SO I THOUGHT I WOULD BREAK THAT TREND AND I START WITH A PICTURE OF ME SO FOR THOSE OF YOU WHO DO KNOW ME, I'M TONY ROTHFUCHS AND THIS IS WHAT I LOOKED LIKE BACK IN THE DAY SO WHEN I WAS A POSTDOC IN ALAN'S LAB I WAS IN BUILDING 50 AND I ACTUALLY SAT WHERE DAMIAN SAT BUT I DIDN'T SIT MUCH ON THAT SEAT. I WAS MOSTLY SACKING MICE AND ONE OF THE THINGS THAT I DID WAS TO DO MICRO BACTERIAL GRAN YOU LOMA WHICH WAS A COLLABORATION WITH RON'S LAB. WHEN I WAS INVITE TODAY AND AND SPEAK. I HAVET BEEN BACK SINCE 2009. I THOUGHT ABOUT THE EXPERIENCES AND HOW THIS HAS SHAPED MY CAREER AND HOW THIS HAS HELPED ME SET UP SHOP BACK IN AND I'VE FOR THIS SHORT TALK, I TRIED TO CHANNEL THIS TO THREE BASIC COMPONENTS THAT CAN GO THROUGH IN THE ENTIRE TALK. SO, NUMBER ONE, IT'S HERE WERE IN ALAN'S LAB WHERE I LEARNED TO DO MICRO BACTERIAL WORK AND THIS IS SOMETHING THAT MY LAB IN STOCKHOLM DOES ON INFECTION X I LEARNED BSL3 WORK AND THIS IS IMPORTANT TO LAND A KEY COMMISSIONS OF TRUST AT KAROLINSKA WHICH BASICALLY HELPED BRING HOME THE BACON SO TO SAY. AND THE THIRD POINT IS THAT IT'S REALLY -- SO, ONE KEY THING THAT I LEARNED WAS LIKE BE TO BE AFRAID TO JUMP INTO THE DEEP END OF THE POOL SO TO SAY. SO TO TRY NEW THING, SET UP NEW METHODS, A REPLY NEW TECHNOLOGY AND ADDRESS IMPORTANT QUESTIONS. AND I AM NOT DOING SO MUCH IMMUNOLOGY ANYMORE SO I AM IN THE FIELD OF TB ARROW BIOLOGY SO THAT IS REALLY IN THE DEEP END FOR IMMUNOLOGY SO I THOUGHT I WOULD START A LITTLE BIT BY INTRODUCING THIS FIELD WHICH IS REALLY A VERY OLD FIELD IN TB AND YET IT'S STILL AT THE SAME TIME A REEMERGING FIELD SO THIS TB ARROW BIOLOGY WAS KICKED OFF BY CARL IN GERMANY AND AND HE GUINEA PIG TRANSMIT COUGHING. THIS KICKED OFF THE AEROBIOLOGY FIELD. SO TODAY, WE'VE PRETTY MUCH REPLACED FLUGGE'S GUINEA PIGS WITH AIR SAMPLERS TO DO THE SAME THING. SO, THROUGH AIR SAMPLING, IT'S SO IMPORTANT FOR THIS BECAUSE IT A US TO COLLECT THE ARROW SOLE AND DETECT AND ANALYZE THE PATHOGEN COMPONENT. WE'RE STUDYING IMPORTANT GAPS IN TB ARROW BIOLOGY AND TRANSMISSION SO I WILL TOUCH SO YOU CAN IDENTIFY HOTSPOTS OF COMMUNITY TRANSMISSION. WHERE IS TRANSMISSION TAKING PLACE IN A CERTAIN COMMUNITY BY SAMPLING THE AIR. YOU CAN USE IT TO IDENTIFY NEW CASES OF TB BASED ON THE INFECTIOUS AEROSOLS OF THE GENERATED BY PATIENTS AND THIS IS WORK THAT KEVIN FENLY HERE AT NIH HAS REALLY BEEN PIONEERING. AND THE THIRD POINT WHICH IS REALLY EASIER SAID THAN DONE, IS THAT THROUGH AIR SAMPLING WE CAN STUDY THE BUG IN ITS TRANSMISSIBLE FORM AND BY THE WAY THE WORD "BUG" IN CONNECTION TO PATHOGENS I JUST REALIZE IS SOMETHING I PICKED UP FROM YOU, ALAN. PROBLEM THE VERY FIRST THING I PICKED UP FROM YOU AND I HAVEN'T DROPPED IT AND IT DOESN'T TRANSLATE WELL IN SWEDISH. IT SOUNDS LIKE A BUG BUT I USE IT. I JUST REALIZED THAT NOW. SO ANYWAY, SO, WE'RE DEVELOPING AIR SAMPLERS TO ADDRESS THESE POINTS. SO, WE'VE DEVELOPED SOMETHING CALLED A TB HOTSPOT DETECT ERROR 4 AND I CAME UP WITH THE NAME THOR. WORE IN SWEDEN AND WE HAVE TO A COOL NAME SO THOR IS A PORTABLE AIR SAMPLER AND IT'S COMPACT AND EASY TO USE AND YOU CAN SEE IT'S HAND PORTABLE AND IT DOESN'T REQUIRE POWER AND THE PARTS ARE CHEAP AND YOU CAN REPLACE THEM. THE WAY IT WORKS FROM THESE HEADS THERE'S A CORONA DISCHARGE AND IT CHARGES PART ALS IN THE CARE AND THEY GET COLLECTED THROUGH A ON TO THIS COLLECT OR PEACE AND WE REMOVE THIS PIECE AND WE CAN PERFORM ANALYSIS. THIS IS WHAT IT LOOKS LIKE WHEN YOUR SUBPOENA POLL SMOKE. THEY ACCELERATE TOWARDS THE COLLECT OR PEACE AND IT'S CHANGING COLOR AND GETTING A LAYER OF SMOKE AND LET'S WATCH THAT AGAIN. SEE IT'S SHINY AND ON AND THEN THERE IT GOES. THE SMOKE GETS COLLECTED. SO, YOU KNOW, HOW DO WE IDENTIFY NEW INFECTION? SO, A COUPLE OF EXPERIMENTS SO WE STARTED THIS OUT WITH EXPERIMENTS IN BSL2 USING BCG SO THE LIVE VACCINE AND SO THE SET UP THAT WE DEVELOPED THAT WAS FUNDED BY THE GATES FOUNDATION WAS WE SET UP A HUGE PLASTIC CAN PEE IN WHICH WE AR SOLLIZED BCG AND PERFORMED EXPERIMENTS WITH OUR DEVISE AND THIS WE STARTED A COLLABORATION FOR THOSE THAT ARE NOT FAMILIAR WITH GONE EXPERT THIS IS YOU BASICALLY A AUTOMATED PCR SYSTEM USED FOR MOLECULAR DETECTION. IT'S A CARTRIDGE AND YOU PUT IT IN AND IT GOES INTO A LITTLE MACHINE AND IT GIVES YOU A RESULT BACK AND WE MANAGED TO GET CT VALUES FOR THAT BECAUSE NORMALLY WE GET HIGH, LOW, MEDIUM DETECTION AND SO WE PERFORMED THIS EXPERIMENT AND BCG ACROSS VARIOUS CONCENTRATIONS AND WITH THE DEVICE TURNED ON AND TURNED OFF, AND WE WERE ABLE TO EXTRACT THE CT VALUES AS WELL AND SO IT TURNS OUT THAT THE COMBINATION OF THOR AND GENE EXPERT IS AWE SHOULD SO IT GIVES YOU QUITE A SENSITIVE DETECTION AND ABOUT .5 COPIES OF BCG PER LITER OF AIR SO THIS IS KIND OF A -- WHEN THE DEVIS IS TURNED ON, THAT'S WHEN YOU GET THE SENSITIVE DETECTION SO IT IT IS ACTIVE COLLECTION REQUIRED FOR THIS AND SO THIS IS WE THOUGHT OK, THIS SOUNDS GOOD AND BECAUSE WE HAVE NO IDEA OF TRUE CONCENTRATION OF TB IN THE AIR SO IT'S GUESSWORK IN THE AEROBIOLOGY AND WE THOUGHT OK, THIS IS LOW ENOUGH SO LET'S TRY TO DO THIS WITH PATIENTS SO WE TURNED TO COLLEAGUES AT KAROLINSKA UNIVERSITY HOSPITAL AND THIS WAS A SURPRISE TO ME AND THERE'S NEVER GOING TO BE ABLE TO DO THIS IN AND JUST FOR A TRY AND IT TURNS OUT THAT YES, WE HAVE TB IN SWEDEN AND UNFORT LOW THERE'S NOT ENOUGH SWEDES IN THE AUDIENCE HERE FOR ME TO MAKE THAT AS A SUCCESSFUL PITCH THAT YES, WE DO HAVE TUBERCULOSIS EVEN IN SWEDEN AND AT ANY GIVEN DAY OF THE WEEK, AND KAROLINSKA UNIVERSITY HOSPITAL YOU WILL HAVE FOUR PEOPLE WITH TB IN THE WARD, AT LEAST. TODAY WE HAVE MORE TB CASES THAN COVID CASES ACTUALLY. AND WE PERFORMED GENE EXPERT ON THIS AND SO, WE, THIS IS REALLY PRELIMINARY SO WE WERE ABLE TO TRY FOUR PEOPLE AND WHERE DOING THIS AND SO IF YOU LOOK HERE FOR EXAMPLE, SO, YOU SEE THIS. WE TRIED TO SAMPLE PEOPLE FOR SIX DAYS SO WE FIND THIS TRIANGLE, THIS INDIVIDUAL WAS POSITIVE BY COUGH TWICE AND THE ROOM AIR WAS ALSO POSITIVE AT THE SAME TIME, AT THE SAME EVENT OF COUGH SAMPLING AND THIS WAS THE CASE FOR ALL OF THESE INDIVIDUALS. THEY'RE POSITIVE, MOST OF THE TIME THEY'RE POSITIVE TWICE OUT OF FOUR OR FIVE SAMPLE OCCASIONS AND WHICH IS REALLY COOL THAT WE ARE ABLE TO MAKE A POSITIVE MOLECULAR DETECTION JUST BY HAVING A PERSON COUGH ONTO THE DEVICE. BUT THIS IS, I MEAN, THIS IS EARLY DAYS AND NEVERTHELESS, THE CONCEPT, WHERE AND HOW COULD WE USE THIS? SO, IN THE ONE END WOULD BE TO USE A POINT OF CARE SO YOU GO OUT AND THIS DEVICE AND YOU ARE ASKING YOU HAVE THIS AND YOU CAN DO IT ANYWHERE. THE OTHER APPROACH AN YOU CAN SAMPLE THE AIR. FOR EXAMPLE, HERE WOULD BE A GOOD EXAMPLE TO SEE IF WE CAN MAKE POSITIVE DOUGH TEXT AND SO THAT WOULD BE MORE MONITORING OR AN ENVIRONMENTAL SURVEILLANCE FOR TB AND EVEN IN THIS SETTING, YOU COULD FOLLOW ON WITH THE POINT OF CARE SETTING TO IDENTIFY INFECTIOUS INDIVIDUALS IN THAT SETTING. IT'S ON GOING HERE AND WE'RE FOCUSING MORE ON THE POINT OF CARE ASPECT AND IN COLLABORATION WITH PARTNERS AT STELLEN BUSH AND IN MOZAMBIQUE AS WELL AND WORE TRYING TO FIGURE OUT WHAT DOES IT MEAN TO BE PCR POSITIVE IN THIS DETECTION METHOD. WHAT IS THE DIAGNOSTIC PERFORMANCE AND HOW LONG DO YOU HAVE TO SAMPLE AN INDIVIDUAL AND WHAT'S THE PROTOCOL TO MAKE THIS. AND THEN LATER ON IT WOULD BE TO SEE IF ONE IS ABLE TO ASSOCIATE WITH ACTIVE TB IN THE HOUSEHOLD BECAUSE IF IT WORKS, IT WOULD BE A VERY SIMPLE WAY THAT YOU COULD JUST, ANYONE COULD MANAGE A THOR. AND THE PROOF OF THAT IS THAT WE'VE DONE THIS PRELIMINARY IN A BRAZILIAN PRISON WHERE WE HANDED OVER THE DEVICE TO INMATES AND THEY WERE ABLE TO SAMPLE THEMSELVES. SO IF IT WORKS IN A BRAZILIAN PRISON, IT WILL WORK ANYMORE AND IT'S KIND OF THE RATIONALE BUT IT'S EARLY DAYS SO THIS IS WHAT IS ON GOING. SO, WHAT ABOUT AIR SAMPLING TO STUDY THE INFECTION BIOLOGY SO THIS IS VERY TRICKY AND AND IN LET'S SAY THE INFECTION TRANSMISSION AND WE KNOW THAT TB IS AN AIRBORN DISEASE AND WE KNOW VERY LITTLE ABOUT THIS PART AND SO YOU CAN FIND OF SPLIT THIS UP. I MEAN, YOU CAN SPLIT IT UP INTO HOW DOES THE DOUG SURVIVE IN THE AEROSOL AND THAT IS IMPORTANT FOR INFECTION PREVENTION AND HOW LONG IF I COUGH AND I HAVE TB AND I LEAVE THIS ALMOST FOR HOW LONG IS THAT BUG GOING TO BE REMAINING IN THE AIR AND BE A RISK FOR TRANSMISSION SO IT'S A KEY QUESTION. ALSO, YOU KNOW, IN WHAT DOES IT MEAN? THAT AGING IN THE AEROSOL, HOW DOES IT IMPACT THE INFECTION EVENT IN THE TARGET CELLS AND THE MACROPHAGE AND OF COURSE IMMUNE RESPONSE, I AM AN IMMUNOLOGIST AND I WOULD LIKE TO SEE HOW THE ARROW S AEROSOL PROG AND THERE ARE QUESTIONS WITH THE AEROSOL AND WHAT'S THE PARTICLE SIZE BECAUSE IF YOU JUST DOUBLED THE DIAMETER OF THE PARTICLE YOU HAVE INCREASED THE HAVE 10 TIMES SO THAT CAN HAVE A MAJOR IMPACT ON MAKING IT MORE COMFY FOR THE BUG AND TO TRAVEL IN THE AIR AND IT CAN ALSO MEAN A MUCH HIGHER PATHOGEN PAY LOAD AND THAT WOULD IMPACT ON THE ABILITY TO INFECT. AND SO TO TRY TO INVESTIGATE THIS REQUIRES MORE THAN JUST AIR SAMPLING THERE AND IT'S MORE LIKE AIR SAMPLING IS A COMPONENT OF A WHOLE SET OF TOOLS AND GADGETS THAT WE'VE JUST SET UP IN OUR BSL3 AT KAROLINSKA SO WE -- IT'S CRAZY JUST LOOKING AT THE PICTURE. IT'S AN AIRTIGHT GLOVE BOX INSIDE AN AIR FIGHT FLEXIBLE CANOPY AND WE CAN GAS THE HELL OUT OF EVERYTHING AND DISINFECT AND HELP A FILTER IT ALL-OUT SO MAKE SURE THAT NOTHING GETS OUT AND IN THIS GLOVE BOX HERE WE'RE USING THE NEGOTIAT NUMBER YOU LT HAS A IT HAS LIVE BASIL EYE SO WE'RE USING A TAKE IT IN THIS BOX AND THEN WE CAN AGE THE AEROSOL IN THE BOX AND WE COLLECT THEM ON SOMETHING CALLED CORYOLIS SO THIS IS A COMMERCIAL HIGH-VOLUME AIR SAMPLER AND WOVE MADE SOME MODIFICATIONS TO MAKE IT BIO SAFE FOR USE IN BSL3 AND THE WAY THAT IT WORKS IS IT SUCKS PARTICLES INTO THIS HEADPIECE THROUGH A CORY OWE LIS EFFECT AND THEY GET STUCK IN THAT CHAMPAGNE CONE AND IT'S LIQUID AND YOU CAN ANALYZE THE PARTICLES SO IT'S A NICE EXTRACTION PROCESS. WE'VE USED THE SET UP AND IN PRELIMINARY EXPERIMENTS I'M GOING TO SHOW YOU USING BCG, WE HAVE AR SOLLIZED BCG AND COLLECTED THEM ON THE CORY OWE LIS AND THIS SPRAY FACTOR IT'S NOT REALLY IMPORTANT IT'S JUST REALLY -- IT'S A MATHEMATICAL KIND OF DATA CRUNCHING FOR YOU TO REPORT THE AMOUNT OF CFUs THAT YOU'VE EXTRACTED FROM THE AIR RELATIVE TO THE AMOUNT THAT WAS IN THE COLUMN SO IT'S DATA CRUNCHING BUT IT'S A MEASUREMENT OF VIABLE BUGS, THE VIABLE FRACTIONION AND OVER TIME THE SPRAY FACTOR IS GOING DOWN AND IT MEANS THE C RECOVERY GOES DO. IF YOU TAKE THE BUGS THAT WERE AT THIS EIGHT-MINUTE AGING, EARLY, AND YOU COMPARE THEM TO BUGS THAT COME STRAIGHT OUT OF BROTH AND YOU CAN ADD THEM TO BONE MARROW MAX, IT SEEMS THAT THE BUGS THAT ARE COMING FROM THE AIR ARE HAVING A HARD TIME ESTABLISHING THE INFECTION IN THE MACROPHAGE AND THIS IS EARLY DAYS. AND SO WE'RE IN THE PROCESS OF REPEATING THIS WITH TB NOW AND THEN SO WE HAVE A COLLABORATION WITH CHAD ROY AT TO YOU LANE UNIVERSITY AND HE IS LIKE THE KING. HE USED THE COLD BERG DRUM THAT KEEPS AEROSOLS, BUGS IN THE AEROSOLS STATE FOR AS LONG AS YOU WANT PRETTY MUCH WHAT HE HAS DONE HERE IS COMPARED IT SEEMS THAT THE BUGS GOING DOWN IN RISK IN TWO AND ONE THEY SEEM TO SURVIVE LESS WELL IN AEROSOL COMPARED TO TB AND IN THIS CASE THE CDC 1551 WHICH IS A REPRESENTATIVE OUTBREAKS STRAINS AND THAT IS COMMERCIALLY AVAILABLE AND SO, WHAT WE WANT TO DO NOW IS TOGETHER WITH CHAD AND TOGETHER WITH OUR COLLEAGUE AT STELLEN BASH, WE WANTED TO INVESTIGATE WHAT THIS MEANS WITH IS THERE THE ABILITY OF A STRAIN THAT IS SUCCESSFUL WITHIN A GIVEN COMMUNITY AND A GIVEN ENVIRONMENT BECAUSE IT'S MORE FIT AND IT'S ADAPTED TO THE AEROSOL TRANSPORT PROCESS AND YOU CAN ADD THIS TO MACROPHAGES AFTER AEROSOL AGING AND SEE WHAT WE GET OUT IN THE END. AND THAT IS WHAT I WANTED TO SHARE WITH YOU GUYS TODAY SO THIS IS US AT OUR NEW BUILDING AND WE'RE IN THE NINTH FLOOR AND I'M AFRAID OF HEIGHTS BUT WE HAVE TO BE ON THE TOP BECAUSE WHERE THE BSL3 SO IT'S NICE I HAVE A BSL3 BY MY OFFICE AND ALL THE COMPLAINTS FROM ALL THE USERS IS NOT SO NICE AND I TAKE THAT AS PART OF A JOB BECAUSE THERE'S TREMENDOUS OPPORTUNITIES AND THE WORK I SHOWED YOU IS DONE BY IN THE LAB BY NUNO RUFINO DE SOUSA, A STUDENT FROM PORTUGAL. THANK YOU, VERY MUCH. [APPLAUSE] THAT WAS WONDERFUL. I DON'T HAVE MUCH OF A QUESTION BUT MORE LIKE A COMMENT SO I HAVE COLLEAGUES BACK AT RUTGERS AND THEY'VE BEEN LOOKING AT HIGH TRANSMISSION AND LOW TRANSMISSION HOUSEHOLDS ISOLATES AND IT SEEMS LIKE I'M NOT SURE IF THEY FIGURED OUT WHETHER THEY'RE AGAIGONEOWE TYPICALLY DT SEEMS LIKE IT'S INTERESTING AND THE HIGH TRANSMISSION STRAINS SEEM TO INDUCE ACTUALLY LOWER BUG COUNTS IN THE LONG AND THE G -- SOTHEN THERE'S THIS PROPULF THESE BASILII WHERE THE LOW TRANSMISSION STRAINS THEY'RE HIGHER IN THE LUNG BUT THEY'RE ENCASED IN MORE LIKE A TH1 GRANULOMA SO I THOUGHT IT WOULD BE SO COOL, RIGHT, IF YOU CAN AGAIN --IF THEY FIGURE OUT A CN IF YOU COULD SAMPLE RIGHT FROM THE AND FROM THE AIR AIR SAMPLING OF WHICH TYPES OF BASIL EYE ARE GETTING HIGHLY TRANSMITTED,. >> IT'S A GREAT POINT. SO, THE COOL THING ABOUT THIS STELLEN BASH STRAINS WE ARE BASICALLY PICKING IN GRANT FREEZERS THEY'VE BEEN SEQUENCES AND THEY FOLLOWED UP THESE TWO COMMUNITIES IN THE WESTERN CAPE THIS IS ROB WARRENS STUFF AND SO THEY'VE BEEN SEQUENCED SO WE WILL HAVE THAT INFORMATION AND THERE'S MANY ASPECT THAT'S YOU'VE POINTED AT THERE THAT I MEAN THERE'S DIFFERENT WAYS TO LOOK AT THIS. OTHER THING THE FIELD IS INVESTIGATING VERY MUCH IS WE HAVEN'T PULLED DOWN ENOUGH BUGS TO BE ABLE TO DO GENE EXPRESSION ANALYSIS BECAUSE IT'S VERY FEW BUGS HERE SO THAT WOULD BE A WAY TO ADDRESS THAT FURTHER AND. >> WONDERING ABOUT TWO SORT OF RELATED USE WITH THAT THOR SAM PLEASURE YOU SHOWED A NICE PICTURE OF IT PULLING SMOKE IN SO, I ALSO THE RELATIONSHIP BETWEEN HOW FAR AWAY YOU CAN PULL PARTICLES AND THE PARTICLE SIZE SO DO YOU SAMPLE DIFFERENT SIZE PARTICLES AND DIFFERENT DEFICIENCIES. >> YEAH, SO THIS IS THE PART OF THE TRY TO UNDERSTAND THE SPECIFICATIONS OF THE DEVOICE VOICE SO BECAUSE IT'S AN OPEN DEVICE SO WE CAN'T MEASURE THE FLOW RATE WHICH SAY MEASUREMENT OF MANY OF THESE AIR SAMPLERS SO CONSIDER SOMETIMES LIKE A BIG VACUUM CLEANER AND IT WOULD SOMETIMES YOU CAN ADAPT THE FILTER TO THE VACUUM CLEANER AND YOU TURN IT ON AND YOU KNOW IT'S GOING TO -- THE FLOW RATE OF THE VACUUM CLEANER IS 2,000-LITER AND THE VOLUME OF THE ROOM SO YOU HAVE AN IDEA HOW LONG DO I NEED TO SAMPLE IT'S AN OPEN DEVICE SO WE WE KNOW THERE'S NO AIR FILTRATION AND THERE'S NO AIR FLOW OTHER THAN THAT GENERATE BID THE DEVICE AND IT COMES TO YOUR OTHER POINT OF HOW FAR AWAY YOU CAN BE SO THE DEVICE ITSELF, THE SAMPLING PROCESS DOES GENERATE AN AIR FLOW OF SEVERAL LITERS PER MINUTE SO IT'S THAN IT WILL ONLY SAMPLE MATERIALS WHICH IS PRESENT BUT IF YOU ARE COUGHING IT WILL BE MUCH HIGHER THAN YOU PICK UP FROM SOMEONE FURTHER AWAY BUT WE'LL HAVE TO DETERMINE THAT BY SEQUENCING THE PATIENT'S BUG IF WE GET IT FROM THE COUGH. CERTAINLY HOW MANY THORS WOULD YOU NEED TO SAMPLE THIS ROOM? IS THE QUESTION. WE KNOW ONE THOR FOR 10,000 LITERS IS OUR ROUGH ESTIMATE. ANOTHER CAVEAT IS THAT IT'S A GOOD CAVEAT BUT BAD FOR THE SCIENCES THE ELECTROSTATIC SAMPLING KILLS TB AS PART OF THE COLLECTION PROCESS SO IT'S ALSO A FILTRATION DEVICE STERILIZING DEVICE AT THE SAME TIME AS IT COLLECTS WHICH IS GOOD. >> IT'S GOOD FOR COMPLIANCE. >> IT'S VERY GOOD. BECAUSE OTHERWISE YOU ARE CONCENTRATING TB INTO THIS TINY -- IT WOULD BE DANGEROUS. WE KNOW IT'S STERILIZING AND SO YEAH WHERE WAS I. >> IT SOUNDS LIKE THE VOLUME ISSUE IS IT'S A GREAT SAMPLE FOR A HOUSEHOLD. YOU ARE NOT GOING FOR A FOOTBALL STADIUM. >> WE'RE NOT GOING FOR THAT, NO. IT WILL BE SOMETHING YOU CAN TAKE WITH YOU, RIGHT SO YOU CAN HAVE MANY OF THESE SO IT CAN BE THE SMOKE DEVICE TYPE OF THING IN A PRISON CELL WHERE THE INMATE DOES THE SAMPLING AND HANDS OFF THE BULLET OR IT COULD BE WHAT YOU GO INTO THE VILLAGE AND ASK PEOPLE TO COUGH ON TO THIS AND THEY CAN EITHER SHIP THE BULLET OR YOU WASH THE BULLET AT HOME SO IT'S THAT TYPE OF -- YEAH. [OFF MIC] [OFF MIC] YES, SO I HAVE TOLD MANY PEOPLE HERE AND THE PANDEMIC HAS REALLY BEEN HORRIBLE AND ESPECIALLY IN SWEDEN BUT THERE'S THE POSITIVE SIDE OF THE HORRIBLE SITUATION THAT IS IT HAS BEEN VERY GOOD FOR INFECTIOUS DISEASE RESEARCH AND WHAT SPILLS OVER IN TB SO YES WE'VE DONE A LOT OF WORK WITH COVID AND ACTUALLY WE'RE USING THOR TO SAMPLE COVID AND THIS WAS DONE IN (INAUDIBLE) SO WE DIDN'T HAVE ETHICAL TO GET PEM TO COUGH ON TO THE DEVICE BUT WE SAMPLED THE ROOM AND ROOM AIR OF PATIENTS AND WE TOOK SURFACE SWABS AND WE ACTUALLY SO WE'RE THE FIRST GROUP TO TO RECOVER PFUs FROM AIR, COVID PFUs FROM AIR SO THERE'S A LOT OF RNA DETECTION OF AIR FROM COVID AND PATIENT WARDS AND ALSO ON THIS WALLS AND SURFACES AND IN SURFACES THAT CAN'T BE TOUCHED WHICH SUGGEST THAT THEY MUST HAVE FLOWN, TRAVELED THROUGH AIR STREAMS THERE AND SO WE ACTUALLY USING THOR WHICH IS SURPRISING BECAUSE I JUST SAID IT STERILIZES BUT IN THIS CASE WE WERE ABLE TO ACTUALLY REGROW PFUs IN CELLS SO IF YOU ACTUALLY DO THE EXPERIMENT INVET ROW AND YOU PUT VIRUS ON TO THE VICE. THE AIR FILTRATION WAS VERY LOW SO IT WAS 2.4 AIR CHANGES PER HOUR SO VERY, VERY LOW AND SO I WILL NOT ANSWER -- FELIX WILL ANSWER A BUT I WON'T ANSWER A QUESTION ABOUT FACE MASKS BECAUSE I IF THERE WAS ANYONE WITH A FACE MASK QUESTION I WON'T ANSWER THAT BUT I WILL SAY SO AIR FILTRATION IS THE KEY THING FOR PREVENTING AIRBORN TRANSMISSION. SARS IN THE I WON'T MAKE A STATEMENT ABOUT FACE MASKS BUT I WILL SAY AIR FILTRATION IS IMPORTANT. I CAN HAVE ONE MADE FOR YOU NO PROBLEM. >> THAT'S REALLY GREAT. THAT'S FUN. IT'S A SUGGESTION RATHER THAN A QUESTION. THEY TAKE TIME AND IT'S A SLOW PROCESS IF YOU MOVE ON TO MOLECULAR SENSORS, DETECTION OF TB IT WOULD BE GOLDEN THAT'S WONDERFUL. >> THANK YOU. ALL RIGHT. [APPLAUSE] SO OUR NEXT SPEAKER IS CHUCK SCANGA FROM UNIVERSITY OF PITTSBURG SO HE IS ONE OF THE THREE MUSKETEERS. >> THANK YOU, CARL. THANK YOU ESPECIALLY TO DRAGANA FOR BOG THE OPPORTUNITY TO SPEAK TODAY AND ALSO FOR ORGANIZING THIS ENTIRE SYMPOSIUM. AS CARL MENTIONED I'M ONE OF THE ORIGINAL TB MUSKETEERS, CARL, DAMIAN AND I SPENT A LOT OF TIME SETTING UP THE BSL3 AND IT ALLOWED ME TO DO WORK IN KNOCK OUT MICE AND THE WORK THAT FEE LEX MENTIONED YESTERDAY AND AFTER OUR BSL3 GOT UP AND RUNNING I PICKED UP A STUDY THAT HAD BEEN STARTED IN THE LAB BY MARCO SHEET OWE USING HIV TRANSGENIC MICE TO LOOK AT CO INFECTION AND I EXTENDED THAT WORK INTO MTB AND THAT'S A -- THAT CO INFECTION WORK IS SOMETHING THIS CONTINUES IN MY OWN LAB TODAY SO FOR THIS AUDIENAUDIENCE IDON'T NEED TO GL HEALTH THREAT AND NEEDLESS TO SAY IT'S I USE IT COMING FROM RUSSIA AND THIS IS A STUDY THAT STARTED WITH A COLLABORATION WITH SHELBY OWE GONER ABOUT EIGHT YEARS AGO AND WE USED THESE MACAQUES THEY'VE BEEN USED TO STUDY HIV USING SIV AS A GARROW GET AND THEY HAVEN'T BEEN USED MUCH TO STUDY TB AND SALLY SHARP PUBLISHED THE FIRST PAPER IN 2017 JUST AS WE WERE GETTING STARTED AND THE REASON WE USED THEM IS THE MACAQUES COME FROM THIS TINY ISOLATED ISLAND OF MAURITIAS AND THEY STARTED WITH A FOUNDER POPULATION OF JUST 12 ANIMALS AND THIS IS GENETIC BOTTLENECK RESULTED IN A LIMITED NUMBER OF MHC HAB LOW TYPES AND IT'S TH HAPPEN LOW TYPES. WHEN YOU LOOK AT THE PROGRESSION OF TB IN THESE TWO SPECIES, YOU CAN SEE THERE'S FIRST THERE'S A SPECTRUM OF SUSCEPTIBLE WITHIN EACH OF THE SPECIES BUT IN GENERAL, THE CHINESE MACAQUES TEND TO BE MORE RESISTANT AND THEY EXHIBIT LESS PATHOLOGY THAN DO THE RECESS AND THEY HAVE LOWER BACK TER Y'ALL ROADS COMPARED TO THE RHESUS AND WE USE PET IMAGING AND REUSE FDG AS A PROBE FOR A NON SPECIFIC PROBE FOR IN FROM MISSION AND IT LIGHTSFOR --THEY ARE MUCH CLOSES AND SO SHELBY AND I STARTED THIS PROJECT QUITE A FEW YEARS AGO AND THE FIRST QUESTION WE ASKED IS DOES PREEXISTING SIV INFECTION AFFECT TB PROGRESSION AND TO DO THIS STUDY WE INFECTED SOME ANIMALS WITH 3,000TDI50 AND WE EFFECT THEM INTER RECTAL' AND SIX MONTHS LATER, WE CO EFFECT THESE ANIMALS WITH LOW DOSE OF TUBERCULOSIS ANDE USE THE STRAIN AND WE DELIVER IT BY DELIVERY AND WE FOLLOWED THOSE ANIMALS AND WE COMPARED THE TB PROGRESSION IN THOSE CO EFFECTED ANIMALS TO ANIMALS INFECTED WITH MTB ALONE AND THE CO EFFECTED ANIMALS EXHIBITED A LOSS OF CD4 CELLS AS A RESULT OF THE SIV INFECTION AND THEY INTENDED TO HAVE MORE TB DISEASE AND HIGHER BACTERIAL BURDEN. AND GIVEN THAT TB IS RESPONSIBLE FOR SO MANY DEATHS IN PEOPLE LIVING WITH HIV AND IT REALLY BEGS TO QUESTION IS THERE A WAY THAT WE CAN PROJECT THESE HIV POSITIVE INDIVIDUALS FROM TB? OF COURSE, THE SIMPLEST WAY IS WITH A VACCINE AND IN DEED THERE'S A VACCINE FOR TUBERCULOSIS WHICH IS A LIFE ATTENUATED STRAIN BCG IS ONE OF THE MOST WIDELY USED VACCINES IN THE WORLD AND ONE OF THE LEAST EFFECTIVE AND IT IS BEEN AROUND FOR 100 YEARS WHEN IT WAS FIRST DEVELOPED BY CALMETTE-GUERIN AND IT'S GIVEN SHORTLY AFTERBIRTH AND THE DOSE IS FIVE TIMES 10 TO THE FIFTH CFU AND IT'S GIVEN INTRA BERMALLY AND IT DOES PROTECT AGAINST CHILDHOOD TB AND AS THEY GROW UP IT IS POOR PROTECTION AGAINST ADULT PULMONARY TB. WE SEE THIS IN MACAQUES. THIS IS WORK WE DID HERE AT THE VACCINE RESEARCH CENTER AND YOU CAN SEE IN THE SUSCEPTIBLE MA CONTACT MODEL IT GIVES YOU ONE LOG OF PROTECTION IN TERMS OF CFU AND MACAQUES BY CHANGING THE DOSE OR ROUTE WE COULD GET BETTER PROTECTION. AND IN THIS STUDY, HE INCREASED BY 100 FOLD THE INTRA DETERMINAL DOSE OF BCG IN MACAQUES AND DELIVERED HIGH DOSE BCG INTRAVENOUSLY AND HE DID INCLUDED ANIMALS VACCINATED BY THE AEROSOL ROUTE AND BOB AND HIS TEAM FOLLOWED THE DEVELOPMENT AND THE AIRWAY RESPONSES USING ABL AND FLOW CYTOMETRY TO LOOK AT CELLS AND REALLY IT WAS ONLY THE INTRAVENOUS ROUTE THAT WAS AWAY FROM THE TYPICAL LVO OR MACROPHAGES THAT ARE MOST PREVALENT IN BAL TO THIS T-CELL PROMINENT PHENOTYPE AND THAT IS MAINTAINED FOR MANY, MANY WEEKS AND MONTHS AFTER VACCINATION. ONLY IN THE INTRAVENOUSLY VACCINATED ANIMALS AND THEY SENT THOSE ANIMALS TO RTB GROUP AT PIT LED BY JOANNE FLYNN AND WE CHALLENGED THOSE ANIMALS AND YOU CAN SEE THAT THE ANIMALS VACCINATED INTRAVENOUSLY 90% WERE PROTECTED FROM M TUBERCULOSIS. IN FACT, SIX OUT OF THE 10 ANIMALS IN THIS GROUP EXHIBITED STERILIZING IMMUNITY AND IN OTHER WORDS, WE COULD NOT CULTURAL ANY MICRO BACTERIAL FROM THOSE ANIMALS AT THE TIME OF NECROPSY SO THIS WAS REALLY DRAMATIC PROTECTION. IT'S CHALLENGES WITH NTB AND NECROPSY 12 WEEKS LATER. THE SECOND GROUP OF ANIMALS, THESE ARE SIV NAIVE ANIMALS VACCINATED WITH HIGH DOSE INTRAVENOUS BCG AND SEVERAL MONTHS LATER CHALLENGED AGAIN WITH M TUBERCULOSIS. AND ONE GROUP OF ANIMALS WAS INFECTED WITH SIV AGAIN, WE USED THE MACK 239 STRAIN GIVEN ININTEREST RECTAL' AND FOLLOWED THESE ANIMALS AND FOLLOWED VIRAL LOADS IN THESE ANIMALS AND FIVES LATER WE VACCINATED THEM WITH A HIGH DOSE OF BCG INTRAVENOUSLY AND THEN CHALLENGES THEM SEVERAL MONTHS LATER WITH M TUBERCULOSIS. NOW MY BIG FEAR WAS IN THESE IMMUNO DEFICIENT ANIMALS SIV POSITIVE ANIMALS, THIS HIGH DOSE LIVE BCG INJECTIO INJECTED INTOD STREAM AND KILL THESE MONKEYS AND THAT WAS MY BIGGEST FEAR AND BECAUSE OF THAT I DECIDED TO TREAT THE ANIMALS WITH TO KILL THE BCG THROW TO FOUR WEEKS AFTER VACCINATION. AND JUST TO KEEP THAT CONSTANT AND ALL THE GROUPS, ALL THE ANIMALS REGARDLESS OF WHETHER THEY'RE VACCINATED, RECEIVED THIS SAME HRA REGIMENT WHICH WE STOPPED ONE MONTH BEFORE CHALLENGE SO THAT THE DRUGS WERE NOT PRESENT WHEN WE ACTUALLY DID THE CHALLENGE. THE FIRST QUESTION IS IT SAFE? THEY WERE NEGATIVE IN THE BLOOD CULTURE AND WE CAN'T GO OUT BCG TWO WEEKS AFTER VACCINATION AND THEY DIDN'T EXHIBIT ANY CLINICAL SCIENCE OF BCG DISEASE AND THEY HAD NO NORMAL ESRs SO JUST BASED ON THOSE KIND OF CURSORY FINDINGS, IVBCG DOES APPEAR TO BE SAFE IN SIV AND THE CAVEAT THOUGH IS THAT AT LEAST WHEN THEY WERE TREATED WITH ANTI BCG DRUGS, STARTING THREE TO FOUR WEEKS AFTER VACCINATION. SO THE NEXT QUESTION BECAME WHAT EFFECT DOES THAT BCG HAVE ON SIV SO AGAIN WE UPON INFECTION, YOU SEE ALL THE ANIMALS EXHIBIT THIS SPIKE IN VI REEMIA THAT IS THEN THEY HAVE A SPIKE IN VIREMIA AND BACK UNDER CONTROL AND ALL OF THESE ANIMALS REESTABLISHED THEIR ORIGINAL VIRAL BASE LINES. WHAT ABOUT IMMUNO. JUST LIKE THE MACAQUES WE SAW IN SIV NAIVE MACAQUES IV BCG HAD A SHIFT FROM MACROPHAGE DOMINATED BAL CELLS TO ONE DOMINATED BY T CELLS AND THAT WAS MAINTAINED FOR MANY WEEKS AFTER VACCINATION. AND QUITE SURPRISINGLY TO ME WE SAW THAT SAME AFFECT HERE IN THE SIV POSITIVE ANIMALS AND AGAIN, EVEN IN ANIMALS INFECTED WITH SIV, IVBCG STILL ILLICITTED THIS DRAMATIC INCREASE IN THE PREPORTION OF T CELLS IN THE BAL. IT WAS T CONTROLS SHOWN IN THE BAL AND THIS IS BUSY LIED BUT THE THIN LINES AND WHETHER OR NOT THEY HAD PREEXISTING SIV INFECTION YOU SEE THE INCREASE IN THE LUNG AND THESE T CELLS THAT COME INTO THE LUNG WERE ACTUALLY PRIME TO MICRO BACTERIA AN THROW AGAINST SO YOU SEE THIS SHOWING CD4 MEMORY CELLS THAT ARE RESPONDING BY MAKING CYTOKINES IN RESPONSE TO PPD RESTIMULATION. YOU CAN SEE WHETHER THEY HAVE SERVE OR NOT FOLLOWING MTB CHALLENGE. IN THE SIV AND VACCINATIONED AND WE SAW COMPLETE PROTECTION FROM DISEASE IN ALL OF THE ANIMALS. WE SAW THAT IVBCG ELICITED STRIKING PROTECTION IN THE MAJORITY OF ANIMALS THIS SEVEN OUT OF THE 10 ANIMALS THEY REALLY HAD NO DISEASE VISIBLE BY PET CT AND ONE ANIMAL WAS PARTIALLY PROTECT AND TWO ANIMALS THAT WERE VACCINE AND WE CAN QUANTIFY THIS PROTECTION BY MEASURING THE TOTAL FDG ACCUMULATION AND YOU CAN SEE THAT IDBCG ANIMALS, WHETHER THEY'RE INFECTED AND WHETHER THEY'RE INFECTED OR NOT AND AND THEY HAVE STERILIZING IMMUNITY TO A MTB CHALLENGE AND SEVEN OUT OF THE 10 ANIMALS SIV POSITIVE ANIMALS WERE SIMILARLY STERILE AFTER CHALLENGE. AND ONE ANIMAL IS PARTIALLY PROTECTED AND TWO ANIMALS WERE NOT. AND I THINK ONE OF THE MOST DRAMATIC FINDINGS FROM THIS STUDY WHEN WE DID PBMC ALLEY SPOTS, AFTER INFECTION AND RESTIMULATED THE CELLS WITH MTB-SPECIFIC ANTIGENS SO WHICH ARE PRESENT ONLY IN MTB NOT PRESENT IN BCG AND AS EXPECTED BEFORE MTB CHALLENGE AND YOU DON'T SEE ANY RESPONSES IN ANY OF THE ANIMALS AND IN THE UNVACCINATED ANIMALS, AFTER MTB CHALLENGE, YOU SEE THE NICE INTERFERON GAMMA RESPONSES IN THE PBMCs IN RESPONSE TO THESE MTB-SPECIFIC ANTIGENS AND QUITE STRIKINGLY IN THE VACCINATED ANIMALS, EVEN AFTER MTV AND YOU DID SEE RESPONSE AND IT SUGGESTS THAT THE VACCINATED ANIMALS AND ELIMINATING CLEARING THE BACTERIA BEFORE AND ADAPTIVE AND IS EVEN GENERATED AND THAT IS QUITE INTRIGUING AND AMAZING SO THE CONCLUSION 0 CON IT PROTECTV NAIVE AND CONFERRED PROTECTION IN 80% OF THE SIV POSITIVE AND SO THOSE STUDIES CHRONICALLY EFFECTED WITH SIV AND IT REALLY OPENS UP SOME EXCITING POTENTIAL FOR EFFECTIVE VACCINES IN THIS REALLY SIS SUSCEPTIBLE POPULATION. SO WITH THAT LET ME ACKNOWLEDGE LOB, TRISHA, WITHIN THE TB RESEARCH GROUP AT PIT LED BY JOANNE FLYNN WHO I KNOW MANY OF YOU KNOW WELL AND THIS. >> WE MIGHT BE ABLE TO USE MUTANT OR SOMETHING LIKE THAT TO BE A MUCH SAFER VACCINE BECAUSE GIVING HIGH DOSE IVBCD IN HIV POSITIVE PEOPLE PROBABLY IS HIGH BAR TO CLEAR BUT THESE DATA CERTAINLY SUGGEST THAT WE COULD USE A SAFER VARIANT OF BCG. >> DEAD BUGS DOESN'T GET SAFER. >> DEAD BUGS SOMETHING LIKE THAT AND DEFINITELY THINGS THAT WE'RE LOOKING AT. >> COOL. >> CONGRATULATIONS ON THAT REALLY IMPORTANT RESULT. WHY DO THE CELLS GO TO THE LUNG IF THE VACCINATION? WHAT'S THE IMMUNO PHENOTYPE AND FUNCTIONALITY OF THE T CELLS LONG BEFORE INFECTION AND CAN YOU GIVE BCG INTRAVENOUSLY TO TREAT ESTABLISHED TB INFECTION? >> ALL GOOD QUESTIONS, WE'VE NOT DONE THE USE OF BCGIVBCG AS A THERAPEUTIC AND CERTAINLY THAT'S SOMETHING THAT COULD BE INVESTIGATED AND WHY ARE THE CELLS TO THE LUNG AND WHAT ARE THEY DOING WHEN THEY GET THERE IS A QUESTION THAT WE'RE ACTIVELY LOOKING AT NOW AND WE'RE USING A LOT OF SINGLE CELL RNC SEEK TO MORE FULLY CHARACTERIZE THE CELL POPULATIONS SO I HOPE TO HAVE SOME BETTER ANSWERS FOR YOU WITHIN THE NEXT YEAR. >> I WAS VERY STRIKING AT THE END WHEN YOU SUGGESTED THAT YOU COULDN'T FIND THE SPECIFIC T CELLS IN THE BLOOD BECAUSE I THINK WE WERE ALL THINKING THAT'S WHAT YOU WERE GOING TO FIND. WHAT IS THE MECHANISM? WHAT DO YOU THINK IS GOING ON? >> THE MECHANISM IS A REALLY, REALLY IMPORTANT QUESTION AND SO WE'RE TRYING IDENTIFY THE MECHANISM AND LOOK FOR COR IMMUNITY. I KNOW STUDIES WITH BOB SEEDER AND JOANNE FLYNN HAS SHOWN AN NIH NAIVE ANIMALS AND PRETEXT IS ASSOCIATED WITH IGM AND IT'S ASSOCIATED WITH AN INFLUX OF T RESIDENT MEMBER RE CELLS AND THESE ARE ALL ASSOCIATIONS WHERE CURRENTLY DOING STUDIES TO KIND OF MORE FIRMLY HAVE MECHANISM PROTECTION. >> YOU ARE NOT ARGUING IT'S NOT T CELLS YOU ARE ARGUING THEY MIGHT NOT BE IN THE BLOOD. >> THAT'S RIGHT. >> THEY MIGHT BE IN THE LUNG. >> ABSOLUTELY. >> IS IT ETHICALLY FEASIBLE TO SPLIN HE CAN TOM EYES THESE NON PRAIRIE HUMAN IT'S A. PRACTICE. >> DO YOU AT THIS NOTICE THE SPLEEN. >> THEY DEVELOP A SPLEENAMOGLIA AND I THINK IT'S THE IDEA. PRIOR TO INFECTION. >> THANK YOU. >> WAS THAT RELATED IN THE MACAQUES? >> YEAH, THE ANIMALS THIS UNPROTECTED BY IVBCG HAD ONE THING IN COMMON, THEY WERE POOR VIRAL CONTROLLER NUMBER ONE AND NUMBER TWO, THEY INTENDED TO HAVE FEWER -- A MORE DRAMATIC CD4 DEPLETION. WE KNOW THAT WITH ART THERAPY THOSE THINGS ARE CORRECTED SO I THINK CERTAINLY IN HIV CONTEXT ART WOULD BE A REALLY IMPORTANT COMPONENT OF THIS. [OFF MIC] THE [OFF MIC] >> ABSOLUTELY, IT'S A REALLY GOOD QUESTION, IT'S TRUE THAT THE AEROSOL DELIVERED BCG JUST DOESN'T PROTECT WELL. OF COURSE IT'S GET TO GO THE LUNG BY A DIFFERENT MECHANISM, RIGHT, IT'S COMING IN FROM THE AIRWAYS RATHER THAN SOME PLACE ELSE AND THE IMMUNE CELLS ARE ENTERING THE LUNG PRANK FROM THE VAAS CUE LATOUR. IF WE CAN TEASE OUT THE DIFFERENCES BETWEEN THOSE TWO ROUTES I THINK IT WOULD BE REALLY CONSTRUCTIVE. [OFF MIC] >> I WAS REALLY HAPPY TO RECAPITULATE THE FINDINGS BETWEEN JOB AND JOANNE IN A MODEL RATHER THAN REHSUS VERY GOOD. >> THANK YOU, ALL. [APPLAUSE] >> WE'RE GOING TO CHANGE THE TOPIC NOW SO OUR NEXT SPEAKER IS DIEGO COSTA FROM UNIVERSITY OF SAO PAULO. >> I WOULD LIKE TO SAY IT'S A HUGE HONOR TO BE HERE PRESENTING MY WORK AND ALSO TO THANK DRAGANA AND DAN FOR MAKING THIS BOTH ABLE SO I'M GOING TO SHOW YOU THE DEVELOPMENT OF THE WORK I'VE DONE AS A TIME AS A TRAINEE FOR ESTABLISHING THE RESEARCH PROGRAM THAT I'M START TO GO DEVELOP IN BRAZIL RIGHT NOW. I HAVE WORKED WITH ALAN AND I CONTINUE TO DO THAT AND I'M GOING TO SHOW YOU DATA FOR TB AND TARGETING HOMEOSTASIS AND LUNG INFLAMMATION SO IT WAS INTRODUCED HERE A MAJOR PUBLIC-HEALTH PROBLEM AROUND THE WORLD AND SO JUST LAST YEAR, AROUND 10 MILLION PEOPLE HAD BEEN INFECTED AND HALF MILLION OF THOSE PEOPLE WITH DRUG RESISTANT BAGS AND 1.5 MILLION PEOPLE DIED FROM THE DISEASE AND SO WHY DID WE CONTINUE TO BE SUCH A MAJOR PROBLEM IF THERE'S A VACCINE FOR IT AND A TREATMENT AND THE VACCINE IS NOT THAT EFFECTIVE IN CONTAINING PREVENTING TUBERCULOSIS AND THE TREATMENT, THE REGULAR TREATMENT, THE DIRECT TREATMENT SHORT COURSE LASTS FOR SIX MONTHS WHERE FOUR DIFFERENT DRUGS ARE ADMINISTERED SO THE PATIENTS PRESENT SEVERAL SIDE EFFECTS AND A LOT OF THEM DO NOT COMPLY WITH THE FULL TREATMENT IN THAT FAVORS THE RELAPSE FOR THE DISEASE AND SELECTION OF DRUG ROW RESISTANT TREATMENT SOS AN URGENT NEED THAT ELIMINATED THE TREATMENT FOR RAPIDLY AND ALSO THEY DO NOT RELY ON ANTIBIOTICS AND IN THAT SENSE, WE COME TO THE THERAPY AS A STRATEGY BECAUSE THEY TARGET HOST BIOLOGICAL FACTORS THAT CAN FAVOR THE DISEASE DEVELOPMENT AND AND HOW DOES HOST IMMUNITY AND WORK AND IT'S A BACTERIAL THAT IS TRANCE MUTED THROUGH THE AIR SO IT EFFECTS MAINLY THE LUNGS AND IN THE LUNGS, IT'S FIRST ENCOUNTERING THEY ARE THE MAIN CELLS BY TB AND TB HAS DELAND THEYARE PART ANT CYTOKINE HOST COULD CONTAIN THE DISEASE AND THESE MACROPHAGES WILL PHAGO SIGHT THOSE BACTERIA AND PHAGO SOME WILL OCCUR AND THE FUSION AND THIS CAN LEAD TO THE BACTERIA AND IF THEY'RE NOT KILLED AND LATER ON AND TH1 IMMUNITY WILL KICK IN WITH THE PREDICTION AND IT'S IMPORTANT FOR ACTIVATING THE EXPRESSION OF INOS AND OX SA DAZING MACROPHAGE AND THEY'RE HARMFUL BUT EVEN SO, ALL ALONG THE EVOLUTION, THE CO EVOLUTION WITH THE HUMAN SPECIES AND THERE ARE WAYS TO SUBVERT THIS IMMUNO RESPONSE AND IT CAN SURVIVE THIS STRETCH FROM THE MACROPHAGE ONE OF THE THINGS IS THEY CAN TRIGGER A MASSIVE PRODUCTION, ESCAPE THIS TRIGGERS A MASSIVE PREDICTION AND WHEN A CHRONIC DEVELOP WE CAN HAVE A PROTECTIVE CYTOKINES AND LEAD TO GO A MASSIVE PREDICTION AND THIS WILL END UP TRIGGERING CELL DEATH OF THE MACROPHAGES BECAUSE THEY WILL TRIGGER TISSUE DAMAGE AND THE BACTERIA AND SO THE THEY SHOW DESTRUCTION IS WELL CHARACTERIZED IN THE GRANULOMA SO THEY HAVE A MASSIVE CASE OF NECROSIS WHICH WILL LEAD TO THE FORMATION OF THIS CAVITIES AND THIS CAN HELP TO SPREAD BACTERIA TO TRANSMIT THE BACTERIA SO CONSIDERING THAT THE IMMUNE RESPONSE IS IMPORTANT TO CONTAIN THE BACTERIA AND ALSO TRIGGER TISSUE DESTRUCTION THE MAIN STRATEGIES FOR DEVELOPMENT OF HOST DISTRICTED THERAPIST FOR TB RELY ON INCREASING THIS IMMUNITY SO WE CAN GET RID OF THE BACTERIA MORE EASILY OR USE ANTIINFLAMMATORY STRATEGIES SO WE CAN CALM DOWN THIS INFLAMMATORY RESPONSE PRESERVING TISSUE AND DISRUPT GRANULOMA SO THEY PENETRATE INTO THE CORE WHERE THE EFFECTED CELLS ARE. SO, WHEN I WAS IN THE LAB, WE WERE TRYING TO TARGET FOR HOST DIRECTED THERAPIES SO HO1 IS ANEN TIME THAT CATALYZES THE DEROGATION AND RELEASING AMOUNTS OF CARBON MONOXIDE AND BILLY VER DIN AND IT WAS A STUDY THAT WAS DEVELOPED BY BRUNO WHEN HE WAS IN THE LAB. THIS IS HUMANS SUCCESSFUL TREATMENT CURSE THERE'S A EXPRESSION OF HO-1. IT'S A GOOD BIOMARKER BUT WE WERE INTERESTED IN THE ROLE OF IT AND WE USED THESE INHIBITOR OF HO-1 THAT IS ALSO CALLED LIPIDS YOU CAN SEE HERE IT'S AN MOLECULE OF HEME AND INSTEAD OF RAILRRIN FROM THE SO WHEN WE IND THE MICE WITH A LOW DOSE OF TB AND WAITED FOR THE INFECTION TO ROW ESTAREESTABLISH AND ONLY TWS OF TREATMENT LED TO A MASSIVE REDUCTION OF THE BACTERIAL LOAD IN THE LUNGS AND NOT JUST THAT WHEN YOU COMBINE THIS WITH THE REAGAN ANTIBIOTICS AND WE FOLLOE MICE IN THE LONG-TERM YOU CAN SEE IN THE GROWN L GREEN LINE AS COMPARED TO THE RED LINE THAT ANTIBIOTICS ALONE SO IN FOUR MONTHS IT COULD CLEAR THE BACTERIA FROM THE LUNGS AND TREATED ONLY WITH ANTIBIOTICS IT SAKES SIX MONTHS SO WE SAVE TWO MONTHS OF TREATMENT. ONE INTERESTING THING THAT WE FOUND WHEN WE STARTED TO TREAT THESE MICE FROM DAY ZERO IS THAT DURING THIS FIRST THREE WEEKS, WE HAD NO CHANGE IN BACTERIAL LOAD. YOU COULD ONLY WORK LATER ON SO THEM IF WE HAVE SIX WEEKS POST INFECTION WE COULD FIND THE BACTERIA LOAD REDUCTION AND ONE THING THAT IS VERY CLAR TERRIZED ALREADY IN TB IS IN THE THIRD WEEK OF INFECTION, IT'S WHEN THE T-CELL RESPONSE KICKS IN SO HERE WE HAVE THIS MASSIVE INCREASE IN PRODUCTION AND WE CAN SEE THE EXPRESSION SO WE WANTED TO KNOW IF THESE EFFECT WOULD BE THE T CELLS SO AND THEY'RE KNOCK OFF MICE AND TREATED THEM AND WE CAN SEE HERE A NICE REDUCTION AND WE HAVE NO REDUCTION IN GAMMA AND SO THIS SHOWS THAT THE BACTERIAL LOAD RESULTING FROM THE HO-1 IS THE IMMUNE RESPONSE AND MORE SPECIFICALLY IN THE REDEX OF EXPRESSION OF IONS SO WE ALSO MADE A NICE CHARACTERIZATION OF WHICH CELLS EXPRESSED HO1 IN THE LONG INFECTED MICE AND I'M NOT SHOWING THE DATA HERE BECAUSE OF LACK OF TIME BUT BASICALLY WE FOUND THAT THIS CELLS WHERE WE HAD THE LARGEST INCREASE IN HO1 EXPRESSION WERE INFLAMMATORY MACROPHAGES THAT WERE RECRUITED FOR MONOCYTES AND THE LUNGS SO, WE USED AN IN VITRO MODEL OF THE MACROPHAGES TO ANALYZE WHAT WERE THE MECHANISMS BEHIND THIS INQUIZ ITS EFFECT WE WERE SERVE WHAT IS WE BROUGHT THE AND AS EXPECTED, WE SEE JUST MEASURING THE EXPRESSION AND IT'S TREATED WITH GAMMA IN THE PRESENCE OR ABSENCE OF THE INHIBITOR, BLOCK H1 WE HAVE AN INCREASED EXPRESSION OF FOLLOWED BY THE INCREASED PRODUCTION AND SO IN THE BLOCK HO-1 THEY HAVE ENHANCED EXPRESSION IN RESPONSE TO GAMMA ACTIVATION AND SO SINCE HO-1, WORKS BY DEGRADING HIM, RELEASING CARBON MONOXIDE IN WE WERE INTERESTED IN SEEING WHICH OF THESE PRODUCTS WOULD BE SO WE REPEATED THE SAME APPROACH AS WE CAN SEE HERE WE HAVE GAMMA AND THE EFFECT WITH GAMMA PLUS INHIBITOR AND WE GAVE BACK CARBON MONOXIDE OR IRON AND AS YOU CAN SEE HERE, CARBON MONOXIDE MAY BE THAT RESULT IN ANY CHANGE AND BILIVERDIN AND WE REVERSED THE EFFECT AND SO THEY WERE ACTIVATED WITH GAMMA IN THE PRESENCE AND THE INHIBITOR AS YOU CAN SEE HERE, THIS INCREASE THE EXPRESSION IN THE PRESENCE OF H-01 INHIBITOR IS REVERTED IF WE HAD IRON BACK TO THE CELLS AND WHEN WE DO THE OPPOSITE APPROACH WE WILL INCUBATE THE CELLS AND WE HAVE A MASSIVE INCREASE IN THE EXPRESSION OF THE VIRUS SO THIS SHOWS US THAT THE IRON THAT IS BEING RELEASED BY ACTIVITY ACTUALLY IMPAIRS THAT EXPRESSION RESULTING FROM IS THE EFFECTIVE TEE OF THE EFFECTED MACROPHAGES AND SO WITH THESE WE WANTED TO FINALLY PROVE THAT IN FACT THE TREATMENT WITH H-O1 THE IRON LEVELS OF THE CELL SO WE DID WITH DEBATED CELLS RESULTING A REDUCTION IN THE INTER