Confirmation Number:261474 Event Started: 9/21/2004 4:59:25 PM Event Ended: 1/1/0001 12:00:00 AM

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TEST TEST TEST A HEART TY WELCOME TO ALL OF YOU. WE HAVE CONSIDERABLY CHALLENGE AND HARD WORK AHEAD OF US FOR THE NEXT TWO DAYS. WELCOME TO THE NIH'S SYMPOSIUM ON RECOMBINANT RESEARCH WITH CHALLENGES MEETING THE PUBLIC HEALTH CHAMPS. I'D LIKE TO GREAT PLEASURE TO INTRODUCE THE NIH DR. ZERHOUNI WE'LL GIVE OUR KEYNOTE OPENING ADDRESS AND WELCOME EACH AND EVERY ONE OF YOU TO THIS CONFERENCE AND WE LOOK FORWARD TO YOUR INSIGHTS AND SMEK TIFFS. DR. PERSPECTIVES. DR. ZERHOUNI.

THANK YOU, AMY. I'D LIKE TO WELCOME, EVERYONE TO THIS VERY IMPORTANT SYMPOSIUM. LET ME ALSO ACKNOWLEDGE THE LONG-STANDING AND OUTSTANDING WORK OF THE RECOMBINANT ADVISORY COMMITTEE. I THINK THIS IS ONE OF THE ACTIONS THAT WAS TAKEN BY NIH MANY, MANY YEARS AGO THAT HAS TRULY PROMOTED SAFETY IN RECOMBINANT DNA RESEARCH BUT ALSO GENERATED A CULTURE OF RESPONSIBILITIES REGARDING NOT ONLY THE SAFETY BUT THE ETHICS AND THE APPROPRIATE MANAGEMENT OF RESEARCH THAT COULD HAVE NEGATIVE IMPLICATIONS AND TODAY AGAIN THE SYMPOSIUM AS WELL COME SYMPTOM BODIESIUM ON RECOMBINANT DNA RESEARCH WITH PATHOGEN NICK VIRUSES BECAUSE WE'RE FACING NEW CHALLENGES AND DIFFERENT CAPABILITIES AND, THEREFORE, WE NEED TO HAVE A THOUGHTFUL PROCESS WHICH I THINK I AM SURE GIVEN THE QUALITY OF THE PARTICIPANTS HERE AT THE END WE'D GET GOOD GUIDANCE FROM ALL OF YOU. AS SCIENCE MOVES FORWARD TOWARD THE DEVELOPMENT OF PREVENTIVE AND THERAPEUTIC TREATMENTS FOR THESE HIGHLY INFECTIOUS AND PATHOGEN NICK VIRUSES WE MUST AS A COMMUNITIES CONSISTENT FRONT THE PUBLIC HEALTH CONCERNS THAT SUCH RESEARCH ABSOLUTELY ENTAILS. YOU HAVE AN IMPORTANT AND CHALLENGING TASK AHEAD OF YOU AND THAT IS THE DEVELOPMENT OF A SET OF STANDARDS AND PRINCIPLES THAT WILL ENHANCE THE BIOSAFETY OF LABORATORY AND MEDICAL PERSONNEL. I'D LIKE TO JUST GO OVER A FEW SLIDES BECAUSE IT IS CLEARLY A TREND WHEREBY AS YOU KNOW ANYTHING WE DO HAS TO END TAIL A PUBLIC COMPONENT AND ENTAIL THE PARTICIPATION OF OUR COMMUNES AND PUBLIC COMMUNITIES AROUND THE RESEARCH INSTITUTIONS BUT ALSO THE GENERAL PUBLIC. AND WE NEED TO CONTINUE TO DO THAT. WE HAVE SIGNIFICANTLY INCREASED FUNDING AS YOU KNOW FOR VACCINES AND THERAPEUTICS AND WE HAVE TO BE A VERY GOOD STEWARD OF THESE FUNDS. I IS THINK IT'S IMPORTANT TO REAL LYLES WHAT THE CONTEXT IS IN TERMS OF PUBLIC HEALTH CHALLENGES AND I'D LIKE TO LIST FOR YOU WHAT WE CONSIDER THE TOP DRIVERS, IF YOU WILL, OF PUBLIC HEALTH IN 2004 AND HENCE FORWARD. FIRST, WE BELIEVE THAT THERE HAS BEEN A TECTONIK SHIFT IN THE LANDSCAPE OF PUBLIC HEALTH CHALLENGES FROM A MORAC KUFT SHORT TERM, MORE LETHAL PATTERN OF DISEASE 30, 40 YEARS AGO TO WHAT WE'RE FACING TODAY, MORE CHRONIC, MORE LONG TERM, MORE COMMUNITY BASED CONDITIONS THAT ENTAIL AN ENORMOUS COST TO SOCIETY. 75% OF OUR HEALTHCARE EXPENDITURES TODAY ARE RELATED TO CHRONIC CONDITIONS. WE'RE DEALING WITH A DEMOGRAPHIC PATTERN THAT WILL ENTAIL AN AGING OF THE POPULATION WITH A DIFFERENT LANDSCAPE OF CONDITIONS AND ILLNESSES AND DISEASES WE HAVE TO DO RESEARCH ON NOW. I BELIEVE THAT HEALTH DISPARITIES REMAIN A VERY IMPORTANT PRIORITY FOR PUBLIC HEALTH POLICY AND THEN WE HAVE -- , --

FRS.

KOURN TER MEASURES, WE NEED FASTER DEVELOPMENT TIMES. WE NEED TO LOOK AT NEW METHODS OF PRODUCTION OF VIRUSES AND VACCINES. WE NEED TO LOOK AT SCALING UP OF THESE METHODS, ALL OF THOSE STEPS, IF YOU LOOK AT THEM CAREFULLY, ENTAIL SAFETY CONSIDERATIONS THAT WE AND YOU NEED TO TAKE INTO ACCOUNTING AND WE NEED TO DO THIS UNDERSTANDING THAT WE NEED TO AVOID NOT ONLY INADVERTENT

FIVE OF THE EIGHT VIRAL GENES, AND WE'RE ACTIVELY COMPLETING THE SEQUENCING OF THIS GENE. SO WHAT IS THE GOAL THAT WE NEED TO DETERMINE. WE NEED TO UNDERSTAND BETTER THE MOLECULAR MECHANISMS NORTH HIGH PATH EFFICACY AND ANTIVIRAL DRUGS AS WE SPEAK. AT THE SAME TIME, WE NEED TO MAKE PREPARATION, THE H 5 ONE VARIANTS THAT YOU KNOW ABOUT IS CERTAINLY SOMETHING THAT WE'RE ALREADY TAKING STEPS. WE HAVE CONTRACTS OUT TO SCALE UP OUR ABILITY TO PRODUCE NOT ONLY TESTING LOTS BUT ALSO EARLY PRODUCTION LOTS SO THAT WE CAN FLESH OUT THE SCALING UP DIFFICULTIES THAT MAY OCCUR WHERE. WE'RE ACTIVELY FUNDING CELL BASED CULTURE METHODS TO TRY TO GET AWAY, -- NOT GET AWAY BUT AT LEAST DECREASE THE NEED FOR EGGS AND OUR ABILITY TO SCALE UP WILL BE DIRECTLY PROPORTIONAL TO THAT ADVANCE. NOW UNH 5 N11997 HONG KONG, 18 CASES, 2003, HONG KONG, THREE CASES AND TWO DEATHS AND VIETNAM IN 2004 AND THAILAND WITH 37 CASES AND 26 DEATHS, WE HAVE OTHER VARIANTS, H 7 AND 7, IN THE NETHERLANDS IN 2003 WITH MAJOR CALLING OF THE RESERVOIR THAT OCCURRED AT THE TIME AND H 9 INTO 1999 IN HONG KONG AND 2003 HONG KONG SO CLEARLY SOMETHING IS ONGOING AND WE NEED TO BE VERY VILLAGE LET'S. WHAT WILL BE THE SOURCE OF THE NEXT PANDEMIC WE NEED TO BE COMPLETELY ALERT BECAUSE THE REREASSORTMENT OF DIRECT INFECTIONS FROM BIRDS IS NOT THE ONLY WAY WE CAN SEE SCENARIOS, WE CAN SEE OTHER SCENARIOS AND THE KEY HERE IS THE ADAPTATION BE OF THE VIRUS WITHIN THE HUMAN HOST AND THE ABILITY TO RECOMBINATION TO CREATE INFECTIOUS TRANSFER FROM PERSON TO PERSON. THOSE ARE THE THINGS THAT I THINK WE NEED TO BE AWARE OF. WE NEED TO DO RESEARCH ON THAT AND THAT ENTAILS, THEREFORE, VERY CAREFUL THOUGHTFUL AND THOROUGH CONSIDERATIONS OF SCENARIOS OF RESEARCH THAT COULD LEAD TO SAFETY ISSUES, TO THE ABILITY FOR US TO ENSURE THE PUBLIC TRUST IN THE RESEARCH WE'RE DOING. THIS IS THE CONNECTION I THINK THAT TRULY NEEDS TO BE BASED ON -- NEEDS TO BE SCIENCE-BASED AND EVIDENCE-BASED BUT WE CAN'T ALSO BE OBLIVIOUS TO MANY SCENARIOS THAT COULD OCCUR IN REAL LIFE. WHEN YOU LOOK AT THE SARS, OBVIOUSLY SARS IS A NOVEL VIRUS, NOVEL AGENT, IF YOU WILL, CLEARLY THE EVIDENCE INDICATES AN ANIMAL SOURCE, ZONEOSIS, THE RESEARCH IS NOT COMPLETE AT THIS TIME. THE SARS OUTBREAK IN 2002, 2003, MANY OF MY COLLEAGUES WHEN I TALK ABOUT SARS WITH THEM, THEY SAY, WHAT DO YOU THINK. TONY FASHEE AND HE LOOKS AT ME AND SAYS WE DODGED A BULLET AND IS QUITE CONCERN ABOUT REEMERGENCE OF THIS. WE HAVE THE SQEENS AS YOU KNOW. WITHIN TWO MONTHS OF I GIVE THE IDENTIFICATION OF SARS OF IMPACT OF NEW METHODOLOGIES ON VIRAL RESEARCH. BASICALLY THE IDENTIFICATION OF THE VIRUS IS IN GREAT PART DUE TO OUR ADVANCES IN, GEE, NO, MA'AM MIX AND OUR ABILITY, GEE, NOMICS AND TO QUICKLY MATCH SEQUENCES IN THE UNKNOWN VIRUS TO KNOWN SEQUENCES IN OUR LIABLE RACE. I LIABLE HIS. WILL BE LIBRARIES AND AS FOR THOSE OF WHO YOU KEEP TRACK OF THAT THERE'S BEEN QUITE A TRACK OF AMOUNT OF PROGRESS IN UNDERSTANDING THE PATH THOUGH PHYSIOLOGY PATHOPHYSIOLOGY OF THE DISEASE BUT WE NEED TO DEVELOP MORE AI SAYS, VACCINE CANDIDATES AND CERTAINLY AGAIN IDENTIFY TARGETS FOR ANTIVIRAL DRUGS SO A COMPLETE RESPONSE, IF YOU WILL, TO THE INFECTION ENTAILING AGAIN A COMPLETE RESEARCH METHODS AND RESEARCH APPROACHES AND LABORATORIES THROUGHOUT THE COUNTRY AND THE WORLD ENTAILING AGAIN A GOOD CONSTRUCT AROUND THIS RESEARCH OF SAFETY GUIDELINES, ORGANIZATIONAL RESPONSES THAT CAN GIVE US COMFORT THAT NABLG WE'RE DOING THIS RESEARCH LIKE, IN FACT, WE'RE DOING THIS LIKE WE DID IN MANY OTHER AREAS LIKE BIOSAFETY AND PUBLIC HEALTH IN MIND. SO RESEARCH INTO THE DEVELOPMENT OF VACCINES AND ANTIVIRAL DRUGS, I CAN TELL YOU ARE A TOP PRIORITY RIGHT NOW IN THE PUBLIC HEALTH ARENA. IT'S CLEARLY RESEARCH THAT'S OF CRITICAL IMPORTANCE BUT IT CANNOT BE PERFORMED UNLESS WE ARE CONFIDENT THAT IT CAN BE PERFORMED RESPONSIBLY TO PROTECT THE HEALTH NOT ONLY OF THE PUBLIC BUT ALSO THE LAB RESEARCHERS AND FIND A CREATIVE STRATEGY BECAUSE I THINK IT IS A FIELD WHERE CREATIVITY WILL BE REQUIRED AS WELL AS RIGOROUS MANAGEMENT OF THESE ISSUES. THE STANDARDS AS YOU KNOW, THEY'VE BEEN ESTABLISHED THE NIH GUIDELINES FOR RESEARCH INVOLVING RECOMBINANT MOLECULES HAVE BEEN LONGSTANDING. WE HAVE AN INFRASTRUCTURE AND IN A CULTURE OF BIOSAFETY AND BIOMEDICAL LABORATORIES I DON'T THINK THAT IT IS ENOUGH TO THINK THAT THOSE GUIDELINES AND THOSE METHODS ARE GOOD ENOUGH. WE NEED TO ALSO HAVE MONITORING SYSTEMS. WE NEED TO HAVE A BETTER WAY AND AN INTERACTIVE WAY TO UNDERSTAND WHAT THE ISSUES ARE IN EACH LOCALE. WE NEED AN INTERPRETATION, AN APPLICATION OF BIOSAFETY PRINCIPLES TO THESE NEW AREAS OF RESEARCH, ESPECIALLY WHEN YOU'RE DEALING WITH POTENTIAL INFECTIOUS AGENTS OF THE RISK AND MAGNITUDE THAT I JUST DESCRIBED TO YOU. SO IN SUMMARY, I'M HERE TO TELL YOU THAT WE HAVE OUR GOAL IS CLEAR. WE WANT TO NORMALLY ENHANCE THE AWARENESS OF THE SAFETY BUT ALSO THE ETHICAL ISSUES THAT GO WITH IT. I THINK WE CANNOT SUCCEED UNLESS THERE IS A CULTURE OF RESPONSIBILITY ACROSS THE WORLD THAT DEVELOPS NOT JUST IN THE SCIENTIFIC COMMUNITY BUT THE REGULATORY COMMUNITIES AND THE ADMINISTRATORS, POLICYMAKERS, AND WE NEED TO UNDERSTAND THAT IN LIGHT OF THE NEW METHODOLOGIES AND THEIR POWER WE NEED TO DEVELOP GUIDANCE FOR VERY RIGOROUS BIOSAFETY REVIEW AND APPROPRIATE BIOLOGICAL AND PHYSICAL CONTAINMENT AND THINK WHAT WE'RE ASKING YOU TO DO IS TO TRULY VALUE THAT IN THAT CONTEXT, BE THOROUGH, BE THOUGHTFUL, BE PRUDENT, BE CAUTIOUS, BUT AT THE SAME TIME REALIZE THAT HEALTH OF THE PUBLIC DEPENDS ON OUR ADVANCES IN SCIENTIFIC UNDERSTANDING. SO WITH THAT I'D LIKE TO WISH YOU GOOD LUCK. I KNOW YOU HAVE A LOT OF WORK AHEAD OF YOU AND I WOULD LIKE TO CONGRESS LATE ALSO DR. PATERSON, DR. SCARBUT, THE OFFICE OF BIOTECHNOLOGY WHO'VE DONE A REAL WORK, A GREAT WORK IN NOT ONLY PUTTING THIS TOGETHER BUT ALSO PUTTING THE FRAMEWORK FOR THE NATIONAL ADVISORY BOARD FOR BIOSECURITY SCIENTIFIC ADVISORY BOARD FOR BIOSECURITY AND I THINK YOUR WORK WILL SURELY INSPIRE THAT COMPONENT OF THE INFRASTRUCTURE THAT WE NEED TO HAVE IN PLACE. SO WITH THAT AGAIN THANK YOU VERY MUCH. I THANK THE ATTENDANTS FOR BEING HERE AND GOOD LUCK .

THANK YOU VERY MUCH, DR. ZERHOUNI. WE'RE SORRY YOU HAVE TO LEAVE. WE APPRECIATE YOUR REMARKS. AND I WOULD LIKE TO ADD OUR OFFICES WELCOME, WE'LL KEEP THE OPENING REMARKS TO A BARE MINIMUM BUT I THINK IT'S IMPORTANT THAT YOU HEAR FROM OUR OFFICE AT THE AGENCY, AND ACKNOWLEDGE THE VERY HARD WORK OF THE STAFF. STEVE ROW, MARINA RILEY AND MANY OTHERS IN PUTTING THIS TOGETHER AND ALSO THE STEERING COMMITTEE COMMITTEE OF SEVERAL DEDICATED SCIENTISTS THAT HAVE HELPED CRAFT THE QUESTIONS AND THE FRAMEWORK FOR THIS MEETING. WE'RE VERY GRATEFUL TO THEM AND REALLY LOOK FORWARD TO THE OUTCOME OF THE MEETING. I WANT TO ALSO UNDER SCORE A COUPLE OF POINTS THAT DR. ZERHOUNI MADE. FIRST OF ALL, THE TIMING OF THIS EVENT IS NO ACCIDENT. PERHAPS THE PARTICULAR DAY IS. BUT THE ERA THAT THIS IS HAPPENING IN IS NO ACCIDENT. HE'S ALREADY ALLUDED TO, WE ARE ENTERING A TIME WHERE CONSIDERATIONS NOT ONLY OF EMERGING INFECTIOUS DISEASES THAT HAVE TREMENDOUS PUBLIC HEALTH IMPACT ARE BECOMING OF INCREASING IMPORTANCE, BUT WE'RE ALSO ENTERING A TIME WHEN THE SHADOW OF POTENTIAL MISUSE OF BIOTECHNOLOGY ALSO HAVE TO BE TAKEN INTO ACCOUNT. SO WITH GROWING CONCERNS OVER NOT ONLY BIOTERRORISM BUT THE EMERGENCE OF NEW INFECTIOUS DISEASE THREATS INSTITUTIONS CONDUCTING THIS RESEARCH ARE REALLY FACING AN ENHANCED SPECTRUM AND DEPTH AND BREADTH OF RESPONSIBILITIES THAT THEY'RE REALLY BEGINNING TO ACCEPT UP TO THE PLATE WITH AND ADDRESS. IN THIS AREA, AS WELL AS MANY OTHER AREAS IN JUST BASIC RECOMBINANT DNA RESEARCH BIOCOMMITTEES ARE GOING TO NEED A LOT OF SUPPORT AND A LOT OF GUIDANCE. IDC'S, THERE ARE OVER 500 OF THEM SPRINKLED ACROSS THIS NATION, THEY VARY ON PROBABLY APPROPRIATELY SO IN TERMS OF THEIR DIVERSITY AND THEIR THE REFLECTION OF THE HEAT ROW GENERAL INNER INSTITUTIONS THEY SERVE BUT THE UNEVENNESS IN THE QUALITIES. OVER THE PAST FEW YEARS, OUR AGENCY HAS DEVELOPED A GROWING RIGOROUS OUTREACH IN EDUCATION PROGRAM THAT IS BEING STEPPED UP IN KEEPING WITH THE ADVANCEMENT OF THE BIODEFENSE AND INFECTIOUS DISEASE PORTFOLIO OF OUR AGENCY AND I JUST WANTED TO SIGNAL A COUPLE OF THE ACTIVITIES THAT ARE PART AND PARCEL OF THIS PROGRAM. ALSO SIGNAL TO YOU SOME NEW THANKS WE WILL BE UNDERTAKING AND INVITE YOUR COMMENTS AND SUGGESTIONS ON HOW WE COULD FURTHER ADVANCE THIS PROGRAM AND REALLY ASSIST THE INSTITUTIONAL BIOSAFETY COMMITTEES AT THE LOCAL INSTITUTIONS THAT ARE ON THE FOREFRONT OF THIS RESEARCH AND DEAL DAY-TO-DAY WITH THE CONTAINMENT ISSUES. HOW WE COULD BETTER ASSIST THEM IN ACHIEVING THOSE GOALS. THIS CONFERENCE IS AN EXCELLENT EXAMPLE OF THE PORTFOLIO OF THINGS THAT OUR AGENCY'S BEEN TRYING TO DO TO HELP THE WORK OF IDC'S. THIS INCLUDES A VARIETY OF TRAINING CONFERENCES AND WORKSHOPS, AT MAJOR MEETINGS, AT THE AMERICAN BIOLOGICAL ASSOCIATION. THE AMERICAN SOCIETY OF GENE THERAPY AND OTHER CONFERENCES OF THAT ILK. WE HAVE A LOT OF INFORMATION ON OUR WEBSITE. WE HAVE POLICE SERVER THAT WE PUSH INFORMATION OUT TO THE COMMUNITIES IN. WE HAVE A WHOLE SERIES OF FREQUENTLY-ASKED QUESTIONS THAT ARE THE EMBODIMENT OF CONCERNS AND QUESTIONS THAT REPEATEDLY CROP UP ON A DAY-TO-DAY BASIS. WE ANSWER MANY QUERIES FROM IBC'S, THEIR BIOSAFETY OFFICERS AND INVESTIGATORS AMOUNTING TO OVER SEVERAL HUNDRED A YEAR IN TERMS OF QUERIES FROM THE COMMUNITIES, AND WE'LL BEGIN LAUNCHING A SERIES OF SITE VISITS OVER THE NEXT YEAR THAT WILL BE DONE IN CONJUNCTION WITH THE OFFICE OF EXTRAMURAL RESEARCH. ALL OF THAT SAID THERE'S STILL A LOT WE COULD DO BUT THE EXACT FORM THAT THOSE ACTIVITIES WOULD TAKE WE WOULD VERY MUCH APPRECIATE YOUR INPUT AND SUGGESTION ON HOW TO BE OF CONCRETE ASSISTANCE TO THE BIOSAFETY COMMUNE AND WITH THAT, I'LL TURN BACK TO TODAY'S EVENT AND JUST SAY THAT WE LOOK FORWARD TO WHAT WE HOPE WILL BE THE PRODUCT OF YOUR DECLARATIONS OF SERIES OF THOUGHT, WELL DELINEATED POINTS TO CONSIDER THAT WILL HELP GUIDE THE DAY-TO-DAY DECISIONS THAT BIOSAFETY COMMITTEES ARE FACED WITH AS THEY EVALUATE ON A CASE-BY-CASE BASIS INDIVIDUAL PROTOCOLS AND WITH THAT, I'LL TURN THE MEETING OVER TO STEVE ROSE AND OUR CO-CHAIR'S DR. DeLUCA AND HOLMES.

THANK YOU, AMY. I AM -- HAVE GONE NOTHING MORE TO ADD OTHER THAN WHAT AMY ELOQUENTLY SAID AND WILL TURN THE MEETING OVER TO DR. DeLUCA AND DR. HOLMES .

MY NAME IS DR. DeLUCA. I'M NOT. MY NAME IS NEAL DeLUCA. I'M A BIOLOGIST, I DON'T WORK ON ANY OF THESE AGENTS BUT I THOUGHT I SHOULD SAY A FEW WORDS. I'M HERE AS A MEMBER OF THE RACK AND CHAIR OF OUR BIOSAFETY COMMITTEE AT THE UNIVERSITIES OF PITTSBURGH. AND I THINK YOU'VE HEARD BOTH FROM DR. ZERHOUNI AND DR. PATERSON FOR THE NEED FOR THIS SYMPOSIUM. I WANT TO UNDERSCORE SOME OF THE SPECIFIC THINGS THAT WE'LL BE TRYING TO ACCOMPLISH HERE. THESE -- AS AN IBC CHAIR AND I'M SURE A LOT OF WHO ARE ON IBC'S WHO ARE INVOLVED IN REVIEWING PROTOCOLS, UNDERSTAND THE PROBLEM WHEN YOU GET SOMETHING NEW THAT NOT MANY PEOPLE IN YOUR COMMITTEE KNOW ANYTHING ABOUT. THE PURPOSE OF THIS MEETING, THE PRIMARY PURPOSE OF THIS MEETING IS TO GIVE SOME DIRECT ADVICE TO PEOPLE TO IBC'S ON EMERGING INFECTIOUS AGENTS THAT HAVE NOVEL BIOLOGY AND HAVEN'T REALLY BEEN REVIEWED BEFORE, AND WE ARE E USING IT AS AN EXAMPLE, THE 1918 VIRUS, THE PATHOGENIC INFLUENZA AND SARS CO-VAND THERE ARE A COUPLE OF THINGS THAT NEED TO BE CONSIDERED THAT WE'RE GOING TO QUERY OUR EXPERTS HERE AND ONE IS A BIOLOGY OF THESE AGENTS. THESE VIRUSES, 1918 VIRUS, THE AVION INFLUENZA BARS AND SARS CO-VHAVE UNIQUE BIOLOGIES OR ALLOW NOVEL STUDIES SO WHAT BIOSAFETY STUDIES ARE USED TO BE RECOMBINANT RESEARCH IN THESE VIRUSES AND HOW DOING THE CONTAINMENT DIFFER WITH NIECE SPONSORSHIPS, RESORTLESS, ANIMAL STUDIES, CLONING, ET CETERA? AND THEN THERE'S THE CONSIDERATION OF BIOSAFETY CONTAIN MANY LEVELS. THESE PATHOGENIC VIRUSES HAVE BEEN STUDIED AT CONTAINMENT LEVELS IN EXCESS OF 3 AND WE'VE SEEN THAT SEVERAL CONTAINMENT BS L3, AND B SL4. SOME ARE NOT DEFINED IN ANY OF THE BASIC, IN ANY OF THE -- OR THE REFERENCE LITERATURE THAT SO WAR THESE LEVELS AND HOW DO THEY DELIVER? AND THE IMPORTANT THING IS HOW DO IBC'S ASSESS THE RISK OF AN INDIVIDUAL AGENT AND THAT'S BROADLY STATED IN THE NHI GUIDELINES HERE WITH THIS STATEMENT, BUT WE'LL BE DISCUSSING IN DETAIL OR WHAT FACTORS SHOULD IBC'S CONSIDER WHEN ASSESSING THE RISK OF RESEARCH FOCUSING ON THE DETERMINATION OF YET UNIDENTIFIED VIR LESSONS FACTORS. THERE ARE MANY CASES WHERE DIFFERENT RESORTANTS MIGHT HAVE DIFFERENT RISKS THAN THE INITIAL BUT THESE ARE ALL QUESTIONS THAT WE'RE GOING TO TRY TO PUT TOGETHER, ANSWER IN A SERIES OF POINTS TO CONSIDER FOR IBC'S TO HELP THEM ASSESS THE RISK. AND WHEN NECESSARY, IBC'S SHOULD CONSULT WITH AD HOC EXPERTS WHO CAN PROVIDE THE APPROPRIATE EXPERTISE FOR SPECIFIC REVIEWS AND THERE WILL BE A DISCUSSION REGARDING HOW CAN WE DO THAT. HOW CAN MORE EXPERT ADVICE GET TO INDIVIDUAL IBC'S WHERE THEY MIGHT NOT NORMALLY HAVE ACCESS TO THIS TYPE OF GUIDANCE. SO I'M GOING TO TURN THIS OVER NOW TO OUR CO-CHAIR DR. KHOLMES DR. K. HOLMES.