CELL LEVELS AND IN VIVO AS WELL WE ISOLATED LUNGS FROM TREATED MICE OR SMMP TREATED MICE WE CAN SEE HERE THIS BROWN SPOTS WHERE SUSTAINED AND SO WE SEE A LOT OF IRON IN THE LUNG OF ANIMALS INFECTED WITH TB BUT NOT TREAT SOD THERE'S IRON COMBINATION IN CELLS AND WE HAVE A DECREASING AND INFECTED ITSELF AND WE SET UP THE IDEA OF HOW THE MODULATION OF HOMEOSTASIS CAN BE MAUD LATING THE RESPONSE OF MACROPHAGES AND CAN THIS BE TARGETED FOR THE DEVELOPMENT OF NEW THERAPY SO WITH THAT, I WROTE A GRANT APPLICATION FOR IN BRAZIL AND SO I GOT THIS CALLED YOUNG INVESTIGATOR GRANT AND I WAS ABLE TO MOVE BACK TO BRAZIL MORE SPECIFICALLY I WENT BACK TO SAO PAULO AND MORE SPECIFICALLY I WENT BACK TO WHERE I DID MY PH.D IN UNIVERSITY OF SAO PAULO SO HERE IS A PICTURE. HERE IS A PICTURE OF THE MEDICAL SCHOOL IN THE UNIVERSITY OF SAO PAULO AND THIS IS A BEAUTIFUL PICTURE OF THIS BUILDING HERE THAT IS WHERE THE BASIC SIGNS LABS ARE LOCATED BECAUSE HERE IS THE HOSPITAL. OUR LAB IS IN THIS BUILDING AND THERE ARE JOINED ABOUT THE GROUP AND MY GROUP WITH THEY WERE PATIO OF THAT BUILDING AND WE HAVE A NICE CAFETERIA HERE SO SHE HELPED ME TO SET UP THIS PROGRAM THERE AND HER HELP WAS CRITICAL FOR ME TO BE ABLE TO DEVELOP IT THERE AND ACTUALLY I'M START TO GO DEVELOP IT AND HERE I HAVE A PICTURE OF MY OWN GROUP SO THOSE ARE MY STUDENTS WITH ME AND THE INTEREST O ENTRANCE OF THAT BUILDING AND AS I MENTIONED TO YOU I'M INTERESTED IN HOW IRON CAN BE MODULATING THIS I'M UNION RESPONSE TO TB SO PARTICULARLY IN MACROPHAGES, WHICH ARE THE CELLS AFFECTED BY TB WE WANT TO ADDRESS HOW IRON METABOLISM IS WORKING IN TB INFECTION SO MACROPHAGES CAN AK PAIR IRON THROUGH MULTIPLE WAYS SO ESPECIALLY BY RED BLOOD CELLS AND THEY CAN ALSO DEGRADE HEMO PROTEINS AND GET HEMOGLOBIN FROM THE CIRCULATION AND THIS IRON AND ALSO ACQUIRE IRON DIRECT LIEU THROUGH OTHER TRANSMITTERS BUT THEY CAN ONLY GET RID OF IRON BY THESE TRANSPORTERS HERE THAT IS FERRO FOR TIN. WE GOT INTERESTED IN THE MODULATION BECAUSE THE REGULATION OF THE EXPRESSION OF FER OWE FOR TIN. THE PRODUCED BY IRON AND THE SER OWN BUT IT CAN BE PRODUCED BY MACROPHAGES IN RESPONSE TO INFLAMMATORY SUCH AS IL-11, IL-6 AND RECEPTORS 4 SO THESE HAVE PRODUCTION WE WILL FERROPORTIN AND ON THE OTHER SIDE TRYING ORING OF RECEPTORS 2 CAN LEAD TO DOWN REGULATION OF FERROPORTIN TRANSLATION SO THESE ARE ALL ACTIVATED AND WE ARE INTERESTING KNOW THE ROLE OF FERROPORTIN DURING THE TB INFECTION AND HOW DOES IT WORK IN THE MODULATION OF HOMEOSTASIS AND MODULATION IN IMMUNE RESPONSE SO THIS IS A WORK IN PROGRESS AND I'M GOING TO SHOW YOU SOME VERY PRELIMINARY DETAILS THAT I'M START TO GO GET IN THE LAB WITH MY STUDENTS. MY MASTER STUDENT IS WORKING ON THIS PROJECT SO SHE JUST MEASURED H-01 EXPRESSION AND AS YOU CAN SEE HERE WHY H-01 GOES UP AND FERROPORTIN GOES DOWN SO THIS IS FLAVORING THE LUNGS SO INTERESTINGLY, THIS IS THE EXPRESSION OF HELP SEW DIN AND IT GOES DOWN SO THE DOWN REGULATION OF FERROPORTIN IN THE CELLS MAY NOT BE RELATED TO HEPSODINE ITSELF BUT AS WELL AS TALL 1 AND 6 ARE BEING WITH ON GOING INFECTION SO WE BELIEVE THAT MAYBE THIS DOWN REGULATION IS MEDIATED BY REACC RETO BE ANE DID MODEL OF INFECTION IN MACROPHAGES SO WE ASKED THE COMPANY TO SYNTHESIZE AND TEST IT IF IT WAS REALLY WORKING TO DOWN REGULATE FARRO PORTIN THERE'S A DOWN REGULATION IN THE FIRST OF THIS BONE MARROW DRIVE AND WHEN I SEE EFFECTED THOSE CELLS WITH TB AND TREATED THEM WITH GAMMA OR HEPCIDIN IT DID ARE NOT HAVE A CHANGE IN THED CELLS AND INTER FUR AN GAMMA WE SEE A NICE REDUCTION BUT PUT HELP SEDIN HERE AND IT WILL DOWN REGULATE IT AND IT'S A WORK IN PROGRESS AND WE HAVE ESTABLISHED A COLLABORATION WITH DAVID FRASIER FROM AUSTRALIA AND GET THIS IN MICE AND WORE BREEDING THEM WITH SOME PRETTY EXPRESSING STRAINS TO KNOCK OUT IT SPECIFICALLY IN THE MIO LLOYD CELLS AND WEREN'T ABLE BECAUSE WE FOUND THAT THESE ONES WERE NOT VIABLE IF WE CROSSED THEM WITH CSFR1 AND LET'S SEE WHAT HAPPENS SO MA PRONE A IS DOING HER FIRST INFECTION THIS WEEK. LET'S SEE WHAT WE HAVE FROM THAT. AND ALSO, WE'RE INTERESTED IN SEEING HOW IS IT MODULATING THESE AND IN PARTICULAR, WE ARE INTERESTED IN SEEING YOU HOW THEY CAN MODULATE THE EXPRESSION OF HIGH POX AND HI F-1 AND HI F-2 AND WHY IS THAT? AND DEGRADING THE HIF OF THE MOLECULES AND IT TURNS OUT THAT IT ALSO UTILIZES IRON TO THIS AND FO HIF-1 WHEN IRON DEPLETION OCCURS IT CAN MIGRATE TO THE NUCLEUS AND INDUCE THE SUB DESCRIPTION OF SEVERAL GENES ASSOCIATED WITH METABOLISM OF GLUCOSE AND A LOT OF CYTOKINES THAT ARE IMPORTANT FOR IT AND ON THE OTHER HAND, THE MODULATION OF IRON WORKS ON THE OPPOSITE DIRECTION AND THAT WORKS BECAUSE THAT HAPPENS LIKE THIS BECAUSE OF THIS IRON RESPONSIVE ELEMENT AND THIS IS A MOTIVE PRESENT ON THE MANAGERRER RNA MOLECULE SO WHEN WE HAVE THE INTERACTION OF THESE PROTEINS THAT THE IRON REGULATORY PROTEIN WITH THESE IRON RESPONSIVE ELEMENTS AND WHEN THE IRON CONCENTRATION OF THE CELLS AND LOW THEY HAVE A CONFIRMATION, THEY HAVE A HIGH AFFINITY FOR THE IRON SO THEY'RE GOING TO MIND AND THROUGH THE PRESENT IN HIF-2 AND BLOCK ITS TRANSLATION AND WHEN IRON ACCUMULATES INSIDE THE CELLS, IT'S GOING TO BIND TO THIS IRP AND ALTERS ITS CONFIRMATION AND LOSES ITS AFFINITY AND RELEASES THE MRNA AND THEN WE HAVE AN INCREASE IN THE EXPRESSION OF HIF-2 SO WHEN IRON IS HIGH INSIDE CELLS WE HAVE A DEM INAUGURATION OF HIF-1 ALPHA AND HIF-2 ALPHA AND WHAT HAPPENS IS HIF-1 ALPHA IS ASSOCIATED WITH M1 MACROPHAGES WHERE WE HAVE INFECTION OF EFFECT AND BETA PRODUCTION AND ALL IMPORTANT FOR CONTAINING TB REPLICATION AND THEY ASSOCIATED WITH MACROPHAGES AND IT'S AN IMPORTANT INDUCER OF RGNA1 WHICH FACILITATES REPLICATION AND MACROPHAGES SO WE'RE INTERESTED IN ESPECIALLY IN ASSESSING WHAT IS THE ROLE OF THIS ALPHA PATHWAY IN INFECTION AND ESPECIALLY FIGURING OUT HOW THE MODULATION OF THESE IRON LEVELS INSIDE THE CELLS CAN BE REGULATING THESE PATHWAYS SO THIS IS OUR WORK THAT IS BEING DEVELOPED BY A PH.D STUDENT IN MY LAB AND SO THIS IS ALSO SOME VERY PRELIMINARY DATA USING BONE MARROW MACROPHAGES TREATED WITH ANY INHIBITOR WE DO SEE A REDUCTION AND WHEN WE TRICK THEM WITH GAMMA HERE IS THE REDUCTION AND WE PUT GAMMA PLUS THE INHIBITOR WE HAVE THIS OF THE ACTIVITIES OF THIS MACROPHAGES AND WE ALSO USE THIS H37RVN TO BE EXPRESSING SO WE COULD SEE IF INVIVO THE INFECTION WOULD TRIGGER THE INFECTION SO WE COMPARED TO THE NON EFFECTED CELLS SO WITH THAT, WE WANTED TO START A SESSION THE ROLE DURING MTB INFECTION AND WE USE IT MODEL OF A LOW DOSE AND TREATED WITH AN INHIBITOR OF HIF-2 ALPHA AND THE MOUSE HAVE A LOWER BACTERIAL LOAD IN THE LUNGS SO WE GOT THIS AND WHAT WE GOT IS N EFFECT THE MICE WERE CONDITIONED KNOCK OUT MICE WITH LOW OR HIGH DOSE OF BOTH OF THEM AND WE HAVE A LOWER BACTERIAL LOAD WITH MORE EVIDENCE IN HIGH DOSE SO EVERYTHING POINTS OUT TO THE FACT THAT HIF-2 ALPHA IS DETRIMENTAL FOR THE CONTROL OF BACTERIAL INFECTION AND WE BELIEVE THIS IS BEING MODULATED BY THE CHANGES IN IRON LEVELS INSIDE THE CELLS AND IN RESPONSE TO OTHER INFLAMMATORY SO, THAT IS ONE PROJECT AND THIS IS ANOTHER PROJECT THAT WE ARE CARRYING ON IN OUR LABORATORY AND THIS IS IMPORTANT FOR ME TO SAY THAT IT'S BEING DEVELOPED IN COLLABORATION WITH Dr. CARLA ROTH LYNN AND HERE AGAIN CAN BE PRESENT BECAUSE I REMEMBER FIRST OF ALL, CARLA IS AN EXPERT IN THE FIELD OF RECEPTORS RIGHT SO THESE RECEPTORS THEY RECOGNIZE IN THE MEMBRANES OF CELLS AND THEY BRIDGE WITH THESE LIE GANTZ TO PRO MOT THE CELLS AND PHAGO TIE SEW SIS IS CALLED OF CELLS AND ALSO I SAID THIS PATHWAY MUST BE VERY IMPORTANT FOR THE REGULATION OF IMMUNE RESPONSES IN TB AND WE MAY HAVE NICE TARGETS HERE TO TRY TO DEVELOP NEW STRETCHES, YOU KNOW, WE SHOULD GET MICE WITH HER AND SEE AND SHE SAID TOTALLY, LET'S TALK WITH HER, SHE'S A VERY NICE PERSON AND ALAN INTRODUCED ME TO CARLA SO I KNEW HER AND GOT TO KNOW HER AND ACTUALLY SHE SENT THE MICE TO ME AND WE ARE DEVELOPING THIS PROJECT RIGHT NOW AND SO THIS IS SOME RESULTS THAT I'M GOING TO SHOW YOU TOO, THESE ARE VERY PRELIMINARY AND BEING DEVELOPED BY THIS STUDENT OF MINE ANDREA, HE IS A MASTERS STUDENT AND HE IS SHOWN THAT THE EXPRESSION OF AXL IN MACROPHAGES IN RESPONSE TO INFECTION THIS IS 24 AND 48 HOURS AS YOU CAN SEE HERE AND HE ALSO FOUND THAT IN VIVO, IN MICE INFECTED WITH A HIGH DOSE, WE STARTED WITH A HIGH DOSE BECAUSE WE WANTED TO SEE HOW IT COMMODE YOU LATE THE LESIONS IN THIS MICE AND THE SIGNIFICANT INCREASE IN IF YOU ARE MACROPHAGES BUT IF YOU ARE MACROPHAGES ARE ALREADY EXPRESSING A LOT OF AXL SINCE THE BEGINNING AND IT'S LITTLE TO DECREASE BUZZ IN PARTICULARLY IN THIS CD11B POSITIVE CELLS AS YOU CAN SEE IT'S REDISTRICTED AND IT'S POSITIVE EVENTS AND WE HAVE A MAJOR INCREASE IN THE EXPRESSION OF AXL THAT AT THE SAME TIME IN THE LONG OF THESE ANIMALS WE HAVE A MAJOR INCREASE IN THE PREDICTION OF THE LAG ANT SO WE HAVE TWO OPTIONS, FIRST, IF WE AXL CAN BE DOWN MODULATING AND IF YOU COULD BLOCK IT WE CAN MAKE IT BETTER OR ON THE OTHER SIDE, IT'S THERE TO MODULATE IT AND IF YOU BLOCK IT IT WILL BE BAD. TO START ADDRESSING THESE WE EFFECTED MICE WITH A HIGH DOSE OF TB AND TREATED THEM WITH A CONTROL OR AN INHIBITOR AND THIS IS A DRUG THAT HAS BEEN USED IN CLINICAL TRIALS FOR CANCER SO THE GOOD THINGS HERE IS IT CAN BE REPRE POSED. AND WHAT I FOUND IS THAT WE HAD TO SPEED OUR EXPERIMENT BECAUSE BY DAY 25 THE MICE TREATED WITH THE AXL INHIBITOR WERE STARTING TO DIE. AND WHEN WE EUTHANIZED THESE MICERS THERE WAS NO CHANGE IN THE LUNGS OF THE MICE AND THE MICE TREATED WITH THE INHIBITORY THEY HAD MILE OWE CELLS AND IN PARTICULAR, NEUTROPHILS SO WE HAD THE KNOCK OUT MICE IN THE LAB AND AGAIN, NO CHANGE IN WHEE HAVE THEM THREE WEEKS POST INFECTION BUT THEY HAVE WAY MORE SPECIALTY IN THE LUNGS AND WHEN WE MEASURED THE CD4 AND CD8 BY ACTIVATING THE CD4 CELLS WITH PEPTIDES AND SPECIFIC PEPTIDES WE SEE HERE THAT CD4s EXPRESSWAY MORE GAMMA AND TNF ALPHA AND THIS IS THE KNOCK OUTS MICE AND CD8 CELLS WHEN ACTIVATED WITH SPECIFIC PEPTIDES AS WELL THEY ARE HIGHER AND THE MAJOR PHENOTYPE IS THAT HUGE INCREASING IN THE LUNGS OF THESE MICE SO YOU CAN SEE HERE. ASSOCIATED WITH THAT, IT'S KNOWN TO BE DID HE TELL INTEREST DETRIMENTAL AND YOU CANSEE BY TD THERE'S MORE KNOCK OUT MICE AS WELL AS MORE KNOW TOE SIS AND AS YOU CAN SEE BY THE GRAPH SO WE'RE CONTINUING TO WORK ON THAT TO ACTUALLY ASSESS WHICH ARE THE MECHANISMS INVOLVED IN THIS HIGHER INFLAMMATION AND IF NETS ARE REALLY CONTRIBUTING TO THE DAMAGE TO THE LUNGS OF THESE MICE AND WITH THAT WE EXPECT TO PINPOINT SPECIFIC MEDIATORS WE CAN TARGET TO DEVELOP NEW STRAT GOES FOR TREATING THESE MICE WITH THAT, THESE ARE VERY PRELIMINARY DATA THAT I'M SHOWING HERE THAT I'M STILL START TO GO DEVELOP IN BRAZIL. I WOULD LIKE TO THANK ALL THE LAB MEMBERS AND MY LAB MEMBERS FOR THEIR SUPPORT AND WITHOUT WHICH I WOULD NOT START THIS AND ALAN SHER AND THE LAB MEMBERS AND BRUNO AND THEY HAVE ALWAYS PARTICIPATED A LOT IN THE DEVELOPMENT IN THIS PROJECT AND ALAN AS EVERYBODY HERE ALREADY MENTIONED HIS SUPPORT WAS CRITICAL FOR ME AND IT SEEMS LIKE I CAME TO THE NIH UNTIL I LEFT IT WAS CRITICAL FOR ME TO CHANGE MY WAY OF THINKING SCIENCE AND ALAN IS VERY CRITICAL ATTENTION OF MAKING YOU LEARN STUFF AND ESPECIALLY TEACHING YOU HOW TO WRITE. SO ALONG THE HOT SEATS I HAD TO WRITE PAPERS OR EVERYTHING, I THINK MY ENGLISH SKILLS AND WRITING SKILLS IMPROVED EXPONENTIALLY SO AS I SAID, ALAN'S TOUCH TO FACILITATE OUR COLLABORATIONS WITH OTHER PEOPLE IN THE FIELD AS WELL AND THEY HAVE BEEN VERY IMPORTANT AND HIS COLLABORATION HAS BEEN AND CONTINUES TO BE VERY IMPORTANT FOR THE DEVELOPMENT OF THIS WORK AND I EXPECT THAT YOU CONTINUE TO HAVE YOUR SUPPORT AND BE ABLE TO CHAT WITH HIM WITH THE RESULTS HAVING HIS INPUT AND EVERYTHING BECAUSE I KNOW THAT'S "REAL LIFE" CCRITICAL FORT DEVEE WORK. THANK YOU, EVERYBODY. [APPLAUSE] >> SO MANY FASCINATING OBSERVATIONS. I GUESS I'LL START FROM THE LAST SECTION. SO IN THE ACTION I WILL KNOCK OUT MICE, DO YOU HAVE A DEFECT IN NITRIC OXIDE WE ALSO TRIED TO SEE IF THERE'S A DIFFERENCE IN THE AXLE POSITIVE AND NEGATIVE CELLS REGARDING THE INS EXPRESSION AND WE FOUND THAT POSITIVE CELLS ARE DOING AND SPECIFICALLY THAT IS HAPPENING AND OR IF IT'S JUST MORE AND MORE CYTOKINES OR IF THERE'S A ROW FOR AXL IN MODULATE SOMETHING KIND OF SPECIFIC NECROSIS TYPE OF DEATH OR SO WE'RE STARTING TO INVESTIGATE THAT NOW BUT I THINK THE MOST LOGICAL THING TO THINK BY NOW IS THAT IT'S A FAILURE TO CLEAR CELLS. DO YOU KNOW IF IT'S CELL AUTONOMOUS? >> MEANING YOU KNOW, IF YOU MADE A CHIMER IT ONLY OCCURS? >> SO I DON'T REALLY KNOW MANY OF THAT'S ANOTHER THING WE'RE TRYING TO SEE. WE ACTUALLY GOT THE AXL FLUXED MICE THAT WE'RE BREEDING WITH SOME LINES SO WE WILL TROY TO DELETE CELLS AND THE MAIN WOULD BE THE MACROPHAGES. >> I AM ASSUMING THAT THE TRYING TO PROTECT THIS CELL FROM THE RIGHT INDUCTION AND SO -- IT'S HANIN ZOABI HYOU CAN COMPLETE T. >> YES. DEFINITELY. THANK YOU FOR THE EXCELLENT TALK. BOTH THE MACROPHAGE AND THE BUG NEED IRON AND I COULDN'T QUITE TELL WHETHER YOU DISTINGUISHED BETWEEN THE REQUIREMENT OF EITHER OF THE BUG FOR IRON IN YOUR SYSTEM? >> RIGHT. YEAH, SO, THAT'S THE PROBLEM OF MISSING WITH IRON METABOLISM AS A WHOLE, RIGHT. SO IRON IS CRITICAL FOR PART OF GROWTH AND BUT TB SPECIFICALLY IS A LITTLE BIT MORE RESISTANCE TO IRON DEPRIVATION THAN OTHER SPECIES OF MICRO BACTERIA FOR EXAMPLE AND MICRO BACTERIA IS VERY, VERY SUSCEPTIBLE AND IF YOU EFFECT MICE LA LACK IT WILL PUT IRON INSIDE THE PHAGO SOME, THE MICE PROSECUTE GOING TO BE RESISTANT TO AVIAN BUT NOT TO MTV FOR EXAMPLE. AND DEFINITELY IT'S A TWO-HEADED SWORD. IF I LIMIT IRON I'LL BE STARVING THE BUGS AND I MAY BE I'M ALSO GOING TO BE MAKING THE MACROPHAGES WORK BETTER SO THEN THERE'S ANOTHER POINT SO IN STUDYING THE EYE OWN HOMEOSTASIS ITSELF, IT'S TRICKY FOR US TO DEAL, THIS IS GOING TO CAUSE OTHERS PROBLEMS, RIGHT. THIS IS GOING TO CAUSE ANEMIA AND SEVERAL OTHER STUFF OR LET'S INDUCE A LOT OF -- IT WILL MESS UP WITH THE WHOLE BODY OUR HOMEOSTASIS SYSTEM AND THE POINT YOU ARE USED TO ACTUALLY FIND WHICH ARE THE MODULATING THIS AND STRETCH THE TARGET THEN AND FOR EXAMPLE, IF TOO OFTEN IT'S A GO AHEAD ONE, BECAUSE ALSO WE HAVE THESE THAT HAVE BEEN USED ALREADY IN CANCER TREATMENT AND IT CAN BE EASILY REPO POSED FOR TB SO WE'RE TRYING TO THINK AND IDENTIFY THAT CAN BE TARGETED. IF YOU REAL WITH IRON ALONE, IT'S GOING TO BE A MESS, I GUESS. >> SO IT SOUNDS LIKE YOU ARE NOT ENTHUSIASTIC ABOUT DEVELOPING SNPP9 AS A CLINICAL AGENT. >> WE HAVE COLLABORATION THAT PEOPLE HAVE BEEN TRYING TO -- OUR INHIBITORS OF H-O01 AND THEY CAN'T USE IT FOR A LONG TIME AND IT HAS TO BE SHORT TERM. >> ONE LAST QUESTION, THERE'S IRON DEFICIENCY IN THE THIRD WORLD, IS THERE ANY DATA ASSOCIATING BETTER OUTCOMES IN TB PATIENTS THAT ARE IRON DEFICIENT? >> YES, SO IT VARIES A LOT. THERE ARE SOME VERY OLD PAPERS THAT ALREADY SHOW IN THE 1800S THAT PATIENTS THAT HAVE BEEN SUPPLEMENTED WITH IRON WOULD HAVE A WORSENING OF THE HE HAVE EFFECTS OR IT WOULD FAVOR THE DISSEMINATION BECAUSE MAINLY YOU HAVE THESE RETENTION OF IRON IN THE LIVER AND IN THE SPLEEN, SO IT FAVORS THE DISSEMINATION OF BACTERIA. THERE ARE NOT SO MANY NEW STUDIES ON THIS CASE THEY'RE MAJORLY OLD STUDIES. >> I AM BUILDING ON PHIL'S QUESTIONS ABOUT IRON REGULATION. IT'S ALSO THOUGHT TO PLAY A ROLE IN THE OBESITY EPIDEMIC IN NASH WITH HELP SIDE AND I THINK LEVELS DECROWS CREASING AND LEAO HIGH GROW SIS SO I'M CURIOUS WHAT IS THE CORRELATION SORRY CONNECTION BETWEEN OBESITY AND NASH AND SEVERITY OF TB? >> IS THAT KNOWN? >> I KNOW LIKE THERE ARE SOME DATA SHOWING THAT OBESITY IS A RISK FACTOR FOR TB, RIGHT. SO ACTUALLY WE JUST SUBMIT AID PAPER IN AN EXPERIMENTAL MODEL WHERE WE FEED HIGH-FAT DIE ITS FOR THESE HIGHS IT'S NOT SO BAD FOR BACK ROLE AND IT TRIGGERED BECAUSE OF THESE LIP ITS AND THERE'S GOING TO TRIGGER INFLAM OWE SOMEONE'S AND IT MAKES IT WORSE SO FOR OBESITY IT'S BA% œFOR TB BUT MAINLY BECAUSE OF THESE HIGH INFLAMMATION THAT IS GENERATED. >> COOL. NICE WORK. >> THANK YOU. >> [APPLAUSE] [OFF MIC] >> IT'S ALWAYS TREATED BY ANTIBIOTICS. >> THANK YOU. >> ALL RIGHT. SO NEXT SPEAKER IS NICOLAS RITEAU HE IS FROM FRANCE AND PLEASE. >> THANK YOU, VERY MUCH. FIRST I'D LIKE TO THANK DRAGANA, LIKE ALL THE OTHERS, FOR ORGANIZING THIS REALLY NICE SYMPOSIUM SO I WAS A POSTDOC IN ALAN'S LAB FROM 2012 TO 2017 AND SOME OF THE PEOPLE I KNEW IN THE LAB AND IT'S A GREAT OCCASION AND IN ANY CASE TO MEET A LOT OF PEOPLE AND IN FRANCE SO ONCE AGAIN THANK YOU VERY MUCH. SO, DURING MY TIME AT ALAN'S LAB I WORK MAINLY ON CONTEXT OF VACCINES AND NOW WHAT I'M GOING TO PRESENT TO YOU IS MORE STEROID INFLAMMATION AND MODEL IN THE CONTEXT OF THE ROLE OF STING IN THIS DAY SO I'M SENIOR SCIENTIST IN FRANCE AND SO IN ADDITION TO THE CLASSICAL MODEL OF INNATE IMMUNITY, AND FIRST DESCRIBED BY JANEWAY WE HAVE PATHOGEN RECOGNITION AND RECEPTORS AND THAT ARE GOING TO DRIVE IMMUNO ACTIVATION AND REGULATION AND MATT SINGER IN ABOUT 28 YEARS AGO SAID. >> AND ATP AND THROUGH RECEPTIVE 2X7 BUT WE'RE NOW IN LINE WITH THE RECENTLY WE TRIED TO SENSING AND AND ESPECIALLY THE STING PATHWAY SO THIS IS BRIEFLY MENTIONED YESTERDAY BY JASMINE AND THIS STING WAS FIRST DESCRIBED IN 2008 AND THE CONTEXT OF INFECTION AND SO THIS BINDS TO FORMED THEY GO THE IN THTHTHE CYTOPLASM AND. >> SO YOU CAN OTHER AND OR AND DEPENDING ON HOW LONG AND YOU ENVIRONMENT AND SIGNAL AND WHERE YOU WANT TO TREAT AND. >> YOU CAN SEE THERE ARE MANY CELLS EXPRESSING STING AND IT IS EXPOSED BY MANY DIFFERENT CELL TYPES AND SO AT FIRST WE SAW THAT IF WE WOULD REMOVE STING, WE WOULD HAVE LESS PATHOLOGY AND IT'S WHAT WE HAVE SEEN WHEN WE REMOVED INNATE MEDIATORS AND PROTEINS THAT ARE IN PATHWAYS SUCH AS THE INFLAMOSOME AND WE HAD A DECREASED PATHOLOGY AND EVERYTHING IS BY HOST IF YOU REMOVED ONE OF THE PATHWAYS YOU HAVE LESS INFLAMMATORY AND FIBROSIS AND WE GOT REALLY EXCITED WHEN WE REMOVED STINGS AND WE USED STING KNOCK OUT ANIMALS WE HAD AN INCREASE IN THE CLINICAL SCORE OF THE ANIMALS AND INCREASED WEIGHT LOSS AND THE AND AND IN THE LUND THEY'RE FREQUENT AND THEIR PHENOTYPE IS ALTERED AND SO WE LOOK AT DIFFERENT MARKERS AND WE WERE ABLE TO SHOW THAT THE IMAGE CLASS 2 EXPRESSION OF NEUTROPHILS IS DECREASED COMPARED TO THE WILD TYPE NEUTROPHILS SO WE DO KNOW EXACTLY WHAT IT MEANS BUT WE CAN SOMEHOW BE RELATED WITHOUT IT AW AN INCREASE THIS TYPE OF EXPRESSION IN COMPARED TO THE WILD TYPES AND THIS CAN BE LINKED MAYBE TO INCREASED EXPRESSION AND THEY'RE PAPERS SHOWINTHAT