THIS IS JUST BY WAY OF LETTING YOU GET TO KNOW THE PEOPLE WHO ARE YOUR ORGANIZERS TODAY. I'M KATHRYN HOLMES AND I'M A VIRUS OBLIGATION. I'VE NEVER BEEN A MEMBER OF A COMMITTEE BUT I'VE I KNOW ACTED A GREAT DEAL OVER THE COURSE OF MY LIFE AS I'VE WORKED ON CORONA HAVE A VIRUSES FOR MORE THAN 25 YEARS. THAT WAS A COMFORTABLE INTERESTING THING TO DO, CORONAVIRUSES CAUSE A LOT OF DISEASES IN ANIMALS BE A COLDS IN PEOPLE BUT ONLY IN THE LAST YEAR DID SARS COME ABOUT AND A TREMENDOUS I AM PACKAGE IS THAT ON OUR RESEARCH AND ON OUR THINKING HAS BEEN REMARKABLE. I WANTED TO POINT OUT A FEW THINGS VERY BRIEFLY. ONE IS THAT SARS HAS THE LARGEST OF ALL THE RNA VIRUS, GEE, HOMES IN. THE CROHN THAT VIRUSES HAVE JEAN HOMES IN OF 30 KILL BASE SO THE RN AITSELF IS INFECTIOUS. IF IT'S TRANSEXPECTED IN ANY TRANSECTED INTO ANY HOST CELL. ANY VIRUS THAT WE'RE CONSIDERING HERE TODAY AND MANY THAT WE'RE NOT CONSIDERING HAS UNIQUE AND DIFFERENT BIOLOGY SO SOME LANDMARK WORK WAS DONE PRIOR TO SARS BY LILY KUO, PETER ROTEER AND PAUL MASTERS THAT THEY CREATED A MH VMOUSE HEPATITIS VIRUS JEAN NO, MA'AM IN WHICH THE EXTERNAL DOME MAIN OF THE SPIKE THAT BINZ TO RECEPTORS WAS CHANGE EXCHANGED WITH A FELINE CORONAVIRUS AND THAT VIRUS WOULD ONLY GROW IN CAT CELLS AND IF THEY INFECTED CAT CELLS WITH THAT, AND THEN TRANSECTED IN R NA WHICH IS ABOUT NINE OR 10 KILL LA BASES OF A MOUSE HEPATITIS VIRUS GENOME WITH SOME SOLUTIONS IN THE REGION AND I GUESS I DON'T HAVE A POINTER HERE. HERE WE GO. HERE. AND IN THE SAME CELL WHAT HAPPENED WAS THAT THE PRELIMINARY THAT'S MADE FROM THIS ENTIRE GENOME OF THE INFECTIOUS VIRUS WOULD THEN COPY THIS SUBGENOMEIC MESSENGER RNA TEMPLATE HERE AND RECOMBINATION OCCURS WHICH WILL GENERATE THEN A NEW INFECTIOUS VIRUS THAT HAS THE NEWER REASON SPEAK NIGHT AND WE PUT A MUTATION INTO THAT BY KEY COME NAT DNA TECHNIQUES SO WE HAVE A WHOLE INFECTIOUS VIRUS NOW THAT ONLY INFECTS MOUSE CELLS SO THAT'S THE KIND OF TECHNOLOGY GOING ON FOR SOME TIME IN CORONAVIRUS AND DR. BARRETT WILL TALK ABOUT THIS AT LENGTH. JUST TO POINTS OUT THAT CORONAVIRUSES ARE UNIQUE IN HAVING THESE HUGE GENOME AND BECAUSE OF THAT HAVING A VERY HIGH FREQUENCY OF RNA COMBINATION WHICH PREVENTS RESEARCH FOR LIKE THIS BUT POTENTIAL OF RECOMBINATION OF STRAINS THAT ARE DEVELOPED WITH UBIQUITOUS HUMAN CORONAVIRUSES. NOW CORONAVIRUSES ARE DIVIDE INTO THREE OR MORE GROUPS. I VANS PUT THE AVION ONES UP HERE BUT THE ALL MARK IS EACH CORONAVIRUS CHARACTERISTIC CLI CAUSES DISEASE IN A SINGLE HOST. THESE ARE ALL GENETICALLY RELATED IN GROUP ONE, BEES IN GROUP TWO, RAT, PIG AND CATTLE VIRUSES THERE AND SARS IS MORE CLOSELY RELATED TO GROUP TWO THAN GROUP ONE. SO WE'VE CALLED THEM HUMAN CORONAVIRUS AND DOUGH VINE BUT WHEN YOU LOOK AT THE DATA WHAT YOU SEE IN MANY CASES OTHER SPECIES CAN BE INFECTED SO CATS ARE SUSCEPTIBLE TO MANY GROUP ONE VIRUSES ALTHOUGH THEY DO NOT GET INFECTED AND GET DISEASE AND WE KNOW THAT SARS HAS MANY HOST IN THE WILD AS MONKEYS, FERRETS, MICE THAT OF COURSE DEVELOPED. THE DISEASE RIGHT SIDE NOT THE SAME IN ALL OF THESE. EMERGING VIRUSES DO NOT OWN ANIMAL CAUSE ANY DISEASE AT ALL. SO IT'S VERY HARD TO PREDICT AND MY POINT HERE IS SIMPLY TO SAY ONE SIZE DOES NOT FIT ALL WHEN YOU'RE CONSIDERING THE BIOLOGY OF NEW AND EMERGING AGENTS. WE CAN'T PREDICT WHAT NEW VIRUS FAMILY WILL EMERGE NEXT BUT WE CAN BE CONFIDENT THAT THERE WILL BE ANOTHER EPIDEMIC AND WE DON'T KNOW WHAT IT WILL BE. AN IMPORTANTLY FOR IBC'S EACH VIRUS FAMILY HAS DIFFERENT REVERSE GENETICS TECHNIQUES AND DIFFERENT RISKS ASSOCIATED WITH THOSE TECHNIQUES. AND AS THESE TECHNIQUES PROLIFERATE IBC'S HAVE TO KEEP UP-TO-DATE AND WHAT WE WANT TO DO IN THIS PANEL DISCUSSION IS TO DEVELOP SOME POINTS TO CONSIDER TO HELP IBC'S EVALUATE THESE VARIATIONS BETWEEN DIFFERENT VIRUS FAMILIES. AND TO UNDERSTAND WHAT SAFETY CONSIDERATIONS ARE IMPORTANT FOR EACH VIRUS BECAUSE THEY'RE NOT ALL THE SAME. AND ANOTHER IMPORTANT POINT WHICH YOU'VE HEARD ALREADY IS THAT WHEN A NEW VIRUS EMERGES LIKE SARS HAS, THE PUBLIC HEALTH NEEDS REALLY PRESS FOR URGENT RESEARCH ON THESE NEW AGENTS SO NOT ONLY DO WE HAVE TO ASHORE SAFETY BUT WE HAVE TO BE ABLE TO MOVE QUICKLY AND PROMPTLY TO GET THE RESEARCH DONE. AND THAT'S THE BALANCE THAT THE IBC'S ARE INVOLVED IN. IN DOING THAT, I AS A RESEARCHER WAS THINKING ABOUT THIS MEETING AND WAS REALLY IMPORTANT FOR ME IS HOW I CAN COMMUNICATE WITH IBC'S AS A RESEARCHER. AND THIS COMMUNICATION MUST BE VERY THOROUGH. EVERY ASPECT OF THE POSSIBLE RISKS MUST BE DISCUSSED. AND IT ALSO MUST BE EFFICIENT, FAST. NOW THAT IN MANY INSTITUTIONS IS NOT TRUE. IN THE EFFORT TO BE THOROUGH, EFFICIENCY IS SOMETIMES SACRIFICED. AND IN THE MIDST OF AN EPIDEMIC LIKE SARS WE CAN'T DO THAT SO WE WANT TO THINK TODAY ABOUT HOW TO MAKE THIS PROCESS OF COMMUNICATION BETWEEN INVESTIGATORS, FUNDING AGENCIES AND IBC'S BOTH EFFICIENT AND THOROUGH. AND ONE OF THE FIRST THINGS THAT WE WANT TO DO IS BE SURE THAT THE INVESTIGATORS KNOW WHAT INFORMATION THE IBC NEEDS IN ORDER TO FILL OUT AN APPLICATION THAT CAN BE FULLY UNDERSTOOD AND HAVE ALL THE INFORMATION THAT THE IBC NEEDS TO CONSIDER. AND THEN THE IBC NEEDS ACCESS TO UP-TO-DATE INFORMATION IN ORDER TO EVALUATE THIS APPLICATION. THIS IS TREMENDOUSLY, TREMENDOUSLY IMPORTANT. HOW CAN WE HELP YOU AS IBC MEMBERS GET THIS INFORMATION ONCE YOU'VE GOT THIS PERFECTLY FILLED-OUT APPLICATION, GET THE INFORMATION TO ALLOW YOU TO EVALUATE THAT? AND FINALLY, THE EVALUATION OF THE RISK OF A SET OF EXPERIMENTS PROPOSED BY THE INVESTIGATOR HAS TO BE ILL STRAIF. SOMETIMES THE INVESTIGATOR WILL DEVELOP SOMETHING THAT HE THINKS, SHE THINKS AS WONDERFUL, LESS VIR LENTZ, TILELY A 10 YAITED BACKBONE IN WHICH FURTHER EXPERIMENTS COULD BE DONE AT A LOWER BIOSAFETY LEFT LANE OF THIS IS SOMETHING WE ALL HOPE FOR AND IT'S SOMETHING THAT'S TREMENDOUSLY EXPENSIVE TO MAKE AND PROVE AND DR. BARRACK WILL TALK ABOUT THIS WITH REGARD TO SARS. SO IF THE INVESTIGATOR COMES BACK ANNOUNCES THIS TO THE IBC, THERE WILL HAVE TO BE SOME GIVE AND TAKE ABOUT DOWNGRADING B SL-3 LEVELS AS NEW DATA BECOMES AVAILABLE ON THEM AND SO I THINK THIS BECOMES AN ITRATIFF PROCESS WHERE THE IBC'S ARE AN IMPORTANT PART OF THE ADVANCE OF THE SCIENCE IN EVALUATING AND DEVELOPING VACCINES, DRUGS AND DIAGNOSE DIAGNOSTICS FOR HIGHLY PATHOGENIC NEW VIRUSES, THANK YOU.

NOW I THINK WHAT WE'RE GOING TO DO IS INTRODUCE MEMBERS OF THE PANEL. SO IF YOU WOULD START PLEASE AT THE END AND INTRODUCE YOURSELF TO THE AUDIENCE AS WE GO AROUND.

OPENLY RY VACCINE CENTER? ATLANTA, GEORGIA. SAM LIPTON FROM THE PUBLIC HEALTH DEPARTMENT IN CAMBRIDGE, MASSACHUSETTS.

BOB KREUG FROM THE UNIVERSITY OF TEXAS AT AUSTIN.

RYAN MURPHY FROM THE NATIONAL INSTITUTES OF ALLERGY AND INFECTIOUS DISEASE HERE IN BETHESDA.

RANDALL MORI KNOW I WORK AT THE NCI CAMPUS IN FORT DEED PRICK.

ROBERT LAMB.

JIM SCHMIDT FROM THE NATIONAL INSTITUTES OF HEALTH.

JANET PETERSON FROM UNIVERSITY OF MARYLAND COLLEGE PARK.

MADISON POWERS FROM INSTITUTE OF ETHI. GEORGETOWN, UNIVERSITY.

LARRY JOHNSON FROM THE UNIVERSITIES OF NORTH CAROLINA.

PHIL JOHNSON FROM COLUMBUS GEORGE HOSPITAL.

TERRY KWAN, I'M A MEMBER OF IBC'S IN THE BOSTON AREA AND A MEMBER OF THE RACK.

AGAIN ENEMY ROWS CALIFORNIA.

ROSENBURG TUFT MEDICAL SCHOOL IN BOSTON.

EMMETT BARKLEY HOWARD HUGHES MEDICAL INSTITUTE. [ INDISCERNIBLE ]

STEVE ROSE OFFICE OF TECHNOLOGY ACTIVITIES.

AMY PETERSON, ALSO BIOLOGY TECHNIQUES.

WISCONSIN AT MADISON.

-- [ INDISCERNIBLE ] MOUNT SINAI SCHOOL OF MEDICINE IN NEW YORK.

ROB WEB SISTER, RESEARCH HOSPITAL, MEMPHIS, TENNESSEE.

INSTITUTION OF ALLERGY AND INFECTIOUS DISEASE.

PUBLIC HEALTH, UNIVERSITIES OF NORTH CAROLINA.

LARRY ARTHUR NCI FREDERICK AT FREDERICK MARYLAND.

-- [ INDISCERNIBLE ]

INFLUENZA BRANCH CBC ATLANTA.

-- [ INDISCERNIBLE ]

OKAY. I THINK WE CAN START WITH OUR FIRST SPEAKER, OUR FISHING FIRST SPEAKER IS .

OKAY. TODAY I'M GOING TO DESCRIBE . NOW, IN FRONT OF US, LINES TWO, I CAN GOING TO THE NUCLEUS AND THE PROTEINS ARE MADE AND LATE PROTEINS ARE ALSO PRODUCED. AND THESE PROTEINS ARE ASSEMBLED AT THE CELL SURFACE AND RELEASED. NOW, AS I MENTIONED THAT, THE GENOME IS FRAGMENTED IN INTO EIGHT SEGMENTS BECAUSE OF THE PROPERTY, WHEN A SINGLE CELL IS INFECTED WITH TWO DIFFERENT VIRUSES, THE GENONLY RNAs ARE PRODUCED IN THE NUCLEUS AND WHEN THE CELL PRODUCE ADVANCES, THERE ARE MANY ADVANCES THAT HAVE THE DIFFERENT COMMINATIONS OF THE GENOME. -- COMMINATIONS OF THE GENOME. THEORETICALLY, 257, 256 DIFFERENT VIRUSES CAN BE PRODUCED. IN 1999, THE TECHNIQUE TO PRODUCE INFECTIOUS -- [ INDISCERNIBLE ] WAS ESTABLISHED CALLED REVERSE ANTICK S. I WOULD -- ANTICS. I WOULD LIKE TO BRIEFLY INTRODUCE THE TECHNOLOGY. HERE WE HAVE EIGHT PLASMIDS FOR THE PRODUCTION OF EIGHT DIFFERENT RNA SEGMENTS OF THE VIRUS AND FOUR PLASMIDS FOR THE PRODUCTION OF FOUR VIRAL PROTEINS THAT ARE REQUIRED FOR THE REPLICATION OF THESE RNA SEGMENTS WITHIN THE NUCLEUS. UPON TRANSFECTION OF THE DISEASE, 12 -- [ INDISCERNIBLE ] TO CELLS, WE OBTAIN INFECTIOUS INFLUENZA VIRUS . ALSO MENTIONED THAT WE HAVE TWO GROUPS OF INFLEW ENZA DRUGS. THE FIRST GROUP IS M2, INHIBITS THE ION CHANNEL ACTIVITIES. RAMATAJENE IS ONE OF THE DRUGS. THESE DRUGS INHIBIT ALL OF THE STAGE OF THE REPLICATION -- [ INDISCERNIBLE ] ANOTHER GROUP OF THE INFLUENZA DRUG IS NA INHIBITORS, WHICH INHINTS THE, INHIBITS THE -- INHIBITS THE PROTEIN AND ALSO OF THE DRUGS AND THESE DRUGS INHIBIT THE LATE STAGE OF THE REPLICATION, THE RELEASE OF THE VIRUS REPLICATION . [ NO AUDIO ] -- AND SYSTEMIC INFECTION AND SOME OF THE GROUPS ADVANCES INSPECT CHICKENS, ONLY RESPIRATORY INFECTIONS AND ALMOST NO DISEASES. FINE. THE DIFFERENCE BETWEEN THESE TWO GROUPS RESIDES, MAINLY RESIDE IN THIS PROTEIN HEMAGLUTIN. THE HEMAGLUTIN EMPHASIZES A SINGLE PENT SIDE -- PEPTIDE INTO HJ1 AND HA2, THE PROTEASE AND THE HA -- [ INDISCERNIBLE ] ESSENTIAL FOR PRODUCTIVITY. IF IT'S NOT CLEAR, THE VIRUS IS NOT INFECTIOUS. PATHOGENIC AND NONPATHOGENIC DIFFER AT THE SITE RIGHT HERE. PATHOGENIC VIRUSES CONTAIN -- [ INDISCERNIBLE ] -- SUCH AS HARDENING, KNOWN PATHOGENIC DON'T CONTAIN A SERIES OF -- [ INDISCERNIBLE ] I AM NOT GOING TO DISCUSS HOW THIS OPERATES, BUT THIS DETERMINES THE AVIAN INFLUENZA VIRUS. NOW, I'M GOING TO DISCUSS THE REPRESENTATIVE EXPERIMENTS WE DID WITH H5 AND 1 VIRUSES, AND THAT PROVIDES SOME INFORMATION ON THE DETERMINED AFFECT IN PATHOGENIC OF INFLUENZA VIRUS. SO IN 1999, THE H5 N-1 VIRUSES TRANSMITTED TO HUMANS AND INFECTED 18 PEOPLE. THE CAUSE OF THE AGENT WAS IDENTIFIED AS HIGHLY PATHOGENIC H5 NEXT 1 CONTAINING -- [ INDISCERNIBLE ] AT THE FEVER SITE. NOW, 17 -- [ INDISCERNIBLE ] -- WERE MADE FROM THESE HUMANS AND THESE VIRUSES, HUMAN, WERE DIVIDED INTO TWO GROUPS IN THEIR PATH O' -- PATHOGENIC IN MICE. THOSE THAT INFECT MICE AND CODE SYSTEMIC INFECTION AND THOSE INFECT MICE AND POSE ONLY RESPIRATORY INFECTION. SO WE WON'T ADDRESS TWOTONICS, TWO ISSUES. -- TWO TOPICS, TWO ISSUES. THE IMPORTANCE OF HIGH HA -- [ INDISCERNIBLE ] FOR THE PATHOGEN ETIN MICE, AND MOLECULAR BASIS AMONG THE -- [ INDISCERNIBLE ] VIRUSES. WE TOOK THE VIRUS THAT CAUSED SYSTEMIC INFECTION IN MICE THAT -- [ INDISCERNIBLE ] AT THE SITE. WE MADE A VIRUS THAT IS FOUND AT THE HA PREVIOUS SITE, AND THEN TESTED A PATHOGENIC OF THE VIRUSES. AS I MENTIONED, THE VIRUS OF THE INFECTED MICE, THE SYSTEMIC VIRUS INFECT MICE. THE SYSTEMIC INFECTION, THE LDA 50 OF THE VIRUS IN MICE IS 1.7 PSU. WHEN WE MUTATED HA PREVIOUS SITE OF THE VIRUS THROUGH THE ONE NOUPD IN KNOWN PATHOGENIC AND INFLEW ENZA VIRUS, WEOUND INFLU AENZ VIRUS, WE FOUND THAT LDA OF THE WAS -- THIS SHOWS THAT IT'S IMPORTANT FOR H5, THE KNOWN VIRUS IN MICE. WE STUDIED THE MOLECULAR VIRUS IN MOUSE AMONG THESE VIRUSES. WE TOOK FROM EACH GROUPS, THE ONE THAT POSED SYSTEMIC INFECTION IN MICE AND ONE ON THE RESPIRATORY INFECTION. AND AS I MENTIONED IN THE BEGINNING, GENOME VIRUSES ARE FRAGMENTED INTO EIGHT SEGMENTS. SO WE MADE ADVANCES BETWEEN THESE TWO VIRUSES, SO WE MADE EIGHT DIFFERENT REASSORTMENT VIRUSES, EACH CONTAINING ONE SEGMENT FROM PATHOGENIC VIRUS AND THE REST OF THE JEEPS FROM KNOWN -- GENES FROM KNOWN PATHOGENIC VIRUS AND TESTED THE PATHOGENICITY OF THE VIRUS IN MICE AND FOUND THAT ONE VIRUS WAS DIFFERENT IN PATHOGENICITY. THIS ONE CONTAINS PV2 SEGMENT FROM PATHOGENIC VIRUS AND THE REST GENES FROM KNOWN PATHOGENIC VIRUS. PV2 IS ONE OF THE PREALMOSTINARY SUBJECT OF THE INFLUENZA RNA. THERE AREATE AMINO ACID DIFFERENCES BETWEEN THE TWO -- EIGHT AMINO ACID DIFFERENCES BETWEEN THE TWO. WE WERE ABLE TO -- [ INDISCERNIBLE ] FOR PATHOGENICITY BETWEEN THE TWO VIRUSES. THAT IS THE AMEANO ACID 627. WE TOOK THE PATHOGENIC VIRUS AND CONVERTED THIS TO THE ONE THAT IS FOUND IN KNOWN PATHOGENIC VIRUS. WE ALSO TOOK THE KNOWN PATHOGENIC VIRUS AND -- TO THE ONE FOUND AND TESTED THE VIRULENCE OF THE VIRUSES AND FOUND THAT THIS METHOD CHANGED THE PATHOGEN IS -- PATHOGENICITY OF THE VIRUS IN MICE. SO THESE RESULTS SHOW THAT. AMINO ACID ALTERATIONS -- [ INDISCERNIBLE ] 627 TO ENHANCE THE PATHOGENICITY OF THE H5 NEXT 1 VIRUS IN MICE, BUT I SHOULD MENTION THAT ALL HUMAN PROTEINS HAVE LE -- LYACINE AT THIS POSITION. BASED ON THESE RESULTS, WE CAN NOW SAY THE DETERMININANTS OF INFECTING PATHOGENICS OF THE INFLUENZA VIRUS. WE NOW KNOW IT'S IMPORTANT IN MICE, AND PV2, AMEANO ACIDS PUSHING 627 AND POSSIBLY THE OTHER AMINO ACIDS IN THIS PROTEIN MAY ALSO BE IMPORTANT. AND I DIDN'T MENTION ABOUT NS1 PROTEIN, BUT TOOK THE WEBSTERS GROUP THAT SHOWED THAT NS1 PROTEIN IS ALSO IMPORTANT, [ INDISCERNIBLE ] -- AND PETER'S GROUP SHOWED THE PROTEIN IS IMPORTANT FOR PATHOGENICITY OF THE INFLUENZA VIRUS. THESE ARE THE REQUIREMENTS FOR -- [ INDISCERNIBLE ] -- I SHOULD ALSO MENTION THE SENSITIVITY OF H5 N1 VIRUSES. SO THIS IS THE IN VITRO SIATIC ACTIVITY, -- [ INDISCERNIBLE ] THE PERCENT INHAS BEENITION, THE -- INHIBBATION CONCENTRATION. THE HUMAN HV AND 2 VIRUSES, THE IC 50 IS BETWEEN 1-10 MINIMAL. THE H5N1 VIRUS WAS 3.4 -- [ INDISCERNIBLE ] SO H5N1 VIRUSES ARE ENGAGED -- [ INDISCERNIBLE ] SO NOW TOI WOULD LIKE TO DISCUSS THE RISK ASSESSMENTS. I WOULD LIKE TO DISCUSS THE MOLECULAR CLONING EXPERIMENT. THE AGENT IS GENETIC MATERIAL, SO INFLUENZA VIRUS GENOME IS NOT INFECTIOUS. AND THEY DON'T INSERT INTO HUMAN GENOME, SO IT'S RISK GROUP 1 AND SINCE THE LOWEST BIOSAFETY LEVEL IN MY LABORATORY IS DSL 2, SO WE PERFORMED EXPERIMENTS ON THAT DSL 2. NOW FOR THE EXPERIMENTS WITH H5N1 VIRES -- VIRUSES -- [ INDISCERNIBLE ] SECTION IN MICE AND CHICKENS. THESE VIRUSES ARE KNOWN TO CAUSE LETHAL INFECTION IN AGENT SPECIES AND -- AVIAN SPICES AND HUMANS AND AERIAL TRANSMISSION IS POSSIBLE, BUT AVAILABLE AS IT SHOWS. AND HUMAN-TO-HUMAN TRANSMISSION IS LOW. SO WE CONSIDER THIS VIRUS IS A RISK GROUP 3, SO WE PERFORMED THE EXPERIMENTS AND EXPERIMENTS WITH H5NEXT 1 VIRUSES AND DSL 3, 4, LABORATORY EXPERIMENTS AND THE ANIMAL DSL 3 FOR THE ANIMAL EXPERIMENTS. WE ALSO EMPLOYED PRACTICES IN ADDITION TO THIS STANDARD DSL 3 PRACTICES. THE PERSONNEL WALKING WITH H5 N1 VIRUSES ARE REQUIRED TO TAKE ANNUAL INFLUENZA VACCINATIONS AND PROOFA LAXIS IS REQUIRED WHEN -- PROFALAXIS IS REQUIRED AND [ INDISCERNIBLE ] PERSONNEL SHELL OUT AND WE TEST AGENTS, THE H5 NEXT 1 VIRUSES FOR THEIR TOXICITY TO THE PROPHLYACTIC DRUG. I SHOULD ALSO MENTION THAT THE LBL 3 FACILITY HAS THE FEATURES AND PROCEDURES ESTABLISHED IN ADDITION TO THOSE REQUIRED FOR DSL 3. THE ENTRY EXITS THROUGH CHANGE, DOUBLE-DOOR -- [ INDISCERNIBLE ] SHOWER FACILITIES, DECONTAMINATION AND WALK SURFACES, ALL PERSONNEL CLOTHING REMOVED IN OUTER CHANGE ROOMS, ESTABLISHED SYSTEM FOR REPORTING AND TREATING EXPOSURES. ANNUAL INSPECTION BY FEDERAL REGULATORY AGENCIES, CONTACT WITH NONEXPERIMENTAL MICE AND CHICKENS AND RATS IS PROHIBITED WITHIN ONE WEEK OF CONTACT WITH EXPERIMENTAL INFECTED ANIMALS. SO NOW, I'M GOING TO DISCUSS THE EXPERIMENTS WE DID WITH THE VIRUSES PO -- WITH THE 1980 VIRUS CHANGE. SINCE THE -- [ INDISCERNIBLE ] 1918 RETAIL DETAIL, I'M GOING TO BRIEFLY DESCRIBE WHAT WE DID. THE SPANISH INFLUENZA VIRUS DOESN'T EXIST. BUT JEPRY TALGENBERGER, AMPLIFIED THE RNA FROM LUNG TISSUES FROM THE PERSON WHO DIED IN 1918 AND 1919 AND REPORTED THE SEQUENCE OF THE HA AND NA JEEPS OF THE I HAVE RUSSES. -- GENES OF THE VIRUSES. SO WE GENERATED THE HA AND NA GENES BASED ON THE PUBLISHED SEQUENCE AND GENERATED AN INFLUENZA VIRUS THAT CONTAINS THESE GENES BY REVERSE ANTICS. I'M GOING TO DESCRIBE THE PROPERTIES OF VIRUSES WITH 1918 VIRUS HA ANGOLA NA IN POLICE, SO WE TOOK A STRAIN CALLED WSN. THIS IS THE MOUSE-ADAPTED LABORATORY STRAIN SUBTYPE H1N1. THE LD 50 SHOWN HERE AND THE MOUSE IS SHOWN HERE. SO WHEN WE REPLACED THE HA AND NA GENES OF THE WSN STRAIN WITH 19, THOSE OF THE 1918 VIRUS, WE DID NOT FIND MUCH OF A DIFFERENCE IN LD50 AND MOUSE LUNG -- [ INDISCERNIBLE ] WE THEN TOOK A VIRUS CALLED MADH, A VIRUS FROM A HUMAN IN 1988, SUBTYPE H3 AND 2. WE REPLACED THE HA AND/OR NA GENES OF THIS VIRUS WITH THE, THOSE OF THE 1918 VIRUS. WE FOUND A DIFFERENCE IN LD50 AND ALSO, THE VIRUS SITE IN LUNG WAS HIGHER THAN THE PARENT ONES. WE ALSO TOOK A STRAIN, K173. THIS WAS THE 2001 HUMAN H1 N1 AND REPLACED THE HA AND NA GENES WITH THE 1918, THOSE OF THE 1918 VIRUS AND, AGAIN, WE SAW THE DIFFERENCE IN LD 50 AND MOUSE LUNG WAS ALSO HIGHER THAN THE PARENT VIRUS. WE ALSO, THE MEASURED VIREGULAR -- VIRAL NEUTRALLIZING -- [ INDISCERNIBLE ] AGAINST VIRUS WITH THE 1918 VIRUS, HA NA GENES. IN THE SERUM, -- SIS SECRET SERVICE OBDENY -- [ INDISCERNIBLE ] OBTAINED IN 1907 AND 2001. AND THE RESULT IS SHOWN IN THIS SLIDE. YX INDICATES THE VIRAL NEUTRALIZING INDICATOR, AXIS INDICATES VAST YEAR FROM HUMANS WHOM WE ARE PERTAINED TO. AND AS YOU SEE, PEOPLE WHO ARE BORN BEFORE 1918 HAD A VERY HIGH -- [ INDISCERNIBLE ] TO THIS VIRUS AND PEOPLE WHO ARE BORN AFTER 1930 HAD LOWER -- [ INDISCERNIBLE ] TO THIS ONE. OKAY. THESE ARE EXPERIMENTS WE DID WITH THE VIRUS THAT COPTIN -- CONTAINS 1918 VIRUS GENES, I WOULD LIKE TO DISCUSS THE BIOSAFETY, THE RISK ASSESSMENT SO THE MOLECULAR CLONING EXPERIMENTS, THE AGENT, GENETIC MATERIAL THAT IS NONINFECTIOUS AND IS NOT INSERTED INTO HUMANS AS I MENTIONED, IT'S RISK GROUP 1 AND, AGAIN, WE PERFORMED THE EXPERIMENT ON DSL 2. FOR THE EXPERIMENTS WITH THE VIRUSES CONTAINING IN 1918 VIRUS, HA ANTIGENS FOR THE VIRUS -- [ INDISCERNIBLE ] INFECTION IN MICE, WE ASSESSED THE RISK OF THESE EXPERIMENTS THAAS FOLLOWS: SO -- EXPERIMENTS AS FOLLOWS: THIS VIRUS HAS A POTENTIAL TO INFECT HUMANS AND THE INTRODUCTION OF THE HA AND/OR NA GENES ENHANCES THE RECOMMINANT VIRUS, ALTHOUGH THE EXPERIMENTS WERE DONE ONLY IN MICE. HUMAN POPULATION MIGHT BE SUSET -- SUSCEPTIBLE AND TRANSMOTION IS POSSIBLE. AND HUMAN-TO-HUMAN TRANSMISSION IS ALSO POSSIBLE, BUT BEFORE WE INITIATED THE EXPERIMENT, WE DID NOT KNOW THE ECSTASY OF HUMAN INFLEW EPZA AGAINST VIRUS CONTAINING, AGAINST THE 1918-LIKE VIRUS. ALSO, WE DIDN'T KNOW SENSITIVITY, EFFICACY OF THE INFLUENZA DRUGS TO THE 1918-LIKE VIRUSES, ALTHOUGH THE SEQUENCE OF THE NA OF THE 1918 VIRUS INDICATE IT'S SENSITIVE TO AMINOEAS INTUITIVES. BASED ON THIS INFORMATION, WE CONSIDERED THE VIRUSES WITH THE 1918 HA AND NA GENE S TO BE EITHER RISK GROUP 3 OR 4. SO WE PERFORMED THE EXPERIMENTS ON THE DSL 4 CONSTRAINED. AND UNDER DSL 4, WE PERFORMED THE, EXAMINED THE EFFICACY OF THE -- [ INDISCERNIBLE ] -- AGAINST THE 1918 VIRUS HA AND NA. SO MICE WERE TREATED WITH AUTO -- [ INDISCERNIBLE ] AND FOUR HOURS LATER, THEY WERE CHALLENGED WITH THIS VIRUS. AND ALSO, GIVEN DAILY ASPIRINS. AND THE RESULT IS SHOWN IN THE NEXT SLIDE. THE MICE TREATED WITH PVS ALL DIED BY DAY 12. MICE TREATED WITH -- [ INDISCERNIBLE ] TOM PETER ALSO STUDIED SINCE TIST OF THE 1918 -- SINCE TEST OF THE 1918 VIRUS, CONTAINING THE 1918 VIRUS NA ALSO -- [ INDISCERNIBLE ] SINCE DR. ADOLFO GARCIAL-SASTRE IS GOING TO DESCRIBE IN DETAIL, I WILL BRIEFLY DESCRIBE IT. IN IN VITRO IN VITRO STUDY WHERE -- [ INDISCERNIBLE ] -- THE PRESENCE OF -- [ INDISCERNIBLE ] THE VASE THAT CONTAINS THE 1918 HA AND NA WAS ABSENT AS THE OTHER TWO VIRUSES TESTED TESTED TO THIS COMPOUN. IN MICE -- COMPOUND N.MICE NEXT PHOTO ACTIVE STUDIES NEXT MICE, AGAIN, THE PROTECTIVE MICE FROM THE CHALLENGE WITH THIS -- [ INDISCERNIBLE ] TOM PETER ALSO STUDIED THE PROTECTION OF MICE AGAINST THE VIRUS WITH 1980 HA AND NA USING INACTIVATED VACCINE MADE FROM A CONTEMPORARY H1 STRAIN. SO WHEN MICE WERE VACCINATED WITH PVS, ALL THE MICE DIED BY DAY 11. A VAX -- VACCINE MADE FROM A STRAIN H3 AND 2, DIFFERENT FROM THE 1918 VIRUS, DIDN'T CONFIRM MUCH. THE VACCINE MADE FROM THE VASE THAT CONTAINS 1918 HA AND NA PROTECTIVE -- PROTECTED MICE 100%, AND WHEN MICE WERE VACCINATED WITH A VACCINE MADE FROM 1999 HUMAN ICE PLATE OF SUBTYPE H1 AND N1, THE SAME AS THE 1918 HUMAN VIRUS, MORE THAN 79% OF THE MICE WERE PROTECTED FROM THE CHALLENGE. SO THE -- NOW I'M GOING TO -- THIS IS THE EXPERIMENT THAT WE. I WOULD LIKE TO DISCUSS THE RISK ASSESSMENTS. THE MOLECULAR CLONING IS THE SAME AS WHAT I DESCRIBED FOR VH5 AND 1. WE PERFORMED -- [ INDISCERNIBLE ] AND FOR THE EXPERIMENTS WITH THE ADVANCES CONTAINED IN 1918 VIRUS AGENT AND AN AGENT FOR THE VAST GENERATION TO EXPERIMENT AND INFECTION IN MICE, WE ASSESSED THE RISKS AS FOLLOWS: SO THE, THESE VIRUSES HAS POTENTIAL TO INFECT HUMANS AND -- SO, AS I DESCRIBED BEFORE, FIRST, WE ESSENTIALLY REASSESSED THE, ASSESSED THE RISKS OF THE EXPERIMENTS BASED ON NEW INFORMATION AND SO THE 5 POINTS REMAINS THE SAME AND BUT THESE TWO POINTS CHANGES. WE NOW KNOW THE KINDS OF VACCINES MAY OFFER PROTECTION, ALTHOUGH I SHOULD MENTION THAT THE EXPERIMENTS WERE DONE ONLY IN MICE AND WE DO NOT KNOW TO WHAT EXTENT THE MOUSE DATA CAN BE EXTRAPOLATED. AND WE ALSO KNOW DEFINITELY THE, THE H5, THE 1918 VIRUS CONTAINS THE GENES SENSITIVE TO THE AMINOASE INHIBITORS. SO, WE NOW ASSIGNED ADVANCES CONTAINING THESE GENES TO RISK GROUP 3 AND WE NOW HAVE THE APPROVAL TO DO THE EXPERIMENT TO, WITH THESE VIRUSES AND DSL 3. WITH THE SPECIFIC PURPOSES THAT I MENTIONED FOR THE H5 N1 VIRUS NEXT A FACILITY WITH THE FEATURES AND PROCEDURES THAT I JUST DESCRIBED PREVIOUSLY. THANK YOU. [ APPLAUSE ]

THANK YOU VERY MUCH, YOSHI. BEFORE WE ASK FOR A COUPLE, WE HAVE TIME FOR A COUPLE OF QUESTIONS FROM THE PANEL ABOUT THE SPECIFIC PRESENTATION. WE WOULD LIKE TO REQUEST THAT EVERYONE TURN OFF THEIR CELL PHONES AND -- UNTIL THE BREAK. SO, IF THERE IS ANYONE THAT HAS QUESTIONS FOR YOSHI?

DO YOU HAVE ANY INFORMATION IN REGARDS TO WHETHER OR NOT THE PROTECTION OFFERED BY THE VACCINE IN THE MICE TREATED WITH THE 1918 HANA RESTRAINT SPECIFIC?

WHEN YOU STRAIN SPECIFIC OF THE VACCINE MADE FROM THAT OR CHALLENGE THAT. WHICH STRAEP ARE YOU --

MOUSE STRAIN. WILL YOU -- DID YOU USE A SINGLE MOUSE STRAIN OR --

FOR THE CHALLENGE?

FOR THE CHALLENGE.

OKAY. THAT IS NOT MY EXPERIMENT. IT'S DR. -- [ INDISCERNIBLE ] 'S EXPERIMENT. WOULD YOU LIKE TO --

WHAT, WE ARE USING THIS EXPERIMENT IN MOUSE BY CONTAINING THE 1918 SEROGLUTIN OF THE CHALLENGES. IS THAT YOUR QUESTION?

NO, MY QUESTION IS -- [ OVERLAPPING SPEAKERS ]

MOUSE?

WHAT MOUSE --

WOULD YOU GET PROTECTION IF YOU CHANGED THE STRAIN OF THE MOUSE.

WE HAVE MOUSE IN THIS EXPERIMENT, WE HAVE NOT TESTED ALL THE STRAINS, BUT USUALLY WHEN WE USE IN DIFFERENT EXPERIMENTS DIFFERENT MOUSES, STRAINS, WE GET BASICALLY THE SAME EFFICACY. NOT USING 1918 CONTAINING VIRUS, BUT THE RESPONSE, DOESN'T CHANGE TOO MUCH OF THE STRAIN OF MICE THAT WE USE. WE PREDICT THAT IT WILL PROTECT US, THE STRENGTH.

DR. MURPHY.

YOSHI, DID YOU DO ANY EXPERIMENTS WHERE YOU COMPARED THE H1 NEXT 1 CURRENT 1999 VACCINE WITH AN INFECTION WITH THE WILD-TYPE VIRUS BECAUSE ALL HUMANS NOWADAYS ARE RUNNING AROUND WITH ANTIBODIES THAT THEY HAVE HAD FROM MULTIPLE INFECTIONS WITH H1 NEXT 1 VIRUSES AND MIGHT BE MORE PROTECTED AGAINST 1918 VIRUS THAN INDICATED BY YOUR EXPERIMENT.

THAT IS NOT MY EXPERIMENT. ADOLFO GARCIAL-SASTRE'S ALSO.

SORRY.

BUT WE DIDN'T DO THAT EXPERIMENT. I DON'T KNOW. ADOLFO DID.

WHAT TYPE, YOU --

A LIFE INFECT.

LIFE INFECTION, I WOULD SAY VACCINE.

JUST AS AN EXPERIMENT TO INDICATE WHAT LEVEL OF WHAT IS HO TOT H1 N1 VIRUSES.

WE DID NOT DO THIS EXPERIMENT, BUT WE DID THE ONSET. WE USE LIFE IN -- INSPECTION OF X31 AND PR8 AND CHALLENGE WITH 1918. AND WE GET SOME LEVEL OF CROSS PROTECTION WITH THE X, WITH THE X 31 STRAIN, THE ONE THAT DOESN'T SHARE THE HEMOGLUTIN. WE GET -- [ INDISCERNIBLE ] WHEN WE DO THE EXPERIMENT WITH LIKE VIRUSES, OR VACCINATIONS OF MICE .

YOSHI, WHEN YOU HAVE A SYSTEMIC INFECTION, THERE VIREMIA?

VIREMIA.

WHAT OTHER ORGANS AND CELL TYPES INVOLVED?

TARGET ORGANS ESSENTIALLY ANY WAY THAT CAN GROW, INCLUDING THE BRAIN. AO.

YEAH.

AS A FOLLOW-UP TO THAT, ARE THERE EXAMPLES OF SYSTEMIC INFECTION IN HUMANS?

THERE IS NO INFORMATION AVAILABLE ABOUT THES ISIC INFECTION IN HUMANS AND -- THE SYSTEMIC INFECTION IN HUMANS AND ALL THE DATA WE KNOW INDICATES THAT IN HUMANS INFLUENZA VIRUS ONLY GROWS IN RESPIRATORY ORGANS .

QUESTION FROM DR. BARKER.

WERE THERE ANY [ INDISCERNIBLE ] DEGS THAT OCCURRED AND IF SO, DID YOU DETECT ANY SERO CONVERSION.

SERO CONVERSION OF HUMANS?

OF STAFF. YEAH.

WE, WE ARE SORT OF ADVICED -- ADVISED NOT TO, FOR THE PERSONAL REASONS OR WHATEVER, WE'RE OTHERWISE NOT TO TAKE PERSONALS HERE, WILL -- [ INDISCERNIBLE ] HAS HERE, BUT WE DON'T TEST THE SILICONE VERSION YET. THE EXPERIMENT WE DID FOR 1918 VIRUS, THE MOUSE EXPERIMENT, EVERYTHING IS CONTAINED IN THE MICROFIELD OF THINGS.

ANYMORE QUESTIONS FROM THE PANEL? OKAY, WELL, THANK YOU, YOSHI. WE'LL MOVE ON TO CHARACTERIZATION OF THE 1918 INFLUENZA VIRUS. FROM DR. ADOLFO GARCIA-SASTRE FROM MOUNT SINAI SCHOOL OF MEDICINE .