THEY'RE CONNECTIONS BETWEEN WHERE THE INDUCTION OF TYPE ONE INTER NEURONS AND A FEW PAPERS SHOWED IT'S HAPPENING IN OTHER COULD BE TEXT THAT IT'S REALLY IMPORTANT ALSO BECAUSE TH17 CELLS AND I17 IS A DRIVER OF THIS MODEL AND IT'S A PAPER THAT TOM WYNN PUBLISHED ABOUT 10 YEARS AGO SHOWING THAT IL-17 IS IMPORTANT TO DRIVE PATHOLOGY BUT HERE WE DIDN'T SEE ANY DIFFERENCE COMPARING WILD TYPE MICE AND KNOCK OUT ANIMALS AND TREATED WITH ROW HIGH SINCE SO IT'S REPRESENTATIVE AND CD4 POSITIVE T CELLS IN THE LUNGS OF SALINE CONTRA MICE SO ALMOST NO IL-17 AND AFTER THERE'S AN INDUCTION OF IL-17 PROGRESS CELLS BUT NO DIFFERENCE COMPARING WILD TYPE AND STING KNOCK OUT ANIMALS AS QUANTIFY HERE BY IL-17 POSITIVE CELLS OR THE TOTAL NUMBERS OF THESE CELLS AND SO THEN WE MOVED AND WE ARE NOW REALLY INTERESTED IN TYPE 3 INTER NEURONS OR IL28 AND 29 AND THEY ARE AT LEAST NOW MORE THAN THAT FIVE OR SIX MORE PAPERS SHOWING YOU CAN HAVE TYPE 3 INDUCTION DOWNSTREAM OF STING SO IT'S LESS KNOWN THAT THEY ARE DISCOVERED IN 2003 AND THEY ARE FOUR INTERFERON IN HUMANS ONLY TWO IN MICE AND THEY ARE REALLY ABUNDANT AND IN TRACT AND THEY SHARE A UNIQUE RECEPTORS AND WITH ONE CHAIN OF THE I L-10 RECEPTORS AND THIS SIGNALING DOWNSTREAM IS VERY SIMILAR TO THAT OF TYPE 1 NEURONS AND THEY CANNOT INDUCE THE SAME ISGs SO IT SOUNDS EXPRESSING AND THE EVERYTHING AROUND REALLY THE ACTUAL DEFERS AND IT'S BEEN MANY STUDY AND IDEA IS WHEN YOU HAVE AND BY MOSTLY THE CELLS AND THEN IF IT'S EFFECTIVE, YOU WOULD HAVE LESS TYPE 1 INDUCTION AND PATHOLOGY SO THESE ARE THE PRELIMINARY DATA BUT WE WERE ABLE TO SHOW THAT THERE IS AN INDUCTION OF I28 IN THE LONG OF WILD TYPE MICE TREATED WITH A PARTIAL REDUCTION WHEN WE LOOKED IN ANIMALS AND THE CASE IN THE TYPE 1 KNOCK OUT ANIMALS AND IT'S SHOWN HERE BY WESTERN BLOCK AND THERE'S LESS EXPRESSION IN THE KNOCK OUT AS COMPARED TO THE WILD TYPES. SO THAT'S OUR WORK HYPOTHESIS AT LEAST IN LINE WITH TYPE 3 INTERFERON WHICH DOWNSTREAM OF STING THIS WITH BIND TO SELECTIVELY EXPRESS RECEPTORS ON THE CELLS ON NEUTRAL AND THIS CAN SOMEHOW RETURN AND AND EVENT LOW THE FIBROSIS AND WE ALSO KNOW WE ARE INTERESTED IN LOOK AT A SO I DIDN'T SHOW YOU DOWNSTREAM BUT IT'S INVOLVED IN MANY DIFFERENT PROCESSES DEPENDING ON THE CELL TYPES AND CONTEXT AND I HAVE THREE ACTIVATION AND MENTIONED BUT ALSO MORE CELL DEATH ESPECIALLY IN AUTO PATH GEE AND ALSO, WE DD THE EXPRESSION LEVEL AND AND IT'S REALLY HARD AND PATHWAY AND WHAT WE SEE HERE IS WE HAVE INDUCTION OF THIS AUTOPHAGY IN THE WILD TYPE MICE TREATED WITH NOT THE CASE IN SING ANIMALS BUT WE ALSO HAVE A STRONG INDUCTION OF P62 IN WILD TYPE MICE THAT WOULD INDICATE MORE LIKE DECREASED OR LIKE A BLOCKED SO IT'S NOT COMPLETELY CLEAR HOW THIS HAPPENS BUT IMPORTANTLY THIS IS ALSO CRITICALLY DECREASING IN ANIMALS SO THERE'S SOMETHING THAT WE WOULD LIKE TO PURSUE AND SEE WHETHER THIS PATHWAY CAN BE WHAT IS DRIVING THE PROTECTION OF DOWNSTREAM OF STING AND THIS IS RELEVANT IN THE TEAM IT'S BEEN SLOAN THAT IT PROTEINS ARE DECREASED IN PATIENTS AND THAT FEARS AND DEROGATION IN MOUSE AND SURVIVAL AND WHEN IT'S LINK THE TO CELL SIGNIFICANCE AND EFFICIENT MICE AND THEN PEOPLE HAVE SHOWED THAT THIS MICE DID I PLAY INCREASED FIBROSIS AND SO, THIS AND MY LAST DATA SLIDE AND WE ARE NOW ALSO TRYING TO SEE WHETHER WE CAN ACT RATE IS STING SO WE'RE USING THIS REALLY STRONG IMPORTANT AND IN THIS PARTICULAR EXPERIMENT, ONE SINGLE ADMINISTRATION OF DIABZ7 WAS ENOUGH TO REDUCE FIBROSIS WITH THE SCORE AND ALSO, THE COLOGIN CONTENT SO IT'S SOMETHING THAT WE ARE REALLY INTERESTED IN AND WE'RE GOING TO PURSUE SO THE GENERAL IDEA IS THAT IT'S KNOWN TO CREATE A LOT OF CELL THERE'S NOT IN DNA DAMAGE AND DNA RELEASE AND SO THIS WOULD NEED TO AN INCREASE OF MIGHT TOE AND IN THE PLUS UM ACTIVATING AND THE OTHER SENSORS AND THE INDUCTION THAT COULD BE BENEFICIAL AND ALSO STING CAN BE PART OF THIS PROCESS AND WE HAVE A COUPLE OF QUESTIONS THAT WE ARE TRYING TO ADDRESS AT THE MOMENT AND THE TYPE OF ACID AND AND MAYBE SOME OF THEM ARE INTERESTING TO LOOK AT AND ALSO I WOULD GAIN ACCESS TO THE SITE AND IT'S SOMETHING REALLY IMPORTANT IN THE FIELD BECAUSE YOU HAVE CELLS AND DNA FLOATING AROUND, HOW CAN IT COME TO THE SO IT'S NOT SOMETHING THAT WELL ASK THERE'S A BIG BLACK BOX HERE AND AND ALSO, IF WE CAN ACTIVATE STING AND SEE IF CAN BE BENEFICIAL OF THE FIBROSIS AS WELL AS THE EFFECT OF TYPE 3 INTERFERON AND THE AUTO IF GEE AND SO THESE ARE THE PEOPLE IN MY LAB THAT ARE LED BY ISABEL AND THE DIRECTORS OF I KNOW ALAN KNOW THEM REALLY WELL AND I USED TO WORK WITH TB FOR MANY YEARS AND HOW I GOT TO KNOW ALAN AT FIRST AND I'D LIKE TO SHARE A FEW PICTURES SO I TOOK WHEN I WAS A POSTDOC AND LIKE MANY OTHERS, I THINK IT HOTSEAT IS SOMETHING THAT SOME WOULD CALL IT PAINFUL AND YOU TOOK MANY TIME AT EVERY OCCASION IT WAS LIKE A 15-LINE ABSTRACT IT WOULD TAKE ONE AFTERNOON TO GO SO I WAS ABLE TO SPEAK TO HIM A COUPLE OF TIMES DURING THE HOTSEAT SO I DON'T HAVE ALL OF THE PEOPLE THAT WERE AROUND BUT I GOT MANY OF THEM SO KEN AND TOSH, PH.D STUDENT IN THE LAB AND I ALREADY SHOWED THIS PICTURE SO THIS WAS THE NATURE PAPER AND IN PROGRESS TOO AND DRAG AN SA, NO EXCEPTION. HOTSEAT IS NOT ONLY FOR FELLOWS IT'S EVERYBODY HAD TO GO THERE SO THIS IS WHEN IN DRAGANA SAME SO YOU CAN -- YEAH. JACKIE. YEAH. IT'S THE ONLY ONE WHERE HE IS SITTING AND WRITING SO I DON'T KNOW WHAT WAS GOING ON HERE BUT SEE THE KIND OF SURPRISE BECAUSE IT WAS NOT UP ANDIST LEAVING. WHAT ARE WE DOING? I'M HAPPY I DID IT AND IT'S THE ONE I TOOK IN (INAUDIBLE). A FEW YEARS AGO AND SO ALTHOUGH I'M FRENCH I'M NOT BRAZILIAN, MY WIFE IS BRAZILIAN AND IT WAS MY GREAT PLEASURE TO BE SURROUNDED BY SO MANY BRAZILIAN PEOPLE AND BE ADOPTED BY THEM SO I'M REALLY GRATEFUL THAT I'VE BEEN IN THIS ENVIRONMENT AND OF COURSE, MANY OTHER PEOPLE ALYSE, THAT WE'RE PART OF THE COMMUNITY AND THE CHILDREN OF ALAN AT THE TIME I WAS THERE AND WE ALSO REALLY GRATEFUL THAT WE HAVE BEEN INVITED AND WONDERFUL BARBECUES AND ALSO THANKSGIVING PARTIES WITH ALAN AND STEPHANIE AND I'VE ALSO KNOWN IN FACT WITH MANY GOOD FRIENDS IN DIFFERENT LABS SO THAT IS ALAN AND YOU KNOW HIM SO LIKE YESTERDAY, THERE WERE TWO OTHER FRENCH PEOPLE IN THIS ROOM, AT LEAST TWO AND ALAN WANTED TO MACK SURE THAT I WOULD INTERACT AND GET TO KNOW THEM SO OH YOU DON'T KNOW, COME! SO NOW I KNOW ISABEL AND DANIEL BUT IT'S SOMETHING THAT ALAN WOULD DO WITH EVERYBODY. IT'S NOTHING SPECIAL. IT IS A PICTURE, A BLURRY ONE, FROM THE TYPE INTERFERON KEYSTONE MEETING AND JUST TO SAY IN ADDITION TO BE A GREAT MENTOR, AND TO BE AND I'M GRATEFUL HE HIRED ME HE IS A GREAT HUMAN BEING AND I'M HAPPY TO BE HERE TODAY. THANK YOU, VERY MUCH. [APPLAUSE] >> DO YOU WANT TO ASK A QUESTION? >> SO, OK. >> SO NUR ANALYSIS OF THE INCREASE DISEASE IN THE ABSENCE OF STING THE NEWT TRA FILL RESPONSE WAS DRAMATIC AND SUSTAINED SO I'M CURIOUS ABOUT HOW YOU ARE FOLLOWING UP ON THAT AND THAT THE INCREASE CURE ARE YOU CERTAIN THAT WAS SAT A SOL I CAN OR MIG MIGHT OW -- THERE'S W APPRECIATION IT'S INCREASE NORTH VARIOUS CELL TYPES SO A CHANGE IN THE METABOLIC CHANGE OF THE CELLS AND IT'S INTERESTING BECAUSE SYNTHESIZES SUBSTRATES THAT CAN INCREASE FIBERROSIS. >> THAT'S A GOOD POINT. WE DID IT BY FLOW. SO I DON'T KNOW WHETHER IT WAS FROM THE MITO IS IT'S SOMETHING. >> IT WOULD BE INTERESTING TO DO A WHOLE MORE SINGLE CELL ANALYSIS OF THAT BECAUSE THERE IS A LOT OF INTEREST TO THE CD9 POSITIVE SCAR ASSOCIATED MAX AND THEY'RE THOUGHT TO BE PROFANE ROT TICK SO THERE'S A NEWT TRA FILL PART OF THAT. >> YEAH, YEAH. >> IT'S SOMETHING THAT WE ARE REALLY SURPRISED WITH AND REALLY INTERESTED BUT I DON'T KNOW WHETHER IT'S THEY'RE JUST IF IT'S ALSO LIKE COMING OF THE PERSIST AND HOW MUCH IT'S RELATED TO OTHER THINGS THAT ARE GOING ON AND THAT LEAD TO THIS PERSISTENCE OR REALLY THEY'RE PARTICIPATING IN THE DISEASE SO IT'S SOMETHING THAT I DON'T KNOW. >> AND YOUR SNAPSHOT OF IL-17 DID YOU LOOK A SINGLE TIME POINT. >> I DID TWO TIME POINTS SO IT'S THE 8 AND I ALSO HAVE A LATER TIME POINT AND IT WAS NO DIFFERENCE. >> THANK YOU. >> I'LL ASK THE LAST REQUEST QUESTION. SO HOW DO YOU THINK THE ROLE OF TYPE 3 INTERFERON REGULATING NEUTROPHIL RESPONSE IN THIS MODEL? >> IN THIS MODEL I DON'T KNOW BUT IT'S BEEN SHOWN IN TWO OTHER MODELS AND ONE PAPER SHOWED THAT IL-28 CAN LIMIT THE NEUTRAL MIGRATION SO THE SKIN AND THE OTHER ONE SAID THAT IN THE GUT IT'S NOT THE CASE AT ALL. IT DOESN'T CHANGE BUT IT'S EFFECTIVE FUNCTION OF NEWT TREE FILLS THAT ARE EFFECTED WITH LESS WHEN YOU HAVE IL-28 SO IT'S NOT SURE WHAT IT'S DOING. I THINK IT HAS MULTIPLE ROLES. >> ALL RIGHT. THANK YOU. >> THANK YOU. >[OFF MIC]>> IN THE MOUSE YOU HS PATHOLOGY. WITH THE GR1 AT LEAST. [APPLAUSE] >> SO, LAST SPEAKER AND KEVIN SCHENDEROV. DRAGON A, THANK YOU FOR IN INVITING ME TO SPEAK. IT'S REALLY A GREAT HONOR TO BE HERE WITH SO MANY LUMINARIES FROM WITHIN LPD OUTSIDE LPD STANDING AT THIS PODIUM IT'S INCREDIBLE TO BE HERE AND SO GRATEFUL TO BE INVITED. YOU KNOW, TONY PUT THIS AS A FAMILY REUNION AND I ACTUALLY, YESTERDAY AND THIS MORNING I WAS THINKING THE EX ACTS SAME PHRASING THIS REALLY DOES FEEL LIKE COMING HOME TO A FAMILY TO BE AMONG AWFUL YOU AND IT'S SO GRATEFUL TO SEE EVERYONE HORNING ALAN AND USUALLY ALAN IS NOT THE CENTER OF ATTENTION BECAUSE AS ALAN'S TRAINEE, I HAVE FELT AND I KNOW SO MANY OTHERS HAVE SAID THE SAME THING THAT HE ALWAYS PUT US FRONT AND CENTER AND THAT HE WAS ALWAYS SUPPORTING US AND HELPING US AND INTERROG INTRODUO OTHERS EVEN YESTERDAY AT RECEPTION, ALAN'S HONOR, HE WAS INTRODUCING US TO OTHER PEOPLE SO I KNOW THAT THIS IS PROBABLY COUNTER TO YOUR NATURAL AN BUT I'M GLAD WE'RE TAKING THE TIME TO REALLY TALK ABOUT EVERYTHING THAT YOU'VE DONE FOR ALL OF US AND THIS IS OF COURSE FROM LAST NIGHT AND AS CARL MENTIONED, I WAS A GRAD STUDENT WITH ALAN FROM '07 TO 2012 I CAME TO HIM BEING INTERESTED IN WORKING ON TH17 CELLS AND HE GRACIOUSLY TOOK ME ON AND EVEN THOUGH HE WAS BASICALLY WITH PTDOCS OR MUCH MORE EXPERIENCED THAN I WAS AT THE TIME IN THE LAB AND HE WAS WILLING TO HUMOR ME AND TRAIN ME AND TEACH ME SO MUCH ABOUT SCIENCE AND HOW TO CRAFT EXPERIMENT AND HOW TO WRITE PAPERS AND EVEN AFTER I LEFT THE LAB, HE HAS BEEN SO SUPPORTIVE I WENT TO HOP TINS FOR MEDICINE SCHOOL, ALAN CAME TO MY GRADUATION AND SINCE THEN I'M BACK IN THE RESEARCH WORLD AS SAY FELLOW AND PULL LYNN ARY CRITICAL AIR AND I'M APPLYING FOR A GRANT AND RECENTLY AND VERY HELPFULLY SUGGESTED THAT I SPEAK TO EDMONTON PEARCE WHEN EDMONTON PEARCE CAME AND MY CURRENT MENTOR IS LEAVING HOPKINS AND ED IS GRACIOUSLY TAKING ME ON SO ALAN HAS BEEN HELPING ME EVERYWHERE ALONG THE WAY WHEN I WAS IN HIS LAB TO CURRENTLY AND MY WIFE AND I HAD A SON 16 MONTHS AGO NOW AND ALAN ALSO IS THERE AT HIS PANDEMIC ZOOM BIRTHDAY PARTY AND GRACIOUSLY PUTTING UP WITH THE BIBBY SHARK VIDEO THAT WE WERE WATCHING AND SO ALAN HAS BEEN SO MUCH MORE THAN JUST A COLLEAGUE AND A MENTOR AND YOU HAVE BEEN REALLY A FRIEND TO US AND THANK YOU SO MUCH, ALAN. GETTING TO THE SCIENTIFIC PART I'M GOING TO TALK ABOUT WORK RELATED TO ACUTE RESPIRATORY DISTRESS SYSTEM SO I'M DOING TRAINING SO I SPENT IN THE ICUs AND THIS IS A CONDITION WE TREAT REGULARLY AND ESPECIALLY RECENTLY WITH THE COVID PANDEMIC AND SO ARDS IS DISEASES CALLED IN L AS AN ACRONYM WAS FIRST DESCRIBED IN 1967 IN THE STUDY IN THE LAND SET IN A DOZEN PATIENTS WITH RESPIRATORY FAILURE FOR VARIOUS REASONS AND SINCE THEN WE'VE REALIZED THAT AIRDS CAN OCCUR DUE TO DIRECTOR INDIRECT LUNG INJURY FOR ANY NUMBER OF CAUSES THAT EFFECT ABOUT 200,000 INDIVIDUALS IN THE U.S. AND A FEW MILLION WORLDWIDE EACH YEAR AND UNFORTUNATELY WE'RE TERRIBLE AT TREATING IT. WE REALLY HAVE VERY HIGH MORTALITY DESPITE BEST ICU CARE 30% TO 40% OF ERS DIE OF MULTI ORGAN FAILURE PRIMARILY AND JUST TO GIVE AN IDEA OF WHAT IT LOOKS LIKE, THIS IS A NORMAL X-RAY. THE LUNGS LOOK DARK AND YOU DO HAVE TO BE A PULL MINUTOLOGIST TO SEE THE LUNGS LOOK WHITE AND SO REALLY DIFFUSED LUNG INJURY AND I ALLUDED TO MANY DIFFERENT CLAUSES BUT JUST TO SPECIFY A FEW OF THEM IN TERMS OF DIRECT LUNG ENTRY AND PNEUMONIA AND SEVERE COVID-19 LEADING TO PNEUMONIA AND AS SIRRATION AND TRAUMA TO THE CHEST AND NEAR DROWNING AND ALSO INDIRECT LUNG INJURY AND NON NA MOAN KA CAN CAUSE ARDS IN ADDITION TO NA KNOWNIA AND THEY CAN CAUSE ARDS AND REALLY ARE CARE FOR ARDS IS SUPPORTIVE SO PEOPLE HAVE SIGNIFICANT HIGH POXEMIA, LOW LEVELS OF OXYGEN SO WE PUT THEM ON VENTILATORS AND BREATHING MACHINES TO HELP SUPPORT AND WE HAVE LEARNED THAT YOU HAVE TO USE A VENTILATOR IN A SPECIFIC WAY. WE CALL IT LUNG PROTECTIVE VENTILATION AND LOW TITLE VOLUMES TO REDUCE TRAUMA FROM PRESSURE AND OVER DIS TENSION OF THE LUNGS AND WE'VE ALSO FOUND THAT CERTAIN PATIENTS BENEFIT FROM PRONING, TURNING THEM ON THEIR BELLY NOW THAT I HAVE A KID I CALL IT TUMMY TIME AND WE LEARNED YOU HAVE TO BE CAREFUL OF THE A FLUID YOU GIVE THESE PATIENTS BUT THERE'S NO PHARMACOLOGICAL THERAPY FOR ARDS. WE DO KNOW PATH OWE PHYSIOLOGICALLY THAT ARDS IS HIGHLY INFLAMMATORY SO THIS IS A FIGURE SUMMARIZING WHAT WE KNOW BUT IN THE INITIAL PHASE OF ARDS, YOU GET ACTIVATION OF A LOT OF THE INNATE IMMUNE CELLS THAT WE ALL KNOW AND LOVE AND THEY USE LOTS OF LOAN CYTOKINES, REACTIVE OXYGEN PEE SEIZE AND YOU GET LOTS OF BARRIER INJURY TO THE BARRIERS AND MORE CELLS COME IN FLUID COMES IN AND THE VLI ARE FLOODED THAT WHY THE LUNG IS WHITE AS I SHOWED YOU ON THAT X-RAY. IF MANY PATIENTS, IF YOU SUPPORT THEM, THEN OVER TIME THEY WILL HEAL AND THIS WILL ROW SOLVE THERE'S A SHIFT IN THE PHENOTYPES OF THE MACROPHAGES IN THE LUNG AND THEY BECOME MORE PRO RESOLUTION RATHER THAN PRO INFLAMMATORY AND YOU HAVE A DROP IN THE PRODUCTION OF CYTOKINES AND BASICALLY THESE PRONE INFLAMMATORY STIMULI RESOLVE AND THERE'S HEALING AND UNFORTUNATELY AS I SAID, 30% TO 40% OF PATIENTS DOES NOT HAPPEN AND EVEN IN PEOPLE WHO DO HEAL A LOT OF THEM HEAL IN A DIS-REGULATED FIE BROTT TICK WAY AND HAVE ARDS, EVEN IF PATIENTS SURVIVE THEY CAN HAVE MORBIDITY LATER ON. PEOPLE HAVE TRIED KNOWING THIS PATH OF PHYSIOLOGY VARIOUS INFLAMMATORY THERAPIES AND UNFORTUNATELY, THEY HAVE LARGELY BEEN FAILURES SO STEROIDS HAVEN'T WORKED OUTSIDE OF COVID-19 AND ANTI-GMCSF, ALL OF THESE HAVE BEEN FAILED CLINICAL TRIALS AND SO THE THOUGHT BEHIND THE WORK THAT I DID, I STARTED THIS WITH JOHNATHAN POWELL AND MAUREEN HORTON AT HOPKINS AND WE'LL CONTINUE ED PEARCE IS MAYBE WE'RE NOT TARGETING INFLAMMATION THE RIGHT WAY SO WE'RE INTERESTED IN IMMUNO METABOLISM AND PEOPLE HAVE REALIZED THAT THERE ARE PROFOUND METABOLIC SHIFTS GOING ON IN IMMUNE CELLS THAT'S AIR ACTIVATED DEPENDING ON HOW YOU ACTIVATE THEM AND THOSE METABOLIC CHANGES ARE VERY IMPORTANT FOR THEIR FUNCTION. SO, WE HYPOTHESIZED THAT MAYBE BY UNDERSTANDING SOME OF THE METABOLIC CHANGES HAPPENING IN ARDS, WE CAN FIND SOME DIS-REGULATED METABOLIC PATHWAYS SO THAT MAYBE WE CAN TARGET TO TROY TO PROMOTE BETTER RECOVER ROW FROM LUNG INJURY AND REDUCED DETRIMENTAL INFLAMMATION SO OUR APPROACH WAS TO SET UP MODELS OF LUNG INJURY BECAUSE WE WANTED TO FIND PATHWAYS THAT OCCURRED IN MORE THAN ONE MODEL AND WE DIDN'T WANT TO JUST FIND ONE MODEL SPECIFIC PATHWAY AND SO WE SET UP A FEW DIFFERENT MODELS AND USING METABALOMICS, WE MEASURED LEVELS OF A FEW HUNDRED METABOLITES IN THE LUNGS OF INJURED VERSUS UNINJURED MICE AND SO JUST TO QUICKLY TELL YOU THE MODELS WE USED, ONE IS A BACTERIA MA KNOWNIA AND AND EARLY ON AND. >> TO MA LAB LOAM I CAN AND IN RED AND THESE AND DIFFERENT BETWEEN THE DAMAGE LUNGS VERSUS THE NAIVE MICE SO THIS IS IN THE MODEL AND SO YOU CAN SEE MANY METABOLITES ARE CHANGING AND IF YOU DO A PRINCIPLE COMPONENT ANALYSIS WE CAN SEE THE MICE REALLY DO CLUSTER BASED ON THEIR STATUS SO THESE ARE THE NAIVE MICE AND THE INFECTED MICE AT DIFFERENT TIME POINTS AND THE MICE AR RESOLVING THE RESOLUTIOO THEY GO BACK TO THE NAIVE MICE. THIS IS A HEAT MAP SHOWING IS THAT MANY OF THE METABOLITES WE LOOK AT START OFF HIGH IN THE NAIVE MICE SO EACH A IS A MOUSE AND THE RED ARE NAIVE SO THEY START OFF HIGH AND NAIVE AND DROVE DURING INJURY AND START TO RECOVER DURING LATER TIME POINTS AND THEN ALSO, SOME METABOLITES START LOW AND GO UP DURING LUNG INJURY AND THEN START TO COME BACK DOWN SO, THERE ARE BOTH PATTERNS THAT WE SEE AND OF COURSE, IT'S HARD LOOKING AT LISTS OF THESE SO ONE THING WE DID WAS LOOK AT PATHWAY ANALYSIS AND YOU CAN SEE VARIOUS METABOLICS PATHWAYS THAT ARE CHANGING DURING LUNG INJURY WITH STREP NOW MANY OWE AND YOU SEE NUCLEOTIDE METABOLISM, AMINO ACID, TCA CYCLE, ET CETERA. WE BECAME INTERESTED IN TWO SPECIFIC METABOLITES, MIO AND WE SAW THAT ACROSS THE THREE LUNG INJURY MODELS THEY HAD SIMILAR CONNETICS SO IN ALL THREE MODELS, BOTH WOULD DROP DURING ACUTE LUNG INJURY AND THEN AS YOU LOOK LATER THEY START TO COME BACK UP. SO, MANY OF YOU MAY KNOW GLUTATHIONE IS A TRY PEP SIDE AND REACTS WITH FRO RADICALS AND THOSE CHANGE IT FROM A REDUCED FORM TO SAY OXIDIZED FORM AND INTERESTINGLY FOR A LONG TIME ACTUALLY, PEOPLE HAD SHOWN THAT THE LEVELS OF GLUTE A THIGH ON O IT'S A LITTLE CHALLENGING TO ACTUALLY USE THIS AS A TREATMENT THOUGH BECAUSE IT'S NOT INEFFICIENT WAY TO INCREASE SO PEOPLE HAVE TRIED ALTERNATIVES AND SO THERE WAS ACTUALLY A COUPLE OF TRIALS IN PEOPLE USING SISTINE KNACK WHICH IS PROVIDING SISTINE IT'S AN AMINO ACID SO IT'S THOUGHT IT WILL HELP INCREASE IN OTHER COMPOUND WE WERE INTERESTED IN IS IT'S A HAS BETTER BIO AVAILABILITY AND CELL PERMEABILITY AND ACTUALLY CAN GET INTO CELLS AND IS CONVERTED INTO REDUCED GLUTATHIONE AND THE OTHER IS A SUGAR ALCOHOL THAT IS A PRECURSOR USED IN INTRA CELLULAR SIGNALING LIKE THE KAI A NAS PATHWAY SO IT'S A PRE CURSER OF A COMPONENT AND SUR FALK TIN IS IMPORTANT FOR THE LUNG AND TO REDUCE SURFACE FUNCTION AND IT HAS A REGULATORY ROLE AND IT'S BEEN SHOWN TO HELP WITH INSULIN RESISTANCE AND ALSO MOST RELEVANT TO WHAT WE SAW WITH GLUTATHIONE IT HAS A ROLE IN REDOX BALANCE. IT CAN HELP WITH THE PHOSPHATE PATHWAY SO WE ASKED A RELATIVELY SIMPLE QUESTION SINCE THE MIO INOS OWE TOLL AND LEVELS ARE LOW IN THE MICE WITH ACUTE LUNG INJURY, WE TREATED MICE WITH EITHER JUST CONTROL WATER OR MIO MONTHS INTOLL OR KNACK AND IT DIDN'T WORK IN THE HUMAN TRIALS AND DAILY BY PUT THESE COMPOUNDS IN THEIR DRINKING WATER SO THEY WERE EXPOSED TO THEM CONTINUOUSLY DURING THAT TIME AS WELL AND WE WERE EXCITED TO SEE THAT WE DID SEE A REDUCTION IN MORTALITY FROM STREP NUMO AND INTERESTINGLY DIDN'T SEE ANY REDUCTION MORTALITY WITH KNACK SO THIS WE HAVE TO SEE WHETHER THIS HAS ANY RELEVANCE TO ANY MODELS