OKAY. SO WHAT I'M GOING TO TRY TO DO TODAY IS TO WALK THROUGH SOME OF THE EXPERIMENTS THAT WE HAVE -- [ INDISCERNIBLE ] CONCERNING THE CHARACTERIZATION OF THE 1918 INFLUENZA VIRUSES FLU AND THIS HAS BEEN DONE, THEY HAVE BEEN DONE NOW UNDER THE SUPPORT OF NIH AND NID WITH THE PROGRAM PROJECT GRANT WHICH, INVOLVES, ACTUALLY, A LAST COLLABORATION OF DIFFERENT INSTITUTIONS AND DIFFERENT PIs. A DOWNFORCE OF PATHOLOGY, BERGER IS THE PERSON THAT IS ACTUALLY ABLE, HAS BEEN ABLE AND IS SEQUENCING THE GENOME OF THE 1918 VIRUS FROM SAMPLES DERIVED FROM 1918 THAT KEPT IN ARCHIVE OR WERE BURIED IN THE PERMAFROST UNDER THE -- THE THERMA FROST, UNDER THE THERMA FROST LINE IN ALASKA. MY GROUP IN NEW YORK, SPLZ THE GROUP OF PETER AND -- [ INDISCERNIBLE ] WE'RE LOOKING INTO MOLECULAR BIOLOGY OF THE ANTIGENS AND THE PROTEINS BY THE VIRUS. WE FELT THIS MOMENT USING ANY LIVE VIRUS. THE USDA IN ATHENS, GEORGIA, THE GROUP OF DAVID SWAIN WHERE IT WAS, AT THE MOMENT THAT WE STARTED THIS STUDIES, HAVE BEEN RESPONSIBLE OF WORKING WITH ALL THE WORK THAT WE HAVE DONE WITH LIFE, VIRUSES CONTAINING 1918 JEEPS. AND -- [ INDISCERNIBLE ] THE CDC AND WE'RE COLLABORATING WITH THE CDC WHERE THESE EXPERIMENTS ARE ALSO BEING CONDUCTED UNDER THE CDC FACILITIES. THEN AT THE UNIVERSITY OF WASHINGTON, SEATTLE, A GROUP -- [ INDISCERNIBLE ] VERY IMPORTANT TO ANALYZE THE HOST RESPONSE USING GENOMICS AND PROTEOMICS TULS -- TOOLS AND ALSO, -- [ INDISCERNIBLE ] USING IN THE FUTURE FOR STUDYING PATHOGENICITY OF VIRUSES CONTAINING 1918 GENES. FINDING OUT THE STREETS -- [ INDISCERNIBLE ] IT'S CRYSTALIZING DIFFERENT ANTIGENS AND PROTEINS FROM THE I HAVEROUS AND TRYING TO FIGURE OUT THE RELATION BETWEEN THE STRUCTURE AND THE PROTEINS AND WHAT COULD THE STRUCTURE TELL US WITH RESPECT TO THE VIRAL OR PATHOGENICITY THAT THIS HAS MADE TO INFLUENCE THE VIRUS. SO, I HAVE JUST A COUPLE OF INTERESTING STORIES. SLIDES, I THINK, EVERYTHING HAS BEEN INTRODUCED VERY WELL BY THE PREVIOUS SPEAKERS. YES, STILL EMPHASIZE SOME POINTS, IT WILL LOOK TO POINTS OF HUMANS. DURING THE PANDEMIC YEARS, THERE HAVE BEEN 20,000 DEATHS IN THE UNITED STATES. THESE ESTIMATES HAVE BEEN INCREASED RECENTLY IN A PUBLICATION IN 2003 TO 36,000 PER YEAR IN THE UNITED STATES. THIS IS, THIS BY THE ABILITY OF -- [ INDISCERNIBLE ] DESPITE THE ABILITY OF VACCINES, MOST OF THIS IS OCCURRING IN THE HIGH-RISK POPULATION, THE VERY YOUNG KIDS OR THE ELDERLY. NOW, DURING PANDEMIC YEAR, HOWEVER, THE HIGHEST MOBILITY OF MORTALITY, AND ONE CRITICAL FACTOR THAT AFFECTS THE HIGH MORTALITY FACTOR IS THE LACK OF -- [ INDISCERNIBLE ] IN HUMANS. BECAUSE OF THE ANTIGENIC SHIFT THAT HAPPENED DURING THE PANDEMIC YEARS WHEN THE VIRUS ANTIGENIC HEMOGLUTIN MAINLY, SOMETIMES ALSO THE CHANGE, THEN IT'S NOT PRACTICE IN IMMUNITY IN BIG PART OF THE POPULATION AND THAT HAS BEEN -- [ INDISCERNIBLE ] WITH THE HIGH MORBIDITY OF MORTALITY IN HUMANS. THERE ARE ALSO SOME FACTORS THAT ARE NOT ONLY CONCERNING THE LACK IN -- [ INDICERNIBLE ] I THINK THAT YOSHI MADE REALLY A NICE CASE ABOUT SOME OF THE INSPECTORS THAT INFLU ENZA PATHOGEN ISITY OF THE INFLUENZA VIRUS IN HUMANS. OF COURSE, THEY'RE THEY'RE HOST FACTORS. FOR EXAMPLE, PLAY A ROLE. THE ELDERLY ARE MOST SUSCEPTIBLE TO THE SEVERE SYSTEM THAN -- [ INDISCERNIBLE ] NOW, WE JUST LOOK TO THE IMMUNOLOGY OF HUMAN INFLU EPZA VIRUSES IN HUMANS. THERE ARE MAINLY TWO TYPES. TYPE B AND TYPE A. TYPE B AS WELL AS TYPE A, THEY'RE ALL UNDER WHAT IS CALLED ANTIGENIC GRIEF, WHICH IS THE VARIATION FROM YEAR-TO-YEAR, THAT HAS TO BE MOST LIKELY BE THE SELECTION, FOR WHAT ELSE, THE SMOKE MUTITATIONS TO BE SELECTED AND THIS ALLOWS FOR INFECTION OF HUMANS WITHIN A STRAIN THAT HAS BEEN VARIED A LITTLE BIT FROM THE PREVIOUS ONE. THAT IS WHY THE VACCINE NEEDS TO BE UPDATED AND HAVE THREE COMPONENTS, TYPE B AND TWO COMPONENTS OF TYPE A. THEY'RE TOO DIFFERENT ANTIGENIC SITES -- [ INDISCERNIBLE ] ALSO SPREAD IN HUMAN. IF WE LOOK TO INFLU EPZA A VIRUSES, THIS IS THE PANDEMIC FROM BEFORE AND GEARED TO WHAT I CALL ANTIGENIC SHIFT. AND IN THE LAST CENTURY, THERE WERE THREE PANDEMICS, THE ONE IN 1918, DUE TO AN H1 N1 VIRUS N.1957, THIS WAS REPLACED BY A COMPLETELY DIFFERENT HEMOGLUTIN VIRUS CAUSING THE PANDEMIC OF 1957 AND REPLACEMENT OF THE H1 AND N1 STRAIN. IN 1968 -- 1968, TWO VIRUSES CAME TO THE HUMAN POPULATION CAUSING THE PANDEMIC OF 1958 AND IN 1977, H1 AND 1 WAS INTRODUCED IN HUMANS. IT'S -- [ INDISCERNIBLE ] WITH HG AND 2. NOW THE INTRODUCTION OF H1 AND 1 DIDN'T CAUSE A PAN DEMMEC EVENT AS WHAT HAPPENED WITH H3 AND 2. IF WE LOOK TO THE SCHEMATIC THAT IS IN THE UNITED STATES, THE PANDEMIC CAUSED 35,000 EXCESS DEATHS. AS WAS MENTIONED, THE TWO SENATES BETWEEN 1990 AND 70,000 -- THE TWO ESTIMATES BETWEEN 1990 AND 70,000 DEGS. IN 1918, THE PANDEMIC INCREASED A MORTALITY, 500,000 DEATHS, WHAT THIS INTERESTING ABOUT THIS VIRUS. STILL THE REASONS WHY THE VIRUS WAS -- [ INDISCERNIBLE ] H2 AND 2 AND FOR HIGH PATHOGENICITY OF THE VIRUS ARE UNKNOWN. AND, OBVIOUSLY, THERE WAS SOMETHING ELSE GOING TO THROUGH THE LACK OF THE IMMUNITY BECAUSE THERE WAS A BIG DIFFERENCE BETWEEN THE 1918 VIRUS AND THE 1957 VIRUS. NOW, THE THING BECOMES MORE COMPLICATED WITH THE SEARCHING OF AVIAN FLU. THERE HAVE BEEN CASES NOW BEING REPORTED WITH H5 AND 1 VIRUSES, H9 AND 2 I HAVEROUSES, H77 AND 1, AND THIS HAS ALSO DELAYED US WHERE THE STUDY -- [ INDISCERNIBLE ] WITH UPWARDS OF 23 WERE FATAL. FORTUNATELY THESE VIRUS HAVE NOT BEEN, HAVE NOT LEARNED YET HOW TO PROPAGATE FROM HUMAN-TO-HUMAN. SO IT SEEMS TO BE CONFINED FROM JUMPING FROM A BIRD SPEEZOS TO A HUMAN. -- SPECIES TO A HOW. ONE OF THE INTERESTING THINGS ALSO ABOUT A UNIQUE -- UNIQUE THING ABOUT 1918 WAS THE MORTALITY CURVE AS COMPARED TO THE AGE DISTRIBUTION IN HUMANS. SO WE TAKE THE DATA FROM BEFORE 1918 AND WE LOOK AT WHAT IS THE PEOPLE THAT ARE ACTUALLY MORE SUSPENDIBLE TO -- SUSCEPTIBLE WITH THE INFLUENZA VIRUSES. WHAT WE FOUND WAS THAT THE YOUNG KIDS ARE MORE SUSCEPTIBLE, AS WELL AS THE ELDERLY AND THIS HAPPENED ACTUALLY FOR EVERY SEASON, EVERY EPIDEMIC AND WE HAVE THIS U-SHAPE IN WHICH THE YOUNG, THE YOUNGEST AND THE HOLDEST -- OLDEST ARE THE ONES MORE SUSCEPTIBLE TO HAVE SEVERE DISEASES WITH THE INFLU ENZA VIRUS. HOWEVER, WITH THE, DURING THE 1918 PANDEMIC, THERE WAS CALLED A W SHAPE. IT WAS A COMPLETE, A QUITE DIFFERENT EXPERIMENT OF MORTALITY IN WHICH THE VIRUS WAS INFECTING THE YOUNGEST AND OLDEST. THERE WAS A GROUP OF PEOPLE ADULT HELPING -- [ INDISCERNIBLE ] BETWEEN 15 AND 35 YEARS OLD. THEY WERE ALSO STRIKE BY W AND HAVING PROPENSET TO HAVE -- [ INDISCERNIBLE ] AND THE VIRUS CAUSED INCREASED DEATHS IN THE POPULATION. THAT WAS UNIQUE FOR THE 1918 VIRUS. IT'S NOT -- IT MAY HAVE HAPPENED BEFORE 1918, BUT WE DON'T HAVE DATA THAT WOULD SUPPORT THAT. THIS IS SOMETHING THAT DIDN'T HAPPEN DURING THE PAN DAMAGIC OF 1918. SO THAT MAKES THIS THAT MAKES IT A USEFUL VIRUS, ONE OF THE REASONS WHY WE'RE INTERESTED IN IT. WHY STUDY THE 1918 VIRUS. AS I MEGGED, THE INCREASED VIRULENCE ARE EVIDENCE. WHEN THEY GET SGENS DATA FROM DIF -- SEQUENCE DATA FROM DIFFERENT, NOT EVEN THE SEQUENCE DATA, TELL US WHAT HAVE BEEN THE REASONS FOR THE VIRUS IS. THERE WAS NO KNOWN DETERMINE NANS OF VIRULENCE THAT WAS KNOWN. WE KNOW THAT INFLUENZA VIRUS HAVING 1918-LIKE VIRAL PROPERTIES MAY HAVE BEEN INVOLVED AGAIN. THIS VIRUS CAN -- [ INDISCERNIBLE ] AND THERE ARE PANDEMICS EVERY 20 YEARS. IT'S DIFFICULT TO PREDICT WHAT IS GOING TO BE THIS PANDEMIC. WE DON'T KNOW IF IT'S GOING TO COME WITH THE VIRUS CONTAINING SOME OF THE VIRULENCE MARKET OF 1918. HAD OF 1918. IF WE DIDN'TDIFY -- [ INDISCERNIBLE ] OF INFLUENZA VIRUS SPECKALLY FOR A COMPANY, 1918, THIS WILL ALLOW FOR ELDERLY RECOGNITION OF POTENTIALLY HIGH VIRULENCE STRAIN. WE FIND IT IN ONE PARTICULAR STRAIN BEFORE IT'S WIDELY DISTRIBUTED IN THE WORLD, WE'RE VERY PREPARED TO KNOW THERE IS A STRAIN THAT MAY COME THAT HELPS THE POTENTIAL, THAT HAS THIS VIRULENCE MARKER. IT ALLOWS FOR AN -- [ INDISCERNIBLE ] SO WE KNOW ACTUALLY WHAT OUR MOLECULAR RESOURCE IS FOR VIRULENCE. WE MAY BE ABLE TO COME UP WITH STRATEGIES, HOW TO ENIFER FIRE WITH THE VIRHUH, ENCE. -- COME COMES FROM SOMETHING WITH THE H5 STORY, THE VIRUS IN WHICH IS KNOWN THE SITE OF THE HEMOGLUTINEN SPACE, IN ORDER TO MAKE NOW VACCINE STRAINS AND ABLE TO GROW WITH TOO MUCH RISK. THE VACCINE OF STRAINS THEN ONE CAN CHANGE THE VIRULENCE MARKET OF THE HEMOGLUTINEN TO MAKE IT LOOK VIRULENT. WE HAVE A VACCINE OF STRAIN THAT CAN BE GROWN WITHOUT SO MUCH WRECK AND THEN BEING ACTIVATED AND BEING USED. WE KNOW WHAT THE VIRULENCE MARKERS. IF THE 1918 VIRUS COMES, WE CAN ROUGH MAY -- ROUGHLY MAKE A VACCINE THAT WE SHOULD BE ABLE TO GROW IT UNDER, WITH LESS RISKS AND THEN BEING ABLE TO GET IT MORE ON TIME. SO HOW WE ARE APPROACHING THE PROBLEM OF 1918. AS YOSHI MENTIONED, 1918 VIRUS DOESN'T EXIST. BERGER, USING PATHOLOGICAL SPECIMANS OF PEOPLE WHO DIED OF 1918 INFLUENZA, COMING FROM THIS YEAR, HAS BEENABLY TO SEQUENCE A LITTLE -- BEEN ABLE TO SEQUENCE A LITTLE BEESES OF RNA, SEQUENCE THEM AND PUT TOGETHER THE SEQUENCE OF THE GENES OF THE VIRUS. WE HAVE SEQUENCE 5 OF THE GENES OF THE 1918 VIRUS. KNOWING THE SEQUENCE AND ONE CAN NOW RECONSTRUCT THE GENE BY USING A DNA PCL TECHNIQUES AND USING THE GENETICS TECH NBC, IT WAS -- [ INDISCERNIBLE ] TECHNIQUES, ONE CAN TAKE THE GENE AND PUT IT INTO THE BACKBONE OF THE EXISTING ENFLU EPZA VIRUS AND -- INFLUENZA VEUS HAVE VIRUS AND THEN THE CHARACTERISTICS OF THE VIRUS, WE STUDY THAT AND IN ANIMAL MODELS AND HOPEFULLY WE WILL BE ABLE TO FIND ONE OF THESE THINGS TO INCREASE VIR UULENCE. IN ORDER, WE NEED TO BE WORRIED ABOUT WHAT ARE THE LEVEL OF CONTAINMENT THAT WE NEED TO USE AND JUST WHAT WAS MENTIONED, WHAT WAS HIS CRITERIA TO WORK WITH THIS VIRUS. ALSO, WE WORK -- WE WORK THROUGH THE RISK ASSESSMENT TO WORK WITH THE RECOMBINANT VIRUS OF GENE CELLS, 1918 AND CONCLUSIONS WERE SIMILAR TO THE ONES THAT YOSHI MENTIONED. SO, IF WE LOOK FIRST TO PATHOGENICITY, HOW IS IT GOING TO BE THE PATHOGENICITY OF THE VIRUS. WE START WITH THE VIRUS, H1 AND N! VIRUS, THE ONE WE'RE USE ING WE'RE USING THE HUMAN STRAIN CLOSE TO WHAT YOSHI MENTIONED. THEY'RE USED ON THE CONTAINMENT. NOW, IF WE KNOW IT STARTS IN 1918 GENES WHAT, ASK CAN HELP HAPPEN WITH THE VIRUS. WILL IT BE LOWER THAN BACKBONE? THIS MAY HAPPEN. IF YOU KNOW, INFLUENZA VIRUS HAS EIGHT DIFFERENT GENES AND THERE IS ALWAYS, THE GENES ARE ADOPTED, ONE TO EACH. WHEN WE SAY IT'S A COPSTILLING A, WE WILL SAY IT'S ONE GENE FROM 1918. THAT WILL INTERACT WELL WITH THE GENE OF H1 N1. THIS IS ONE OF THE TYPES OF RESEARCH. HOW WE CAN DEAL WITH THE AFFECTS, MEASURING VIRULENCE OF THE VIRUS. THERE IS ONE OF 1918. THIS MAY NOT POP OUT. WE PUT IN THIS GENE INTO AN H1 N1 VIRUS. MAYBE THIS JEEP DIDN'T INTERACT WELL WITH THE OTHER GENES OF H1 N1. MOST LIKELY, WHAT WE'RE LOOKING FOR IS SOMETHING BETWEEN THE BACKBONE AND NEXT 2. THAT WILL PROBABLY HAPPEN. COULD IT BE WE MAKE THE REASSORTMENT WITH AVERULENCE HIGHER THAN 1918? I THINK MOST LIKELY -- LIKELY THERE ARE NO EXAMPLES LIKE THAT IN THE INFLUENZA VIRUS. NOW, THAT WILL BE PROPORTIONAL TO A NUMBER OF GENES. IF NOT, THIS WOULD PROBABLY BE, EVEN IF THERE IS A MEASURE OFVERULENCE OF 1918 BECAUSE OF THE CONSTELLATION AFFECT THAT I MENTIONED IF WE PUT THE VIR UULENCE MARKETER IN THE CONTENTS OF 1918, WE WILL GET IT CLOSE TO 1918. THE WHOLE VIRUS WILL BE 1918, ANYWAY. NOW, WHAT OTHER FACTORS WE SHOULD HAVE IN CONSIDERATION AS YOSHI MENTIONED, IMMUNITY TRAITS ONE I HAVEROUSES. THIS MIGHT BE ONE THING THAT MAY REDUCE THE RESULTS OF WHAT CAN -- WORKING WITH THESE TYPES OF STRAINS. NOW, WE LOOK TO THE ROUTE OF TRANSMISSION OF THE VIRUSES. THE VIRUSES TRANSMILL THEIR RESPIRATORY BY AEROSOLS. THE STABILITY INFLUENZA VIRUSES FOR A LONG TIME ON SURVASE -- SURFACES. IT WON'T SURVIVE ON DRY SURFACES AND IS SUSCEPTIBLE TO A NUMBER OF INFECTS. IT'S -- [ INDISCERNIBLE ] CONTAMINATE. IN TERMS OF THE INFECTIOUS ADULTS, IT'S BETWEEN TWO TO 1,000 VIRUS PARTICLES WILL BE INFECTIOUS THROUGH THE RESPIRATORY ROUTE AND THE CONCENTRATIONS I USE IN THE LAB, -- [ INDISCERNIBLE ] -- THIS HIGH COMPARED TO INFECTIOUS DOSE. NOW, HOW WE CAN INCREASE NOW THE RISK AND WHAT TYPE OF FACILITIES WE NEED TO USE SAW, IN TERMS OF DECREASE, THE RISK TO PERSONNEL, THEN ONE COULD, ONE SHOULD USE PRIMARY BARRIER SUCH AS PROTECTIVE CLOTHING AND THE USE OF PAPERS, AIR PRESSURE RESPIRATORS THAT YOSHI WAS MENTIONING ALSO, AND BIO -- BIOLOGY -- [ INDISCERNIBLE ] I PUT THIS IN RED IN TERMS OF THESE ARE PICTURES THAT WILL BE ENHANCED PICTURES FOR A BSL 3 FACILITY. SO, BSL 3 FACILITY HAVE ALL BIOLOGICAL SAFETY COVENANTS BY THE USE OF FULL CLOTHING OR THE USE OF -- [ INDISCERNIBLE ] -- ENHANCED. NOW TO THE ENVIRONMENT DECREASED BY USING A -- [ INDISCERNIBLE ] FACILITY. I ON PURPOSE PUT HERE ENHANCED BSL FACILITIES. WE USE IN USDA, WE USE AGRICULTURAL LEVEL. BECAUSE OF THE DIFFERENT ISSUES, WHAT IS ENHANCE, WHAT IS CULTURE, I THINK IT'S IMPORTANT FOR US TO CONSIDER IF A FACILITY CAN OR CAN'T CONTAIN THE VIRUS WHAT. I MEAN BOO BYE CAN OR CAN'T COPTAPE THE VIRUS IS ANYTHING THAT LEAVES THE FACILITY HAS A POSSIBILITY TO HAVE AN INFECTIOUS SITE. THAT'S BASICALLY WHAT WE'RE LOOKING FOR. EVERY BSL 3 HAS PRESSURE. NOW, SOME OF THE MAJOR ENHANCEMENTS NOW MAKING YET IMPOSSIBLE FOR AN INFECTIOUS VIRUS TO LEAVE THE FACILITY IS KEY CONCENTRATION SO THAT THERE IS KEY CONCENTRATION OF THE AIR COMING OUT OF THE FACILITY, ALLOWS FOR NO INFECTIOUS AGENT LEAVING THE FACILITY AND RIGOROUS CONTAMINATION WITH THE PROCEDURES THAT WOULD ALLOW US AN INFECTIOUS AGENT LEAVING THE FACILITY. THAT'S A DOUBLE-DOOR AUTOCLAVE FOR EXIT OF EVERYTHING AS WELL AS THE CONTAMINATION. NOW, TO BOTH PERSONNEL AND ENVIRONMENT RISK IS MINIMIZED BY USING, BY MINIMIZING OUR -- [ INDISCERNIBLE ] ONE OF THE IMPORTANT THINGS THAT ALSO ENHANCES THE BSL 3 FACILITIES IS HAVING THE LAB PERSON SHOULD BE APPROPRIATELY TRAINED. EVEN IF THE FACILITIES ARE COMPLETELY UNABLE TO, IT TO LIFT ANY AGENTS FROM THE FACILITY, IT'S IMPORTANT THAT THE PERSON LAB PERSON IS WELL-TRAINED. [ INDISCERNIBLE ] -- BY HAVING THE TRAINING, REG ROUSE TRAINING OF THE PERSONNEL, DEMONSTRATE PROFICIENCY, RECEIVE PROPER TRAINING, STANDARD OPERATING PRACTICES, REPORT ENSY DENS OF CAUSE AND SUPPORT -- [ INDISCERNIBLE ] -- THIS IS HOW SOME WILL HAVE IN PLACE IN ORDER TO WORK WITH THIS, WE THINK, THAT IS VERY IMPORTANT FOR WORKING WITH THIS TYPE OF AGENT. AND THIS IS TERRY, ONCE HE WAS WORKING IN THE USDA IN 1918, WITH THE VIRUS CONTAINING ONE OF THE 1918 GENES. AS YOU SEE, WE HAVE HERE THIS TOP PERSON, RESPIRATORS COMPLETE CLOTHING, DOUBLE CLOTHED AND THAT'S HOW THE PERSON LOOKED LIKE IN THIS FACILITY. YOU IN, PART OF THE RISK ASSESSMENT IS WHAT, WHAT WE KNOW ABOUT THE DATA FROM STUDIES. AS YOSHE MENTIONED IN THE BEGINNING, WE DIDN'T KNOW ANYTHING ABOUT 1918, HOW VIRULENT IT WOULD BE FOR MICE OR DIFFERENT ANIMALS STUDIED. I WANT TO DESCRIBE SOME OF THE STUDIES WE HAVE ALSO IN OUR GROUP. CONCERNING THE QUESTION, HEMOGLUTIN OF THE 1918 VIRUS JEEPS CONTRIBUTOR. FOR THAT, WHAT WE DID IS WE EXPERIMENTED, SIMILAR TO THE ONE DESCRIBED BY YOSHI. WE TAKE IT AND WE REPLACE THE HEMOGLUTINEN BY THE VIRUS IN THIS VIRUS BY THE 1918 -- [ INDISCERNIBLE ] THEN WE ASK HOW VIRULENT IS IT IN MICE. THEN H1 N1 GENES COMING FROM THE CURRENT STRAIN, 1999, ALSO SEQUENCE -- [ INDISCERNIBLE ] SO IF WE LOOK TO THE Ws VIRUS, CONTAINING OBVIOUSLY THE W H1 N, THE 50 IN MICE IS TO THE, AROUND 10 TO THE THREE, INFECTION FROM ITALY, WE HEAL APPROXIMATELY 50% OF THE MICE AND THIS IS BECAUSE THE HAVE US HAVE -- VIRUS -- [ INDISCERNIBLE ] AND IT WILL LOOK TO THE MOUSE ONE, 10 TO THE SIX, 10 TO THE SEVEN BY DAY 3. WE TAKE W AND PUT THE H1 N1 FROM A NEW CATEGORY, ONE OF THE 1999 STRAINS IN HUMAN. NOW, THIS VIRUS GOES TO ALMOST CLOSE EDENTICAL -- [ INDISCERNIBLE ] BUT THIS VIRUS IS HARDLY RATED IN MICE. THERE IS, WE CAN'T INFECT MICE WITH THE ADULTS THAT WE KILL MICE. AND THIS VIRUS GROWS AROUND -- [ INDISCERNIBLE ] -- SO IT'S AROUND 3, A LOT SLOWER THAN THE VIRUS AND THAT'S TYPICAL FOR HUMAN VIRUSES, UNLESS THERE IS MOUSE ADAPTATION. THEN THE VIRUSES ARE -- [ INDISCERNIBLE ] WE WE TAKE NOW THE 1918 GENES AND WHAT WE FOUND AGAIN IN THE CELLS CLOSE TO -- [ INDISCERNIBLE ] BUT NOW WE HAVE IN ELDERLY PEOPLE SIMILAR TO IT FOR NOW. THIS HEMOGLUTININ, THEY'RE CAPABLE TO HAVE, TO MAKE THE VIRUS VERAL IN MICE. WE TAKE -- [ INDISCERNIBLE ] TO THE 4. ALSO THE -- IT GOES QUITE WELL. WE TAKE A VIRUS CONTAINING THE HEMOGLUTINEN AND HELPS ALSO TO AN INTERMEDIATE TO THE 50, INCREASING THE OF THE COMPARED TO THE CONTROL, BUT IT'S INTERMEDIATE FROM THE ONE IN 1918 AND ACTUALLY, THIS ONE DOESN'T GROW VERY WELL IN LENGTH. WHEN SEPARATE, THESE RUTS ARE A CORP OPERATION BETWEEN THE HEMOGLUTINEN OF 1918 IN ORDER TO ACHIEVE MUCH MORE EFFICACY OF THE VIRUS. SO THIS DATE, I WOULD SUGGEST ALSO WHAT YOSHI CONSENTED THAT BOTH 1918 AND -- [ INDISCERNIBLE ] ARE LIKELY TO CONTRIBUTE TO IT IN MICE. AT LEAST IN MICE. NOW, IN TERMS OF AVAILABILITY OF INFECTED PROPHLYAXIS OF INTEGRATION FOR INFLUENZA VIRUS, THERE ARE EXISTING ASPECT VIRALS AND THIS IS THE PSYCHE -- ANTIVIRALS AND THIS IS THE CYCLE OF THE VIRALS. I WOULD GO THROUGH THE CYCLES YOSHI WAS MENTIONING, AND REPRESENTATION IN THE NUCLEUS, BUT TO MENTION THE TWO TYPES ARE AVAILABLE THERE IN BETWEEN INHIBITORS, THE INHIBITOR PROTEIN OF THE VIRUS OR VIRAL LOCATION OF THIS LEVEL AND THE NEUROMASEP HECTORS THAT BLOCK VI -- INHIBITORS THAT BLOCK AT THIS LEVEL. WE ASK WITH THE VIRUSES CONTAINING 1918 GENES WILL BE SUSCEPTIBLE TO THIS ANTIHAVE LEGS AVAILABLE, THIS OTHER DATA WE HAVE WITH -- [ INDISCERNIBLE ] AND INHIBITOR, IT WILL LOOK TO IN VETRO AND WE LOOK TO ABILITY TO INHIBIT THE NEUROMINEASE IN VITRO. WE FOUND OF THE VIRUS, THE HUMAN STRENGTH OR OF THE 1918. SUGGESTING THAT THIS VIRUS IS A VIRUS CONTAINING 1918 NEUROMENACY WILL BE SUSCEPTIBLE TO NEUROMENACE INHIBITORS. IT WILL LOCATE IN VITRO USING VIRUSES. WHAT WE FOUND IS THAT, AGAIN, THERE IS MORE EFFICACY OF THE -- [ INDISCERNIBLE ] -- P HINTING BLOCK FORMATIONS OR REPLICATIONS IN VOTE ROW DIFFICULT FOR W SEN CONTAINING THE 1918 NA AND HA OF W SEN OR CONTAINING BOTH HA AND NEUROMENASE OF 1918. NOW, FINALLY, WE LOOK IN THE ANIMAL. NORMALLY IN MOUSE WHAT WE FOUND IS THAT LIVE WE TAKE MICE -- [ INDISCERNIBLE ] -- AND INFECT IT WITH THE VIRUS CONTAINING THE 1918 HNA, HOWEVER, IF WE TAKE THE -- [ INDISCERNIBLE ] MICE, WE PROTECT THE MICE FROM DEATH, FROM THE VIRUS AND -- [ INDISCERNIBLE ] -- WHAT THIS DATA WILL TELL US IS THAT A VIRUS CONTAINED IN 1918 WILL BE SUSCEPTIBLE TO THE MUROMINASE INHIBITORS. ALL THE COMMON INFLU ENZA VIRUS. NOW NEXT TERMS OF THE INHIBITORS, WE MADE THE VIRUS CONTAINING THE M GENE OF 1918 AND THE ABILITY TO BE INHIBBED BY -- [ INDISCERNIBLE ] -- COMING FROM THE SEQUENCE WE THOUGHT WOULD BE SUSCEPTIBLE AND THE DATA OF THE -- [ INDISCERNIBLE ] -- SO, IF WE DECIDE THE DATA WITH MICE, AND THE DATA -- [ INDISCERNIBLE ] -- UNINFECTED WITH THE VIRUS CONTAINING THE 1918 GENE. SO, ABLE TO PREVENT ALSO LOTTAL INFECTION IN THE MOUSE MORE -- LETHAL INFECTIONS IN THE MOUSE -- [ INDISCERNIBLE ] USING 1918. NOW FINALLY, I WANTED TO MENTION SOME OF THE STUDIES WE HAVE DONE IN TERMS OF THE POSSIBILITY OF PREVENTING THE VIRUS BY VACCINATION. ONE OF THE THINGS THAT CAN ALSO BE USED TO, FOR PROPHLYAXIS INTERVENTION IS VACCINE. WE NEED TO UNDERSTAND MORE OF THE AUTHENTICITY. MOST OF THE VACCINES, MOST OF THE VACCINES, AT LEAST THE KILL VACCINE THAT ARE AT LEAST IN HUMANS FOR INFLUENZA FOR PREVENTIVE INFLUENZA VIRUS INFECTION IS, IS BASED ON THE -- [ INDISCERNIBLE ] -- ANTIBODY RESPONSE OF HEMOGLUTINEN. IN THIS EXPERIMENT, WE'RE LOOKING TO THE HEMOGLUTTINNIN. I WILL WALK THROUGH THE SPERMS WHAT. WE DID IS TOOK WE -- [ INDISCERNIBLE ] AGAINST DIFFERENT H1 AND N1 RESTRAINTS, 1918, 1930s, 1933, WS, N IN 1934, 1987, 1983, 1984, AND 1989. WE LOOK FOR THE AVAILABILITY OF THE SERA TO INHINT THE HEMOGLUTTINATION OF THE VIRUSES ALSO CORRESPONDING TO THE -- SECRET SERVICE SIS -- [ INDISCERNIBLE ] IF WE LOOK TO THIS PART, THIS IS THE AVAILABILITY, 1918, '30, '33, '34, '73, '71, 1999. TO NOTE THAT INHIBITOR GLUTTINNATION OF NO ONE 18. AND, OF COURSE, WE USE THE SAME -- [ INDISCERNIBLE ] AND WE GET A NICE INHIBITION. WE JUST, A VIRUS COMING FROM THE 1930s. WE GET SOME ENHIBBING A -- INHIBBITIONS, SOME WITH B 33 -- W33 AND START TO LOSE THIS IN THE '30s. WE'RE STILL GETTING INHAS BEENATION BUT IT'S LOWER COMPARED TO -- [ INDISCERNIBLE ] WE DO THE OPPOSITE SPERM . THIS IS ABLE -- OPPOSITE EXPERIMENT. THIS IS -- [ INDISCERNIBLE ] I'M NOT SO WELL NOW AFTER THE 30s FROM THE STRENGTH. THIS WILL TELL US THAT ONE OF THE CLOSEST RELATIVES OF 1918 HEMOGLUTINEN IS THE VIRUS THAT WAS ISOLATED IN 1913. WE ALSO -- A STUDY SIMILAR TO THE ONE PRESENTED WITH NEUTRALIZING ANTIBODIES IN HUMANS AND HERE ARE DIFFERENT SAMPLES TAKEN FROM PEOPLE BORN IN 1910, 1911, 1928, '32, '33, '34 AND SO ON. WHAT WE FOUND IS PEOPLE BORN BETWEEN 1910 AND 1928, THEY HAVE NEUTRALIZED ANTIBODIES AGAINST 1918. THIS, THIS WAS BORN IN 18EN IN -- 1910 AND 1911. PROBABLY THEY WERE, THEY EXPERIENCE INFECTION WITH 1918 VIRUS. THE PERSON BORN IN '28 SHOULD HAVE BEEN INFECTED, EXPOSED TO A VIRUS THAT IS NOW A DERIVATIVE OF 1918. AROUND THIS TIME, THE VIRUS WAS CLOSER TO 1918. THEN AT THE, AFTER THE '30s, MOST LIKELY THE -- [ INDISCERNIBLE ] OF THE HEMOGLUTINEN WAS BIGGER AND WE START TO LOSE THEABILITY TO NEUTRALIZE IT. -- THE ABILITY TO OUT INERALIZE IT. IN THIS NEW -- [ INDISCERNIBLE ] WITH THIS EXPERIMENT THAT YOSHI MENTIONED THAT WE LOOK TO THE PROTECTION USING THE ACTIVATED VACCINE AGAINST 1918 NAHA AND THE VIRUSES, IF WE TAKE THE MICE AND VACCINATE IT AND CHALLENGE WITH THE 1918 MATERIAL-COPTAPING VIRUS, THE MICE HAVE A LETHAL INFECT. WE IMMUNIZE IT WITH THE KILL VIRUS. WE GET COMPLETE PROTECT. WE USE A KILL X31 VIRUSES TO CONTROL THE STRAIN 1 AND TWO. IT HAS A DIFFERENT HEMOGLUTINEN. IF WE USE THE HEMOGLUTINEN FROM THE H1 N1 STRAINS, 1999, '91, PR834 OR 30, WE GET THIS WITH THE CLOSEST HEMOGLUTINEN. WE GET THE COMPLETE PROTECTION AS IT THE HUMIDITIO GLUTTENEN IS STARTING -- [ INDISCERNIBLE ] FROM 1918. WHAT WE'RE GETTING IS NOT COMPLETE PROTECTION BUT PARTIAL PROTECTION . I JUST SUMMARIZED NOW IF WE KNOW ABOUT THE 1918 VIRUS. FIRST OF ALL, THE GENES OF THE VIRUS MIGHT HAVE CONTRIBUTED, WHAT THE MOUSE DATA WOULD SUGGEST AND WHAT WE'RE INTERESTED IN IS LOOKING TO ALL THAT ANIMAL MODELS TO VALIDATE THAT THAT'S THE CASE. -- [ INDISCERNIBLE ] -- VIRULENCE AND WHAT ARE THE MOLECULAR -- [ INDISCERNIBLE ] THESE ARE IMPORTANT QUESTIONS. KNOWING WHAT ARE RESPONSIBLE AND KNOWING WHAT IS THE MOLECULAR MECHANISM IS RESPONSIBLE FOR VIRULENCE. WE CAN NOW TACKLE MUCH BETTER WHAT I MENTIONED IN THE BEGINNING, IN THE CASE THAT A VIRUS COMES WITH THE VIRULENCE SIGNATURE SIMILAR TO 1918. WE CAN TACKLE THE VIRUS MORE EASILY. WE SHOULD BE ABLE TO PRODUCE VACCINES MORE RAPIDLY AND WE CAN THINK ABOUT WAYS TO INTERVENE WITH THE VIRUS LATER. WE ALSO KNOW THAT VIRUSES CONTAINING 1918 GENES ARE -- [ INDISCERNIBLE ] TO VIRALS AND KNOW THAT H1 N1 VACCINES ARE ABLE TO PROTECT IT AT LEAST -- [ INDISCERNIBLE ] SO I JUST WANT TO FINISH MY PRESENTATION WITH THIS QUESTION. WOULD THE 1918-LIKE H1 AS VIRUS AS IN 1918 AND IF WE GO BACKEN TO TO THE W MORTALITY SHAPE, ONE OF THE FUNNY THINGS IN THIS CURB IS WHY THIS PEOPLE DIDN'T HAVE SEVERE DISEASE AND PRIOR TO 1918, THERE WAS ANOTHER PANDEMIC IN 1918, '19. WHAT WAS THE VIRUS SCINTILLATING BETWEEN 1819 IS NOT KNOWN. IF WE THINK THAT H1 WOULD CONFIRM THE PORTION, THE H1 VIRUS WOULD CONFIRM POSSIBLE PROTECTION AGAINST A 1918 LEAK VIRUS, ONE OF THE WAYS TO EXPLAIN THE W SHAPE IS THE PEOPLE BORN BEFORE 1890, THERE WAS AN H-1 SECRET -- AND THEY HAVE H1 IN ONE COMMUNITY. THE REASON WHY MORTALITY DECLINED IN THIS PARTICULAR AGE GROUP. I JUST WANT TO FINISH HERE.

THANK YOU.

THANK YOU VERY MUCH. [ APPLAUSE ] WE HAVE TIME FOR MAYBE ONE OR TWO QUICK QUESTIONS.

SO IS THERE ANY THOUGHT TO PROTECTING PEOPLE WORKING WITH 1918 RECOMBENANCE, USING THE 30 STRAIN VIRUS, THE 30 STRAIN VIRUS. IS THAT A POSSIBILITY TO -- .

THAT'S A POSSIBILITY TO CONSIDER. NOW WHAT WE ARE DOING IS ON VACCINATION, THE SAME THING AS YOSHI'S DOING AND WE ALSO HAVE AVAILABLE ANTIVIRALS AVAILABLE TO THE PEOPLE DOING THE RESEARCH. IN THE CASE IT HAPPENS, THIS WILL BE ABLE TO TAKE ANTIHAVE RAW -- VIRALS PROPHLYACTALLY. I DON'T KNOW HOW DIFFICULT -- DIFFICULT IT WILL BE TO MAKE A VACCINE BASED ON THIS PARTICULAR VIRUS FOR PEOPLE. BASICALLY.

I HAVE A QUICK QUESTION FOR YOU; THE VIRUSES CONTAINING THE 1918 HA AND NA GENES ARE SENSITIVE TO THE TWO ANTIBIOTICS THAT YOU HAVE TESTD. ARE THE VIRUSES BEARING THE FIVE-CLONE GENE FROM THE 1918 ALSO, JUST AS SENSITIVE TO THE ANTIBIOTICS.

YEAH. TRY TO REMEMBER. I THINK A MONTH IN THE EXPERIMENT WAS ALSO DONE WITH THE FIVE GENES AND THAT WAS ALSO PROTECTED. BUT I CAN'T RECALL NOW EXACTLY THE DATA. THAT WAS DONE BY TERRY TEMPE. I AM ALMOST SURE THERE WAS WITH THE FIVE GENES BUT I CAN'T COULDN'T.

OKAY, THANK YOU VERY MUCH. THE NEXT TALK, DR. ROBERT WEBSTER FROM ST. JUDE CHILDREN'S RESEARCH HOSPITAL. HIGHLY PATHOGENIC AVIAN INFLUENZA VIRUS RESEARCH .

GOOD MORNING. I WAS ASKED TO SPEAK ABOUT HIGHLY PATHOGENIC A RAIN AND INFLUENZA JEYOU -- VIRUSES. I WOULD COPFINE MY TALK TO THAT. I WOULD LIKE TO PREEMPT MY TALK WITH ONE OR TWO COMMENTS IS THAT THIS IS A VERY, VERY IMPORTANT MEETING TO TALK ABOUT VIRUS SAFETY. I ALSO WANT TO REMIND YOU THAT WE'RE CURRENTLY FACED WITH AN ENORMOUS THREAT IN THE AVIAN INFLUENZA SITUATION, SO WE HAVE TO BALANCE THE RISK VERSUS THE SAFETY ISSUES. AND WHILE WE MUST GIVE GREAT ATTENTION TO SAFETY, WE MUST ALSO CONSIDER THE RISK THAT IS CURRENTLY FACING US. SO A RAIN -- AVIAN INFLUENZA VIRUSES. JUST AS A PRIMER FOR THOSE NOT WORKING WITH INFLUENZA VIRUSES, THERE, AMONGST INFLUENZA VIRUSES, THERE ARE 15 HEMOGLUTINEN AND 19 AMINE -- AMINO URAC, S. THEY'RE FOUND IN 15 AQUATIC BIRDS OF THE WORLD, WHERE THEY LIVE IN EQUALINIUM AND NATURAL RESERVOIRS. ALL OF THE SUBTYPES CAUSE NO DISEASE AND THEY'RE IN EVOLUTION RESTASES. THIS IS AN UNUSUAL SITUATION. WE HAVE AN IRONY VIRUS LIVING IN STASES AND NOT EVOLVING. NOW TO TRANSMISSION TO OTHER HOSTS THEY EVOLVE RAPIDLY. THE H5 AND H7 BECOME HIGHLY PATHOGENIC. AND ALSO, THESE ARE THE 15 HUMIDITIO GLUTTINNEN SUBTYPES WE'RE PARTICULARLY CONCERNED WITH H5 AND H7. YOU NOTICE I, WE'RE NOT H 1. WE HAVE JUST BEEN HEARING ABOUT 1918 IS NOT CONSIDERED AN AVIAN VIRUS. IT PROBABLY WAS ORIGINALLY. SO, THE POSSIBILITY FOR 1918 TO REAPPEAR, THESE VIRUSES ARE ALL IN THE AQUADDIC BIRD RESERVOIR. WHICH WILL WILL COME ACROSS AND CAUSE THE NEXT PAN DEMMEC, WE REALLY DON'T KNOW. -- PAN DEMMEC, WE REALLY DON'T KNOW. AT THIS TIME, THE GREATEST THREATS ARE H5, H7, H9, H6, AND H3, PROBABLY IN THAT ORDER. THAT'S WHAT WE'RE THINKING ABOUT AT THIS TIME. SOCIETY, I'VE ALREADY MENTIONED THAT -- SO, I'VE ALREADY MENTIONED IN THE NATURAL RESERVOIRS, THESE VIRUSES IN EVOLUTIONARY STASIS. WHERE THE VIRUS IS REPLICATING MAINLY IN THE INTESTINAL TRACT, THE BIRDS POOP THE VIRUS INTO THE WATER AND IT'S A FECAL ORAL TRANSMISSION. THESE ARE MIGRATING BIRDS AND NATURE HAS EVOLVED A WONDERFUL WAY TO SPREAD THE VIRUSES AROUND THE WORLD, BUT IN A BENIGN FASHION. AND IT'S ONLY AFTER THEY ENTER THE POULTRY OF THE WORLD, THE BACK YARD FLOCKS WHERE WE HAVE THE DUCKS AND THE GEESE AND THE CHICKENS, EVERYONE MIXED TOGETHER, AND THEN THEY GET INTO THE CHICKEN HOUSES, GO THROUGH HUNDREDS OF BILLIONS OF RIPICATIONS AND BECOME HIGHLY PATHOGENIC. -- RIPICATIONS AND BECOME HIGHLY PATHOGENIC. THIS IS AN EXAMPLE WHAT HAVE HAPPENS WHEN THE MEXICO VIRUS, THE H5 NEXT 2 EVOLVED IN 1995. A BENIGN VIRUS CIRCUMSTANCEULATING IN THE NORTH AMERICAN AQUATIC BIRDS GOT INTO THE POULTRY OF MEXICO THE MAIN CHANGE, AS -- AS YOSHIRO KAWAOKA MENTIONED, IS IN THE HEMOGLUTINEN AND THE PEPTIDE REGION DUE TO, PRESUMABLY REPLICATION. AND SO THE SERIES OF AMINO ACIDS AND THE VIRUS SPREADS AND REPLICATES IN EVERY TISSUE OF THE BODY . THE RECENT HIGHLY PATHOGENIC INFLUENZA OF THE H5 AND THE H7 WOULD BE THE HONG KONG VIRUSES, THE VIETNAM VIRUSES. ACROSS THE WHOLE OF ASIA AND MORE RECENTLY, H5 N2 IN TEXAS AND H7 AND 7 IN HOLLAND AND MORE RECENTLY IN CANADA. THE H7 VIRUSES MAINLY CAUSE CONJUNCTIVITIS. THERE HAS BEEN ONE DEATH IN HOLLAND IN HUMAN AND SO THAT GOES INTO THE HIGH-RISK CATEGORY FOR HUMANS. MOST OF THE CASES IN HUMANS HAVE BEE CONJURVGHT -- CONJUNCTIVITIS, BUT A LARGE NUMBER OF PEOPLE HAVE BEEN INFECTED IN THE HOLLAND INCIDENT AND PROBABLY MORE THAN WE REALIZED. I WANT TO SPEND THE REST OF THE TIME ALLOCATED TO ME ON THE H5 N1 SITUATION. BECAUSE THIS IS THE RISK FACING EVERY ONE IN THIS ROOM TODAY. IN THE BEGINNING OF THIS YEAR, HAPPENS OF MILLIONS OF CHICKENS WERE CULLED ACROSS ASIA TO STOP THE SPREAD OF THE VIRUS. IN VIETNAM AND THAILAND, THERE HAS BEEN A TOTAL OF 39 HUMAN CASES, 28 DEATHS OF THESE 20 IN VIETNAM AND EIGHT IN THAILAND. SO THIS IS AN UNPRECEDENT, VENT IN THE HISTORY OF INFLU EPZA. PEOPLE ARE IN -- INFLUENZA. PEOPLE ARE INCLINED TO SAY THIS IS ONLY BECAUSE THE CDC, NIH AND EVERYONE IS DOING MUCH MORE SURVEILLANCE. THAT'S PARTLY TRUE, BUT ALSO, THIS IS SOMETHING THAT WE HAVE NEVER SEEN BEFORE AND IS PROBABLY THE KIND OF THING THAT HAPPENED IN 1918. WE SAW PEOPLE FROM -- [ INDISCERNIBLE ] 'S WORK, PEOPLE BORN BEFORE 1918 HAD ANTIBODIES. THIS IS PROBABLY THE RUN-UP TO WHAT IS GOING TO HAPPEN IN THE WORLD. SO TO WALK YOU THROUGH THE GENESIS OF THIS VIRUS. THIS VIRUS FIRST APPEARED IN GEESE IN GUAN DONG IN 1996 AND SPREAD INTO THE POULTRY MARKETS IN HONG KONG AND THIS VIRUS MATED WITH AN H9 IN QUAIL AND A DUCK VIRUS AND THIS TRIPLE REASSORTMENT WENT INTO HUMANS IN HONG KONG AND KILLED SIX OF 18 PEOPLE. THAT REVIRUS WAS GOTTEN RID OF BY -- THAT VIRUS WAS GOTTEN RID OF BY SLAUGHTERING ALL THE POULTRY IN HONG CONK, BUT THE PARENTAL VIRUS WAS STILL OUT THERE IN GEESE, AND FROM 1997 TO 2002, THIS GOOSE H5 N1 MATED WITH MULTIPLE DIFFERENT VIRUSES, BUT IN THIS PERIOD OF TIME, THE VIRUS WAS ANTIGENICALLY CONSERVED. THERE ARE MULTIPLE GENOTYPES FROM REASSORTMENT AND THE VIRUS WASN'T PATHOGENIC IN DUCKS. THIS VIRUS, AS DR. CHEN SHOWED, THE GOOSE VIRUS THEN SPREAD INTO THE DUCKS OF CHINA AND ALL OF THE DUCK ISOLATES FROM THE COASTAL PROVINCES OF CHINA FROM 1996 TO 2002 WERE ALL PAGOGENIC FOR POULTRY -- PATHOGENIC FOR POULTRY. AND THEN THIS H5 N1 IN 2002 CHANGED. THE VIRUS CONTINUED TO MATE WITH VIRUSES IN THE REGION AND PRODUCED VIRUSES THAT WERE THEN ISOLATED FROM WILD MIGRATED BIRDS. THIS IS A FIRST TIME FOR MANY, MANY YEARS THAT A HIGHLY PATHOGENIC VIRUS HAS BEEN ISOLATED FROM WILDER ABOUTS. THIS VIRUS -- WILD BIRDS. IT'S SHOWING DRIFT IN THE HEMOGLUTINEN AND HAS BECOME PATHOGENEC FOR AQUATIC BIRDS. IN 2003, A FAMILY VISITING -- [ INDISCERNIBLE ] -- A DAUGHTER DIED. THE VIRUS WASN'T ISOLATED. THE FATHER AND SON RETURNED TO HONG KONG AND THE FATHER DID -- DIED. THAT'S THE PRECURSOR OF THE VIRUS OUT THERE CAUSING TROUBLE AT MOMENT. THIS IS A DIAGRAM OF THE GENESIS. THE GOOSE VERACE -- VIRUS MATED IN 2000 WITH ANOTHER VIRUS N.20 WHERE ARE 1, IT MATED WITH YET MORE VIRUS PEACE AND SO ON. THIS RES -- REASSORTMENT IS VERY, VERY ACTIVE WITH THIS VIRUS. FROM 2003 ON, WE HAD SELECTION DOWN TO WHAT WE CALL THE Z GENOTYPE, THAT IS DOMINANT AND PASSING TO HUMANS BUT NOT PASSING HUMAN-TO-HUMAN. SO HOW PATHOGENIC IS THIS VIRUS? THIS VIRUS, THE VIETNAM 2004, 1203 KILLS CHICKENS IN LESS THAN ONE DAY. IT TAKES ONE INFECTIOUS UNIT TO KILL A CHICKEN OVER NIGHT. IT'S ENORMOUSLY PATHOGENIC. IT KILLS DUCKS IN ONE TO TWO DISTRICT ATTORNEYS AND HAS A HIGH RISK OF DEATH IN HUMANS, APPROXIMATELY -- DAYS, AND HAS A HIGH RISK OF DEATH IN HUMANS IN 40 YEARS WITH DIARRHEA AND RESPIRATORY SYSTEMS. THE MODEL, THE AVAILABLE MODEL WE HAVE FOR H5 N1 IN MAMMALS IS THE FERRET, THE CLASSICAL FERRET MODEL. IN THE FERRET MODEL, THE VIRUS CAUSES RESPIRATORY SYMPTOMS, GAINED DIARRHEA, HIND-LEG PARALYSIS AND THEY ALL DIE. OR THE MAJORITY DIED. DR. HE, M LOOKED AT THE FIRST -- HEIM LOOKED AT THE FIRST 10 CASES IN VIETNAM. THE -- [ INDISCERNIBLE ] DEATH USED IN 27 YEARS. NO PRE-EXISTING MEDICAL CONDITIONS. HIGH FEVER, LYMPHOPENIA, 7 OUT OF 10 HAD DIAROWA, AND 7 OUT OF 10 DIED. THIS IS A VERY, VERY BAD VIRUS. THIS, IF WE DO POLYGENETIC ANALYSIS, THIS VIRUS IN VIETNAM IS MOST CLOSELY RELATED -- RELATED TO A VIRUS IN DUCKS AND TO CHICKENS, SO THAT THE HUMAN AND DUCK VIRUSES ARE GENNATE GENETICALLY CLOSELY RELATED AND THE NEXT MOST CLOSELY-RELATED GROUP ARE FROM WILD MIGRATING BIRDS THAT HAVE DIED. WHAT ABOUT PIGS? PIGS ARE CONSIDERED THE INTERMEDIATE HOST WHAT. DO WE KNOW ABOUT PIGS AND THIS VIRUS? ONE ISOLATE HAS BEEN REPORTED FROM HANOE. WE DID SYROLOGY IN HONG KONG, 139, AND MORE RECENTLY IN CHEAPA, CHINA REPORTED THE ISOLATION OF H5 N1 ON A COUPLE OF OCCASIONS FROM PIGS, BUT THIS IS NOT THE X GENOTYPE. ON THE Z GENOTYPE. WE PUT THIS VIRUS INTO PIGS IN THE THREE AUGMENTED CONDITIONS AND LOOK FOR REPLICATION. THE VIETNAM HUMAN ISOLATE, THE CHICK ISOLATE, THE DUCK ISOLATE, AND THE GOOSE ISOLATE IN THAILAND ALL REPLICATED VERY WELL IN PIGS BUT DID NOT TRANSMIT TO CONTACT PIGS. AND FURTHER SYROLOGY IN ASIA INDICATES THERE ARE SOME POSITIVE PIGS. THIS VIRUS IS NOT WIDESPREAD IN PIGS. THE MESSAGE IS THE VIRUS DOES HAVE THE CAPACITY TO REPLICATE IN THE PIGS, BUT LIKE IN HUMANS, IT HAS NOT YET LEARNED TO TRANSMIT PIG TUG PIG. -- PIG-TO-PIG. WE ALSO KNOW THAT THIS VIRUS HAS TRANSMITTED TO THE HIGHER CATS, LEOPARDS AND TIGERS AND THE GROUP IN HOLLAND, AUSTER HOUSES GROUP SHOWN EXPERIMENTAL TRANSMISSION IN CATS. DOMESTIC CATS IN EVERY HOUSEHOLD. THIS IS A SERIOUS PROBLEM AND THEY MADE THE MISTAKE IN THAILAND OF FEEDING DISEASED ANIMALS TO THE TIGERS AND CATS. I ALSO HAVE TO IMPRESS ON YOU THAT THE PEOPLE IN THIS REGION OF THE WORLD ARE VERY POOR THE LAST CASES OF HUMAN INFECTION IN VIETNAM WERE CAUSED BY THE PARENTS FEEDING DISEASED CHICKENS TO THE FAMILY. THESE PEOPLE ARE SO POOR THAT THEY ONLY GET CHICKEN TO EAT WHEN THE CHICKENS ARE DYING. SO THIS IS THE SITUATION IN THIS PART OF THE WORLD. VERY POOR PEOPLE LIVING CLOSE PROXIMITY AND SO THEY DO THESE KINDS OF THINGS THAT WE DON'T ACCEPT. THE SEASONALITY OF THE H5 N1, WE LOOK BACK TO HONG KONG AND FIND THE OUTBREAKS HAVE OCCURRED MAINLY IN THE FALL AND WINTER MONTHS FIRST IN THE AQUATIC BIRDS, THEN IN THE TERRESTRIAL BIRDS LIKE CHICKENS. AND SO WE NOTICED THAT IN 2001, '2, '3, '4, THEY HAVE OCCURRED IN THE WINTER MONTHS. WHAT IS GOING ON NOW? THERE'S AN OUTBREAK GOING ON CURRENTLY IN VIETNAM AND THAILAND IN THE SUMMER MONTHS. THAT'S WORRYING. SO, WERE MIGRATING BIRDS INVOLVED IN THE SPREAD OF THIS VIRUS? MAYBE. THESE VIRUSES WERE ISOLATED FROM BLACK HEADED GULL, PIGEONS AND LEGRETS, BUT THESE ARE SCAF EACHERS. THESE ARE NOT SCAFENGERS, GRAY HERON, PREREGRIN FALCOPS, NOT REALLY, WE DON'T KNOW IF THEY'RE OUT THERE IN MIGRATING BIRDS. PREVENTION AND CONTROL. THE PRACTICE OF WET MARKETS AS WE CALL THEM, WHERE YOU HAVE RAISED, YOU HAVE CHICKENS, LIVE CHICK KNOWS AND DUCKS AND EVERYTHING -- CHICKENS AND DUCKS AND EVERYTHING TOGETHER FOR SELL TO THE POULTRY IN ASIA AND IN NEW YORK IS ONE OF THE PROBLEMS THAT WE FACE. THIS IS A CULTURAL PREFERENCE TO THE WAY YOU BUY POULTRY. IT'S ONE OF THE PROBLEMS FOR INFLUENZA. BECAUSE THESE ARE PERFECT BREEDING GROUNDS FOR INFLUENZA AND IN HONG KONG OVER THE YEARS SINCE 1998, WE HAVE SHOWN, WE HAVE TAKEN THE DUCKS AND THE GEESE OUT OF THE MARKETS, WE HAVE CLEAN DAYS TO IMPROVE SAPPITATION. WE'RE TAKING THE QUAILS OUT. -- SANITATION. WE HAVE TAKEN THE QUALS OUT AND WE KEEP CHANGING THINGS. VACCINES HAVE BEEN USED TO VACCINATE THE FARMS IN THE NEW TERRITORIES, ADDITIONAL CLEAN DAYS AND NOW WITH ALL OF THESE STEPS HONG KONG DNTS HAVE H5 IN -- DIDN'T HAVE H5 IN 2004. THERE ARE WAYS TO IMPROVE THIS. IMPROVE SANITATION, USE OF VACCINES. I DON'T HAVE TIME. THE AGRICULTURAL VACCINES ARE TO ME THE MAIN PROBLEM. IF WE GO ACROSS THE BORDER TO CHINA, THIS IS WHAT IS GOING ON. NONE OF THESE THINGS HAVE BEEN LISTENED TO. IN CHINA, THEIR TRADITIONAL MARKETS STILL GO ON AND THROUGHOUT ASIA AND SO CHANGES IN THE TRADITIONAL MARKET ARE CONSISTENT AND PERSISTENCE IS GOING TO TAKE A LONG TIME. VACCINE WE HEARD ABOUT REVERSE GENETICS FROM YOSHIRO KAWAOKA, AND THIS IS PROBABLY THE FIRST IMPORTANT USE OF REVERSE GENETIC STRATEGY. THIS VIETNAM VIRUS WITH A VARIANT AND SO THE, THE YOUNG PEOPLE IN MY LAB MADE, MAY USE REVERSE GENETICS TO MODIFY THE HUMAN GLUTTINNIN, WHICH IS ONE OF THE MARKETS FOR PATHOGENICITY, CLIPPED OUT THE NUCK LEER -- NUCLEOTIDES AND MADE A MODIFIED HEMOGLUTINEN AND THE EUROMINASE PLUS THE SIX GENES FROM CLASSIC AT -- CLASSICAL PR8 INFECTED INTRAVIRAL CELLS AND THEN PRODUCING A VIRUS THAT WE CAN THEN GROW IN EGGS. THESE VIRUSES, GO BACK A BIT. THESE VIRUSES ARE NONPATHOGENIC IN FERRETS, MICE, AND CHICKENS AND IMMUNOGENIC IN FERRETS. AND THIS IS THE VACCINE THAT IS UNDETESTING BY NIAID AT THE CURRENT TIME. ANTIVIRALS, UNFORTUNATELY, THE H5 N1 VIRUSES ARE NOT SENSITIVE TO AMENTADINE DIMENTADINE. THE ORIGINAL 1997 VIRUSES WERE SENSITIVE BUT FROM 2003 ON, ALL OF THE VIRUSES OR THE MAJORITY OF THE VIRUSES ARE RESISTENT. AND WE'RE NOT AT ALL SURE WHY THIS RESISTANCE HAS OCCURRED. MY HYPOTHESIS IS THIS IS A PHENOMENON, SIMILAR TO THE VIRUSES AROUND IN HUMANS IN THE PR8 TO 1940 PERIOD. SO WHAT ABOUT TENSE TIST TO OSILTAMADIRE? THE GOOD NEWS IS YES THEY'RE SENSITIVE TO OSILTAMIBIRE AND THE VIRUS DOESN'T SHOW UP IN THE LUNGS, BRAIN OR SPLON WHEN THE POLICE ARE TREATED WITH OSILTAMIBIRE. SO WE HAVE A SUITABLE DRUG FOR THESE VIRUSES. SO "TIME" MAGAZINE HAD YET RIGHT. ASIA IS HATCHING, PROBABLY HATCHING THE NEXT HUMAN PANDEMIC AND IN ADDITION, THERE ARE MANY OTHER THINGS GOING ON WITH INFLUENZA. IN BRITISH COLUMBIA, WE JUST HAD H7 AND 3 GENERATED BY RECOMBINATION, NOT REASSORTMENT. IN THE UNITED STATES, IN THE EAST COAST, WE HAVE H7 AND 2 BREWING, A VERY HIGHLY PATHOGENIC STRAIN. IN A YEAR OR TWO, H5 AND 2 IN SIX SIX -- TEXAS, H6 AND 2 NON-PATHOGENIC IN CALIFORNIA. MANY THINGS GOING ON. AND HAVE SOME OF THESE -- HAD SOME OF THESE GENES GONE BACK INTO THE WILD BIRDS. THAT'S WHAT WE HAVE TO KNOW. WHAT ABOUT THE FACILITIES WE USE TO DO THESE EXPERIMENTS. WHAT ABOUT THE SAFETY WE USE IN ALL OF THESE SPERMS THAT WE HAVE DONE HAVE BEEN APPROVED BY -- EXPERIMENTS THAT WE HAVE DONE HAVE BEEN APPROVED BY A SAFETY CONTAINMENT AND IN CONJUNCTION WITH OUR BIOSAFETY OFFICES. THE FACILITIES ARE STATE-OF-THE-ART THAT WE HAVE AT ST. JUDE CHILDREN'S RESEARCH HOSPITAL AND THE SAFETY FEATURES THAT YOSHIRO KAWAOKA DESCRIBED ARE EXACTLY THOSE USED FOR THE EXPERIMENTS. THE STAFF ARE PROTECTED AND THE STAFF ARE EXTREMELY WELL-TRAINED. WE HAVE A MANAGER OF OUR FACILITY AND THE YOUNG PEOPLE COMING IN ARE ALL THOROUGHLY TRAINED. SO, THE NONPATHOGENIC AVIAN INFLUENZA WORKING IN DL 2, THE PATHOGENIC A RAIN VIRUSES THAT DON'T TRANSMIT TO HUMANS, DL 3. THE PATHOGENIC AVIANS THAT TRANSMIT TO HUMANS, BL 3 AUGMENTED WITH THE USE OF PAPR AND IN ISOLATEORS. AND THE PATHOGENIC AVIAN VIRUSES WITH GENE EXCHANGE OR GENE MODIFICATION ALLOW THE PRODUCTION OF THE VACCINE THAT IS CURRENTLY NECESSARY PRODUCED IN VL 3 AUGMENTED WITH PAPR WITH ISOLATEORS AND THEN -- THEN WE HAVE TO GO THROUGH THE PROCEDURES OF REMOVING, HAVING THAT STEP DOWN TO ALOWER LEVEL AND -- A LOWER LEVEL AND -- [ INDISCERNIBLE ] -- WILL PROBABLY DEAL WITH THAT INFERTILL. SO, I WILL STOP THERE. I WANT TO ACKNOWLEDGE THE SUPPORT OF NIAID, OF THE, MY COLLEAGUES AT ST. JUDE CHILDREN'S RESEARCH HOSPITAL, RICHARD WEBBY AND ERIC HOFFMAN MADE THE VACCINE STRAINS, THE STUDIES IN FERRETS AND THE COLLABORATORS IN THE HONG KONG MALIKA TERRACE AND GUAN DONG. THESE STUDIES WOULDN'T BE POSSIBLE WITHOUT THE COOPERATION OF THE INDONESIAN, VIETNAM AND THAILAND BUREAUS OF MINISTRIES OF AGRICULTURE AND PUBLIC HEALTH. SO, I, I JUST STOP AND RETURN TO MY THEME THAT THERE IS A GREAT THREAT OUT THERE IN ASIA AT THE MOMENT. JUST TO REITERATE THE MAIN POINTS. THERE IS A LACK, TOTAL LACK OF ANTIBODIES OR IMMUNITY IN THE POPULATION TO THIS VIRUS AS DISTINCT WITH THE 1918. AT THAT -- THAT RATE IN MY MIND MAKES THE H5 N1 MUCH MORE DANGEROUS VIRUS TO BE WORKING WITH THAN 1918 THIS SUMMER OUTBREAK GOING ON IN ASIA NOW, IS IT A HERALD WAY FOR WHAT IS GOING TO HAPPEN THIS FALL? WE HAVE BETTER GET OUR DUCKS IN A ROW, AS IT WERE. AND GET OUR OSILTAVIM, R STORED IN SUFFICIENT QUANTITIES BECAUSE THE ONLY THING WE HAVE GOT UNTIL THE VACCINES ARE TESTED AND PREPARED ARE, IS OSILTAMAVIR AT THIS TIME. AND WE HAVE TO WORK THROUGH QUICKLY, MAKE USE OF THE BIOSECURITY FACILITIES TO PRODUCE THE VACCINES TO GET THEM TESTED SO THAT COMPANIES CAN WORK OUT, CAN THEY MAKE VACCINES? DO THEY HAVE FACILITIES IF THIS KIND OF VIRUS SPREADS? SO, I WILL STOP THERE. AND THANK YOU. [ APPLAUSE ]

WE HAVE TIME FOR SOME QUESTIONS.