OF LUNG INJURY BUT WE WERE EXCITED TO SEE THIS AT LEAST IN ONE MODEL. AND WE'RE WORKING ON MECHANISTICALLY SEEING WHAT IS GOING ON BUT ONE THING WE HAVE SEEN IS REDUCED BACTERIAL LOADS SO FASTER BACTERIAL CLEAR NOT IN ANCE IN THEBAL AND IN DAY FIVE R INFECTION SO IT SEEMS THAT TREATING THE MICE WITH THESE COMPOUNDS DOES SPEED UP BACTERIAL CLEARANCE SO OF COURSE, WE'RE WONDERING WHETHER THIS IS AN ALL RELEVANT TO PEOPLE SO THIS MIGHT JUST BE A PHENOMENON OF MICE SO TO GET AT THIS, WE'VE DONE SOME METABOLOMICS IN HUMANS SO IN OUR ICUs OF COURSE, ESPECIALLY EARLY IN THE PANDEMIC WE HAD A LOT OF COVID-19 PATIENTS AND SOME OF THEIR FAMILIES CONSENTED FOR US TO DO NON LAVAGE WHERE WE PUT A SUCTION CATHETER DOWN IN THE TRACK Y'ALL TUBES AND PULLED IT BACK AND WITH OUR COLLABORATORS WE'VE DONE -- COMPARED TO PATIENTS UNDER ANESTHESIAIA FOR ELECTIVE SURGERY AND AGREED TO LET US DO THE SAME PROCEDURE, COMPARED TO THESE HEALTHY CONTROLS, THE COVID PATIENTS WITH ARDS HAVE LOWER LEVELS OF MIO INOS TOLL AND OF COURSE WAS EXCITING SINCE THAT CORRELATED WITH WHAT WE SAW IN THE MOUSE MODELS. SO, TO CONCLUDE, WE'VE SEEN THAT THERE ARE SOME METABOLICS SHARED BETWEEN VARIOUS LUNG INJURY MODELS AND BOTH DIRECT AND INDIRECT LUNG INJURY AND IT SEEMS THAT THEY ARE INDEPENDENT OF THE EXACT MECHANISM OF INJURY AND SO WE'RE HOPING THAT THIS ACTUALLY IS RELEVANT TO HUMAN ARDS AND LOOKING AT OTHER LUNG INJURY MODEL AND MODEL THAT NICO SHOWED LOOKING MORE AT MECHANISM AND WHAT THEY'RE DOING AND ALSO ALL THE DATA I'VE SHOWN YOU IS LOOKING AT WHOLE LUNG AND WE'RE INTERESTED IN UNDERSTANDING WHETHER THESE METABOLICS CHANGES ARE HAPPENING IN THE IMMUNE OR NON IMMUNE AND AND METABOLOMICS ON THOSE SO INTERESTED IN LOOKING AT REGULATION OF GONE EXPRESSION METABOLIC ENZYMES AND SEEING IF THEY'RE CHANGING WHETHER IT UNDERLIES THE METABOLITE CHANGES WE'RE SEEING AND LOOKING AT METABOLIC FLUX THROUGH THESE PATHWAYS AND LOOKING AT NON COVID-19 ARDS PATIENTS TO SEE WHETHER THIS IS RELEVANT TO DISEASES OTHER THAN COVID-19 SO A LOT OF WORK TO DO SO WE HAVE SO FAR SO, I WANT TO THANK MY ADVISERS AND AS I MENTIONED I STARTED THIS WORK WITH MAUREEN AND JOHNATHAN AND ED IS GOING TO TAKE ME ON AS JOHNATHAN AND MAUREEN HAVE LEFT AND WE HAVE A LOT OF HELP FROM PEOPLE AT MAUREEN'S LAB AND LEON AND ED IS REALLY THE KEY METABOLOMICS COLLABORATOR AND WE HAD A LOT OF HELP AND SO THANK YOU GUYS SO MUCH FOR YOUR ATTENTION AND LOVE TO TAKE ANY QUESTIONS. [APPLAUSE] >> TERRIFIC TALK. THANK YOU. YOU KNOW, MORE OF A COMMENT THAN ANYTHING ELSE. GIVEN THE HUGE AMOUNT OF LITERATURE ON OXIDATE TIVE STRESS, AND THE FACT THAT CLINICAL TRIALS BASICALLY HE HAVE ALWAYS FAILED, , IF YOU LOCK AT THAT LITERATURE THE THING THAT IS MOST A PARENT IS AS PEOPLE MOVE TRANSLATIONALLY COMPLETE LACK OF ATTENTION TO PKPD RELATIONSHIPS AND SO AS YOU F. THIS GOES FORWARD AS YOU DO THIS I THINK IT'S THE CRITICAL THING WHEN YOU START THINKING TRANSLATIONALLY AND BECAUSE THAT LITERATURE IS JUST RIFE WITH DISAPPOINTMENT AND RIFE WITH POOR TRANSLATION. >> THAT'S A GREAT POINT, THANK YOU. >>LET'S START WHERE WHERE STOPPED BEFORE LUNCH SO WE'LL JUST FINISH THE LAST SECTION AND WE MENTION SEVERAL TIMES IN THIS MEETING, ALAN'S LAB ALWAYS HAS A LOT OF BRAZILIANS SO I THINK THIS IS INGREDIENT FOR BRAZILIAN SCIENTISTS EVEN ENRICHED IN THE TV PROGRAMS OR FIVE OR SIX BRAZILIANS AND SIX OR SEVEN OR 10 YEARS AND SHE SPENT SIX YEARS AND IN TUCSON AND ALL RIGHT -- I HOPE EVERYBODY IS WELL FED. SO, I WANT TODAY REALLY EXPRESS MY GRATEFULNESS TO BE HERE AND I'M REALLY GRATEFUL, NOT ONLY TO BE HERE NOW BUT GRATEFUL BY ALL THE TIME THAT I SPENT IN THE IMMUNO BIOLOGY SECTION. NOW I'M BACK IN BRAZIL AND I SET UP MY SHOP SOUTH OF BRAZIL ALTHOUGH AS A BIT COLD SO ALAN VISITS THERE BUT TAKES A WHILE BECAUSE HE REALLY, REALLY LIKES THE NORTHEAST PART OF BRAZIL AND THAT IS HIS FAVORITE AND LOOK AT THAT. THAT WAS NATAL AND SO LOOK AT HIS FACE AND HIS SMILE. HE IS RELAXED THERE. HE IS ALWAYS LIKE THAT WHEN HE IS IN NORTHEAST, I CAN TELL YOU. I AM GOING TO READ SOMETHING THAT I WROTE SO I DON'T -- I DON'T MISS ANYTHING THAT I WANT TO SAY. ALAN, FIRST EVER ALL, I LOVE YOU. I LOVE YOU GATO, A LOT. SERIOUSLY, 22 YEARS AGO, HOW CAN I NOT LOVE THIS GUY. 22 YEARS AGO, ALAN TAKES A RIDE WITH ME BECAUSE MY FORMER PROFESSORS GINA BA HAW ASKED ME TO TAKE ALAN, STEPHANIE AND BOB COFFMAN TO SHOW THEM A GOOD TIME. OF COURSE, THAT'S WHAT I DID. SO, WE WENT TO PILLOW RIO AND ALISON AND LAURA WAS ALSO THERE SO WE WENT AND DID A LOT OF DANCING AND SALVADOR IF YOU ARE NOT FAMILIAR WITH THE AREA. A LOT OF CYPERONIAS. IT WAS AN OLD CAR AND I FELT REALLY RESPONSIBLE FOR THESE GUYS, I BECAUSE NOTHING COULD HAPPEN LIKE AN ACCIDENT OR ANYTHING SO IT WAS KIND OF A VERY BIG RESPONSIBILITY. SO, I DID MY JOB, I DELIVERED THEM VERY HAPPY TO THE HOTEL. SO THE NEXT DAY, ALAN ASKED ME WHAT I WANT TO DO AFTER MEDICAL SCHOOL AND I SAID, I WANT TO DO RESEARCH AND HE SAID, I LIKED YOU. I WANT TO WORK ON A MECHANISM FOR YOU TO COME TO THE IMMUNO BIOLOGY SE SECTION AND WORK WITS SO IT WAS MY DREAMT AND I FELT REALLY, REALLY HAPPY ABOUT IT. SO I GUESS ALAN SAW SOMETHING IN ME THAT I WAS NOT EVEN SEEING BACK THEN. SO I ARRIVED HERE IN 2001 AND IT WAS A VERY COLD WINTER BUT COMING FROM BRAZIL BUT I WAS STAYING AT CHIEFERS PLACE, REST IN PEACE, AND HE INTRODUCED ME TO A LOT OF THE THINGS, EVEN THE CHIEFERENI WHICH IS A WAY TO MAKE HYPER INIA. I LOVE THIS RIGHT AWAY. I MET THESE GUYS ARE THE THREE MUSKETEERS AND I REMEMBER THEM IN IT TOOK ME LIKE IN THE HAND AND GAVE ME A TOUR AT THE NIH, WE CAME TO FAS BACK THEN WE HAD TO SHOW THE IAP66 FORM BUT NOW THEY CHANGED TO HOLD THE NUMBERS SO IT WAS REALLY FUN. AND THAT WAS -- [LAUGHTER] SO I HAVE LEARNED ANYTHING FOR YOU. CHOW CHUCK, DAMIAN AND ALSO I WORKED A LITTLE BIT AND I LEARNED A LOT EVEN THOUGH I DON'T THINK I'M A TOXO PERSON BUT I LEARNED A LOT FROM THE MEETINGS AND WITH DRAGANA, ROMINA AND JULIO BACK THEN. AFTERWARDS, I WORKED WITH TWO CRUISE IN THE LAB AND HOW TO PAG STUDENT AND HE CAME SO HE WORKED HARD EARL ON. SO HERE, I DID SOME PADDING AND I DID A LOT OF THE BARBECUE FOR ALAN AND I DID A LOT OF LAB ENTERTAINMENT. AND I CAN TELL YOU FROM MY HEART, I LIKED THAT EACH ONE OF YOU CAN FEEL MY LOVE AND MY HUG FOR THE SPECIAL TREATMENT I RECEIVED TO HEAR AND ACTUALLY, IT WAS VERY IMPORTANT FOR ME NOT ONLY AS A SCIENTIST BUT AS A SCIENTIST BUT AS A HUMAN BEING. ALAN, I NEVER TOLD YOU GUYS I WON THE LOTTERY HERE BUT SOMETHING WAY, WAY BETTER THAN MONEY, NEW SISTERS AND BROTHERS CAME INTO MY LIFE WHICH I STILL HAVE TODAY SO MY STUDENTS, FORMER STUDENTS IN THE LAB, THEY CAME TO CHUCK'S LAB AND THEY EVEN WENT TO AUSTRALIA AND ACROSS GROUP AND THEY DID A LOT OF COURSES WITH BIO FORMAT TICKS SO I'M TRYING TO GIVE BACK TO THE PLANET WHAT YOU TAUGHT ME. AND THANK YOU VERY MUCH FOR THE OPPORTUNITY. SO, JUST THAT'S ALAN I WAS A VISITING PROFESSOR IN 2017 AND I ALWAYS COME HERE AND ALAN WOULD ALWAYS TAKE ME AND DRAGANA AND EVERYBODY WAS, I WAS VERY HAPPY AND I FELT REALLY AT HOME AND THAT DRAGANA, MAYBE I CAN DO THIS ALL ON HERE. THERE'S A SOUND THERE WE CANNOT HEAR SO HE IS MAKING A COFFEE AND HE IS LIKE, OH, I GOT TO DIG UP THIS OLD PICTURE OF ME ON MY SURF BOARD BECAUSE I WAS DOING SUFFERING BACK THEN SO, I DON'T THINK THAT'S THE PICTURE BECAUSE THE SKIN COLOR DOESN'T MATCH BUT IT COULD BE YOU I STILL WAIT ON THAT PICTURE. OF COURSE, THE ONE TO ONE HOTSEAT WRITING THE PAPER AND I HAD MY CURLY BIG HAIR DURING MY SA BAD CAL. I CAME HERE AND WORKED ON THE HOTSEAT. I TOOK OUT THE ADDRESS BUT I KNOW YOU ARE INVITING PEOPLE SO YOU CAN REMOVE IT AND PEOPLE CAN SEE THE ADDRESS OVER HERE. SO, AND THAT IS ALAN'S INVITATION. HE LIKED THE LITTLE WORD DOCK GUYS AND I THINK THAT WAS BEFORE THE PANDEMIC SO YOU SEE ALSO DIEGO HERE, EDUARDO, THAT IS PEDRO AND THAT IS -- WHAT'S HIS NAME. AND THAT IS ALSO -- WHAT'S HIS NAME? SO, I DIDN'T BRING ANY DATA. I WANT TO SHOW YOU WHAT I HELPED PEOPLE AT MY UNIVERSITY. SO WE WORKED ON THIS NEW HERE AND THAT IS MY RIDE I USED TO DO EVERYDAY DURING THE PANDEMIC AND HERE THAT'S OUR IF STYLE THAT I'M GOING TO BE RESPONSIBLE IN THAT INSTITUTION NOW. DEDICATING TO OUR BRAZIL VERSION OF THE LAB OF IMMUNO BIOLOGY THAT'S NOT OF COURSE STOPPING HERE AND IT'S DAVID SACKS PUT IT REALLY NICELY BEFORE. SO, IN THIS 13 AND 14 YEARS A LOT OF PEOPLE STUDENT HE'D IN THE LABS ABLE TO CREATE THREE P.I.s AND EVERYTHING THAT ALAN CAUGHT ME AND AS I PROMISED, THAT'S ISABELLA MY DAUGHTER AND WE USED TO GO SURFING TOGETHER AND OVER HERE AND SHE WAS LITTLE SHE STARTED PAINTING AND NOW SHE'S GRILL AND SHE'S BECOMING AN ARTIST WITH THE WATER PAINTING THAT SHE'S DOING A LOT NOW AND ALAN IS ALWAYS ASKING HOW IS SHE. SHE'S NOW 14-YEARS-OLD SO NOW THAT SHE'S GOT HER MOTHER'S LOOKS, I GUESS, BEAUTIFUL. AND THAT IT. THANK YOU, VERY MUCH. [APPLAUSE] I FORGOT TO TELL YOU GUYS, I WAS GOING TO DID A GEORGE YO PLAY SO HE INVITED YOU TO GO TO HIS HOUSE IN ITALY. THAT'S NOT AS BEAUTIFUL AS ITALY HOUSE BUT YOU CAN COME TO MY HOUSE. IT'S AVAILABLE TO YOU TOO. THANK YOU. [APPLAUSE] >> GOOD AFTERNOON, EVERYONE, I'M FROM THE NATIONAL CANCER INSTITUTE AND TOGETHER WITH NIKKI AT NIDCR I'M CHAIRING THE NIH FDA CYTOKINE CENTER GROUP AND. >> IT'S AN 800-MEMBER STRONG COMMUNITY OF SCIENTISTS AT NIH AND FDA INTERESTED IN SIGN CYTOKINE BIOLOGY AND WE DO TWO OR THREE MINI SYMPOSIUMS ON CYTOKINE RESEARCH EVERY YEAR AND WE WATER ANNUAL BEST PAPER IN SIGHT OWEICYTOKINE AND WE HAVE R WINNER FROM MIKE AND THE REASON WHY WE ARE HERE ACTUALLY IS BECAUSE NIH CIG HAS FINISHED TELEVISION 1992 AND HAS BEEN CO FOUNDED, IT'S ACTUALLY A BRAINCHILD OF ALAN, WHO ORGANIZED AT IT AT THAT TIME AND HAS BEEN STILL RUNNING IT ON TO THIS TIME TOGETHER WITH US AND SO IN 2017 THE CIG CELEBRATED ITS 25th ANNIVERSARY AND I REMEMBER IT WAS A COMMITTEE IN GEORGE YO'S LAB AT BUILDING 37 AND WE WERE THINKING ABOUT HOW TO ORGANIZE IT AND MAKE A POSTER FOR IT AND ALAN SAID I THINK WE SHOULD HAVE A BALLOON? I SAID YOU WANT A BALLOON FOR A POSTER. I WANT A BALLOON WITH THIS ONE HERE AND I SAID, YOU WANT A CONGRATULATIONS BALLOON, NO, I WANT MANY BALLOONS. SO, I WAS ASKING ALAN S. THAT ENOUGH? YEAH BUT THEN IT NEEDS TO BE CONNECTED TO THE NIH AND I REALLY HAD NO IDEA WHAT IS GOING ABOUT AND LISTENING TO ALL THE TALKS, ALAN HAS AN ALREADY AND HE LIKES TO DRAG IT OUT AND THEN AFTER TALKING WITH ALAN, I FINALLY FOUND OUT WHAT HE WANTED. AND HE WANTED, HE GOT INSPIRATION FROM HIS FAVORITE MOVIE WHICH IS THIS -- AND WHAT HE WANTED IS HAVING A LOT OF BALLOONS AND LIFTING UP A HOUSE WHICH IS THE NIH. SO, WE ENDED UP WITH THIS POSTER. AND AS WE CAN SEE, IT'S THE BALLOON AND ALAN WAS HAPPY AND HE WAS LIFTING THE NIH THAT WAS ALSO GOOD AND THEN HE SAYS NOW LET'S ALL DECIDE TO TEST COVERED BY NIH INVESTIGATORS WHO ARE AT THE NIH. SO IT WAS GREAT. I THINK IN HINDSIGHT WE WERE WRONG BECAUSE THE SCIENCE AT NIH IS LIFTED BY THE PEOPLE WHO DO THE SCIENCE AND IN HAD I THINK THERE'S A BIG BALLOON WHICH IS MISSING WHICH IS ALAN AND THIS IS JUST CELEBRATING THAT HE REALLY HAS WON THIS ALL TOGETHER TO CYTOKINE FOR THE LAST 30 YEARS AND AS YOU CAN REMEMBER, THIS YEAR IT'S GOING TO BE A 30th ANNIVERSARY OF THE THE NIH FDA CYTOKINE INTEREST GROUP AND ALAN HAS A LOT OF FREE TIME NOW -- [LAUGHTER] >> I HOPE IT'S GOING TO HELP US TO ORGANIZE AND I'M NOT SURE WHAT MOVIES HE IS WATCHING BUT AT LEAST WE'LL HAVE DECIDE ABOUT WHAT THE POSTER LOOKS LIKE SO WITH THAT, I'LL GIVE IT TO KAREN. [APPLAUSE] THANK YOU SO I'M KAREN I'M FROM THE CENTER FOR BUY LOGICS OF THE FD A AND CONTINUING THE THEME OF INTEREST GROUPS REPRESENTATION I'M HAPPEN TO BE THE CURRENT CHAIR OF THE IMMUNOLOGY INTEREST GROUP OF WHICH ALAN HAS BEEN A SUPPORTER EVERY MANY, MANY YEARS. I HAVE TO DIVERT AND BLOW DRAGANA FIVE MINUTES TO ALSO ACKNOWLEDGE THAT I THINK I'M A REPRESENTATIVE OF ONE OF THE' NUMEROUS ABLE PEOPLE OUT THERE BEYOND THE IMMEDIATE TRAINEES WR I WAS YOUNGER AND STUPIDER THAN I AM NOW, ALAN WENT OUT OF HIS WAY TO TAKE AN INTEREST IN ME AND MY CAREER AND MY SCIENCE AND FOR MANY YEARS, HE HAS BEEN A SOUNDING BOARD, A SOURCE OF WONDERFUL IN A SCIENTIFIC INSPIRATION, PERSONAL SUPPORT AND ADVISE, I AM STANDING IN MY CURRENT JOB ALMOST ENTIRELY BECAUSE OF A PIECE OF SAGE ADVICE HE GAVE ME WHEN I WAS CONSIDERING THIS RELATIVELY NEW POSITION FOR ME AND WHICH I CANNOT REMEET A PUBLIC MEETING. THANK YOU SO MUCH FOR IN CONCLUDING ME AND MY LAB AND EVERYTHING THAT IS HAPPENED OVER THE LAST MANY, MANY YEARS. YOU KNOW, WE HAVE ALL TALKED THROUGHOUT THE LAST TWO DAYS ABOUT ALAN'S BUILDING OF COMMUNITIES EVERYWHERE WITHIN THE LPD, OF COURSE BUT THROUGH OUT THE SCIENCE WORLD AND SO THE REST OF MY TASK IS TO RECOGNIZE HIS ROLE IN FORMAL COMMUNITY BUILDING AND HYUN TOLD US ABOUT THTHTHE SIGH COC SIGH CO -- HE D THE KNIT BRITS AND THAT COMMUNITY TOO IS DRAWING TOGETHER PEOPLE WITH COMMON INTERESTS IN TUBERCULOSIS AND IT IS GOING STRONG. ON BEHALF OF ALL OF THOSE NORMAL GROUPS THAT YOU HAVE GONE OUT OF YOUR WAY TO CREATE AND NURTURE, AND TO MOVE ALONG TO THE NEXT GENERATION OF PEOPLE, WE HAVE A VERY SMALL TOKEN OF APPRECIATION WHICH THIS CAN GO TO THE BATHROOM AND MOVE DAVID'S MUCH BETTER POSTER UP. [APPLAUSE] [APPLAUSE] >> THANK YOU. >> HELLO, I'M CATHY ZOON I'M THE FORMER SD OF NIAID AND I HAVE A DIVISION OF INTRAMURAL RESEARCH AND SO I WOULD LIKE TO FIRST OF ALL THANK THE ORGANIZERS. >> GENEROSITY IN SO MANY WAYS AND I'LL ELABORATE ON TWO THINGS THAT HAVE GOTTEN SOME ATTENTION ALREADY. HOPEFULLY WE'LL ADD A FEW MORE TIDBITS TO ALAN'S REMARKS OVER THE PAST TWO DAYS. I'D LIKE TO TALK A LITTLE BIT ABOUT ALAN'S LEADERSHIP SKILLS AND THEN ALSO HIS MENTORING SKILLS AND YOU KNOW, AS A LEADER IN ANY ORGANIZATION YOU CAN ONLY DO SO MUCH WITHOUT THE SUPPORT OF OTHER LEADERS IN YOUR ORGANIZATION AND ALAN HAS ALWAYS BEEN ONE OF THE FOLKS WHEN I WAS SCIENTIFIC DIRECTOR I COULD COUNT ON. HE WAS ALWAYS THERE AND ASKING WHAT HE COULD DO TO HELP. SOMETIMES IT WASN'T HELP BUT -- ME WAS THERE AND YOU KNOW, I HAVE TO SAY ALAN, AND THIS IS PARTICULARLY INTERESTING BECAUSE ALAN DOES ALWAYS SMILE, ALWAYS SMILES. BUT ALAN ALSO WORRIES ALL THE TIME. AND YOU KNOW, NO ONE BROUGHT THAT UP. ALAN, YOU WOULD COME INTO MY OFFICE AND YOU WOULD HAVE A SMILE AND YOU WOULD SAY WELL, I'M CONCERNED ABOUT THIS AND I'M CONCERNED ABOUT THAT AND WE WOULD TALK FOR ANOTHER HOUR AND FINALLY WE WOULD GET TO THE NITTY GRITTY OF THE PROBLEM AND I APPRECIATED THAT BECAUSE YOU EXPANDED MY HORIZONS AND WHAT TO THINK ABOUT AS WELL AS MAKE SURE I DIDN'T OVERLOOK ANYTHING AND WE ALL RECOGNIZE ALAN'S GREAT SCIENTIFIC ACHIEVEMENTS AND I HAVE TO SAY, SINKEL BSE THE LABORATORY HAD OUTSTANDING REVIEW AND PROBABLY THAT I SHOULDN'T SAY THAT BECAUSE IT'S PROBABLY CONFIDENTIAL INFORMATION BUT TOO BAD AND HE DID A MARVEL US JOB PREPARING HIS FOLKS FOR THE BSCs, ALWAYS OUTSTANDING PRESENTATIONS AND HE WENT OVER EACH AND EVERY ONE OF YOUR SLIDES AND MADE SURE IT WAS ALL PERFECT AND I BELIEVE IT HAD A GREAT CONTRIBUTION TO THE SUCCESS OF LPD AND WHEN WE HAD NEW INITIATIVES AND I COULD SAY FOR SURE, THERE WAS A TIME AT NIH WHEN ACTUALLY WE DIDN'T HAVE AN INCREASE IN OUR BUDGET WE ACTUALLY HAD A RELATIVE DECREASE IN OUR BUDGET AND IT WAS GOING TO BE PRETTY TRAUMATIC FOR THE INTRAMURAL RESEARCH DIVISION SO WE GOT TOGETHER TO COME UP WITH SOME IDEAS AND ONE OF THEM WAS TO MAKE SURE THAT WE PRESERVED THE RESOURCES FOR OUR TENURE TRACKS AND ALAN WAS A BIG SUPPORTER OF THAT TO REALLY MAKE SURE OUR YOUNG PEOPLE COULD DO THEIR RESEARCH AND BE ABLE TO MOVE ON. SOME OF THE OLDER FOLKS WERE NOT ENAMOR WITH THAT BUT THEY UNDERSTAND THE IMPORTANCE OF ITG TO ALL OF THAT. I ALSO WANT TO SAY THAT YOU KNOW, TOM WYNN IS HERE BUT WHEN I LEFT NIH I WENT TO BE ON THE BOARD OF THE INTERNATIONAL BIOMEDICAL RESEARCH ALLIANCE AND THAT IS A NON-PROFIT ORGANIZATION WHICH HELPS SUPPORT THE NIH OXFORD, CAMBRIDGE SCHOLARS PROGRAM AND TOM, WHO WAS LEADING THE PRO VERSE MICHAEL LENARD OWE AND TOM WYNN AND TOM HAD AN OFFER HE COULDN'T REFUSE TO GO TO PFIZER SO ALAN STEPPED IN TAKING ON THE LEADERSHIP ROLE THERE AND I HAVE TO SAY ALAN, WITH THE HELP OF CHRISTIE PORTER, WHO IS NOW YOUR RIGHT HAND PERSON, YOU'VE DONE AN OUTSTANDING JOB IN HELPING THIS PROGRAM TO GROW AND CONTINUE TO BENEFIT ALL OUR GRADUATE STUDENTS WHO ARE INVOLVED IN THAT PROGRAM SO, THANK YOU AND I HAVE EVERY CONFIDENCE THAT YOU ARE GOING TO BE CONTINUING TO HAVE A ROLE IN THAT PROGRAM AND HELPING MENTOR THOSE STUDENTS BECAUSE THEY REALLY LOCK FORWARD TO ALL YOUR SAGE ADVICE. WELL WITH MENTORING, SO MANY PEOPLE HAVE TALKED ABOUT ALAN AND HIS MENTORING SKILLS. HE EVEN MEN FORWARD TO ME SO THANK YOU, ALAN. FOR PEOPLE WHO DON'T KNOW, ALAN IS SO GENEROUS WITH HIS TIME AND WE HAVE COMMUNITY AND PREPARATION FOR THEIR TENURE AS WELL AS FOR THEIR BSCs AND HE WAS ALWAYS THERE GIVING THEM ADD VOICE FOR ALL THE COMMITTEES HE WAS ON SO ALAN, YOU'VE DONE SUCH A GREAT SERVICE TO ALL. AND JUST A NOTE. WHEN I RETIRED, IT WAS ALWAYS A PUSH-PULL. YOU NEVER KNOW WHAT THE RIGHT TIME IS IT TO -- NOT EVEN RETIRE RETIRE BUT EVEN TO STEP AWAY FROM THE BENCH I WOULD SAY AND SO ALAN, I KNOW YOU HAVE COME TO THE FORK IN THE ROAD AND AS YOGI BAR A SAYS, TAKE IT. AND I THINK TAKE SOME TIME NOW TO DO THE THINGS YOU LOVE AND BEING WITH YOUR FAMILY AND BEING WITH VISITING THE CHILDREN AND GRANDCHILDREN AND TRAVEL AND DOING THE THINGS YOU LOVE AND I WILL BE HERE TO MENTOR HERE ON HOW TO RETIRE. [LAUGHTER] AND SO, YOU ARE DEVOTED AND WILL WANT SCIENTIFICALLY BECAUSE IT'S INAND I CAN NEVER GUARANTEE SO I WANT TO CLOSE IN SAY IN THE WORDS OF ALBERT EINSTEIN. KEEP YOUR BALANCE ON A BICYCLE YOU MUST KEEP MOVING AND ALAN, I'LL BE THERE FOR YOU AND IT'S BEEN WONDERFUL WORKING WITH YOU AND WARM REGARDS ON YOUR NEXT STEPS IN YO YO JOURNEY. THANK YOU. >> THANK YOU, VERY MUCH. THANK YOU FOR STICKING AROUND FOR THE LAST SCIENTIFIC SESSION OF THE AFTERNOON AND BEFORE I START, I'LL JUST MAKE A COUPLE OF QUICK COMMENTS. I THINK OVERWHELMING MAJORITY OF THE PEOPLE IN THE ROOM ARE ALAN'S SCIENTIFIC GRANDCHILDREN AND CHILDREN AND I FALL IN THE CATEGORY OF NEPHEW OR FIRST COUSIN ONCE REMOVED AND I SAY THAT BECAUSE IT TURNS OUT THATTAL ANNE AND CRITICAL ROLE TO PLAY AND IF AND HAD ANY RESEARCH JOB WITH WITH A COLLEAGUE OFFALLANCE WHO SORT OF ALLUDED TO BY AND THERE WERE THREE PH.D STUDENTS AT THE IN SAN DIEGO AND ALL TOGETHER AND MADE A DECISION TO DITCH BASIC IMMUNOLOGY AND MOVE TO NEGLEC NEGLECTED. >> WHILE I WAS THERE I WENT TO HARVARD WHERE ALAN AND ANTHONY BUTTERWORTH