I WONDER IF YOU COULD COMMENT A BIT MORE. YOU MENTIONED TRAINING THAT INDIVIDUALS COULD WORK WITH THESE AGENTS, AND I WONDER IF YOU COULD JUST ELABORATE ON WHAT YOU FEEL IS APPROPRIATE FOR SPECIAL TRAINING AND WHETHER THERE ARE SPECIFIC GUIDELINES THAT YOU USE.

WELL, WE HAVE A MANAGER OF OUR BL 3 FACILITY AND WE HAVE SOP 8 AND THE -- SOPs AND EVERY PERSON BEFORE THEY ENTER THE BL3 IS TRAINED AND HAS A LITTLE EXAMINATION AND IS, THEY MUST FULLY UNDERSTAND THE PROCEDURES IN USE. AND THEN THEY GO THROUGH A PERIOD OF APPRENTICESHIP. YOU JUST DON'T PASS THE EXAMINATION AND GO IN. THEN YOU WORK, AS AN APPRENTICE WITH THE PEOPLE IN THE BL3 FACILITY&UNTIL THEY ARE APPROVED -- FACILITY UNTIL THEY ARE APPROVED. THAT'S THE KIND OF TRAINING, FIRST THEORETICAL, THEN PRACTICAL AND THEN YOU'RE SIGNED OFF AND THEN WITH THESE AGENTS, OF COURSE, YOU MUST BE APPROVED AHEAD OF TIME TO BE ABLE TO WORK WITH A SELECT AGENT. YES, SIR.

ARE THERE ANY SPECIAL PRACTICES IN THE BL3. YOU MENTIONED BL3 BEING APPLIED IN THE LAST THREE CATEGORIES OF WORK, AND I'M WONDERING IF THERE ARE ANY SPECIAL PRACTICES THAT YOU HAVE INTRODUCED INTO THE LAB BEYOND STADDARD BL3A PRACTICE. -- STANDARD BL 3 PRACTICE.

YES, WITH THE HUMAN VIRUSES, WE'RE USE THE PAPRs AND WE ARE USING ALL OF THE STRATEGIES THAT YOSHI MENTIONED THAT THE STAFF HAVE TO BE VACCINATED. ONE OF THE THINGS THAT I DID, STRONGLY DISAGREE WITH IS THE REMOVAL OF THE PRACTICE OF TAKING SERUM FROM STAFF. I ARGUE VERY STRONGLY AGAINST THAT. IF THE STAFF ARE WILLING TO HAVE SERUM TAKEN, WE NEED TO KNOW IF WE HAVE ANY RISKS AND THIS BUSINESS OF NOT HAVING SERUM TAKEN IS A TERRIBLE MISTAKE. IN ADDITION, THE STUDIES ON, IN ANIMALS ARE DONE IN THE ISOLATED SYSTEMS THAT HAVE THEIR, THAT ARE INSIDE BL 3 AIR-CONTROLLED, WATER-CONTROLLED SYSTEMS, EACH WITH THEIR OWN FILTER SYSTEMS AND, IN OTHER WORDS, A SEPARATE UNIT INSIDE A BLESS -- BL THIRTY-TWO. THOSE ARE THE ADDITIONAL LEVEL -- BL3. THOSE ARE THE ADDITIONAL LEVELS WE HAVE PUT INTO PLACE. YES.

YOU MENTIONED SEVEN OUT OF EIGHT PEOPLE GOT THE H5 N1 AND DIARRHEA. WAS VIRUS EXCLUDED IN THE STOOL?

YOU KNOW, ONE OF THE DIFFICULTIES IN CHINA IS TO GET THE INFORMATION LIKE THIS THAT YOU NEED. THE CHINESE FOLKS DO NOT LIKE TO GIVE PERMISSION FOR POST MORTEMS. WE, THESE THINGS ARE BEING LOOKED AT. WE DON'T HAVE ADEQUATE INFLAMMATION. I WOULD SUGGEST THAT FROM THE RESULTS THAT WE'RE OBTAINING IN FERRETS TO THE QUESTION YOU ASKED PREELS, I SUSPECT THE -- PREVIOUSLY, I SUSPECT THE VIRUS IS GETTING OUTSIDE THE LABORATORY. THAT WOULD BE MY GUESS. I THINK WE HAVE TO WORK UNDER THE ASSUMPTION BASED ON THE FERRET MODEL THAT THE, THESE VIRUSES ARE CAUSING NEUROTROVIC DISEASE IN THE FERRETS. IT'S GOING, IT'S HAPPENING. YES.

IS ANY INFORMATION IN TERMS OF THE CLINICAL SYMPTOMS, THE SYMPTOMS FIRST, DIARRHEA WAS FIRST. WAS THAT RECORDED IN.

THE DIARRHEA IS MOCKED IN THE HUMANS AND AND THE FERRET. ONE OF THE INTERESTING THINGS THE PATHOLOGY DOESN'T GO ALONG WITH THAT WE DON'T SEE ANY PATHOLOGY, SO IT'S ONE OF THOSE INTERESTING THINGS THAT YOU CAN HELP WORK OUT, PERHAPS. YES.

BOTH OF YOU MENTIONED THAT DUE TO SOME FACTORS, YOU'RE NO LONGER, OR YOU ARE NOT CHECKING SERUM FROM STAFF. CAN YOU SAY HOW THAT CAME ABOUT AND WHAT THE AUTHORITY THAT PREVENTS YOU FROM DOING THAT IS? IS THAT LOCAL OR IS THAT --

I THINK IT'S A RECOMMENDATION FROM HIGHER UPS SOMEWHERE IN THIS COUNTRY OF ETHICS. IT COMES FROM AN ETHICS COMMITTEE SOMEWHERE THAT SAYS THAT IT WAS BASED ON AN ETHICS COMMITTEE, A RECOMMENDATION TO A BIOSAFETY COMMITTEE AND I, I ARGUED WITH THE BIOSAFETY OFFICER AND DID NOT LET THEM REMOVE THIS FROM MY FACILITY. I, MY ARGUMENT WAS THAT IF MY STAFF IS WILLING TO HAVE THIS AND NOT HAVE THEIR NAMES ASSOCIATED WITH IT, LET'S DO IT. IT'S ABSOLUTELY ESSENTIAL FROM MY POINT OF VOW THAT THIS IS DONE. BECAUSE WE NEED TO KNOW WITH ALL OF THESE LEVELS OF CONTAINMENT THAT WE USE, ARE THEY UNDER RISK AND THIS IS INFORMATION THAT IS VITAL FOR EVERYONE.

SO THAT'S NOT REGULATIONS, BUT RECOMMEND -- RECOMMENDATION, WHICH YOU ARE ABLE TO NEGATE AT YOUR FACILITY?

YES, I, I DID. MAYBE I SHOULDN'T SAY THAT. [ LAUGHTER ]

RIGHT.

DR. WEBSTER, IN THOSE SEVEN OF 10 PATIENTS, I BELIEVE WHAT MY NOTES SAID, SEVEN OF 10 PATIENTS WHO DIED.

IN VIETNAM.

IN CHINA?

IN VIETNAM.

YES.

SEVEN OF 10 PATIENTS HAD DIAROWA SYMPTOMS. WERE THOSE THE SAME PATIENTS?

YES.

THE SAME PATIENTS WHO DIED WERE THE ONES THAT HAD THE DIARRHEA?

I UNDERSTAND THAT WAS THE SAME. THAT'S NOT NOT MY WORK, BUT HEINZ WORK THAT IS PUBLISHED. CAN YOU GO BACK AND CHECK IT IN "THE NEW ENGLAND MEDICAL JOURNAL." THAT'S MY UNDERSTANDING.

OKAY, THANK YOU VERY MUCH. I THINK WE NEED TO MOVE ON. THE NEXT TALK IS FROM DR. KANTA SUBBARAO. FROM THE NATIONAL INSTITUE OF ALLERGY AND INFECTIOUS DISEASES AND SHE'S GOING TO TACK ABOUT VACCINE APPLICATIONS. -- TALK ABOUT VACCINE APPLICATIONS .

I THANK YOU. I'M GOING TO SPEND THE NEXT 20 MINUTES OR SO TALKING ABOUT THE APPLICATION OF REVERSE GENETICS TECHNIQUES TO VACCINE DEVELOPMENT FOR INFLUENZA. AS YOU HAVE HEARD BEFORE, THE SCHEMATIC OF THE ENFLU EPZA VIRUS, I WANT TO DRAW YOUR ATTENTION TO TWO PROTEINS, THE TARGETS OF THE PROTECTIVE IMMUNE RESPONSE N.DEVELOPMENTING VACCINES, THERE ARE TWO LICENSED VACCINES FOR HUMAN INFLUENZA. THAT BOTH ACT ON THE PRINCIPAL OF INDUCING PROTECTIVE IMMUNE RESPONSE AGAINST THE HEMOGLUTINEN PROTEIN. THE VACCINES LICENSED FOR USE ARE TRIVALENT VACCINE, THAT CONTAINS STRAINS THAT ARE REPRESENTATIVE OF WHAT ARE CURRENTLY CIRCULATING AND CURRENTLY BILOZAR, THE H1 N1, H2 IN INFLUENZA A AND B COMPONENTS. THE CURRENTLY-LICENSED CONVENTIONAL AND ACTIVATED VACCINE CONTAINS REASSORTMENT VIRUSES THAT CONTAIN THE HEMOGLUTTINNEN AND EURO MENNACY GENES WITH INIT WEREAL GENES FROM VACCINE DONOR STRAIN CALLED PR8s AND INFLUENZA B COMPONENT IS REPRESENT -- REPRESENTED BY WILD-TYPE INFLUENZA B VIRUS. IN THE CASE OF THE LIVE ATTENUATED ADAPTIVE VACCINE RECENTLY LICENSED, THE VACCINE IS, AGAIN, A TRIVALENT VACCINE CONTAINING REASSORTMENT VIRUSES CONTAINING THE HEMOGLUTINEN AND -- [ INDISCERNIBLE ] OF INFLUENZA A AND B VIRUSES AND FROM MASTER DONNOR STRAINS THAT HAVE THE COLD ADAPTATION AND ACCUMULATION PHENOTYPES CALLED RA AND ARBOR 160 AND 166 RESPECT ELF. THE REASON THAT INFLUENZA VACCINES ARE UPDATED ANNUALLY IS BECAUSE VANEXT, -- ANTIGENIC DRIFS. YOU HAVE BEEN HEARING ABOUT THE ANTIGENIC SHOULD OFS. IT'S THE -- [ INDISCERNIBLE ] MAINLY THE HEMOGLUTINEN AND THE NEUROMINNOWACY AS WELL, RESULTING IN THE ABILITY TO ANTIBODY TO PREVIOUS STRAINS OCCURRING AS A RESULT OF A NATURAL INFECTION OR PREVIOUS INHUMANIZATIONS TO NEUTRALIZE NEW VIRUSES. THIS DRIFT RESULTS FROM POINT MUTATIONS IN THE HEMOGLUTINEN AND NEUROMINOASE GENES. THEREFORE, IT HAS TO BE UPDATED ANNUALLY, THE VACCINE. IN THIS DIAGRAM OF THE, OF ONE OF THE MONOMERS FORMING THE TRIMERS OF THE HEMOGLUEDINNEN, THERE ARE FIVE COMBINING SITES IDENTIFIED IN AND AROUND THE RECEPTOR BINDING POCKET OF THE HEMOGLUTTINNEN. WE KNOW THAT AMINEO ACID SUBSTITUTIONS IN TWO OR MORE OF THESE ANTIBODY COMBINING SITES RESULTS IN AN ANTIGENIC DRIFT VARIANCE. SURVEILLANCE DATA TELLS US OUR LOGIC SURVEILLANCE TELLS US WHERE CHANGES ARE OCCURRING IN CIRCULATING INFLUENZA STRAINS THAN ARE, THE INFORMATION IS TAKEN INTO CONSIDERATION IN UPDATING VACCINE RECOMMENDATIONS. THE GENERATION OF REASSORTMENT INFLUENZA VIRUS AS THE CONVENTIONAL APPROACH HAS BEEN TO GENERATE VACCINE STRAINS BY REASSORTMENT, AND HERE WE TAKE A VACCINE DONOR VIRUS WITH THE PHENOTYPE IN THE CASE OF A COLD ADAPTED VIRUS IN ATTENUATION OR PR8 IN HIGH GROWTH IN EGGS. THAT IS THEN MATED WITH THE CIRCULATING WILD TYPE VIRUS TO CREATE WHAT IS CALLED A 6-2 REASSERT -- REASSORTMENT WHERE THE HEMOGLUTTENEN AND NEUROMINOASE STRAIN ARE DERIVED FROM THE SIX INNER TENNAL SEGMENTS FROM THE VACCINE DONOR STRAIN N.THE CASE OF THE INACTIVATED VACCINE COMPONENTS OF THE INFLU AEPZ A GROUP, THE PR8 GENES WITH A HIGH GROWTH IN EGGS AND THE COLD ADAPTED VACCINES IN INFLUENZA A AND B CONFER ATTENUATION. NOW, REVERSE GENETICS CAN BE APPLIED TO THE RAPID GENERATION OF THESE VACCINE STRAINS INSTEAD OF RELYING ON REASSORTMENT AND ANTIBODY SLICKS. THESE STRAINS CAN BE GEN -- SELECTIONS. THEY CAN BE FAR MORE RAPTLY THAN REVERSE GENETICS. YOU HEARD ABOUT THIS FROM THE PREVIOUS SPEAKERS. BUT THE TECHNOLOGY THAT YOSHI FIRST DESCRIBED EARLIER HAS BEEN MODIFIED TO THE USE OF EIGHT PLASMIDS WHERE YOU CAN USE TWO PLASMIDS IN COATING THE GENES FOR THE HEMOGLUTTINNEN AND THE EURO MINNOW ASE JEEBS AND THE SIX GENES THAT ENCODE THE INTERNAL GENES FROM A VACCINE DONOR STRAIN. IF THEY'RE COTRANSVECTED INTO VACCINE GRADE CELLS, CAN YOU RECOVER VIRUS WITH THE DESIRED GENOTYPE. SO THE BIOSAFETY MEASURE THAT WOULD BE USED TO GENERATE SUCH REASSORTMENT INFLUENZA VACCINES WHEN WE'RE WORKING WITH CONVENTIONAL HUMAN EN -- INFLU ENZA VIRUSES, THEY WOULD BE REGENERATED AT THE BIOSAFETY LOCALS RECOMMONED FOR WORK AND THAT, IN THE CASE OF HUMAN VIRUSES, IS THE SL2, AND THE VACCINES AS MANUFACTURED, THE C VIRUSES THEN PROVIDED TO MANUFACTURERS. THE VACCINES, VACCINE VIRUSES CAN BE GENERATED WITH THE TECHNOLOGY, BUT I DON'T BELIEVE ANY OF THEM HAVE BEEN LICENSED. THE CONVENTIONAL VACCINES ARE GENERATED BY REASSORTMENT. BUT THE APPLICATION, THE APPLICATION OF REVERSE GENETICS TO THE GENERATION OF VACCINES IS ALSO VERY IMPORTANT&IN PREPARES -- PREPARING VACCINES AGAINST POTENTIAL PANDEMIC STRAINS AND YOU HAVE HEARD, YOU HAVE HEARD THIS FROM A NUMBER OF SPEAKERS, THE RESERVOIR OF A VIRUS IN WATERFOWL AND SHORE BIRDS, THERE ARE 15 HEMOGLUTTINNEN AND 9 NEUROMINOAS AND OVER THE LAST CENTURY, THREE PANDEMICS RECORDED, THE 1918 OF H1 N1 VIRUSES, THE H2 N2 VIRUSES IN 1957, THE H3 AND N2 IN 1968 AND H1 N1 REAPPEARED IN 1977. WE KNOW THAT THE, THAT THE RESERVOIR VIRUSES IN NATURE ARE WATERFOWL AND SHORE BIRDS AND AS YOU HEARD, THEY'RE THE SOURCE FROM COME NOVEL INFLUENZA SUBTYPES ARE INTRODUCED INTO THE HUMAN POPULATION. YOU HEARD, AGAIN, FROM DR. WEBSTER ABOUT THE NUMBER OF RECENT OUTBREAKS OF AVIAN INFLUENZA IN POULTRY AND THIS IS BY NO MEANS A COMPLETE LIST STARTING FROM 1983, THE OUTBREAK IN PENNSYLVANIA, THE H5 N2 INFECTS IN POULTRY, THE 1997 -- 1995 H5 H2 IN MEXICO AND FROM 1997 ONWADS, YOU HEARD OF MORE INSTANCES OF H5 AND H7 INFECTS CAUSING OUTA -- INFECTS CAUSING OUTBREAKS OF INFLUENZA IN POULTRY AROUND THE WORLD. OUR CONCERN FROM THE HUMAN INFLUENZA STANDPOINT IS THE RISK OF THE B VIRUSES INFECTING HUMANS. AGAIN, YOU HEARD ALL OF THESE DATA BMPLT H5 VIRUSES TRAPPED MITTED TO HUMANS IN 19 MECHE HAD HONG CONK WITH 18 CASES AND SIX DEATHS. IN 2 VERY 0 -- 2003, A FAMILY WAS INFECTED WITH THREE CASES AND TWO DEATHS AND VIETNAM AND THAILAND, COMBINING THE SPRING OUTBREAK WITH THE CURRENT OUTBREAK, UM, TO DATE, THERE ARE 39 CASES AND 28 DEATHS. IN, WITH THE H7 EPFECKS, THERE HAVE BEEN A NUMBER OF CASE REPORTS OF CONJUMPGHT VITIS AND THE MORE REVENT LARGE OUTBREAK IN 2003 SPREADING FROM THE NETHERLANDS TO A NUMBER OF OTHER COUNTRIES AND THERE WERE A NUMBER OF CASES OF CONCONJUNCTIVITIS AND ONE DEATH. H5 VIRUSES TRANSMITTING TO HUMANS HAVE NOT BELONGED TO THE PATHOGENIC GROUP OF VIRUSES BUT THERE HAVE BEEN DOCUMENTED CASES OF TRANSMISSION IN HONG KONG AND SOUTHERN CHINA AND THEN MORE RECENTLY IN 2003, IN HONG KONG, TWO CASES OF H9 AS WELL. THE REASON WE'RE CONCERNED ABOUT THESE AS BASED ON A MATHEMATICAL MODEL BY MARTIN MELTZER AT THE CDC, THE ESTIMATES FOR THE FIRST YEAR OF A PANDEMIC IN THE ABSENCE OF INTERVENTION ARE THAT WE COULD ANTICIPATE BETWEEN 89 AND 207 DEATHS IN THE UNITED STATES ALONE -- AND 300 TO 733,000 HOSPITAL SITUATIONS WITH A VERY LARGE NUMBER OF OUTPATIENT VISITS AND A TREMENDOUS ECONOMIC IMPACT. SO IT'S IMPAIR TIF -- IMPAIRATIVE WE ACTUALLY HAVE VACCINES AVAILABLE TO PROTECT AGAINST THE SUCH EVENTS. SO THE APPLICATION OF VEVERSE GENETICS OR THE APPLICATION TO THE CURRENT TECHNOLOGY TO DEVELOPING PANDEMIC VACCINES COULD BE BASED ON THE PLASMID-BASED TECHNOLOGY THAT YOU HEARD B. THE SAME PRINCIPLE COULD APPLY WHERE YOU COTRANSVECT PLASMIDS, ONE FROM AN AVIAN INFLUENZA VIRUS AND SIX PLASMIDS FROM A DONOR STRAIN. THE STRATEGY COULD BE, OF COURSE, THE CONVENTIONAL TECHNOLOGY OF CONVENTIONAL REASSORTMENTS IF YOU DIDN'T HAVE TO MODIFY THE GENE SEGMENTS. YOU COULD RELY ON REVERSE GENETICS, I MEAN ON REASSORTMENT. WHAT I'M GOING TO DO IS TALK TO YOU, GIVE YOU EXAMPLES OF USE OF BOTH OF THESE TECHNOLOGIES THAT WE HAVE BEEN INVOLVED WITH IN DEVELOPING VACCINES AGAINST THE DIFFERENT SUBTYPES AND THE GENERAL APPROACH THAT WE HAVE APPLIED TO THE VALUATION OF CANDIDATE VACCINE VIRUSES BECAUSE THE, OBVIOUSLY THE, THE AVIAN INFLUENZA VIRUS PARENT IS SOMETHING WE WORKED WITH AT THE APPROPRIATE LEVEL OF CONTAINMENT AND GENERATE THE REASSORTMENT OR RECOMBENANT VACCINE VEUS HAVE -- VIRUS AND GO THROUGH A NUMBER OF STEPS TO ESTABLISH THE PROPERTIES OF THE VIRUS. ONE OF THE THINGS WE DO IS TO LOOK AT THE GENOTYPE AND SEQUENCE OF THE HUMAN GLUTTINNEN AND EURO MINNOW DAES GENE AND SEQUENCE THE ENTIRE GENONLY. WE COMPARE THE ANTIGENICITY OF THE VACCINE REASSORTMENT WITH THAT OF THE PARENT VIRUS, WE COLLABORATE WITH THE USDA TO LOOK AT THE PATHOGENICITY OF THE CANDIDATE VACCINE IN CHICKENS. WE LOOK AT THE PATHOGENICITY AND THE LEVEL OF REPLICATION OF THE CANDIDATE VACCINE VIRUS IN MICE OR OTHER SUITABLE ANIMAL MODELS AND LOOK AT THE EFFICACY OF PROTECTION FROM CHALLENGE WITH THE WILD-TYPE VIRUS IN THE ANIMAL MODELS. TO GO OF YOU AN EXAMPLE OF AN H9 N2 VACCINE THAT WE GENERATED IN THE ANN ARBOR COLD ADAPTED LIVE ATTENUATED VACCINE BACKGROUND, WE GENERATED THIS VIRUS BY CONVENTIONAL REASSORTMENT. WE DIDN'T HAVE TO MODIFY ANY OF THEVERULENCE MOTIVES IN THE HEMOGLUTINEN OR EURO MINODASE GENES IN THE CASE OF THE ANN HASH ON -- ARBOR, THERE ARE PROPERTIES TO LOOK FOR. TEMPERATURE SENSITIVITY AND COLD ADAPTATION IN VOTE ROW, WHICH ARE PROPERTIES CONFERED BY THE SIX INTERNAL GENE SEGMENTS IN THE RES -- RE REAORT -- ASSORTMENT VIRUS. WE'RE LOOKING FOR THE H2 NEXT NINETEEN VEUS HAVE, THE SOLID BARS ARE THE ANANARBOR COLD ADAPTED VIRUS AND THE OPEN BARS ARE THE REASSORTMENT VACCINE VIRUS. YOU CAN SEE THE VACCINE VIRUS REPLICATES AS WELL AT 25 DEGREES AS IT DOES AT 35 DEGREES WHICH, IS DESCRIBED AS A COLA, DAPTATION FENNO TYPE AND IN THE LOW -- PHENOTYPE, IN THE LOWER GRAPH, CAN YOU SEE THE WILD-TYPE H9 N2 VIRUS EQUALS AT 39 DEGREES, NOT TEMPERATURE SENSITIVE, THE ANANARBOR-COLD ADAPTED IS IN THE SOLID BARS AND THE OPEN BARS ARE THE VACCINE VIRUS, TEMPERATURE SENSITIVE. WHEN WE THEN TAKE THIS, THESE CANDIDATE VACCINE AND THE TWO PARENT VIRUSES AND ADMINISTER THEM INTRANASALLY TO MICE AND HARVEST THE TISSUES FROM THE UPPER AND LOWER TRACT TO BASE 2, THREE, AND 4 RESPECT ELF, IF YOU LOOK AT THE LEVEL OF REPLICATION IN THE VIRUS IN THE UPPER RESPIRATORY TRACT AND THE LOWER, CAN YOU SEE THE WILD-TYPE VIRUS REPLICATES EFFICIENTLY IN BOTH THE UPPER AND LOWER TRACT WHERE THE VACCINE VIRUS HAS ACQUIRED THE PROPERTIES OF ATTENUATION FROM THE ANN ARBOR COLD-ADAPTED VIRUS. WHEN WE THEN LOOK AT THE EFFICACY OF PROTECTION WE FIND THAT THE H9 N2 REASSORTMENT VIRUS IS ABLE TO PROTECT MICE AGAINST SUBSEQUENT CHALLENGE WITH BOTH THE HOMOLOGIST AND HETEROLOG, ST CHALLENGE. THE CHALLENGE IS THE YELLOW OR REASSORTMENT VACCINE STRAIN, PBS IS THE CONTROL WHERE THESE ARE ANIMALS THEN NOT PROTECTED AT ALL. SO WHEN, WHEN THE VIRUSES ARE ADMINISTERED IN A -- INTRANAZALLY AND A MONTH LATER THE MICE ARE CHALLENGED WITH THE WILD-TYPE VIRUS, CHALLENGED WITH A HOMOGOLOUS VIRUS AND A HETEROOLOGOUS VIRUS, WE SEE REPLICATION OF BOTH OF THE VIRUSES IN THE WHITE BARS AND THE UPPER AND LOWER TRACT WHEREAS THE VACCINE VIRUS AND THE WILD-TYPE PARENT VIRUS ARE ABLE TO PROTECT COMPLETELY FROM REPLICATION. AS AN ADDED MEASURE OF SAFETY IN TERMS OF THE SAFETY FOR THE ENVIRONMENT GO&AGRICULTURE, WE HAVE COLA -- COLLABORATED WITH DAVID SWAINFROM THE USDA AND LOOKED AT THE SHEDDING OF THE VIRUSES WHEN ADMINISTER -- ADMINISTERED TO THE CHICKENS. WE DID THEM INTRAVENOUSLY AND INTRANAZALLY. IN THE YELLOW LINES IN BOTH SETS, THE VACCINE VIRUS DOESN'T SHED IN EITHER THE ORO PHARYNX OR CLOACAL IN CHICKENS WHEN ADMINISTERED INCENTER -- INTRAVENOUSLY. IT'S NOT LIKELY TO REPRESENT THE MAJOR RISK FOR AGRICULTURE. THAT'S AN EXAMPLE OF A VIRUS WE GENERATED IN THE ANN ARBOR-COOLED ADAPTED BACKGROUND. IT WAS AN EXAMPLE OF A VACCINE VIRUS GENERATED BY REASSORTMENT. IF WE CHANGE TO LOOKING AT THE HIGHLY PATHOGENIC INFLU EPZA VIRUSES, YOU HAVE HEARD ABOUT THE IMPORTANCE OF THE MULTIBASE AMEANO ACID MOTIVE AND THE CONNECTING PEPTIDE OF THE HEMO GLUTINEN -- I'M SORRY, SHOULDN'T SAY HONG KONG. VIETNAM AND THAILAND ISOLATES HAVE MULTIPLE BASIC AMINO ACIDS AT THIS SITE. IT'S IMPORTANT FOR US TO ENGINEER THOSE, THOSE, THAT VIRULENCE MOTIF OUT OF THE HUMIDITIO GLUTINEN IN PART BECAUSE AS YOSHI'S PRESENTED DATA, THEY REPRESENT A VIRULENCE MOTIVE AND WOULD ALLOW US TO GROW THE VIRUS IN EGGS -- EGGS WITHOUT THE EMBRYO -- [ INDISCERNIBLE ] IN WORK I CAN AT THE CDC, WE GENERATED A VACCINE CANDIDATE AGAINST THE H5 N1 FROM 1997 WHERE WE ENGINEERED OUT THE MULTIPLE BASIC AMINO ACIDS IN THE HEMOGLUTINEN IN GENES AND PRODUCED A MODIFIED H5 N1 CANDIDATE VACCINE BY PLASMID-BASED REVERSE GENETICS WHERE WE TOOK THE HEMOGLUTINEN, MODIFIED IT OF THE H5, TOOK THE GENE AND COLD TRANSPICTURED WITH PLASMIDS FROM EITHER THE ANN ARBOR COOLED ADAPTED PUBLISHED BY LOWE AND PRAT THAT WE PUBLISHED IN VIROLOGY LAST YEAR. NOW NEXT JUST THE SHOWING YOU OF THE CHARACTERISTICS OF THE H5 N1 PR8 CANDIDATE VACCINE, WE SEQUENCED THE GENES, SHOWED THAT THE MULTIBASIC AMINO ACID CLEAVAGE SITE MOTIVE WAS REMOVED AND CONFIRMED THE SEQUENCE OF THE REMAINING GENES BY RFLP AND SEQUENCE ANALYSIS AND SHOWN IN VITRO THE REMOVAL OF THE SITE MADE THE BUYERS DEPEND ON TRIP SIN FOR CLEEFAGE OF THE HEMOGLUTTINNEN. WHEN WE ADMINISTERED THIS VIRUS INTRAVENOUSLY TO CHICKENS. YOU HEARDARD, -- ALREADY THE STANDARD TEST FOR HIGH I -- HIGHLY PATHOGENIC AVIAN VIRUS IS TO INSERT IT INTRAVENOUSLY TO CHICKEN ANDS OBSERVE MORTALT. WHEN WE ADMINISTERED THE WILD TYPE VIRUS TO CHICKEN, THERE IS 100% MORTALITY AND VIRUS IS SHED FROM THE ORO PHARYNX. THE CHICKEN ARE NOT LIVE LONG ENOUGH IS TO ESTABLISH THE SERO CONVERSION DATE AN. WHERE AS THE H5 N1 PR8 CANDIDATE VACCINE DIDN'T KILL ANY OF THE CHICKEN OR SHED IN EITHER THE UPPER, OR THE RESPIRATORY OGE TRACT. WHEN WE ADMINISTERED THE VIRUS THEN TO MICE, WE CAN SHOW THAT THE H5 N1 WILD-TYPE AS YOU HEARD IS LETHAL FOR MICE AT LESS THAN 1 INFECTION -- [ INDISCERNIBLE ] WHEREAS THE REASSORTMENT WITH THE MOFCKZ AND THE HEMOGLUTINEN GENE, WE COULD NOT ACHIEVE A LETHAL DOSE WITH THAT VIRUS. BUT YET, THE VIRUS REPLICATES EFFICIENTLY IN THE RESPIRATORY TRACT OF MICE. WE PREPARED FORMAL AND INACTIVATED MAXEP -- VACCINES WITH THIS VIRUS COMPARING IT WITH A FORMAL AND ACTIVATED VACCINE MADE WITH THE WILD-TYPE VIRUS OR WITH A LIVE INFECT AND WE COULD SO THAT THE VACCINE, THE FORMAL AND INACTIVATED REASSORTMENT VIRUS LISTED AN HI RESULT THAT IS -- [ INDISCERNIBLE ] AND THERE IS COMPLETE PROTECTION FROM LETHALITY AS WELL AS REPLICATION OF THE CHALLENGE VIRUS. SO, I'LL JUST CLOSE WITH TELLING YOU GOODBYE TO SAFETY MEASURES THAT WE HAVE USED FOR GENERATING THESE PANDEMIC INFLUENZA VACCINES, THE REASSORTMENT VIRUSES ARE RECOMMENDED TO WORK WITH THE WILD-TYPE AVIAN INFLUENZA VIRUS PARENT. WE GENERATE RISK ASSESSMENT DATA REGARDING INVOTE ROW AND IN VIVO PROPERTIES OVER THE RECOMBINANT MODIFIED IN THE CASE OF THE HIGHLY PATHOGENIC VIRUSES OF THE CANDIDATE VIRUSES. BASED ON THE RISK ASSESSMENT DATA, WE WOULD THEN LOOK TO LOWER THE BIOSAFETY LEVELS THAT IS REQUIRED FOR MANUFACTURE. IT'S CRITICAL THAT WE GENERATE VACCINES THAT WE CAN, AS DR. HOLMES SAID EARLIER, THE PROCESS HAS TO BE ITERATIVE. WE HAVE TO BE ABLE TO REASSESS THE RISKS ASSOCIATED WITH THE MODIFIED REASSORTMENT VIRUSES SO THEY CAN BE GIVEN TO VACCINE MANUFACTURERS TO PRODUCE VACCINES. I WILL CLOSE THERE AND SEEIVE IF YOU HAVE QUESTIONS. [ APPLAUSE ]