WERE SHARING A CUBICAL AND AND I MET HIM AND I WAS ABLE TO PARLAY THAT INTERACTION AND INTO A AFTER THE FIRST YEARS OF MEDICAL SCHOOL AND KENYA AND IT WAS THAT EXPERIENCE THAT FIELD WORK AND DISEASE AND WANTED TO DO AND SO FOR THAT, INDIRECTLY, ALAN REALLY WAS ALREADY AN INFLUENCE AND FOR THE PROGRAM AND AND IN 1982 AND PRIVILEGE TO WORK ALONGSIDE AND AND CAREER AND WE WERE COMBATING ARMS AND FIGHTING OFF THE ONSLAUGHT AND OF MALARIA AND MAKING BEGINNING FOR AND RESEARCH AND FOR THE PAST 20 SO YEARS AS SORT OF A PARTNER IN AND HELPING TO OVERSEE THE OPPORTUNITY AND I VALUE EVERY MINUTE AND ALAN WAS AND SO THANK YOU. [APPLAUSE] WE'RE GOING TO THE PROGRAM AND IT'S ED PEARCE, IS THAT -- >> THANK YOU FOR GIVING ME AN OPPORTUNITY TO SAY A FEW WORDS ABOUT ALAN. I THINK AS YOU GET OLDER, I HAVE REALIZED THE IMPORTANCE THESE EVENTS AND SUE GET TO SAY THINGS THAT YOU MIGHT NOT SAY TO OTHER PEOPLE AND YOU SAY THEM PUBLICLY AND I DON'T KNOW WHY THAT'S EASIER BUT PERHAPS IT IS. ANYWAY, I'M -- I WAS GOING TO SAY EXACTLY WHAT TOM SAID ABOUT ALAN BEING ONE OF THROW THREE MUSKETEERS WHO WERE BASIC IMMUNOLOGISTS WHO TURNED TO WORKING -- WHO DECIDED IDEALISTICKLY TO WORK ON NEGLECTED TROPICAL DISEASES AND THAT USE MADE AN IMPRESSION ON ME AND I GOT TO MEAT PAUL AND DONATO THE OTHER TWO GUYS THAT WERE ALAN'S FRIENDS AND THEY MADE AN IMPRESSION ON ME AS WELL. SO, THE THING THAT ALAN LOVES SCIENCE AND HE IS INTERESTED IN EVERY SCIENTIFIC QUESTION AND ONE OF THE FANTASTIC THINGS ABOUT ALAN IS HIS ENTHUSIASM AND AS A TRAINEE IN HIS LAB, YOU COME UP WITH AN IDEA AND HE WOULD ALWAYS PUT A POSITIVE SPIN ON IT AND TRY AND MAKE IT GO FORWARD AND SEE THE GOOD SIDE OF IT AND YOU KNOW, BE SUPPORTIVE AS YOU TRIED TO MAKE SOMETHING COME OF THAT IDEA THAT YOU'VE HAD AND THAT WAS AND INVALUABLE AND SUCCEED AS WELL AS ALAN DID AND THAT IS SOMETHING THAT WAS REALLY AND WHEN I WAS IN HIS LAB AND IT'S SOMETHING THAT I REALLY APPRECIATE. SO A POINT THAT WAS MADE, MAYBE IT WAS IT'S SOMETHING WE'LL ALL AWARE OF HOW PROLIFIC AND IT'S PUBLISHED AND OVER 500 PAPERS AND IT'S A TESTAMENT TO HIS WORK OATH I CAN AND ALAN WORKS REALLY, REALLY HARD AND IT'S SOMETHING I RECOGNIZED AS WELL AND YOU KNOW THAT IS AN AMAZING ROLE MODEL IN THAT RECORD. SO, WHEN I THINK ABOUT MY TIME IN ALAN'S LAB IT CERTAINLY IS THE TIME WHEN I LEARNED TO BE A SCIENTIST. ALAN TAUGHT ME HOW TO BE A SCIENTIST AND THROUGH HIS ENTHUSIASM AND THROUGH TAKING THE TIME TO SIT DOWN AND TALK TO ME AND THE EXPERIMENTS I WAS DOING AND HOW I SHOULD INTERPRET THE DATA AND WHAT WE SHOULD SEE IN IT. HE TAUGHT ME HOW TO WRITE. THE STORY THAT EVERYBODY HAS HAD ABOUT BEING IN THE HOTSEAT, I WAS IN THE HOTSEAT MANY, MANY TIMES AND IT'S AMAZING WE WERE DOING IT IN A LONGHAND ON A YELLOW PAD AND HE WOULD WE REWRITE IT AND IT'S HARD TO THINK THAT'S THE WAY WE DID IT. THAT'S HOW I LEARNED TO WRITE AND IT'S INVALUABLE. AS A SCIENTIST, IF YOU CAN'T WRITE YOU ARE DONE. SO, HE NOT ONLY TAUGHT ME HOW TO BE A SCIENTIST AND HOW TO WRITE BUT HE TAUGHT ME OTHER THING. HE TOOK ME SAILING ONCE AND TRIED TO TEACH ME HOW TO SAIL. >> THAT SPEAKS TO ALAN'S CONNECTIONS AND HE IS CONNECTED TO SO MANY PEOPLE AND OBVIOUSLY THE EMPHASIS TODAY IS ON IMMUNOLOGISTS AND HE IS WIDELY RESPECTED AND CONNECTED IN THE IMMUNOLOGY FIELD AND HE IS AND HE IS CONNECTED TO HARDCORE PATHTOLOGISTS AND THAT'S ANOTHER THING AND HE IS MEETING THESE FANTASTIC WHO COME THROUGH AND VISIT AND ALL SORTS OF PEOPLE WERE COMING THROUGH AND VISITING IN THE LAB ALL THE TIME AND HE WOULD GO OUT OF HIS TO INTRODUCE US AND I GOT TO KNOW PAUL ENGLAND AND JOHN HOPKINS WHO WAS ONE OF THE PEOPLE THAT WORKED OUT HOW GPI ANCHORS ARE CONSTRUCTED AND INSIDE A CELL AND I GOT TO MEET A OCC WANG FROM UCSF WHO DISCOVE INEVER MEC TIN AND ALAN IS THE CENTER OF THIS SCIENTIST OF PEOPLE THAT I'M A PART OF AND THIS IS FANTASTIC BECAUSE IT OPENS THE DOOR TO US ALL GOING TO EACH OTHER AND I FEEL LIKE AND THAT'S BECAUSE OF ALAN AND THAT AND THE WAY HE CAN BEHAVE AS A CHEERLEADER AND TO BE HONEST WITH YOU, IT'S PRETTY INTOXICATING WHEN HE IS WORKING WITH YOU TO GET THAT PAPER TOGETHER AND AND IT WAS BETTER WHEN HE WAS THINKING ABOUT MY WORLD TO WORK HARD AND HE DID THAT IN A GREAT WAY AND AND THE WAY HE TALKED ABOUT MICHAEL AND MICHAEL GOOD WAS MANY OF YOU MAY KNOW AND HE ENDED UP AND BEING CURE IMR AND HE REALLY IS A GOOD SCIENTIST AND WHEN ALAN WERE TALK TO US ABOUT MICHAEL GOOD HE WAS A PARADIGM OF VIRTUE AND THIS IS HOW YOU GET AND MICHAEL. GOOD. MICHAEL EXCELLENCE AND SO I JUST THINK ABOUT IT AND I NEVER THINK OF MICHAEL GOOD WITHOUT THINKING ABOUT HIM AS MICHAEL EXCELLENT. THANK YOU, ALAN, IT'S BEEN MY PRIVILEGE TO BE HERE TODAY. THANK YOU. AND SO MOVING ON FROM THERE, I HAVE THE PRIVILEGE ALSO OF INTRODUCE PAUL BRINDLEY, MY OLD COM PADRES FROM ALAN'S LAB AND WE SPENT A LOT OF TIME TOGETHER AND I'M LOOKING FORWARD TO HEARING WHAT YOU HAVE TO SAY, PAUL. [APPLAUSE] >> THANK YOU, ED. AND THANK YOU TO DRAGANA AND THE OTHER FOLKS WHO INVITED ME INTO PUTTING ON THIS WONDERFUL SYMPOSIUM. OK. SO, DRAGANA ASKED ME OR I SAID I COULD TALK ABOUT SOMETHING TO DO WITH GENOMES AND RELATED FLUKES SO I WANT TO MAKE A FEW SIMILAR REMARKS TO THAT PROFESSOR PEARCE AND OTHERS WHO HAVE MADE TODAY BECAUSE I ALSO WAS A POSTDOC IN ALAN'S LAB AND SO I'M GOING TO TALK ABOUT THE SCIENTIFIC ASPECTS A LITTLE BIT FURTHER ON AND I'M GOING TO CONSUME TOO MUCH TIME AND I'VE BEEN OFFERED LESS BUT I WANT TO MAKE A COUPLE OF PERSONAL REMARKS. SO, THAT IS THE INTERVAL WHEN I WAS IN ALAN'S LAB IN 1985 TO 1991 AND WHERE AS ALAN MIGHT HAVE KNOWN Dr. MICHAEL EXCELLENT HE DIDN'T KNOW ME AND EVEN THOUGH MICHAEL AND I KNEW EACH OTHER AND WE WERE BOTH FROM AUSTRALIA I WANT TO TELL YOU HOW I CAME TO A ARRIVE HERE SO, I JUST FINISHED MY PH.D DEGREE AT AN INSTITUTE ALAN NEVER HEARD OF AND THE DOCTOR GOOD WAS ALSO A GRADUATE AND SO I WROTE A LETTER AND PUT IT A LETTER IN A BOTTLE THAT I TOSSED INTO THE PACIFIC OCEAN AND I FORGOT ABOUT IT AND I HAD WRITTEN THIS LETTER TO THE DIRECTOR OF NIAID WHICH I TAKE IT OUT WAS PART OF THE NIH AND SOMETIMES LATER, I RECEIVED A LETTER BACK IN THE MAIL AND AND I SAVED AND HOW LONG AND AND I OPENED IT UP AND IT JUST ALLUDE TODAY AND AND SEVERAL PAGES OF THE CURSE TOMORROW EXPLAINED WHO HE WAS AND THAT MY LETTER HAD BEEN RECEIVED BY THE NIAID AND IT MOVED Dr. GALAN'S OFFICE AND HE TALKED TO Dr. FRANKLIN THE CHIEF OF THE LPD AND SOMEHOW Dr. NEVER DECIDED THAT I MIGHT BE A PERSON WHO COULD HELP HIM OUT BECAUSE THIS GUY BRINDLEY CLAIMED HE KNEW SOMETHING ABOUT WORMS IN HIS DOCTORATE AND Dr. NEVER HAD AN INTEREST IN WORMS BUT HE TALKED TO ALLEN AND SAID, YOU'VE BEEN BADGERING ME FOR A POSTDOC BUDGET SALARY AND HE SAID I'VE GOT AN INTEREST IN THIS LETTER THAT Dr. GALLON HAS PASSED ON TO ME AND WE BETTER DO SOMETHING ABOUT IT BUT I WILL TAKE HIM ON IF YOU WILL TAKE HIM ON TOO. I THINK IT'S HOW THE STORY WENT AND SO ALAN EXPLAINED TO ME IN THIS LETTER ON THE YELLOW NOTE PAD THAT YES, THAT I'M INTERESTED IN HIRING ME BUT I WOULD HAVE TO WORK PRETTY HARD, 12 HOURS FOR Dr. NEVER EACH DAY AND 12 HOURS FOR Dr. SHER. THE UP REPO SHORT OF THAT, SO A LITTLE WHILE LATER, AFTER IF I SAT IN THE HOTSEAT WITH ALAN WE MANAGED TO PUBLISH THIS PAPER AND THIS PAPER DEALT WITH EXPOSURE OF MICE TO THE ANTI- PARASITIC DRUG AND OTHERWISE MY SURFACE THAT Dr. HALL BY OTHER ANTIGENS WERE EXPOSED TO SERUM ANTIBODIES AND SOON THE MICE WERE EXPELLED SO THE LINKAGE WAS THERE THAT EFFICACY OF THIS DRUG HAD AN IMMUNE DEPENDEN SEE COMPONENT AND THE OTHER THING I WOULD LIKE TO SAY ABOUT THIS PAPER AS PROUD OF IT I WAS, THE DAY THE LETTER FROM THE JOURNAL OF IMMUNOLOGY SAID IT WAS ACCEPTED WE WERE FEELING CHUFFED AND ANOTHER LETTER APPEARED ON MY DESK AND THIS LETTER EXPLAINED THAT THEY MADE A TERRIBLE MISTAKE. AND THAT THERE WAS A CLERICAL ERROR AND ACTUALLY THE PAPER HADN'T BEEN ACCEPTED AND THEY WERE WITH DRUG THEIR OFFER OF ACCEPTANCE AND YOU WOULD HAVE TO TROY SOMEWHERE ELSE BUT Dr. CHIEFER WAS RESPONSIBLE FOR THAT. THAT WAS MY THE LAB IN IMMUNO BIOLOGY AND IMMUNE SERUM AND ALAN PROBABLY PRESENTLY DECIDED THAT THIS GUY BRIND LIKE UNLIKE Dr. EXCELLENT, KNEW NOTHING ABOUT IMMUNOLOGY AND THAT'S THE LAST I WORKED ON ANYTHING IMMUNE LOGICAL SO ALAN SUGGESTED I WORK ON THE WORM THEMSELVES OR THEIR GENOMES, I'M GOING TO SHOW THE WORK I DID TO ALAN'S ADVICE TO STICK TO THE PARASITE. SOME FOLKS MENTION Dr. GEORGE YAP INDICATED THAT IF YOU HAD A FORK IN THE ROAD AND Dr. SHER SAID YOU CAN WORK ON TOXO YOU WOULD SOON BELIEVE YOUR CAREER WAS DEAD ENDED IF YOU WORK ON SISTER SO YOU BETTER WORK ON TOXOPLASMOSIS AND HE DIDN'T OFFER ME THAT ADVICE, I DIDN'T GET THE OFFER SEW SAID YOU WORK ON SHISTER SEEMS. SO ANYWAY, THE PROBLEM WITH S THEY'RE GENETICALLY NOT VERY ATTRACT ABLE AND THEY'RE WORK TO WORK ON AND THEY'RE UNLIKE SINGLE CELL PATHOGENS THEY HAVE A COMPLEX GENOME AND THEY SEX I'LLLY REPRODUCE AND THEY HAVE A COMPLEX LIFE CYCLE AND SOME OF THEM ARE MORE COMPLEX AND THEY HAVE A SMELL INVOLVED. SO IN SUBSEQUENT YEARS, YOU KNOW, IN A PARTICULAR SINCE THE CRISPR REVOLUTION AND THE NOBEL PRIZE TO RELATED TO THAT, PEOPLE NOW HAVE ADAPTED CRISPR GENE NOAM ETIQUETTING AND WE'VE BEEN INVOLVED IN THAT OURSELVES AND I'LL MENTION THAT AND WHAT WE REALLY WOULD LIKE TO DO IS IT'S A CHARGE BUT IT'S WHAT WE DO IN SOME RELATED WORMS NOWADAYS. TO GET INTO CRISPR EDITING, A COUPLE OF YEARS A. I CAME BECOME TO SEE ALAN AND DRAGANA JUST BEFORE THE COVID AND I WAS A WAIVER THEIR PAPER FROM THE MIDDLE, WELL ABOUT 2009 AND I QUICK WILL HE CHECKED PUBMED TODAY AND AMONG THOSE 500 PAPERS THAT ALAN PUBLISHED THIS WAS THE LAST ONE THAT HAD THE NAME OF SHER AND SCHISTOME IN THE SEARCH THAT I PICKED UP. I WAS AWARE THAT THE EGG OF THE SCHISTOSOME WAS RESPONSIBLE FOR THE DISEASE AND SO WE DECIDED IN MY LAB, PARTICULARLY WITH HOTSHOT YOUNG ASSISTANT PROFESSOR WHO WORKED WITH ME THAT WE WOULD EDIT SOMETHING AND GET A PHENOTYPE AND WE KNEW THE EGGS HAD A PHENOTYPE OF THE GRANULOMA AND THE TH2 SHIFT AND SO WHAT COULD WE EDIT IN A EGG AND THERE WAS THIS PEOPLE AND THEY PUBLISHED SIMILAR PAPERS THAT THE EGG SECRETS A OMEGA 1 PROBABLY NAMED DISCOVERED LONG BEFORE BY DAVID DUNN AS WELL. THE OMEGA STANDARD FOR THE FRACTION FROM THE CROW MATGRAM SO WE'RE ABLE TO LOOK AT THE GENOME. THE GENOME IS IN GOOD SHAPE NOWADAYS BUT NOT TOTALLY COMPLETE. UNFORTUNATELY THIS PARTICULAR ENZYME HAS THREE GENES BUT THEY'RE ALL SIMILAR AND I WANT TO POINT OUT WHAT IS SOMETHING ON THE BOTTOM OF THE PANEL THERE. DON'T WORRY ABOUT THE GENE STRUCTURE. THAT PARTICULAR ENZYME HAS A VERY SPECIFIC DEVELOPMENTAL EXPRESSION AND IT'S ONLY EXPRESSED IN THE EXPRESSION LEVELS OF THE TRANSCRIPT AT VARIOUS STAGES OF THE LIFE CYCLE OF THE THREE DIFFERENT GENES. ONLY SEEN WITHIN THE EGG AND THE EGG PROBABLY TAKES SEVEN DAYS TO DEVELOP WHEN THE FEMALE LAYS IT UNTIL IT COULD REACH THE BELL AND ONLY EXPRESSED? THE LAST THREE OUR FOUR DAYS OF THAT SO IN THE MATURE EGG SO WE DECIDED TO TRY AND EDIT THAT THIS THE UP SHOT OF WHAT WE SAW THAT ON THE LEFT HAND PANEL WE CAN GET A REDUCTION THAT WE CAN MEASURE IF WE TOOK RNA FROM THE EGGS AND WE DELIVERED IT AND WE ALSO CAN DELIVER BY THE VIRUS AND IT'S INFECTIOUS THOUGH OR OTHER RETRO AND SO AND I DON'T WANT TO SHOW YOU THE EDITING PAD ENTER AND A SMALL PERCENTAGE OF THE GENOMES SO WE TAKE 10,000 EGGS AND EDIT THEM AND RECOVER THE FEW DAYS AND, I WANT TO TALK ABOUT JUST AS I FINISH UP ON THIS RUN RIGHT NOW. BUT THOSE MANIPULATED EGGS IF WE PUT THEM INTO LUNG GRANULOMA WHEN YOU LOOK AT THESE 10 DAYS LATER IN THE MOUSE, THAT IS THE VOLUME OF THE GRANULOMA SURROUNDING THE EGG. THE EGG IS RIGHT THERE. THE GENOME EGGS IS JUST GOBSMACKEDDINGLY ABSENT AND THE SIZE OF THE GRANULOMA AND ROB THOMPSON WHO WORKED FOR TOM WYNN HELPED US SCAN THOSE AND HE LET US USE BUT THIS IS JUST THE VOLUME AND THE AMOUNT OF VOLUME TO AND THAT IS THE EDITING CAN DO ON THIS PARTICULAR ENZYME AND IT WAS CURIOUS WHEN WE SAW THAT BECAUSE WE GOT THIS 100% REDUCTION IN TRANSCRIPT LEVEL AND AT LEAST 50% REDUCTION IN THE RNA ACTIVITY AND AT THE GENOME LEVEL A SMALL PERCENTAGE WERE EDITED AND WHY IS THAT BECAUSE THIS IS THE STAGE, THE EGG STAGE IS WHAT WE EDITED AND THIS IS NOT OUR WORK THIS IS FROM THE LITERATURE AND THE EGG ITSELF WHEN IT'S DEVELOPED AND YOU CAN SEE THE SILLIA ON THE OUTSIDE OF THIS WITHIN THE EGG SHELL IN THE MOUSE LIVER, THIS EXAMPLE. AND THERE'S ACTUALLY SOME TISSUE JUST INSIDE THE INNER COAT OF THE EGG AND IN AN ENVELOPE WHICH IS ANOTHER THING WHEN TISSUE GROWS OUT AS THE EGG DEVELOPS DD IT'S THE ONLY PLACE IN THIS ORGANISM WHERE OMEGA IS EXPRESSED AND WHEN WE INTRODUCE THOSE EGGS IN CRISPR IN THOSE EGGS CLEARLY JUST PHYSICAL LIE WE ONLY REACH THAT TISSUE SO VERY WE KNOCKED OUT THIS VACCINES OF PHENOTYPE AND WE CHOSE AN INTEREST GENE TO SO IT HAPPENED THERE. I MENTIONED THAT THE SISTER SEEMS ARE DIFFERENTLY AND TO MANIPULATE SO YOU CAN WELL MARRIAGE OTHER PEOPLE ARE INTERESTED IN THIS GERM LINE EDITING AND THAT WAS AN EXAMPLE OF TISSUE EDITING DONE NOW FOR THE TREATMENT OF HUMAN HEMOGLOBIN FOR INSTANCE. AND WE WOULD LIKE TO GERM LINE AND CHANGE THE ORGANISM AND TURN IT BLUE OR GROWN TO MAKE THE COVER EVER A MAGAZINE OR SOMETHING BUT TO GET IT TO GERM LINE AND THE GERM LINE IS A VALUABLE IN A NUMBER OF WINDOWS IN THE DEVELOPMENTAL CYCLE BUT IT'S AVAILABLE AS SOON AS THE EGGS LAY BECAUSE THEY'RE THAT IS WHAT HAPPENS IN THE LIVER WHERE THEY'RE TRAPPED SO WE WANTED TO LAY CRISPR MATER MATERIAL AND TS OUR PLAN SO WE DECIDE THAT BEFORE CRISPR BY VIRAL INFECTION OF EGGS AND MAKE GERM LINE WORMS AND WE PUBLISHED THIS A FEW YEARS AGO AND I WENT GO THROUGH THE PATENT BUT THESE THE GENE HERE WE CAN FIND IT IN THE F-1 AND THE DIFFICULTY IS THAT THAT IS A SINGLE SEX. IT'S HOW THE MALE OR FEMALE AND THAT'S WHAT WE GOT AND WE MADE A NUMBER OF AND WE CAN TRANSIT AND GOWN THE GENERATIONS AND AND THE REPORTED SOON WAS SOONLY ACTIVATED WHEN USED RETRO VIRUS AND THIS INVOLVED AND THEY'RE OFTEN DEPENDING ON THE LAND AND NONE THE LESS I WANT TO TRY TO TELL YOU THAT WE CAN MAKE AND WE CROSS THEM AND IT WILL BE IN CONTAIN THE TRANCE GENE AT THIS LOCUS AND THEN IT WILL BE PASSED DOWN THE LINE AND IN VERY RECENT TIMES THESE DATA ARE UNPUBLISHED AND WE ACTUALLY IDENTIFIED WHAT WE CONSIDERED GENOME SAFE HARBORS IN THE GENOME SAYING TO THE ROSE 26 SIDE AND OTHERS THAT ARE USED IN HUMAN GENE THERAPY OR MOUSE MODELS OF HUMAN GENE THERAPY AND SO WITH A NUMBER OF MANIPULATIONS AND GRUNT WE CAN DELIVER CRISPR REPORTED TRANCE AGAINST WE WANT TO PUNE NA TIVE SAFE HARBOR SO IF WE LOCK TO THE EGG IT WAS FLORES ANTS WE CAN GO 70% OF THE EGGS WILL BE DARPHA CONTACTED US AND CALLED ABOUT THE BIO SYSTEM AND I WON'T GO INTO THIS AND I WILL PROBABLY BE BREAKING SOME CONFIDENCE WITHIN AND I'LL BE ARRESTED BUT, THEY'RE INTERESTED IN CHANGING THE MICROBE OF SOLDIERS AND MEDICAL FIRST RESPONDERS BY A NUMBER OF WAYS THEY SAID COULD A MACRO BUY OT I CAN ORGANISM LIKE A SCHISTOSOMES OR A HOOK WORM BE SOLVED AND AND IT'S MUCH MORE GENETICALLY ATTRACT ABLE. AND IT'S DIFFICULT PROJECTS AND SOUTHEAST ASIA IN THE RIVER DELTA AND THEY'RE A CURSE AND. >> THE FISH THE LARVA AND AND ARRIVES IN THE UPPER SMALL INTESTINE AND LIVES IN THE TRACT AND IT'S A HUMAN AND RELATIVELY RESEARCH IN MEDICAL SCHOOL IN NORTHEAST THAILAND AND YOU CAN SEE A COUPLE OF THE FLUKES ARE JUST POPPED OUT AND SEVERED WILD AND THIS TISSUE WAS SECTION BY THE PATHOLOGIST IN THE LAB OF A COLLEAGUE OF MINE AND THE TUMOR THE SURGEON WAS ADDRESS WHAT IS THIS SECTION IS UP IN THE TOP CORNER AND IT'S A DIRE DISMAL PROGNOSIS AND MOST OF THE PEOPLE WOULD BE AND DEAD AFTER THE DIAGNOSIS. THE WORMS LIVE IN THE BILE DUCT AND LONG LIST LIKE SCHIS. >> Toni-Marie: AND THEY EAT IN THE BIO DUCT IN THIS TISSUE HERE AND THIS IS EXPERIMENT FROM OUR GROUP WHERE WE JUST CULTURE HUMAN CHOLANGIOCYTES AND YOU CAN SEE THE VEHICLES ARE UPTAKEN BY THE CHOLANGIOCYTES AND WE GEE OWN EDIT THIS FLUTE AND AFTER WE MA LIP NATURED THE LAVA INFECTING THE HAMSTERS AND YOU LOCK AT EXPERIMENTALLY INFECTED WITH A FLU AND RIGHT THERE THIS ALREADY HYPO PLASIC LINING IS SEVERAL CELLS THICK IN NATURE IT'S ONE CELL THICK HOWEVER, THE PLACE WE HAD THIS GENOME IN CODES OF GROWTH FACTOR KNOWN AS GRAN YOU LYNN AND IN THE ABSENCE OF GRANYOU, LYNN THERE'S NO PROLIFERATION OF THE TRACT AND THAT IS THE CELL THAT IS SELL OF ORIGIN OF THIS LIVER CANCER AND WE HAD BEEN PLAYING AROUND WITH A GROWTH FACTOR AND I DON'T HAVE TIME TO TELL YOU ABOUT IT. WE COULD GET RID OF SOME OF THE DISEASE AND SO, CHANGE TRACK A LITTLE BIT, THIS IS NOT MY WORK BUT WORK BY CONCORD UNIVERSITY. THE GEE OWN SEQUENCED CARCINOMA FROM HUMANS AND IN THAT PART OF THE WORLD SO LIVER FLUKE IT'S A GREAT ON CARCINOGEN AND WHEN THEY PROFILED THE GENETIC CHANGES, ON THE FEWTATION THAT'S PROGRESSIVE IN VERSUS NON LIVER FLUKE AND IT MIGHT TURN BORING I WANT TO POINT OUT THAT TP5 3 IS SEEN AND THIS IS A CHRONIC INFECTION SO WE'RE IN A GENERAL WAY THIS IS A CHRONIC INFECTION ASSOCIATED MALIGNANCY AND SO BACK TO MY STORY. SO WOAD LIKE TO MODEL THAT. IF WE INFECT A HAMSTER IN THE LAB AND THESE WORMS ARE WILD ANIMALS AND THESE ARE NOT MAINTAINED IN THE LAB. THE COLLEAGUES HAVE TO GO TO THE MARKET OR HAVE SOMEONE FISH TO FIND THESE SO WE ONLY GET THE WORMS FROM THE WILD. AND THEN YOU INFECT THE LAB RODENTS OR WHATEVER HAMSTER AND YOU DO AROUND EXPERIMENT IN THE LIFE OF A HAMSTER IN THE LABS A YEAR OR SO, THE TUMOR DOESN'T DEVELOP AND IT DOESN'T DEVELOP UNTIL THE MID DECADES OF LIFE IN PEOPLE BUT IT WAS WELL-KNOWN FOR 30 OR 40 YEARS, PARTICULARLY FROM PEOPLE AT THE UNIVERSITY IN BANGKOK, THAT THEY COULD INDUCE A MODEL OF COL ANGIE OWE C CARCA BY ADDING IT RELEVANT BECAUSE THE DIET OF THE PEOPLE IN NORTHEAST THAILAND IS RICH CAN YOU INDUCE IN A FEW MONTHS SO DURING COVID WE TRIED TO DID THAT EXPERIMENT AND WE HAD TO GUESS WHICH OF THESE LEVELS SO WE ACTUALLY DIALED RIGHT UP AND WE USED 100 NUCKS AND 12.5 PARTS PER MILLION AND WE INFECTED A NUMBER OF HAMSTERS AND VERY BRIEFLY TO FINISH UP, WHEN WE DID THAT, AND I ONLY JUST PLEASE LOOK AT THE TWO LEFT HAND BARS THERE, THE PUNCH LINE IS THIS AND THIS IS UNDER REVIEW AT PRESENT PEER ROW VIEW BUT IT'S IN YOU WANT TO LOOK AT IT AND ALL THE HAMSTERS GOT CARCINOMA BUT THERE WAS SIGNIFICANTLY LESS DISEASE AND LESS MARKET DISEASE WHEN WE KNOCKED OUT GRANULE LAR AND INFECTED LIVER FLOCKS SO FL. IT'S OVER LEVEL FLEW BUT WE LOOKED AT MANY OF THE KIND OF HALLMARKS OF CANCER THINGS, CANCER EXPERTS LOOK AT INCLUDING PROLIFERATION AND THERE'S A LOT MORE PROLIFERATION OF THE SITES AND IN THE CONTROLS VERSUS THE ONES THAT WHERE IT'S KNOCKED OUT OR KNOCKED DOWN AND I THINK LET ME SEE, YEAH, AND P53 THIS IS LESS OF MUTANT P53 THAN IN THE CONTROLS SO, I THINK I'M FINISHING UP NOW IS CRISPR EDITING IS ACTION TIVE IN LIVER FLOCKS AS SOME OF THE THINGS YOU CAN DO WITH IT SUCH AS REPAIR AND IN TERMS OF THE TWO EXAMPLES OF SHOWED YOU, WE IN SCHISTOSOMES WE KNOCKED OUT THE MEGA RESPONSIBLE FOR THE TH2 AS AN EXAMPLE AND WE'RE TRYING TO DERIVE A LINE BUT IT MAY BE A LETHAL MUTATION AND I SHOWED YOU THAT WE CAN GET SIMILAR THINGS TO WORK IN LIVER FLUKES SO WE THINK FUNCTIONAL GENOMICS IS ACCESSIBLE FOR PARA SID I CAN ELEMENTS FOR THE MAJOR AND HOPEY IT'S LOWER. THESE ARE THE FOLKS I'VE COLLABORATED. JUST TOO MANY PEOPLE TO GO INTO. I WILL MENTION MY COLLEAGUES IN CON CANE UNIVERSITY AND JAMES COOK UNIVERSITY. I WANT TO FINISH UP WITH THIS SLIDE BECAUSE BLACK AND WHITE, ALL WE COULD AFFORD IN THE PAPER WAS A BLACK AND WHITE PHOTOGRAPH. I NOW HAVE A COLOR CAMERA AND I STILL COME DOWNTOWN HERE AND TAKE ELLEN'S SAGE ADVICE ABOUT OMEGA AND OTHER THINGS AND IT'S USUALLY IN AN IRISH BAR SO THANK YOU, VERY MUCH, ALAN. [APPLAUSE] [OFF MIC] WE KNOCKED OUT A SECOND GENE ENCODING A BAN ON THE WORMS DIDN'T SURVIVE. THE GRANULAR WORMS ARE RECOVERABLE AT THE END OF THE SIX MONTH EXPERIMENT AND WE DON'T THINK THERE'S FIT BUT IT'S TRACT ABLE TO DO THAT KIND OF MALIGNANCY ASSOCIATED RODENT EXPERIMENT. WE CAN LOOK AT MARKERS BUT DEFINITELY FIT ENOUGH TO DO THIS KIND OF EXPERIMENT. WHAT I WOULD SAY ABOUT LIVER FLUKE THOUGH, WE DON'T HAVE ANY PRE TENSION THAT'S WE COULD MAKE A GERM LINE TRANSGENIC ANY TIME SOON BUT WE CAN DEFINITELY DO THAT WITH (INAUDIBLE) NOW. [OFF MIC] >> OH, BECAUSE THE -- NO, THE BLOCK IS, WELL, THE BLOCK IS HUMAN GET AROUND ABLE WITH MY POOR EXAMPLE OF ENGLISH. IF YOU DON'T HAVE OMEGA, YOU WON'T GET A GRANULOMA AND THE GRANULOMA IS NEEDED FOR THE TRANSIT ACROSS THE BALL WALL. WE PRESUME WE WOULD RECOVER EGGS FROM THE LIVER, THERE WOULDN'T BE A GRANULOMA AND THEY VERY WELL MAY HATCH. THANK YOU. >> THE NEXT SPEAKER IS Dr. P'NG LOKE. NIAID WAS LUCKY ENOUGH TO RECRUIT HIM RECENTLY AND WHO IS TALK TO TALK ABOUT HETEROGENEITY MACROPHAGES. >> WELL, FIRST OF ALL IT'S REALLY SUCH A PLEASURE AND HONOR TO BE SPEAKING AT ALAN AND IT IS DRAG AM A FOR PUTTING TOGETHER THIS AMAMAZING EVENT. LONG BIFF METAL AN I WAS READING HIS PAPER SO I AM ABOUT THE CELLS THAT WOULD HAVE MAJOR EFFECTS ON THE WAY THAT I THOUGHT ABOUT PARASITE IMMUNE RESPONSES SO AND I USE IT A LOT IN MY PRESENTATION AND IT WAS ALREADY THERE IN THIS ORIGINAL REVIEW ARTICLE. AND YOU KNOW, DURING THE COST OF THE LAST FEW DAYS, WE'VE HEARD A LOT ABOUT THE TCH AND HE PASSED AND TALKING ABOUT THE ASPECTS OF IMMUNE RESPONSE AND THE CONTRIBUTION AND I DON'T HAVE ALAN AND UNDERSTANDING AND AND LONG BEFORE HE WORKED ON THE TB GRANULOMA AND IT WAS A KEY COM PHONE AROUND OF THE PARK IN SOMEONE THEY GET STUCK IN THE LIVER AND CAUSE THESE GRANULOMA TO FORM AND IN DEED THE GRANULOMA IS THE PASSAGE FOR THE INTESTINE AND THIS CAME OUT YESTERDAY THAT I OFTEN THINK OF THIS AS BEING A GOOD MODEL FOR TUMOR METASTASIS AND TUMORS HAVE GRANULOMA STRUCTURE THAT MIGRATE THROUGH DIFFERENT TISSUE WALLS AND AS WE HEARD FROM BOB COFFMAN EARLIER THERE WAS A REMARKABLE PERIOD OF WORK FROM ED PEARCE AND TOM AND ALAN TO REALLY DESCRIBE THE FUNCTION OF AND IL4 AND IL5 AND IL-12 INTERFERON GAMMA AND THE ROLE OF ALL THESE CYTOKINES AND FUNCTION OF THESE CYTO TO INITIATE THIS ENTIRE CASCADE AND I THINK THE WORK THAT ALAN'S GROUP HAS DONE ON ALL THESE CYTOKINES ARE INCREASINGLY IMPORTANT IN OUR UNDERSTANDING OF THE ROLE OF THE TYPE 2 IMMUNE RESPONSE AND MAINTAINING TISSUE HOMEOSTASIS AND IT BECOMES MORE AND MORE CLEAR THAT OVER THE LAST, FROM WORK OVER THE LAST FEW YEARS THAT THIS TYPE 2 RESPONSE IS VERY IMPORTANT FOR THE PROCESSES OF WOUND REPAIR AND WOUND HEALING AND TISSUE REPAIR AND ALSO, ADD A POST TISSUE AND METABOLISM AND I USED TO BE IN THE CAMP THAT BELIEVED THE TYPE 2 IMMUNE RESPONSE EVOLVED TO PROTECT US FROM HELL MINUTE INFECTIONS TO MAINTAIN AND WAS CO OPTED BY THE IMMUNE SYSTEM AND THE SAME PROCESS OF WOUND HEALING AND TISSUE REPAIR AND VERY IMPORTANT ALSO IN RESISTANCE AND TOLERANCE MECHANISMS TO HELMINTH INFECTIONS SO FOR ME PERSONALLY AND I THINK MY INTEREST START TODAY OVER HAL THAT WAS DONE AND DEACTIVATED AND MAKE ROW PAGES AND THIS CONCEPT I THINK THAT AND AND THE TEEN SIGHT AND HE DIFFERENT PHONY TYPE AND ACTIVATION AND NOW WELL SEPTEMBER AND BY EVERYONE AND IT'S CLEAR THAT THE THESE ALTERNATIVE MACROPHAGES AND WITH HIGH CONCENTRATIONS O THEY'RE NT REPRESENTATIVE PERHAPS OF WHAT IS FOUND IN THE TISSUE AND IT'S CLEAR SEEK EXPERIMENTS AND MACROPHAGES IN THE TISSUE AND FALL MORE DIFFERENT ACTIVATIONS STATES OF WHICH AND THE OTHER IS THESE ARE ACTIVATION STATES SO MACROPHAGES ARE MUCH MORE PLASTIC CELL TYPES AND THE TRANSCRIPTION FACTORS ARE MORE SNAPS DEPEND AND LINEAGE DEPENDING AND THAT YOU CAN GET WITH T CELLS. MY INTEREST IN THESE ALTERNATIVELY ACTIVATED MACROPHAGES STARTED WHEN I WAS A PH.D STUDENT AND AND IT'S ANOTHER FAVORITE ORGANISM OF THE LPD AND ALTHOUGH I THINK TOM THOUGHT THESE EXPERIMENTS WHERE WE IMPLANTED THESE PARASITES INTO THE PERITONEAL CAVITY WAS NOT PHYSIOLOGICAL BUT NONE THE LESS, WHILE I WAS PH.D STUDENT IN JUDY ALLEN'S LAB, WE FOUND THAT THEY INDUCE ACCUMULATIONS AND THEY HAD A DISTINCT PHENOTYPE OF BEING ABLE TO SUPPRESS T-CELL PROLIFERATION AND EXPRESS THESE GENES THAT AT A TIME THE FUNCTION OF WHICH WAS VERY MUCH UNCHARACTERIZED AND HAS BECOME REALLY USEFUL MARKERS FORT PRESENCE OF IL-4 ACTIVATED MACROPHAGES OVER THE LAST FOUR YEARS AND WE KNOW FROM THE WORK OF TOM WYNN THAT JUDY ALLEN'S LAB HAS CONTINUED TO WORK ON THE CHITON ACE LIKE MOLECULES AND ANOTHER TRAINEE IN JUDY ALAN'S LAB HAS CONTINUED TO WORK ON ALL OF THESE MOLECULES PLAY VERY IMPORTANT ROLES IN IMMUNE REGULATION AND IMMUNE REGULATION IS A VERY IMPORTANT FUNCTION OF THESE ALTERNATIVE ACTIVATED MACROPHAGES AND NOW, I WENT AND DID MY POSTDOC TO BAL WORK IN JIM WALL SON'S LAB AT BERKELEY SO I SEARCHED AWAY AND STARTED TO WORK ON MOLECULES AND ONE OF THE THINGS I FOUND AT THE TIME WAS IF YOU PAR TENILLEAL MACROPHAGES IF YOU HAVE IL-4 OR TH-2 CELLS THEY WOULD UP REGULATE SPECIFICALLY THE PDL2 SO THE TH1 CELLS AND THE TH2 CELLS UP REGULATED PDL2 BUT THE THING THAT BOTHERED ME ABOUT THE WORK THAT WE DID AT THE TIME WAS NO HARD I LOOKED THEY WERE INDUCED NEVER UP REGULATED PDL2 AND THIS IS SOMETHING THAT REALLY BOTHERED ME AT A TIME AND DIDN'T MAKE IT INTO THE MANUSCRIPT OR THE PAPER, AND IT WAS REALLY ONLY MANY, MANY YEARS LATER THAT I UNDERSTAND WHY THIS WAS THE CASE AND IT COMES BACK TO SOME OF THE SUBJECT THAT FELIX WAS TALKING ABOUT THAT WE KNOW NOW THAT TISSUE MACROPHAGES COMES FROM DIFFERENT ORIGINS AND YOU HAVE INFLAMMATORY MACROPHAGES THAT ARE DERIVED FROM MONOCYTES AND YOU ALSO HAVE TISSUE RESIDENT MACROPHAGES DERIVED FROM EMBRYONIC PRE CURSERS THAT CAN PROLIFERATE AND SELF-RENEW WITHIN THE TISSUES AND IT TURNS OUT THAT DEPENDING ON THE TYPE OF PARASITE, HELMET PARASITE YOU ARE LOOKING AT YOU CAN GET ACCUMULATION OF ALTERNATIVELY ACTIVATED MACROPHAGES THAT COME FROM DIFFERENT ORIGINS AND SO I HAD THIS FUN VISIT TO EDINBURGH AIR STARTED MY LAB AT NY YOU AND JUDY AND RICK INVITED ME BACK TO GIVE A SEMINAR AND I WAS CHATTING WITH JUDY AND I WAS SAYING, OH, I KNOW WHERE THESE ALTERNATIVE MACRO RATE ISED COME FROM MAN OWE SIGHTS AND JUDY ALAN WAS LIKE NO, NO, NO, THESE COME FROM PROLIFERATING TISSUE AND MACROPHAGES AND WHAT IT TURNED OUT AND SO WE AND ALONGSIDE, WE KNOW THAT IT INDUCES THE ACCUMULATION OF TISSUE RESIDENT ALTERNATIVELY ACTIVATED MACROPHAGES THAT ARE DERIVED FROM FETAL PRE CURSERS WHEREAS, IN OUR LAB, AT THE TIME, WE WERE STUDYING THE SHIR SOMA AND THE MACROPHAGES YOU GET AROUND THE ARE DERIVED FROM MONOCYTES AND PHENOTYPICALLY THEY CAN ALTERNATIVE MACROPHAGES DO NOT EXPRESS AND THEY CANNOT UP REGULATE PDL2 AND THEY EXPRESS HIGH LEVELS OF F-4 08 AND UNCOUPLING PROTEIN UCP1 AND THE FUNCTION OF WHICH WE STILL DON'T FULLY UNDERSTAND AND SO THIS ENDS UP BEING A VERY NICE MODEL BECAUSE DEPENDING ON WHICH LINEAGE OF MACROPHAGES YOU ARE INTERESTED IN STUDYING, YOU CAN USE DIFFERENT HELMET PARASITES FOR THE DEVELOPMENTAL BIOLOGY AND ONE OF THE REASONS I HAD READ UP ON ALL OF THE WORK IN AND THE CELLS WERE INVITE RALLYE GRANULOMA AND THE PRODUCTION OF IL-4 AND IT ALSO BECAME CLEAR TE IL-4 WERE ACTING ON THE MACROPHAGES IN ORDER TO HAVE THE PROPER GRANULOMA AND SO THEY WERE ESSENTIAL FOR HOST SURVIVAL AND WERE INCORPORATED INTO THESE GRANULOMA INDUCED BY THE TYPE 2 CYTOKINE. WHEN WE STARTED THIS EXPERIMENT, THERE WAS A QUESTION OF WHETHER THESE MACROPHAGES WERE DERIVED FROM LY60 HIGH OR HIGH MONOCYTES AND IT WAS CLEAR FROM A POSTDOC IN MY LAB, ESPECIALLY TRUE MONOCYTE TRANSFER EXPERIMENTS IT WAS THE LY60 HIGH MONOCYTES THAT WERE THE SOURCE OF THESE GRANULOMA MACROPHAGES AND THIS WAS AROUND THE SAME TIME I WORK FROM ED PEARCE'S LAB AND ALSO PAUL CUBES' LAB AND IT ALL POINTED TO THE DIRECTION OF THE LY6C HIGH MONOCYTES AS BEING THE PRIMARY SOURCE OF THE ALTERNATIVELY ACTIVATED MACROPHAGES AND WHAT WAS INTERESTING THOUGH IS THAT AS THE LY60 HIGH MONOCYTES ENCOUNTER THIS INFLAMED TISSUE THEY WOULD DOWN REGULATE EXPRESSION OF AND ADOPT PET TROLLING BEHAVIOR THAT WAS REMINISCENT OF THE LOW MONOCYTE POPULATION BEFORE UP REGULATING EXPRESSIONS OF PDL2 AND WE HYPOTHESIZED THIS IS A IMPORTANT PART OF ROW MODELING PROCESS TO REMOVE THE EGGS OUT OF THE BLOOD VESSELS. OF COURSE, BEING IMMUNOLOGY, THINGS GET MORE AND MORE COMPLICATED AND ONE OF THE THINGS THAT TASHA IN THE LAB AT THE TIME WAS THINKING AND WHAT TASHA REALIZED WAS THAT THIS INFLAMMATORY MACROPHAGES, WHEN THEY SPENT ENOUGH TIME IN THE TISSUES, BEGIN TO ADOPT THE PROPERTIES OF THE TISSUE RESIDENT MACROPHAGES AND IN THE GRANULOMA, ONE COULD SEE THAT AS THE GRANULOMA MATURE, THE MACROPHAGES AROUND THE FOR FRINS ADOPT PROPERTIES OF THE TISSUE RESIDENT AND AND AT THE SAME TIME, WAS STUDY THE VITAMIN A ON THE THERE WAS A FAILURE FOR THESE INFLAMMATORY MACROPHAGES TO CONVERT INTO THIS TISSUE RESIDENT PHENOTYPE. AND IN DEED, IN VITAMIN A MICE THEY CONDUCT SOPPED T -- LEADINC PHONY TYPES TO ONE OF THE AND IS THAT AND AT NYU ONE OF MY COLLEAGUES AND DARWIN AND POINTED OUT TO ME SOME WORK AND WHAT WAS REMARKABLE ABOUT THIS EPIDEMIOLOGICAL STUDY WAS THAT THEY FOUND THAT BASELINE VITAMIN A DEFICIENCY WAS ACTUALLY ASSOCIATED WITH A 10 FOLD INCREASE RISK OF TUBERCULOSIS DISEASE AND SO IT'S POSSIBLE THAT THIS CONVERSION PROCESS TO TISSUE RESIDENT MACROPHAGES HEY BE IMPORTANT FOR A WIDE RANGE OF INFECTIOUS DISEASES AND ESPECIALLY SITUATIONS WHERE YOU GET FORMATION OF GRANULOMA. NOW, BEING IMMUNOLOGY, THINGS GET EVEN MORE COMPLICATED. BECAUSE AS WE SHOWN WHAT WE REALIZE AND LATER EVENT AND SUBSEQUENT THAT IN ADDITION VITAMIN A DEFICIENCY AND REGULATE THIS CONVERSION AND MACROPHAGES PHENOTYPE IN DIFFERENT INBRED STRAINS OF MICE AND THE RATE OF A CONVERSION TO A TISSUE RESIDENT MACROPHAGES IS DIFFERENT AND EVEN BETWEEN DIFFERENT SEXES OF MICE AND IS THIS REALLY THE WORK FROM JUDY AND AND THEY STRUGGLE THAT MALE MICE MORE RAPIDLY CONVERTED BON MARROW MACROPHAGES INTO TISSUE RESIDENT MACROPHAGES THAN FEMALE MICE SO IN THE CASEF JUDY ALAN'S LAB I WANTED TO SHOW YOU THIS PARTICULAR PIECE OF DATA FROM THEIR LAB WHERE THEY SHOWED THAT THERE IS A BIG DIFFERENCE BETWEEN THE BLACK SIX MOUSE AND THE C MICE WHERE AND THE EXPANSION OF THE TISSUE MACROPHAGES DURING THE INFECTION PRIMARILY THESE LARGE PARROT TENILLE MACROPHAGES AND THERE'S SOME EXPANSIONS OF THE MONOCYTE DERIVED MACROPHAGES BUT IN THE C MICE, THERE'S MUCH LESS ACCUMULATION OF THE TISSUE RESIDENT AND MACROPHAGES AND THEY HAVE NOW DONE EXPERIMENTS TO INDICATE THAT INFLAMMATORY MACROPHAGES GET STUCK IN THE MICE INTO A TISSUE RESIDENT MACROPHAGES AND THIS IS INTERESTING IN THE CONTEXT OF INFECTION AND JUDY THINKS THIS MIGHT BE RELATED TO THE DIFFERENCE IN SUSCEPTIBLE BECAUSE UNLIKE THE INTESTINAL AND AND COMING BACK AND IN ABOVE SEA MICE AND THERE'S LESS CONVERSION OF THE MONOCYTE DRIVE MACROPHAGES INTO THE TISSUE AND YOU ALSO ZIZI GREATER ACCUMULATION OF PDL2 POSITIVE MACROPHAGES WHICH PREVIOUSLY NOT SEEN IN THIS C57 BLACK SIX MICE SO THIS MADE UP BEGIN TO REALIZE AND AND BIOLOGY AND DIFFERENTIATION IS BASED ON WORK AND ALL THE LINEAGE TRACKING SYSTEMS AND ALL THE LABS HAVE UTILIZED BASED ON THE BLACK SIX SYSTEM, SO NOT ONLY IS DEVELOPMENTAL BIOLOGY POTENTIALLY EFFECTED AND BY THE GENETIC BACKGROUND OF THE MICE AND CHRIS GLASS HAS DONE AND THEY MADE BONE MARROW MACROPHAGES FROM FIVE DIFFERENT STRAINS OF MICE AND STIMULATED THEM WITH IL-4 TO LOOK AT HOW THESE GENETIC BACKGROUND OF THE BONE MARROW MACROPHAGES AND IN VITRO SETTING AND IT WOULD EFFECT THE TRANSCRIPTIONAL RESPONSE TO IL4 AND ONLY 27 GENES AND MACROPHAGES AND SO THERE'S A HIGHLY STRAIN SPECIFIC TRANSCRIPTIONAL RESPONSE TO IL-4 AND MEDIATED SIGNALING AND SO I COME FROM A SELL EYE LAR IMMUNOLOGY BACKGROUND AND IT WAS REALLY A POSTDOC IN THE LAB MAYSON TANG WHO KIND OF BROUGHT US DOWN BIT ROAD OF TRANSCRIPTIONAL REGULATIONS AND I'M SURE THAT ALAN AND ALL THE OTHER PEOPLE HERE HAVE APPRECIATED WHEN THEY HAD A TRAINEES THAT TAKEN DOWN A AND BROUGHT THE IDEAS OF TRANSCRIPTIONAL REGULATION TO A LAB AND ONE OF THE THINGS THAT SHE FOUND WAS THAT IN DEED, YOU KNOW, IN THE PAR TA KNEAL CAVITY THAT WAS CONSISTENT WITH THE WORK THAT JUDY AND WE HAVE THIS VERY LARGE THOSE STRINGS AND AND ACTIVATION AND THIS WORK IS BEING CONTINUED IN OUR LAB HERE AT NYU, AT NIH. [LAUGHTER] I SAID THAT, I'VE BEEN USING NYU FOR SO MANY WORDS IT'S HARD TO TRANSITION. AND SO THIS IS A VERY SIMPLE SIL AND THE COMPLICATED HELM AND THE MACROPHAGES FOR IN THIS CASE RNA-SEQ AND WHAT IS REALLY INTERESTING HERE IS THAT THE IS THE STRAIN OF MICE IS ACTUAL WILL HE EXPLAINS 61% OF THE VARIANTS WHEREAS THE EFFECTS OF IL-4 IS 34% OF THE VARIANTS INDICATING THAT THE STRING SPECIFIC EFFECTS ARE GREATER THAN THE EFFECTS OF THIS STIMULATION ITSELF AND WHAT WE'VE ALSO BEEN REALLY INTERESTED IN IS THAT DESPITE THE FACT THAT WE THINK OF C MICE AS BEG MORE TH-2 BOY AS WHEN IT COMES TO THE MACROPHAGES TRANSCRIPTIONAL RESPONSES TO THIS IL-4IP TREATMENT, IT IS THE BLACK SIX MACROPHAGES THAT ARE MORE RESPONSIVE IN TERMS OF TRANSCRIPTIONAL CHANGES IN RESPONSE TO IL-4 AND JUST AS WAS SHOWN IN CHRIS GOSSE'S WORK THAT'S ACTUALLY QUITE A SMALL OVERLAP IN TERMS OF THE GENES THAT ARE INDUCED IN BOTH OF THE BLACK 6 AND C MICE SO SHE'S BEEN USING THESE DIFFERENT DENOAM I CAN APPROACHES TO UNDERSTAND THE DIFFERENCE THIS IS THIS TRANSCRIPTIONAL REGULATION RESPONSE TO IL4 IN THE BLACK SIX MACROPHAGES. SO I JUST WANT TO SUMMARIZE SOME OF THE OBSERVATIONS SHE'S MADE THAT ESSENTIALLY AS I SAID THE BLACK SIX MACROPHAGES ARE MORE RESPONSIVE TO IL-4 STIMULATION AND IT DOES SEEM TO BE ENRICHMENT FOR THE ACTIVITIES OF THE PU1 AND NF FACTORS WHEREAS IN THE MICE WE SEE MORE ENRICHMENT IN THE AP1 FAMILY OF TRANSCRIPTION FACTORS AND FOR THE BLACK SIX GENES, THERE ARE INDUCED BY IL4 THEY'RE ASSOCIATED WITH SUPER ENHANCERS WHICH MIGHT EXPLAIN WHY THEY ARE MORE RESPONSIVE TO I L-4 STIMULATION AND THE SEA MICE IN THE SINGLE CELL RNA-SEQ THOSE HETEROGENEITY AND HOW IT MIGHT APPEAR TO BE LESS TRANSCRIPTIONALLY RESPONSIVE AND HOWEVER WHAT WE WERE HOPING TO DO IS MAP THE DIFFERENCES IN THE CHROMATIN DIFFERENCES WITH SPECIFIC SEQUENCE IN THOSE SITES AND IN THIS ESSENCE WE WOULD LIKE TO GO IN AND DID CRISPR REGIONS RIGHT AND WE WOULD FIND WE WOULD FIND THE REGION IN THE GATA6 LOCUS THAT WOULD THE LEVEL THAT DIFFERENTER BETWEEN THE NOT ASSOCIATED WITH DIFFERENCES IN SEQUENCE IN THOSE PARTICULAR REGIONS AND SO THERE IS CERTAINLY STILL A LOT MORE WE DON'T UNDERSTAND ABOUT WHY YOU HAVE THESE STRAIN-SPECIFIC DIFFERENCES IN EPIGENETIC AND AS WELL AS TRANSCRIPTIONAL RESPONSES TO IL-4 AND BUT WE COULD GOING BACK TO THE MODEL AND I CONTINUE TO HAVING BRAZILIAN POSTDOCS IN THE LAB AND A NEW POSTDOC FROM BRAZIL IN THE LAB AND SHE HAS DONE EXPERIMENTS WHERE WE ARE LOOKING AT GOING BACK TO IN BRED STRAINS OF MICE AND THIS IS THE GOING BACK TO THE ROOTS OF PARASITE IMMUNOLOGY AND THE BLACK 6 MICE AND DIFFERENCES IN THE RESPONSES AND WHAT CAMILLA HAS FOUND IS THAT IN SOME OF THIS IS REPEATING WORK THAT HAD BEEN DONE IN THE 1980s AND 1990s THAT THE DIFFERENCES IN SUSCEPTIBLITY BETWEEN DIFFERENT INBRED STRAINS OF MICE SO SCHISO INFECTION AND WHAT IS INTERESTING IS THAT SP LE NOMEGALY THAT IS POORLY UNDERSTAND IN THE CONTEXT OF SCHISTO INFECTION AND AN INDICATION OF EXTRA (INAUDIBLE) IS MUCH GREATER IN THE STRAINS OF MICE AND PARASITE INFECTION AND IN DEED THIS IS NOT ACTUALLY REFLECTED IN NECESSARILY IN THE LIVER AND IN THE LIVER SHOWN BY MIGGAL GROUP SUFFERED MUCH GREATER AND SOMEHOW THEY SURVIVED AND SEA MICE AND IT REPRESENTS THE WORK I'VE BEEN TRYING TO PURSUE AND IT REFLECTS HOW HOW INFECTIONS DELIVER IN TERMS OF PATHOGENESIS BECAUSE PEOPLE AND IN AN ENDEMIC SITUATION, MOST PEOPLE ARE ASYMPTOMATIC FROM THE HELMINTH INFECTION AND PEOPLE THAT DEVELOP PATHOLOGY FALL ON THE EXTREMES AND EITHER THEY ARE SUPER INFECTED OR THEY SUFFER FROM IMMUNO PATHOLOGY AND THIS KIND OF DISTRIBUTION IS COMMON AMONG ALL AND THIS KIND OF ENTER INDIVIDUAL VARIATION IN IMMUNE RESPONSES BETWEEN PEOPLE AND SOMETHING THAT I THINK AND THEY HAVE TAKEN REDUCTIONIST APPROACHES OVER THE LAST FEW DECADES TO TRY AND UNDERSTAND SPECIFIC CELLULAR AND MOLECULAR MECHANISMS BUT WHEN WE WANT TO TRY AND UNDERSTAND VARIATIONS IN IMMUNE RESPONSES WE MIGHT HAVE TO TURN TOWARDS TOOLS THAT ECOLOGISTS AND COMPUTATIONAL BIOLOGIST USE TO MEASURE THE EFFECT SIZES AND OF THE DIFFERENT COMPONENTS AND OF COURSE, IT'S GOING TO BE A COMBINATION OF GENETIC AND ENVIRONMENTAL FACTORS THAT DETERMINE THIS INTER INDIVIDUAL VARIATION BUT HOW THIS GENETIC AND IT'S A KEY THAT FAULT IN THE GENOME IN OFTEN FOUND THAT WILL BE THE LARGEST CONTRIBUTORS TO THE INDIVIDUAL VARIATION AND IN SOME WAYS THE MICE ACTUALLY A GOOD MODEL FOR THAT BECAUSE ONE CAN TRACK, ONE CAN ESSENTIALLY AND AS FORM YOU MUTATION ALANAL SIS AND TO STUDY SEQUENCE VARIANTS THAT FALL INTO THESE NON CODING REGIONS. SO, I FEEL -- SO THIS WORK OF COURSE HAS BEEN DONE BY -- THIS IS THE FIRST COHORT OF PEOPLE IN THE LAB AND OF THIS COHORT, TASHA, MICHAEL AND THAT IS AN PLAYED CRITICAL ROLES IN OUR STUDY ON MACROPHAGES BIOLOGY THAT WAS ALWAYS BEEN A CLOSE COLLABORATION WITH JUDY ALLEN AND HER GROUP BECAUSE NOW IN THE UNIVERSITY OF MANCHESTER