THANK YOU. ANY QUESTIONS FROM THE PANEL?

[ INDISCERNIBLE ] I WOULD LIKE TO MAKE ONE COMMENT IF I COULD, RATHER THAN ONE REAL QUESTION. THAT IS, I THINK NEEDS TO BE STRESSED FOR THE NONINFLUENZA VIRUS PEOPLE HERE AND THE FDA, THE NEWSPAPERS AND THE GENERAL PUBLIC AT LARGE THAT AN INFLUENZA VIRUS IS GENERATED BY RECOMMINANT DNA TECHNOLOGY, REVERSED GENETICS. THE SO-CALLED 6-PLUS TWO VIRUSES YOU SPOKE ABOUT TO MAKE A VACCINE AND A VIRUS THAT YOU COULD HAVE MADE, WE HAVE BEEN DOING BY 50 YEARS BY REASSORTMENT TO GENES IS EXACTLY THE SAME. THAT IS A ROSE IS A ROSE. ONCE YOU HAVE GOT THIS VIRUS, IT DOESN'T MATTER HOW IT WAS PRODUCED. AND I FIND THIS, IN FACT, A LOT OF PEOPLE THINK THERE IS SOMETHING INCREDBLY INCREDIBLY SPECIAL ABOUT HOW THE INFLUENZA VIRUS IS BEING USED FOR VACCINE PURPOSES, MADE BY REVERSE JEPPETICS VERSUS ONE DONE THE OLD-FASHIONED WAY. I THINK SOMEHOW WE HAVE TO COPHAVE A TO THE PUBLIC IN A BETTER WAY THE FACT THAT TWO THINGS ARE ABSOLUTELY IDENTICAL. IT'S INFLU EPZA VIRUS AND IT HAS THE RIGHT HEMOGLUTINEN AND THE RIGHT EU -- UROMINADASE ON IT.

I WOULD LIKE TO FOLLOW UP ON THAT A BIT. DR. WEBSTER MENTIONED THE POLY, THE POLL, THE BASIC TRACT THAT MAKES THE VIRUS SUSCEPTIBLE, THAT MAKES ITABLY TO GROW WITHOUT TRIP SIN WAS GENERATED BY SLIPPAGE OF OUR -- [ INDISCERNIBLE ] ERRORS. OKAY, NOW IN THE CASE OF THE GENNETICALLY-ENGINEERED VIRUS WHERE YOU TOOK THAT SEQUENCE OUT, WAS IT TAKEN OUT IN A WAY SUCH THAT YOU COULDN'T GET THAT SLIPPAGE TO REPRODUCE THOSE BASIC AMINO ACIDS SO THAT IT WOULD REVERT TO A VIRUS THAT IS, REVERT TO A VIRUS THAT CAN GROW INDEPENDENTLY WITH TRYWPSIN. THE FOLLOW SUP HAVE YOU CHECKED TO SEE IF THE VIRUSES CAN GROW INDEPENDENTLY WITH TRIP SIP. WHAT IS THE FREQUENCY TO PATIENTS WITH TRIP SIN INDEPENDENCE?

WHAT WE HAVE DONE IS THE PRELIMINARY SLIPPAGE OCCUR WHEN IS THERE ARE RUNS OF As AND Gs. WE HAVE REPLACED IN THE, IN THE CODE-ON USEAGE, WE HAVE REPLACED THE NUCLEOTIDES IN, WHEN REMOVING THE MULTIBASK MEETIVE -- MOTIF, WE MOST THE RESIDUES PRECEDING THAT WITH RESIDUES WE THINK SHOULD MAKE IT HARDER FOR PRELIMINARY SLIPPAGE TO OCCUR, WHETHER THAT WILL ACTUALLY TRANSLATE INTO PRACTICALLY UTILITY, WE DON'T KNOW. WE KNOW WE -- PASSAGE THE VIRUS AND I BELIEVE DR. WEBSTER'S LAB HAS PASSAGED THEIR REUS HAVE -- VIRUS MULTIPLE TIME TIMES SOEE SEE IF THE VIRUS CAN RE-INSERT IN THE SITE. WE HAVEN'T SEEN THAT HAPPEN. DR. WEBSTER, DO YOU -- HAVE --

WE TRIED TO MAKE THE VIRUS REREVERT AND WE CAN'T. WE HAVE DONE THE SAME THING. WE PUT IN THE SEQUENCE OF A NON-PATHOGENIC STRAIN AND THIS, NO MATTER WHAT WE DO, WE CAN'T, PAST IT 15, 16 TIMES IN HIGH MULTIPLICETEES AND CAN'T GET IT TO REVERT.

FOR PEOPLE REVIEWING THESE THINGS, CAN YOU PUT AN UPBER -- UPPER LIMIT ON THESE THINGS, USING 46ES TO SPITE STOCKS OR SOMETHING LIKE THAT.

I THINK WE REALLY HAVE TO, SHOULD THINK OF WHAT HAPPENS IN NATURE. THESE VIRUSES EXIST OUT THERE IN NATURE IN THE NONPATHOGENIC FORM. THAT'S THE STANDARD STRAIN WHERE THEE VIRUSES HAVE EXISTED FOR MILLIONS OF YEARS AND -- THE HIGHLY PATHOGENIC STRAIN IS ANAABBERENT STRAIN AFTER IT'S BEING GENERATED THROUGH HIGH-DENSITY POULTRY. THE ABRENT STRAIN. THE STANDARD STRAIN IS THE NONPATHOGENIC STRAIN.

YOU'RE ALSO PROPOSING TO MAKE IT VERY EFFICIENT IN VITRO BY GENERATING LARGE QUANTITIES OF THE STRAIN AS WELL.

RIGHT.

THAT'S THE ISSUE. DOES IT, DOES SOMETHING HAPPEN AND CAN YOU PUT A 46O IT. DOES IT GENERATE -- A FREQUENCY ON IT. DOES IT GENERATE MORE PATHOGENIC VIRUSES THAT CAN GROW INDEPENDENT OF TRYP, CIN. WHEN YOU GROW IT IN A SITUATION. ARE YOU TRYING TO GET A LOT FOR VACCINE PURPOSES.

WE HAVE TRIED TO MIMIC THAT AND CAN'T DO SO.

OKAY .

WELL, IT'S TIME FOR A 15-MINUTE BREAK. 11:35 S.WHEN WE'LL START UP AGAIN. I WOULD LIKE TO THANK THE SPEAKERS THIS MORNING. VERY INFORMATIVE PRESENTING AS. PRESENTATIONS.

OKAY. A COUPLE OF ANNOUNCEMENTS. THERE ARE THREE BY FIVE CARDS OUT ON THE TABLE OUTSIDE IN A BOX. IF ANYBODY HAS QUESTIONS OR ISSUES THAT THEY WOULD LIKE THE PANEL TO TAKE INTO CONSIDERATION IN THE DEVELOPMENT OF THE POINTS TO CONSIDER THIS EVENING, PLEASE PUT THOSE QUESTIONS ON TO THE 3 BY 5 CARDS AND WE WILL TAKE THOSE INTO ACCOUNT THIS EVENING IN DEVELOPMENTS -- DEVELOPING THE POINTS TO CONSIDER. WE WOULD APPRECIATE YOUR PROVIDING THAT. THE SECOND THING I WOULD LIKE TO DRAW THE PARSE TIS PANTS AND THE AUDIENCES' -- AUDIENCE'S ATTENTION TO, ON AUGUST 26th, THE DEPARTMENT OF HEALTH AND HUMAN SERVICES ISSUED THE NATIONAL PANDEMIC INFLUENZA PREPAREDNESS PLAN AND THAT IS OPEN FOR PUBLIC COMMENT FOR 60 DAYS FROM THE AUGUST 26th ANNOUNCEMENT AND THIS IS POSTED ON THE HHS WEBSITE AND IF ANYBODY NEEDS THE URL, I WILL HAVE IT UP HERE, BUT IF YOU GO TO THE DHHS WEBSITE, I AM SURE RIGHT ON THE FRONT PAGE THERE IS A LINK TO THAT PLAN. SO IF YOU HAVE COMMENTS ON THE NATIONAL PREPAREDNESS PLAN, PLEASE BE SURE TO GET THOSE IN. IT'S DRAFT AND IT'S OPEN FOR PUBLIC COMMENT. WITH THAT, I WILL TURN IT BACK OVER TO THE MODERATERS -- MODERATORS. KAY.

IT'S MY GREAT PLEASURE TO INTRODUCE DR. RALPH BARIC, ONE OF THE PIONEERS IN DOING REVERSE GENETICS IN CORONA VIRUSES AND RALPH IS GOING TO TALK ABOUT RECOMBINANT SARS CORHONE -- CORHONE -- COROPEA VIRUS RESEARCH. -- CORONA VIRUS RESEARCH.

THANK YOU, KAY. SO WE'RE GOING TO SWITCH GEARS AT THIS POINT AND NOW TALK ABOUT SARS CORONA VIRUS AND OTHER CORONA VIRUSES. THAT THIS IS A MICROGRAPH WHAT HAVE IT LOOK LIKE. THE WORK I'M GOING TO TALK ABOUT TODAY WAS DONE BY BOYD YOUNG, CHRIS CURTIS AND AMY SIMMS IN MY LABORATORY. WE'RE GOING TO TACK ABOUT CORONA VIRUS REVERSE GENETIC REVERSES IN GENERAL AND FOCUS SPECIFICALLY ON THE CORONA VIRUS. THE CORONA -- COVONA VIRUSES ARE -- [ INDISCERNIBLE ] THE GENOORGANIZATION. THEY'RE DIVIDED INTO THREE GREWS OF ONE, TWO, THREE, AND THE IMPORTANT CORONA VIRUSES FOR TODAY'S TALKING TO, TRANSMISSAL GASTROITIS, IMPORTANT, HUMO I HAVE RISE, CAUSING THE COLDS IN THE WINTER. THE HEPATITIS VIRUS CAUSING THE ANIMAL MODEL FOR HUMAN DISEASE AND THE SARS CORONA VIRUS CAUSING SEVERE LOWER RESPIRATORY INFECTION IN ADULTS. THE GROUP 3 CORONA VIRUSES INCLUDE INFECTIOUS BRONCHITIS VIRUS. THERE ARE MOLEC -- MOLECULAR FORMS FOR THE CORONA VIRUS, AND INFECTIOUS BRONCHITIS VIRUS, AND TARGETED RRCK NA RECOMBINANT APPROACHES TO DO GENETICS WITH MP -- MHE AND FELINE INFECTIOUS CARDOITIS VIRUS. MOST CORONA VIUSES WERE BUSY ASLEEP IN 2002 WHEN THE SARS CORHONE -- COVON -- CORONA VIRUS IS WEEK US UP AND BROUGHT US INTO THE NATIONAL LIMELIGHT, UPFORTUNATELY. SARS EVENTUALLY INFECTED ABOUT 8,000 PEOPLE AND ABOUT 33 DIFFERENT COUNTRIES WORLDWIDE RESULTING IN 800 DEATHS. THE OVERALL MORTALITY RATE WAS 10 TO 12% WITH 20% OR SO REQUIRING INTENSIVE CARE TREATMENT. ONE OF THE GREAT MAJOR RISK FACTORS FOR THE SEVERE DISEASE IS THE AGE OF THE INDIVIDUALS THERE. IS A 50% MORTALITY RATE IN INDIVIDUALS 65 YEARS OR OLDER. WHAT IS THE FUTURE OF THE SARS CORONA VIRUS. WELL, THERE WERE THREE LABORATORY-ACQUIRED INFECTIONS IN 2003 AND 2004. THE TAIWAN AND SINGAPORE INCIDENT WERE CONFINED TO THE INDIVIDUALS IN THE LABORATORY WHO WERE INFECTIOUS -- INFECTED. THE TWO INDIVIDUALS INFECTED IN CHINA, ONE SPREAD THOSE TO HEALTHCARE WORKERS AND CLOSE FAMILY CONTACTS, RESULTING IN ABOUT NINE PEOPLE INFECTED IN THAT OUTBREAK IN THREE ROUNDS OF TRANSMISSION. THERE WERE ALSO FOUR CASES OF SARS CORONA VIRUS INFECTIONS REPORTED IN CHINA THAT HAD GENETIC ORIGINS, BUT IF YOU LOOK AT SERUM BANK SAMP OLE -- SAMPLES FROM 2001 AND 2002 TWR-FROM-HONG CONK AND THE PROVINCE, 1 OR 2 ARE POSITIVE TO THE SAR SAYS CORONA VIRUS. IT'S LIKELY THE SAR SAYS COVON -- CORHONESA VIRUS HAS BEEN INTRODUCED PLENTY OF TIMES AND IS PROBABLY NOT DONE WITH US YET. THE OBJECTIVES, BECAUSE I'M THE ONLY SPEAKER ON CORONA VIRUS IS TO GIVE A BRIEF REVIEW OF THE LIFE CYCLE AND REPLICATION STRATEGY WITH SPECIFICALLY TRYING TO DIRECT THIS TO UNDERSTANDING HOW MOLECULAR GENETIC APPROACHES WERE DEVELOPED FOR CORONA, TO TALK ABOUT TARGETED APPROACHES AND CORONA VIRUSES GENETICS, USING OTHER SYSTEMS BESIDES THE SYSTEM WE DEVELOPED. THE DEVELOPMENT OF THE SARS CORONA VIRUS SYSTEM IN OUR LABORATORY AND ITS APPLICATION TO UNDERSTANDING PATHOGENIS REPLICATION AND VACCINE DEVELOPMENT. AND THEN I WANT TO BRIEFLY TALK ABOUT REPLICATION CONFIDENT PROPAGATION DEFECTIVE VIRUSES THAT CAN BE BUILT WITH THIS TECHNIQUE AND FINALLY I'LL TALK ABOUT SAFETY MEASURES THAT WE HAVE IN PLACE TO PREVENT INFECTION. SO, THIS IS A SCHEMATIC CAR TOON OF THE CORONA VIRUS PARTICLE, CONTAINS A SINGLE-STRANDED GENOME OF 29,000 NUCLEOTIDES, AS KAY SAID, THE LARGE OF THE RNA GENOME IN NATURE, WRAPPED IN MULTICOPIES OF PROTEIN TO FORM A HELICAL CAP STRUCTURE WRAPPED BY A NEW CLEEID LAYER. THE ENVELOPE CONTAINS THREE VIRUS SPIKES. THE S-LIKE PROTEIN IS A PREDOMINANT VIRUS SPIKE GIVING THE VIRUS ITS UNIQUE APPEARANCE IN THE MICROSCOPE INTERACTING WITH THES A 2 OR THE RECEPTOR MOLECULE THAT ALLOWS THE VIRUS TO INFECT CELLS IN CULTURE AND IN THE HOST. AND IT'S ALSO A MAJOR DETERMININANT FOR HOST STRAINS AND NEUTRALIZING ON THE SPIKE LIKE A PROTEIN. THE E AND M PROTEINS SHOWN HERE, THIS IS E AND THIS IS M. THEY'RE ABSOLUTELY IMPORTANT IN ASSEMBLY AND RELIEF. THE E AND M PROTEIN TARGET AN INTERMEDIATE COMPARTMENT BETWEEN THE ROUGH ER AND THE GOLD G AND THE CAPSULE STRUCTURES AND THE S-PROTEIN GETTING INCORPORATED INTO THE PARTICLES AS THE MATURATION AND RELEASE OCCUR. SARS IS TRANSMITTED BY RESPIRATORY DROPLETS, LARGE DROPLETS AND READILY INACTIVATED 65 DEGREES BY HEAT BECAUSE OF THE BY LAYER, INACTIVATED BY 70 TO 100% ALCOHOL AND PHENOLOIC BASE DIIN -- DISFEP -- DISINFECTANTS. THIS IS THE CORONA VIRUS GENOME. THE FIRST 21,000 NUCLEOTIDES AND CODES, RIP CASE GENES DIVIDED INTO ORPH 1-A AND ORPH 1-B, ECPRESSED AS A SINGLE POLYPROTEIN OR A LARGE POLYPROTEIN SUBSEQUENTLY PROCESSED BY TWO PROTEASES, THE PLP PROTEASE MAKES THIS CLEAVAGE, VENT OCCUR. [ LAUGHTER ] -- EVENT OCCUR. DON'T LEAVE ME UP HERE TECHNOLOGICALLY HELPLESS. I THINK I HIT SOMETHING HERE. YEAH

THIS PLP PROTEASE MAKES THE CLEAVAGE EVENTS IN THE ORPH 1-A POLYPROTEIN SHOWN IN YELLOW AND THE PROTEASE MAKES THE CLEEF AM EVENTS SHOWN HERE WITH THE WHITE ARROWS. DOWNSTREAM OF THE RIPLY CASE ARE A SET OF SUBGENOWNIC ORPH S. THE E PROTEIN, ORPH NOON-A ENCODES THE NUCKLYAR CAP SID PROTEIN. IN ADDITION TO THE STRUCTURAL JEEPS OF OVERS OR GROUPS OF SPECIFIC JEEPS, THIS WORLD 3-A, 3-B, 6, 7-B, 8-A, COME&19-B. THE FUNCTION AND CORONA VIRUS IS NOT KNOWN. UPON ENTERING CELLS, CORONA VIRUS HAS EXPRESSED A SERIES OF SUBGENOMIC RNAs RANGED -- ARRANGED IN FORMS ALL THE SEQUENCES ARE PRESENT IN THE NEXT LARGER MESSAGE. THIS ALLOWS DIFFERENT OPEN READING FRAMES -- FRAMES TO BE PLACED AT THE FIVE ENDS OF MESS EDGE EVERY RNAs WHERE THEY WILL BE EXPRESSED. EACH MESSAGE CONTAINS A LEADER MESSAGE OF SEVEN NUCLEOTIDES INTO THE GENOME AND JOINED TO THE BODY SEQUENCES OF THE HIGHLY -- HIGHLY CONSERVED SEQUENCES SHOWN IN RED, THE SAGAC. IN ADDITION TO THE PRESENCE OF BOTH FULL-LENGTH AND SUBGENOMIC LENGTH NEGATIVE STRANDS. IN ADDITION TO THE PRESENCE OF THE FULL-LENGTH AND SUBGENOMIC LENS POSITIVE STRANDS, THERE ARE FULL-LENGTH AND SUBGENOMIC LENGTH STRANDS AND STAN HAS PROPOSED -- PROPOSED A MODEL TO EXPLAIN THE RNAs BY A MECHANISM CALLED TRANSCRIPTION ATTENUATION N.THIS MODEL REPRESENTING TO THE PREPRIMED END OF THE STRAND, DRIVING THE TRANSCRIPTION OF AN INCOMPLETE NEGATIVE STRAND WHICH TERMINATES IN THE HIGHLY-CONSERVED CONSECUTIVE SEQUENCES IN THE GENE. SOME WILL TERMINATE HERE, SOMEONE WILL TERMINATE HERE AND THEN THROUGH A JUMPING MECHANISM THAT IS NOT COMPLETELY UNDERSTOOD, THE SEQUENCES TRANSLOCATE TO THE PEOPLE END OF THE GENOME AND ANGAS A PRIMER TO DRIVE THE SYNTHESIS OF ANTILEADER SEQUENCES TO MAKE SUBGENOMIC NEGATIVE STRANDS THAT SERVE AS TEMP ITS FOR THE SYNTHESIS OF POSITIVE STRANDS. IN ESSENCE, THE CORONA VIRUS REPLICATE BY A MECHANISM OF SITE-ASSISTED, I'M SORRY, TRANSCRIBE RNA BY RECOMNATION. IT WAS THIS OBSERVING A AND THE OBSERVATION THERE ARE FULL-LENGTH AND SUBGENOMIC STRANDS IN CORONA VIRUS CELLS THAT LED TO THE OBSERVATION THAT THE CORONA VIRUS UNDERGO A HIGH-FREQUENCY RNA DURING MIXED INFECTION. IF YOU IMAGINE A CELL INFECTED WITH THE YELLOW CORONA VIRUS, IT WAS PINK ON MY COMPUTER BUT LOOKS LIKE IT'S GRAY HERE. A GRAY CORONA VIRUS DURING THE SIS OF EITHER POSITIVE OR NEGATIVE STRAGHTS, THERE WILL BE TEMP LATE SWITCHING EVENTS RELATING IN GENOMES WITH CHARACTERISTICS FROM BOTH PARENTS N.THIS CASE, IT OCCURS BETWEEN TWO FULL-LENGTH MOLECULES. BECAUSE OF THE PRESENCE OF SUBGENOMIC LENGTH NEGATIVE STRANDS, TEMP LATE SWITCHING CAN OCCUR TO PRODUCE RECOMBINANT MOLECULES. WHAT PAUL MASTERS DID WAS REALIZE THE BEAUTY OF THIS AFFECT TO DEVELOP A, THE FIRST MECHANISM TO DO GENETICS OF CORONA VIRUSES AND SO HE DEVELOPED AN APPROACH CALLED TARGETED RECOMBINATION WHICH I'M GOING TO SUMMARIZE HERE N.THIS CASE, IMAGINE THE MHA 59 GENOME SHOWN AT THE TOP. HE MADE A SYNTHETIC DONOR RNA, CONTAINING THE FELINE INFECTIOUS PERREAULT TINNITIS. IT CONTAINS -- [ INDISCERNIBLE ] THE PRIME END THAT ALLOWED IT TO REPLICATE WITH THE HELPER REPLICATES PROTEINS PRODUCED BY THIS PARENT. BECAUSE RECOMBINATION CAN OCCUR BETWEEN FULL-LENGTH AND SUBGENOMIC TEMP ITS, DURING RIPICATION, THE FELINE SPEAK IN THE WHOLE SEGMENT FROM THE DONNOR RNA GETS TRANSLOCATED OR RECOMBINED TO PRODUCE A VIRUS TO GROW IN FELINE CELLS. NOW WITH THE FELINE OR FMHV VIRUS IN HAND, WHICH INFECTS FELINE CELLS BUT NOT MA -- MOUSE CELLS, CAN YOU INFECT FELINE CELLS TRANSFECKING A SECOND TOPPOR RNA CONTAINING A FULL LENGTH OF THE MHG GENOME WITH THE PRIME END SEQUENCES TO ALLOW IT TO REPLICATE AND THEN WITHIN THAT DONOR RNA, INCORPORATE WHATEVER CHANGES YOU WANT. THE ONE I WILL MENTION IS THE SARS 3 PRIME END. THERE ARE THREE PRIME END OF GROUP TWO CORONA VIRUS ARE HIGHLY CONSERVED. ONE QUESTION YOU COULD ASK ARE THE THREE PRIME END SEQUENCES OF THE CORONA VIRUS, DO THEY FUNCTION IN THE MHA 59 BACKBONE. THIS IS TRANSVECTED INTO CELLS AND INTO THE FELINE VIRUS. YOU SELECT WITH THE MHV HOST SPECIFICITY WITH THE SPIKE AND GET VIRUSES CONTAINING THE THREE PRIME END CORONA VIRUS, WHICH ARE SHOWN HERE. NOW, CAN YOU USE THE APPROACH TO INCORPORATE BASICALLY WHATEVER SARS OR IF YOU WANTED, THE THREE PRIME END OF THE 59 GENOME N.THIS PARTICULAR INSTANCE, PUBLISHED LATE SUMMER OF THIS YEAR, THIS RS WAS DEBILITATED AND REPLICATED ABOUT A LOG LESS EFFICIENTLY THAN A VIRUS. IF YOU WANT TO DO TARGETED RECOMBINATION WITH THE SAR SAYS VIRUS, YOU WILL HAVE TO DEVELOP A KYMEREC OF THE CORONA VIRUS CONTAINING THE MHV SPIKE-LIKE PROTEIN IN THE BACKBONE OF THE SARS CORONA VIRUS AND THE DONOR RNA TO ALLOW THE SARS SEQUENCES TO BE INTRODUCED BACK IN. CAN YOU DO REVERSED GENETICS OF THE ENTIRE END OF THE GENOME. TO MY HANDLE, IF THIS HAS BEEN DONE, IT HASN'T BEEN PUB ISSUE LEAD -- PUBLISHED IT. I WOULD GUESS THE VIRUS EXHIBITS IN THE WORLD SOMEWHERE. THERE ARE CONCERNS ABOUT THE APPROACH. AND ACTUALLY ALL OF THESE TYPES OF RECOMMINANTS CAN BE BUILT WITH FULL-LENGTH INFECTIOUS CLEANS -- CLONES AS WELL. YOU CAN END UP WITH RODENT VIRUS BACKBONES HUMANIZED WITH THE SARS SAYS S--LIKE PROTEIN IN THEM, TARGETING AN MHV VIRUS INTO HUMAN CELLS. YOU CAN HAVE WEEKLY PATHOGENIC CORONA VIRUSES IN THIS CASE OR -- [ INDISCERNIBLE ] LESS OF CONCERN. THEY WOULD TARGET MICE INSTEAD OF HUMANS. CAN YOU MAKE RECOMBINANT SARS CORONA VIRUS JEEPS. THIS IS ONE APPROACH THAT WILL PROBABLY BE DEVELOPED IN THE NEAR FUTURE TO MANIPULATE THE SARS CORONA VIRUS GENOME. NOW, THE PROBLEM WITH DEVELOPMENTING CORONA VIRUS ENFCIOUS CLONES ARE SUMMARIZED HERE. YOU HAVE THE LARGE SIZE OF THE GENOME, STABLE CLONING VECTORS, YOU HAD REGIONS OF CRIMESOMAL TOXICITY, WHICH MADE IT DIFFICULT TO CLONE A CORONA VIRUS SEQUENCES IN BACTERIA. YOU HAD THE 60 SIZED TRANSCRIPTS IN VITRO AND YOU HAD TO HAVE A SYSTEM YOU COULD MANIPULATE. I THINK IT WAS A TESTAMENT TOLD -- TO THE FIELD THAT THREE DIFFERENT GROUPS MAD THREE DIFFERENT APPROACHES TO SOLVE THE PROBLEM. THE FIRST MOLECULAR CLONE FOR THE CORONA VIRUS WAS USED WITH TRANSMISSAL GAS. I'M NOT GOING TO GO THRUL THE DETAILS. HE CLONED THE TGBE CLONE TO THE PROMOTER ELEMENT IN A BACTERIAL ARTIFICIAL CHROMOSOME. THIS IS THEN ELECTROPARADED INTO CELLS. THE CNV PROMOTER DRIVES USING, DRIVING THE SEPGHT SIS OF FULL-THREPGHT TRANSCRIPT BOOTING INFECTIVITY AND THE VIRUS IS RELEASED FROM THE CELLS. THE SECOND APPROACH, OTHER APPROACH DONE AFTER US WAS IN STEW ART'S LAB. HE CLONED THE HUMAN CORONA VIRUS 229 E GENOME INTO VACCINEA VIRUS. THE ENTIRE GENE ONLY. THIS IS ELECTROPARADED IN THE CELLS INFECTED WITH CELL POX ACTING AS A HELPER VIRUS AND YOU END UP WITH VIRUSES CONTAINING THE FULL-LENGTH CORONA VIRUS IN VET. THIS CAN BE CUT WITH A RESTRICTION ENZYME AND IS A T-7 TO DRIVE FUN-LENGTH TRANSCRIPTS THAT CAN BE ELECTROPARADES INTO THE CELLS AND THE VIRUS CAN COME OUT. BOTH OF THE SYSTEMS HAVE FULL-LENGTH GENOMES AND SELF-REPLICATING VICTOR -- VECTORS. IT'S AN EASILY REMOVABLE RESOURCE AND THE FEATURES ARE DIFFERENT IN THE VECTOR SEGMENTS I'M GOING TO DESCRIBE NEXT. OUR APPROACH WAS TO DEVELOP A SYSTEMATIC ASSEMBLY APPROACH OF A SET OF ADJOINING SUBCLONES AND WE HAVE DONE THIS NOW WITH MOUSE HEPATITIS RSTVJ AND THE CAR SAYS CORONA VIRUS. BASED ON THE USE OF CLASS 2-S RESTRICTION ENZYMES LEAVING UNIQUE AND INTERCONNECTING JUNCTIONS. THE ENZYME BAGLE 1 CUTS THE SIMMETICAL SYSTEM CDG CDC BUT LEAVES ANnd. THERE ARE 64 DIFFERENT ENDS TO BE GENERATED WITH BAGLE 1, MOST WHICH ARE INCOMPATIBLE WITH ANY GENERATED WITH BAGLE ONE. BECAUSE OF THIS, CAN YOU USE THE RESURS -- RECURSIVE STRATEGY. YOU CAN ASSEMBLE TWO TO THE 64th FRAGMENT OF 4,000 BASED OR SO AND IF YOU USE AN ENZYME SSI1, YOU CAN ASSEMBLE TWO TO THE 64 FRAGMENTS OF ABOUT 64 KB. SO WHAT THE TECHNIQUE REALLY DOES ALLOW IS THE DIRECTIONAL ASSEMBLY OF RECOMBINANT DNAs INTO THE MICROBEUAL GENOMES, THEORETICALLY. SYNTHETIC TO CDNA IS THE 5 TO 8 KBs READILY AVAILABLE COMMERCIALLY. THE APPROACH WITH THE SARS CORONA VIRUS IS SHOWN HERE. WE BROKE THE GENOME INTO SIX PIECES. DESIGNATED A-THROUGH -- A THROUGH 6. THE T CLONE HAS -- [ INDISCERNIBLE ] AT THE PEOPLE END. THE F CLONE HAS A POLYT-TAIL TO DRIVE A POLYA-TAIL AT THE GENOME. AT THE THREE PRIME END OF THE A-CLONE, WE USE A BAGLE SITE WITH A TA 8 OVERHANG AND WE START SAME RESTRICTION LEAVING AN ATT COMPLEMENTARY OVERHANG. NOTICE THESE TWO ENDS CAN NOT INTERACT WITH THEMSELVES. RATHER, THEY CAN ONLY INTERACT WITH EACH OTHER. WE USE A DIFFERENT BAGLE SITE TO PRODUCE THE PRIME END OF THE B-CLEAN AND THE -- CLONE AND THE SAME SAME SITE TO PREINTRODUCE THE NUCKO -- [ INDISCERNIBLE ] WHEN THESE THINGS GO TOGETHER IN TINGER TOY FASHION, IT RE-FORMS THE EXACT SARS CORONA VIRUS SEQUENCE. SO IN THIS WAY, WE CAN USE IN VITRO LIGATION TO ASSEMBLE THEM TOGETHER LIKE TINKER TOYS. THE BASK APPROACH IS SHOWN HERE. WE HAVE SIX PLASMIDS ENCODING THE SIX FRAGMENTS OF THE SARS CORONA VIRUS GENOME. WE USE BAGLE 2 TO CUT OUT THE PIECES AND THE RESULTS ARE IN A SET OF CONTIGUOUS 5 KB PIECES OR SO. THESE ARE LENLYINGATED TOGETHER BY T-14, AND I SHOULD POINT OUT THIS RESULTS IN A FINITE SOURCE OF NON-REPLICATING FULL-LENGTH CDNA THIS IS CONTIMED -- CONSUMED IN THE REACTION, OKAY, IN THE FOLLOWING STEPS. THIS IS NOT INFECTIOUS. YOU HAVE TO DRIVE FULL-LENGTH TRANSCRIPTS OF THE RNA AND ELECTROPARADE THIS INTO CELLS. IT HELPS BOOT THE INFECTIVITY OF THE CORONA VIRUS GENOME FROM 10 TO A THOUSAND FOLD AND FROM THAT TRANSVECKZ MIX, CAN YOU GET VIRUSES OUT. AND SO THIS IS JUST THE FIRST EXPERIMENT WE DID WHERE IN THE PRESENCE OF END TRANSCRIPTS, YOU GET REUS HAVE GROWING 10 TO THE SIXTH IN 48 HOURS AND IN THE ABSENCE OF END TRANSCRIPTS IS A LOG, LOG AND A HALF LEFT. AGAIN, VIRAL ANTIGEN PRESENT IN THE CELLS. THERE ARE MORPH LOGICALLY AND DISTINGUISHAGE BETWEEN THE WILDFIRE AND THE RECOMBINANT FIRES -- VIRUS. THE RECOMMINANT VIRUS HAS WHATEVER MARKER MUTATIONS WE PUT INTO IT. IF YOU WANT TO UNDERSTAND WHY SARS IS DIFFERENT FROM THE OTHER HUMAN CORONA VIRUSES IN TERMS OF THE PATHOGENIS, YOU HAVE TO BE ABLE TO DEMONSTRATE THE VIRUS IS PAGOGENIC AS THE PARENTAL VIRUS FROM WHICH IT WAS ISOLATED N.COLLABORATION WITH JASON AND PETER, WE INOCULATED MACAQUES WITH THE WILD-TYPE FOR BONNIE, MONITORED THEM DAILY FOR CLINICAL SIGNS OF DISEASE, THE CHEST X RAYS AND LOOKED AT VIRUS IN A VARIETY OF CLINICAL SPECIMENS. OUR RESULTS WERE ENDIS TING WISHABLE BETWEEN RECOMBINANT IC SARS AND WILD-TYPE. BONNIE, WE DIDN'T SEE ANY DISEASE OR CHANGES IN BLOOD CHEMISTRY. TEMPERATURE CHANGES OR OXYGEN SAT RAGE. WE JUST SAW RADIOGRAPHIC EVIDENCE OF MULTIFOCAL PNEUMONIA AND THE IC SARS CORONA VIRUS INFECTED AND THE WILD-TYPE BONNIE INFECTED ANIMALS. THIS IS DAY 8, DAY 0 AND DAY 8 AND YOU CAN SEE THE LOWER RIGHT LOBE PNEUMONIA. WHICH EVENTUALLY RESOLVED BY DAY 18. SO IN SUMMARY, THE IC SARS DID PRODUCE A RADIOGRAPHIC PNEUMONIA, REMINISCENT OF THE CLINICAL COURSE OF SARS IN CHILDREN. IT WAS VERY, VERY MILD. NEUTRALLIZATION FIGHTERS WERE RELATIVELY ROBUST AND THE IC SARS INOCULATED ANIMALS, PROTECTED FROM CHALLENGE. OUR RESULTS WERE CLEARLY NOT AS DRAMATIC AS REPORTED BY -- [ INDISCERNIBLE ] WHO PRESENTED DATA SHOWING THAT SARS PRODUCED A MORE SEVERE DISEASE IN MACAQUES. I THINK OUR RESULTS ARE MORE IN LINE BY WHAT WAS REPORTED BY OTHER INVESTIGATORS IN THE STATES AND FINALLY IN COLLABORATION WITH CONTA, THE IC SARS AND THE WILD-TYPE VIRUS WERE PUT INTO MIDDLE SCHOOL AND REPLICATED. AS FAR AS WE CAN TELL WITH THE EXISTING ANIMAL MODELS IN HAND, THE IC SARS VIRUS AND THE IV -- IRVANI ARE COMPARABLE. WE WOULD LIKE TO DO SOMETHING WITH MOLECULAR CLONE NOW. SO, WE HAVE A FAIRLY INTENSE INTEREST IN THE FUNCTION OF THE ACCESSORY OR THE SARS COCRONE -- CORONA VIRUS INFECTION. BECAUSE OF THE STUDIES WITH OTHER CORONA VIRUS, GROUP 1, 2, AND 3 CORONA VIRUS, IF YOU REMOVE THE ACCESSORY ORPHS, IT HAS LITTLE IMPACT ON THE IN VITRO REPLICATION OF SARS BUT ATTENUATES IT IN A NATURAL HOST. WE THINK IF YOU REMOVE THE ACCESSORY ORPHS FROM SARS, IT SHOULD ATTENUATE THE VIRUSES. STARTING WITH ORPH 7-A AND B, THE X-4 PROTEIN, WE DELETED THE X-4 PROTEIN OR DELETED THE X-4 PROTEIN AND PUT IN GREEN FLORESCENT PROTEIN, IN ESSENCE, MAKING A SARS CORONA VIRUS TO PRODUCE GREEN-LOOKING CELLS IN THE MICROSCOPE. WE GOT LOTS OF ROBUST GROWTH AND THE X-4 DELETION VIRUSES, REPLICATED AS WILD-TYPE VIRUS. I'LL SHOW THAT DATA IN A MINUTE. WE INCORPORATED LUVIFHEROASE IN THE 7- -- 7 A-B-POSITION. [ INDISCERNIBLE ] YOU CAN SEE A CLEAR EVIDENCE OF -- [ INDISCERNIBLE ] EXPRESSION BY WESTERN AND IF DO YOU, IT LUCIPHERATES ACTIVITY, PARENTAL VIRUS. I CAN'T SEE TOO WELL FROM THIS LOCATION, BUT THE MOCK INFECTED AND THE WILD-TYPE FOR BONNIE HAS LITTLE LUCIPHOO -- OUS ACTIVITY, ABOUT YOU IT HAD SIX LOGS INCREASED IN THE EXPRESSION. WE CAN USE THE LUCIPHERATE IS CONTAINING VIRUS AND THE GFT VIRUS AS PLATFORMS FOR SCREENING, RAPID SCREENING OF COMPOUNDS THAT HAVE ANTISARS CORONA VIRUS ACTIVITY. WE'RE ALSO INTERESTED IN THE OTHER ACCESSORY ORPH, 3-A AND 6 WERE DELETED. AGAIN, THIS IS A WESTERN BLOT USING ANESERE. YOU CAN SEE WITH BONNIE, THE WILD-TYPE. I SEE SARS AND THE X-3 DELETED VIRUSES, THE X-1 IS MADE. IN THE X-1, IT'S NOT PRESENT. URBANI AND VIRO CELLS GROW TWO TIMES 10 TO THE 7, PFU IN 30 HOURS. ALL OF OUR RECOMBINANT VIRUSES GREW TO EQUIVALENT OF ONE TIMES 10 TO THE SEVENTH OR HIGHER. THE DELETION OF THE THREE ACCESSORY ORPHS HAD LITTLE IMPACT -- EMPACT ONP VOTE ROW REPLICATION AS WOULD HAVE BEEN PREDICTED FROM STUDIES OF OTHER CORONA VIRUSES. WE'RE USING THE MOLECULAR CLONE AS A PLATFORM TO BUILD ATTENUATED VIRUSES OR VIRUSES THAT CAN BE USED SAFER IN A LABORATORY SETTING AND THIS DATA IS HEAVILY BASED ON COLLABORATIVE WORK WE'RE DOING AT VANDERBILT UNIVERSITY. WHAT MARK'S GROUP HAS SHOWN IS THAT WITH MOUSE HEPATITIS VIRUS, CAN YOU DELETE THE CLEAVAGE SITES, CS1 AND 2 POSITIONS, REGULATING IT, AND MHV GROWS FINE IN VITRO BUT THE VIRUS ATTENUATED. THE CLEEF AM SITES ARE CONSERVED IN SAR SAYS CORONA VIRUS AND CAN BE DELETED IN SARS CORONA VIRUS AS WELL. OUR LAB, AND AGAIN NEXT COLLABORATION WITH MARK'S LAB IDENTIFIED A SECOND MUTATION IN ANOTHER ORPH 1-B CALLED XON, COMPLETELY ATTAINIATING IN NH -- MHB. THIS CAN BE INCORPORATED INTO THE SARS CORONA VIRUS AS WELL, BECAUSE IT'S GOING TO CONSERVE LOCATION, AND WE'RE HOPING THE COMBINATIONOR EITHER SINGLELY OR IN COMBINATION OF THE MUTATIONS WILL RESULT TO THE ATTENUATION OF SARS. WE CAN MAKE A VARIETY OF COMBINATIONS BECAUSE OF THE COMPONENT CLONES, CAN YOU ACTUALLY EASILY MIX AND MATCH COMPONENT PIECES. THEY HAVE MADE MUTANTS WITH X-1, X-2, X-3 DLEEGS. THEY GROW EX -- DLEETIONZ. THEY GROW A HALF LOG LESS SUFFICIENTLY THAN WILDLIFE -- [ INDISCERNIBLE ] WE HAVE MADE MUTATIONS IN THE MIDDLE OF ORPH 1-B AND THE GENOME THAT REPLICATE EFINISHIENTLY. WE CAN THINK THEY CAN BE LIVE VIRUS CANDIDATESOR SEED STOCK FOR KILLED VACCINES. CURRENTLY IN COMMON, IN COLLABORATION WITH CONTA, THE X-1 DELETION VIRUSES ARE BEING STUDY IND HAT STOMACHSTER MODEL THAT LOOKS AT THE SARS CORONA VIRUS PATHOGENESIS. I DON'T HAVE RESULTS TO TALK ABOUT AT THIS TIME. FINALLY, I WANT TO TALK A LITTLE ABOUT REPLICATION CONFIDENT PROPICATION DEFISHIENT CORONA VIRUSES, PREVIOUS RECOMBINANT DNA COMMITTEES MIGHT SEE IN THE NEAR FUTURE. THE IDEA IS BASED ON THE IDEA THAT THE E AND M PROTEIN ARE ABSOLUTELY, SENSUAL FOR MATURATION AND RELIEF OF CORONA HAVE US -- VIRUSES. IF YOU DELETE THE E AND M PROTEIN ENCODED IN ORPH 4 AND 5, THIS MOLECULE WILL REPLICATE 5 AND PRODUCE SUBGENOMIC MESSENGER RNAs TO ENCODE THE R AND S PROTEIN BUT NO VIRUSES WILL BE MADE BECAUSE THERE ARE NO E AND M PROTEINS TO BE THE FUNCTION IN THE PRODUCTION OF VIRUS PARTICLES. -- HOWEVER, CAN YOU PROVIDE E AND M ON HELPER RNAs AND TRANS. IT WILL COMPLEMENT THE DEFECTS LACKING IN THE GENOME AND YOU CAN GET VIRAL RIPLYCON PARTICLING IN 10 TO TO THE FIVEth OR 10 TO THE SEVENTH. THIS WAS DONE WITH TGZ AND E IN MY LAB AND MORE ELEGANTLY BY LOUISE WHO MODIFIED IT TO GET UP TO THE 10 TO THE 7th. THIS FUNCTION OF SINGLE HIT RIPLYCON PARTICLES THAT CAN INFECT CELLS BUT BECAUSE THEY DON'T HAVE THE GENES, THEY WON'T BE ABLE TO SPREAD AMONG THE CELLS. THEY WILL BE MUCH SAFER, THEORHETT KEY, THAN TO TO USE IN A LABORATORY SETTING THAN THE TRADITIONAL CORONA VIRUS. THERE ARE THINGS WE NEED TO UNDERSTAND WHAT. IS ESSENTIAL FOR SARS PARTICLE PRODUCTION. THIS IS AN IMPORTANT QUESTION. E, PROTEIN IS ABSOLUTELY ESPIRITUAL FOR PACKAGING AND RELEASE OF TGVE PARTICLES, BUT NOT ESPIRITUAL FOR PACKAGING AND RELEASE OF MHB. MOST CORONA VIRUSES WILL AGREE IF YOU MAKE REPLICATION COPFIDDEN, PROPAGATION, FISHIENT VIRUSES WITH TWO OR MORE STRUCTURAL JEEPS, THIS WILL BE SAFE. THE ONE CAVEAT IS RECOMBINATION REPAIR TO WORRY ABOUT, EVEN WITH THE HELPER RNAs THAT WE HAVE NEVER SEEN AND LOUISE'S GROUP HAS NEVER SEEN OR WITH EXOGENOUS CORONA VIRUS THAT MAY INADVERTENTLY CONTAMINATE CULTURES. IT'S NOT CLEAR WHETHER OTHER CORONA VIRUS E&M PROTEINS COULD COMPLEMENT THE DEFECT OF THE VIRUS AND THAT NEEDS TO BE LOOKED AT. THE CORONA VIRUS THE RIP CASE CHAINS SO YOU CAN'T DO ANYTHING UNLESS YOU WORK WITH THAT. FINALLY, SAFETY PRECAUTIONS. SAY THAUL INTO THREE CATEGORIES: FACILITY, PERSONNEL AND COMMUNITY. PERSONNEL, COM