AND THIS WAS THE SECOND COHORT OF PEOPLE IN MY LAB AND IT WAS AN NYU JUST BECAUSE WE JOINED THE NIH AND I'M REALLY EXTREMELY EXCITED TO BE HERE AND WE NOW HAVE A GREAT DEAL OF PEOPLE AND I WANT TO REALLY THANK ALAN FOR EVEN I THINK SEVERAL YEARS BEFORE THE POSITION WAS OFFICIALLY OPENED, IT WAS ALREAENCOURAGING ME TO THINK ABT THE POSSIBILITY OF AND THIS AND AND MUCH LATER AND MOST OF YOU BECAUSE HE WAS HE WAS A LEGEND AND WHEN I MET HIM IN-PERSON AND HE REALLY DIDN'T FIT WITH MY INITIAL CONCEPTIONS OF THIS LEGEND OF THE FIELD AND AND I THINK THAT THE THING THAT HASN'T BEEN MENTIONED BECAUSE EVERYTHING HAS ALMOST EVERYTHING HAS ALREADY BEEN SAID, I HAD THE PRIVILEGE OF ORGANIZING THE IMMUNOLOGY MODULE FOR THE PARA SITTISM COURSE AND AND THIS IS A LONG HISTORY AND WE INVITED ALAN TO GIVE THE PRODUCTION TO IMMUNOLOGY COSTS AND I WOULD SAY 80% OF STUDENTS ON THIS CAUSE ARE BASIC AND YOU CAN SEE THE EYES LIGHT UP AS HE EXPLAINS TO THEM EXPANSION AND THEY'VE NEVER THOUGHT ABOUT THAT WHOLE PROCESS BEFORE AND SO ALAN IN ADDITION TO ALL THE THE THINGS THAT YOU GUYS HAVE SAID IS ALSO A REMARKABLE TEACHER AND OF OF NEW COMMERCE TO IMMUNOLOGY AND I THINK IT'S REALLY A PRIVILEGE TO NOW HAVE YOU AS A COLLEAGUE AND OF COURSE HE IS DONE ALL THE THINGS YOU HAVE SAID AND HE HAS INTRODUCED ME TO EVERYONE IN THE NIH, AND CONNECTED ME TO BASED ON THE SCIENCE, YOU KNOW I THINK THAT'S THE AMAZING FEATURE OF ALAN IS HE IS ABLE TO IDENTIFY THE RIGHT FIT OF PEOPLE BASED ON THE WORK THAT THEY'RE DOING SO KIND OF BRING THEM TOGETHER AND SO WITH THAT, THANK YOU, ALAN FOR ADOPTING ME INTO THE KIND OF LPD/NIH FAMILY. [APPLAUSE] >> SO, THANK YOU AND VERY INTERESTING. I WANTED TO ASK ABOUT TWO OTHER GENETIC DIFFERENCES BETWEEN VALVE C AND 6 THAT MIGHT PLAY A ROLE. ONE IS THAT B6 EXPRESSES HIGHER LEVELS OF ONE CHAIN OF THE IL-12 RECEPTORS WHICH MAKES B6 MORE RESPONSIVE TO IL12 OF COURSE LEADING TO INTERFERING GAMMA PRODUCTION. AND THE OTHER IS DIFFERENCES IN THE NK LOCUS THAT WAYNE HAS STUDIED EXTENSIVELY BETWEEN THE TWO STRAINS WITH REKUM BENT STRAINS THAT ARE PART OF ONE STRAIN AND PART OF THE LOCUS FROM ANOTHER STRAIN AND THAT INFECT THINGS LIKE THE MCMV INFECTION AND OF COURSE NK CELLS MAKE ANOTHER GAMMA INTERFERON CELLS SO THAT'S A LOT OF COM ANALITY AND YOU FOCUSED ON IL-4 BUT I WONDER IF THOSE GENETIC DIFFERENCES MIGHT PLAY A ROLE HERE? >> I THINK IT'S VERY IMPORTANT POINT THAT HAS BEEN POINTED OUT TO ME BY DRAGANA AND DAVID SACKS IS THAT THE DIFFICULTY IN SEPARATING THE ENVIRONMENT FROM THE CELL INTRINSIC EFFECTS OF THE MACROPHAGES THEMSELVES SO WE PLAN TO TRY AND ADDRESS THIS QUESTION AND WHETHER THE ENVIRONMENT AND THE TISSUE ENVIRONMENT AND AND THE COMPETITION OF THE IMMUNE CELLS AND ALTHOUGH I GUESS, THE CASE AND ADDRESS THAT AND TRUE USING, DOING A. >> GREAT TALK. SO M. WHAT RELATED TO THE COMMENTS (INAUDIBLE) DUE TO OTHER CELLS AND THE VARIATION CAUSED THE BEHAVIOR OF OTHER CELLS AND I THINK DIRECTLY THE MACROPHAGES BUT WHEN YOU THINK ABOUT IT ALL THOSE PROBABLY WOULD BE SOMEWHAT MARKETED GREN ET TICK LEVEL SO HAVE YOU ACTUALLY TRIED TO LOOK AT BEFORE OURS AND THERE'S AND IT'S THAT WERE PRONE THEM TO BE MORE AND AS I SAID BEFORE AND STIMULATION AND AND IT'S SOME OF THE WORK THAT MING HAS BEEN DOING AND WE LOOKED AT. EXIST EXAMINING DIFFERENCES AND EPI GENETIC LEVEL AND WHAT IS INTERESTING AND THE RESPONSE TO I-L4 BETWEEN TWO DIFFERENT STRAINS OF MICE AND INDUCED CROW A TIN CHANGES AND AND WHAT IS AND THE REGENTS THAT ARE AND STATUS AND THOSE TEND TO FALL IN THE NON CODING REGIONS AND THE NAIVE SO THE NAIVE CON AND AND PROMOTE REGIONS AND SO THERE IS LESS OF THOSE DIFFERENCES BUT THEY'RE INDUCED CHANGES TEND TO BE AND AND IN BETWEEN THE STRAINS AND SO -- >> IS IT FAIR TO SAY THAT THOSE FUNCTIONING MOSTLY IMPORTANTLY THE GENES AND ACTUALLY VERY CONSERVED WHERE LESS IMPORTANT GONE MAY REPRESENTING AND INDUCING MAY NOT BE FUNCTIONING AND CAN YOU ACTUALLY A GOOD EXAMPLE THAT DEMONSTRATES AND FUNCTIONALITY. >> THAT'S A GREAT POINT AND IN FACT, I WAS TALKING TO JUDY ALLEN ABOUT THAT AND I WAS SPECULATING SO THAT HUMANS, THE LONG BEEN KNOWN THAT ALTERNATIVELY ACTIVATED GENES IN MICE ARE NOT REFLECTIVE IN HUMANS, RIGHT AND SO ONE OF THE HYPOTHESIS WAS MAYBE IT'S BECAUSE WE'RE STARTING BLACK SIX MICE AND MAYBE WE NEED TO LOOK AT THE CONSERVED GENES THAT ARE CONSERVED AND ALTHOUGH AND THEY PHASE AND THOSE ARE HIGHLY CONSERVATIVE OF BEING INDUCED AND WHERE THOSE ARE THE ONES THAT ACTUALLY AND AND IT MIGHT BE SPECIFIC CORE KIND OF EFFECTS THAT ARE PULLING. >> THAT'S BEAUTIFUL AS ALWAYS. A REMARE DIFFERENCE IS BETWEEN NAIVE BOB C AND B6 MACROPHAGES BEGGING TO QUESTION IF YOU AND AND MACROPHAGES AND STRAINED CROSS TO CHAIN DEFICIENT MICE AND TO ELIMINATE THE FACTS OF INNATELY AND T CELLS PRIMING IN IDEALLY EVEN DERIVED TO GERM FREE ENVIRONMENT WHAT DO YOU THINK WOULD BE THE DIFFERENCES? WOULD IT BE REDUCED? >> I MEAN, YEAH, THAT'S AGAIN A GREAT IDEA TO TRY AND REMOVE AS MUCH AS POSSIBLE THE CONTRIBUTIONS OF THE OTHER CELL TYPES AND INTO THE SYSTEM. THE GROUP HAS LOOKED AT THAT FROM THE PERSPECTIVE OF BONE MARROW MACROPHAGES AND WE KNOW THAT TISSUE AND MACROPHAGES ARE NOT THE SAME AS BONE MARROW MACROPHAGES SO THAT'S A GREAT IDEA. >> BONE MARROW MACROPHAGES THE HEAVY INFLUENCES AND JASMINE SHOWED BEAUTIFULLY AND IT'S A WAY TO START. >> YEAH. YEAH. >> GOOD. >> THANK YOU. [APPLAUSE] >> WE'RE GOING TO TURN THIS OVER TO TOM WYNN WHO IS GOING TO HAVE TO PENT ULTIMATE REMARKS. >> I SEE THE BIG BOSS IS HERE, HI, STEVE. SO I PROBABLY DON'T HAVE AS MUCH TIME AS I THINK OF THE THANK YOU FOR ORGANIZING IT'S BEEN A REALLY FUN TWO DAYS AND MY CAREER AT NIH STARTED 31 YEARS AGO AND I SPENT 26 YEARS HERE BEFORE DECIDING TO LEAVE ABOUT FIVE YEARS AGO AND IT WAS A VERY HARD DECISION. IT WAS A JOY TO WORK WITH ALAN FOR SO MANY YEARS. AND IT WAS A GREAT CHOICE TO COME HERE AND I WAS DECIDING BETWEEN HIS LABORATORY AND ROBERT MAUD LYNN WHO WORKED ON HELP RA SEE AND TB AND THE TH2 AND RICHARD AND THERE'S A LOT OF FUNNY STORIES I WANT TO SHARE. AND IT'S GREAT TO SEE LPD AND AND INFECTION MORE THAN I STILL HAVEN'T FOUND IT AND I KNOW IT'S OUT THERE SOMEWHERE. DAVID SACKS FOR LETTING ME SIT IN HIS LAB WHEN I FIRST CAME. ALAN'S LAB WAS OVERFLOWING WITH PEOPLE NOT SURPRISINGLY AND DAVID LET ME SIT IN HIS LABORATORY AND I SAT IN THE AREA WHERE THEY KILLED THE MICE AND THAT WAS AN INTERESTING EXPERIENCE AND SO I WAS SITTING RIGHT BEHIND HIS REALLY AWFUL OFFICE AND I STILL HAVE AND SO THANK YOU AND THANK YOU. WE HAD A LOT OF THIS AND A LOT OF PEOPLE TOUCHED ON THE AMAZING EDUCATION AND CAREER AND I'M GOING BACK FURTHER BACK TO COLLEGE AND THIS IS MY FIRST CONNECTION FOR ALLEN AND AND ALAN WOULDN'T TO OBSERVER LAND AND FANCY AND ELITE LIBERAL ARTS COLLEGE AND BEAUTIFUL COLLEGE AND NEAR CLEVELAND AND SOME OF YOU DO REALIZE AND CORRECT ME IF I'M WRONG BUT YOU WERE AN ENGLISH MAJOR. >> WHAT IS IT? >> CLASSICS. BUT OK CLASSICS. HE IS A GOOD WRITER BECAUSE OF HIS EDUCATION AT THE SCHOOL I THINK. THERE WAS A REAL FOCUS ON WRITING. AND WE'VE ALL TALKED ABOUT THAT TODAY HOWEN INFLUENTIAL THAT'S BEEN AND AS WE MOW WE MOVED ONTO THE MILL HILL, ASSIST APARTMENT PROFESSOR AT HARVARD AND THE NIH AND THIS IS WHAT WE'RE LEFT WITH. ALAN WAS ALWAYS A LOT OF FUN AND HE WAS NOT ONLY A GREAT MENTOR BUT HE WAS A REALLY FUN MENTOR TO WORK WITH AND WE'VE GONE THROUGH ALL OF THIS AND THE HISTORY AND I THOUGHT I WOULD TURN IT BACK ON ALL OF YOU BECAUSE THIS IS WHAT ALAN CARES ABOUT THE MOST. I'M GLAD WE'RE ABLE TO DO THIS NOW BEFORE YOU ARE 100-YEARS-OLD AND WE'RE CELEBRATING YOUR 100th BIRTHDAY SO YOU STILL REMEMBER WHO WE ARE. AND SO IT'S GREAT YOU CAN BE HERE AND CELEBRATE AND PUBLISH SEMINOLE PAPERS AND WE'VE TALKED ABOUT THE EARLY WORK WITH PHIL SCOTT AND MOB MAUDELYNN AND IT'S AN AMAZING AND AND ALAN AGAIN, A LOT OF AND I THINK THIS IS ALAN, ROB THOMPSON SENT ME THESE PHOTOGRAPHS AND I THINK THAT'S ALAN ON THE LEFT. HE REALLY LIKED HALLOWEEN AND HE LIKED TO I COULD NOT FIND A PHOTO OF THAT AND AND BENEFITED FROM AMMAN WORKING ON TOXO SO IT FREED THAT AREA UP FOR THEM TO EXCEL AND BE RECOGNIZED FOR THEIR OWN SEMINOLE RESEARCH. HE MOVED ON TO TALK AND I'M SURE THE HIV EPIDEMIC HAD A LOT TO DO WITH THAT PARTICULAR INTEREST AND IT'S AN AMAZING GROUP OF COLLEAGUES THAT WORKED WITH HIM OVER THE YEARS AND SOME OF THE NICE PAPERS THAT GROUP PUBLISHED AND SOME OF THE MOST CITED PAPERS ARE SHOWN HERE AND ONE THAT WAS PERSONALLY INSPIRING FOR ME AND TO A DIFFERENT WAY AND AND IT'S CRITICAL AND I THOUGHT THAT WAY IN MY OWN RESEARCH FOR MANY YEARS AND IT GUIDES THE RESEARCH THAT I DO. HE ALSO MOVED ON ANOTHER MAJOR AREA AND I HAVE TO SAY A PET AREA OF RESEARCH IDENTIFYING THE PERSON PARASITE FACTORS THAT WERE DRIVING AND WHAT WERE THE FACTORS WHAT WERE THE RESPONSE AND AND THAT WAS A PROJECT THAT ALAN ENJOYED HERE AND HE IS STANCING AFTER THE DISCOVERY OF PRO FILL AND INTERACTING WITH RECEPTORS 11 AND THEN DRAGANA'S BEAUTIFUL WORK ON THE OMEGA 1 THAT WE HEARD ABOUT EARLIER. AND A BIG MOVE AGAIN THIS CAME AROUND THE TIME WHERE I WAS BECOMING A 10-YEAR TRACK INVESTIGATOR AND ACTUALLY ANOTHER FUNNY STORY IS THAT I NEVER REALLY WORKED IN ALAN'S LAB WHEN I FIRST CAME. I SAT IN DAVID'S LAB AND WE ACQUIRED SPACE IN THE SUB BASEMENT WHERE THERE ARE NO WINDOWS AND BUILDING 4 AND I WAS TOWN THERE WITH LENA AND DRAGANA AND OUR OWN LITTLE TRIO AND NO WINDOWS AND WE WOULD BE THERE UNTIL 8:00 AT NIGHT BECAUSE WE HAD NO IDEA WHAT TIME IT WAS BUT IT WAS A GOOD THING AND IT WAS A FUN TIME AND I REMEMBER ONE STORY THAT RICARDO COMING DOWN. IT WAS A PLACE WHERE WE KILLED MICE AND COLLECTED PARASITE MATERIALS AND RICARDO COMES DOWN COLLECTING MOUSE BRAINS TO GET TOXO AND HE IS SITTING RIGHT NEXT TO ME. I WAS A MOLECULAR BIOLOGIST DOING PCR AND RICARDO IS RIGHT NEXT TO ME, LAB COAST, GLOVES, MASK, EYE FIELD, AND I SAT HERE WITH NOTHING AND NOTHING AND RICARDO, DO I NEED TO BE WORRIED ABOUT THIS. TOM, TOM, NO! YOU DON'T HAVE TO WORRY. SURE ENOUGH, WHEN WE WENT TO THE COURSE LATER THAT YEAR ONE OF THE STUDIES THAT WE DID WAS TO DO A SERUM AS SAY FOR EXPOSURE TO AND I WAS THE POSITIVE CONTROL AND I WAS SCREAMING AND THANK YOU, RICARDO. BUT THIS WAS AN AMAZING RESEARCH AND I LIKE THE PAPER AND AND BY ANNE REALLY AND BEAUTIFUL WORK AND THE TV WORK IS CONTINUING IN GREAT HANDS HERE AT THE NIH WITH CAT AND BEGAN SO T DAN SO IT'S T OF CHARACTERS THAT PLAYED A KEY ROLE IN THE WORK AND I CAME INTO ALAN'S LAB IS A PAPER LOOKING AT THE THI AND TH2 DICHOTOMY TO JUST TO SOME INFECTION AND IT WAS LIKE SO I JUST THOUGHT I WAS GOING TO FREE ASSOCIATE WHAT WERE THE WORDS I ASSOCIATE WITH THAT AND BE RELENTLESS AND ALAN WAS RELENTLESS WHEN I DECIDED TO COME TO HIS LAB. AND IN FACT, WHEN I WAS DECIDING BETWEEN BOB AND RICH, HE LITERALLY AND HE CALLED AND SHE SAID IT'S YOUR NEW BOSS ON THE PHONE AND SO HE WAS REALLY RELENTLESS ABOUT ME DECIDE TO GO COME TO HIS LAB AND I ALWAYS APPRECIATED THAT BECAUSE I KNEW THAT HE WANTED ME TO COME AND THAT MEANT A LOT TO ME AND I THINK ABOUT THAT A LOT WHEN I WOULD HIRE PEOPLE THAT MAKE IT CLEAR THAT YOU ARE REALLY WANTING THEM. BE SEEN AND WOVE HEARD A LOT ABOUT THAT AND GO TO MEETINGS AND BRAG ABOUT YOUR WORK AND YOU ARE THE CHEERLEADER AND I HAVE THAT BELOW. ALAN WAS AN AMAZING CHEERLEADER AND ED USED THAT WORD AND ALAN IS A CHEERLEADER. I REMEMBER THE FIRST YEAR IN THE LAB, NO ONE KNEW WHICH WAS AND . PEOPLE WERE COMING UP TO ME SAYING WOW WE LIKE YOUR WORK AND I WAS LIKE HOW DO YOU KNOW. I HAVE NOT PUBLISHED ANYTHING AND ALAN WAS OUT THERE BEING MY CHEERLEADER AND BE A STORY TELLER AND A FANTASTIC AND STRIVING FOR EXCELLENCE AND WE'VE HEARD ABOUT TEAMWORK AND THANK GOD FOR SARAH HEINE'Y AND PAT CASPER SO WE CAN EXCEL AT OUR WORK AND OF COURSE COLLABORATION BOB COFFMAN AND GORGIO AND HE HAD A GREAT COLLABORATORS AND THAT ALSO ELEVATED THE KIND OF WORK THAT HE DID AND HE WAS ALWAYS GRACIOUS AND THANKED US AND PATIENTS AND NOT SO MUCH. AND HE WOULD I'VE LEARNED TO BE PATIENT IN MY OWN LIFE. HE WAS COURAGEOUS AND LET'S TRY WITH THEM AND LET'S SAY AND SITTING IN THAT AWFUL SEAT AND WE ALL LEARNED A LOT AND HE ALWAYS CELEBRATED OUR ACCOMPLISHMENTS AND THERE WAS A BOTTLE OF CHAMPAGNE AND LOVED THAT AND MADE YOU FEEL SPECIAL AND LOVED TO PARTY AND ALWAYS HAD GREAT PARTIES ALAN CHIEFER WAS A PESSIMIST AND WHAT A GREAT SITUATION. ALAN SAID EVERYTHING IS EXCITING AND YOU CAN DO THAT AND I COULD TELL ALAN AND SAID YOU CAN DO THAT FIVE YEARS AGO HE WOULD GO TO HIS COMPLEX FILE SYSTEM AND PULL THE PAPER OUT AND SHOW ME BUT THERE'S ALWAYS A NEW ANGLE ON IT AND SO I HAD TWO GREAT MENTORS HERE AND HE WAS A GREAT COOK AND I HAD INDIAN DINNERS AND HE WAS VERY GOOD AT LAMB AND THAT WAS HIS CONTRIBUTION AND THE HOTSEAT, THIS IS LIZ WHO COULDN'T BE HERE AND SO SHE WOULD HAVE LOVED TO HAVE BEEN HERE AND WE ALL HATED THIS HOTSEAT AND I LEARNED A LOT ABOUT WRITING AND I ALSO LEARNED A LOT ABOUT WHAT FAVORITE WORDS ALAN HAD AND YOU CAN THROW OUT HIS FAVORITE WORDS AND I REMEMBER NEVERTHELESS. HE WAS SO HAPPY IF HE USED IT. >> SO WILL YOU, ALAN. I IT'S AN IMPACT AND THIS IS A REVIEW ABOUT COLLEAGUES ABOUT TH1 AND T H2 PARADIGM WHICH NEVER DIES AND HOPEFULLY THIS WILL ILLUSTRATE WHY IT'S IMPORTANT AND ALAN WORKED ON ALL OF THESE AND THE FLIP SIDE AND TH-2 RESPONSE AND WHEN YOU THINK ABOUT OTHER DISEASES THAT ARE IMPACTED BY THIS PARADIGM, DISEASES THAT I NOW WORK ON, PSORIASIS, ARTHRITIS, CROWNS, RA, ALL DRIVEN BY WHAT LOOKS LIKE A TYPE ONE INFLAMMATORY RESPONSE AND THEN ON THE OTHER SIDE, THESE DISEASES, FOOD ALLERGY, ASTHMA AND DRIVEN BY THAT TH2 SIGNATURE AND THERE'S SOME DISEASES THAT DON'T FALL NEATLY INTO THESE CATEGORIES THAT HAVE A MIXED INFLAMMATORY RESPONSE AND WHAT IS TRULY AMAZING IS THE NUMBER OF NEW MEDS THAT ARE BEING MADE AND BASED OFF OF THE SITE OWE KEEN WORK THAT A LOT OF YOU HAVE DONE IN THIS ROOM AND HUMARA IS A OTHER BILLION DOLLAR A YEAR INDUSTRY CREATING MANY DIFFERENT DISEASES, ANTI-IL-6 AND IL-1 AND WE THINK ABOUT IL-17, IL-23, P-19, IL-12. AND MULTI BILLION DOLLARS DRUGS AND EVERY YEAR AND TREATING THESE DISEASES AND ON THE FLIP SIDE, THE TH2 RESPONSE AND ANTIBODY IL4 RECEPTORS AND IL5, MORE RECENTLY TSLP ARE START TO GO TREAT THESE TH2 DISEASE AND THE OVER ALL IMPACTS IS IT'S A $1 BILLION A YEAR INDUSTRY SO A LOT OF THAT CAN COME BACK TO YOU AND THE TRAINEES YOU'VE TRAINED OVER THE YEARS, I'LL STOP THERE. THANK YOU. [APPLAUSE] >> WE'RE COMING TO THE END SO I'M GOING TO TURN IT >> THE SCIENTIFIC DIRECTOR OF THE DIVISION OF INTRAMURAL RESEARCH RIGHT HERE. >> ALL RIGHT, ALLAN. ALAN. TOM, I WAS VERY HAPPY TO HEAR YOU TALK ABOUT $100 BILLION COMING BACK TO ALAN. I WILL HAVE MY PEOPLE TALK TO YOUR PEOPLE. THANK YOU. ALAN, HAVE YOU HAD A CHANCE TO STAND UP AND LOOK AT EVERYBODY HERE? STAND UP. JUST STAND UP. TAKE A TURN AROUND. THIS IS REALLY REMARKABLE. [APPLAUSE] I MET ALAN ABOUT 33 YEARS AGO IN THE BASEMENT OF BUILDING 4, AND I SAW THAT PIPETTE TIP. I THINK IT WAS THE SAME ONE FOR SEVERAL YEARS. HE WAS AN ENERGETIC OMNIPRESENT GUY, ABSOLUTELY NEVER AT REST. THAT POISE THAT YOU CAUGHT DANCING, IT HAD TO BE A FREEZE FRAME BECAUSE IT DIDN'T HAPPEN ANY OTHER TIME. SO I HAD A CHANCE OVER THE YEARS TO COLLABORATE WITH ALAN TO LEARN A LOT FROM HIM, BOTH AS A YOUNG PRE-TENURED TRACK, TENURED TRACK, AND THEN A MORE USELESS INVESTIGATOR. AND ALAN HAS ALWAYS BEEN REALLY EXTRAORDINARY IN TERMS OF THE ABILITY TO GIVE ADVICE, TO GIVE ENCOURAGEMENT, AND JUST TO GIVE WISDOM. ALAN IS A REALLY REMARKABLE GUY. AND THIS MORNING, AS I WAS THINKING ABOUT THIS, I HAD REMEMBERED THAT ALAN HAD GIVEN AN ORAL HISTORY. I DON'T KNOW IF ANY OF YOU HAVE READ HIS OR ANY OF THE ORAL HISTORYS THAT HAVE BEEN DONE HERE AT THE NIH FOR INVESTIGATORS. THEY'RE REALLY REMARKABLE. AND ALAN'S HISTORY WAS WONDERFUL. I HAD A GREAT TIME READING THAT. I WANTED TO SHARE A COUPLE OF QUOTES FROM IT. I'M NOT ORIGINAL ENOUGH TO COME UP WITH MY OWN, AND THESE, I PROMISE, ARE VERBATIM. A GOOD LEADER FINDS OUT WHAT PEOPLE ARE GOOD AT, AND THEN LETS THEM DO THAT THING. SEEMS PRETTY STRAIGHTFORWARD. NOT MANY PEOPLE ARE ABLE TO DO THAT. I HEARD ABOUT THE HOT SEAT. I READ ABOUT THAT TOO. BUT IT IS THAT ABILITY TO TRY AND FIGURE OUT, WELL, WHAT IS SOMEBODY GOOD AT? AND THEN MAKE SURE THAT THEY GET A CHANCE TO DO THAT AND PROVIDE THAT OPPORTUNITY. AS PUN GWAS POINTING OUT, IT'S SORT OF BEING CAUGHT IN A SPIDER'S WEB BUT RATHER THAN THAT SPIDER THAT SUCKS ALL THE JUICE OUT, IT'S THE OPPOSITE. YOU GET CAUGHT AND THEN HE'S LIKE, HERE, COME ON, DO THIS, DO THAT, WHAT ARE YOU DOING, DO THIS, DO THIS. ALAN IS A SPECIAL INVIGORATING ENLIVENING SOURCE. HE ALSO SAID AS YOU POINTED OUT, MY PHILOSOPHY IS SIMPLE. I'M A CHEERLEADER. AND THERE'S NO QUESTION THAT THAT IS -- YOU KNOW, THERE ARE A LOT OF PEOPLE WHO ARE CHEER LEERS. THERE AREN'T A LOT OF PEOPLE WHO ARE OUTSTANDING CHEER LEERS. LEERED -- CHEERLEADERS. ALAN REALLY HAS BEEN THAT THROUGHOUT THE 33 YEARS THAT I'VE KNOWN HIM. IN THE LAST FEW YEARS, WE AS AN INSTITUTION HAVE BEEN ENORMOUSLY BLESSED TO HAVE HIM LEADING THE PROGRAM, HELPING TO TAKE YOUNG PEOPLE AND PERVERT THEM TO OUR WAY OF LIFE. [LAUGHTER] AND ALAN DIDN'T COME TO THIS BY ACCIDENT. AND ONE OF THE THINGS THAT YOU ALSO LEARN IS THAT ALAN, AS YOU ALL KNOW, HE'S BRAVE, HE'S INDEPENDENT, ICONICLASTIC, WISE, AND THAT WAS SHOWN WHEN HE LEFT BASIC IMMUNOLOGY TO GO AFTER, EWW, PARASITES. TOM, I KNOW WHERE YOUR FOOT IS, DON'T WORRY. TO GO LOOK AT PARASITOLOGY, AND THEN WHEN HE MADE THAT HORRIBLE, HORRIBLE BLUNDER TO LEAVE HARVARD AND COME HERE. MY GOD, WHAT A BACKWATER, YOU KNOW? WHO GOES THERE EXCEPT TO DIE? BUT ALAN KNEW THAT THIS WAS SMART, HE KNEW IT WAS TIME TO GO INTO SOMETHING NEW, LIKE PARASITOLOGY. THE IMPACT HE'S HAD ON THESE FIELDS AND OUR LIFE AND WORLD IN WHICH WE LIVE HAS BEEN EXTRAORDINARY. SO I JUST WANTED TO CONCLUDE BY THANKING ALAN FOR COMMITTING THIS EXPERIMENT IN LEAVING THE BEATEN PATH AND IN COMING HERE AND CARVING OUT NEW AREAS AND NEW PARADIGMS, SHARING THEM WITH US AND SHARING THEM WITH THE WORLD. YOU HAVE DONE AN EXTRAORDINARY JOB, AND MADE US RICHER AND BETTER FOR IT. AND I JUST WANT TO EXTEND MY PERSONAL AND PROFESSIONAL GRATITUDE FOR YOUR SERVICE HERE. THANK YOU. [APPLAUSE] >>SO WITH THIS, THIS CONCLUDES THIS INCREDIBLE TWO-DAY SYMPOSIUM IN ALAN'S HONOR. ONE MORE CHEER FOR REALLY PUTTING THIS TOGETHER. [APPLAUSE] IT WAS ABSOLUTELY -- IT COULD NOT HAVE HAPPENED. AND I KNOW THIS WON'T BE THE LAST TIME WE'RE ALL TOGETHER. SO ANYWAY, ENJOY THE REST OF THE FESTIVITIES.