IT'S TIME TO GET
GOING AND I'M JUST THRILLED TO
BE HERE, EVERYONE, ON THIS
BEAUTIFUL AFTERNOON.
NICE COZY AFTERNOON TO BE IN
HERE LISTENING TO AMAZING
SCIENCE IT IS MY PLEASURE TO
INTRODUCE OUR SPEAKER FOR
TODAY'S ROLLA E. DYER LECTURE
DR. JASON Mc LELLAN, AND I
UNDERSTAND THIS IS A WELCOME
BACK WHICH IS WONDERFUL, HE IS
AT THE DOCTOR OF BIOSCIENCES AT
THE UNIVERSITY OF TEXAS AT
AUSTIN.
BEFORE I TELL YOU ABOUT HIS
BACKGROUND, I SHOULD TELL YOU
THIS STUFF, IF YOU ARE ATTENDING
THIS CODE FOR CME, THE CODE IS
50666.
IF YOU ARE WATCHING ONLINE
PLEASE CLICK ON THE SEND LIVE
FEEDBACK BUTTON AND YOUR
QUESTION WILL BE RELAYED TO THE
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PICTURE.
YOU CAN SUBMIT QUESTIONS AT ANY
TIME DURING THE LECTURE.
SO NOW THE IMPORTANT THINGS.
WE ARE DELIGHTED TO HAVE
DR. Mc LCERTAINLY--CERTAINLY
LLAN HERE TODAY, WE IS A
STRUCTURAL BIOLOGIST
SPECIALIZING IN VIROLOGY AND
VACCINES.
AS A POST DOC IN THE
LABORATORIES OF PETER KD--SALLY
MEN AND WOMEN ASK BARNY GRAHAM,
FROM 2000-2013 HE WAS
RESPONSIBLE FOR A PRAIK THROUGH
TOWARD DEVELOPING AN RSV
VACCINE.
HE DETERMINED THE STRUCTURE OF
THE RSV-F PROTEIN AND ENGINEERED
A PREFUSION STABILIZED ANTIGEN
WHICH ELICITED VERY HIGH TITERS
IN ANTIBODIES AND THEN IN
HUMANS, THE MULTIPLE APPROVE
WILL OF VACCINES LAST YEAR AND
THIS YEAR ENDED A 50 YEAR QUEST
TO DEVELOP AN EFFECTIVE VACCINE
AGAINST RESPIRATORY STITTIAL
VIRUS.
THIS HAS BEEN A GREAT BURDEN TO
SOCIETY OVER THE YEARS
ESPECIALLY TO VULNERABLE
POPULATIONS NEOINATES FOR
ESPECIALLY FOR WHOM IT'S BEEN
THE LEADING CAUSE OF
HOSPITALIZATION.
BUILDING ON HIS PIONEERING RSV
STUDIES, DR. Mc LELLAN'S
COLLAB HELPED DETERMINE THE
STRUCTURE OF THE HUMAN COONA
VIRUS, THEY RESEARCHED
S-PROTEINS INCORPORATED INTO THE
COVID VACCINES IN DEVELOPING
COUNTRIES.
WITH 150 PUBLICATIONS, PULT PEL
PATENTS AND STILL IN HIS EARLY
40S DR. Mc LELLAN IS REGARDED
AS THE LEADING SCIENTIST IN THE
FIELD OF STRUCTURE BASED VACCINE
DEVELOPMENT.
CAN'T WAIT TO SEE WHAT YOU DO
NEXT.
AMONG OTHER HONORS HE RECEIVED A
NATIONAL ACADEMY OF SCIENCES
AWARD IN MOLECULAR BIOLOGY IN
THE UT AUSTIN PRESIDENT'S
RESEARCH IMPACT AWARD BOTH LAST
YEAR AND A TRIPLE AAS GOLDEN
GOOSE AWARD IN 2020.
THOSE ARE GREAT.
YEAH, THOSE ARE GREAT.
HE EARNED A BACHELOR OF SCIENCE
IN CHEMISTRY FROM UNIVERSITY OF
DETROIT, AND Ph.D. IN
CHEMISTRY FROM JOHNS HOPKINS
SCHOOL OF MEDICINE.
PLEASE YOIN ME IN WELCOME BEING
DR. Mc LELLAN.
[ APPLAUSE ]
>> ALL RIGHT, WELL THANK YOU
VERY MUCH FOR THAT GENEROUS
INTRODUCTION, I THOUGHT I WOULD
TALK ABOUT THE ROLE THAT NIERK H
AND NIH FUNDING HAS PLAYED IN
THE DEVELOPMENT OF INFECTIOUS
DISEASE, GOING THROUGH AND
HITTING SOME OF THE HIGHLIGHTS
ABOUT RSV AND CORONA VIRUS AND
ENDING ON SOME OF OUR RECENT
WORK ON CONGO FEVER VIRUS, I DO
HAVE CONFLICTS OF INTEREST WHICH
WERE ALREADY SHOWN ABOUT I AM AN
INVENTOR TO PATENT AND RECEIVE
PATENT ROYALTIES RELATED TO WHAT
I WILL SPEAK ABOUT.
SO FOR STRUCTURE BASED ANTIGEN
DESIGN, IT'S 1 COMPONENT OF THIS
REVERSE VACSINNOLOGY APPROACH TO
VACCINE DESIGN AND WE SAY
REVERSE BECAUSE HISTORICALLY YOU
WOULD START WITH THE PATHOIEN
AND ISOLATE THE PATHOIEN IS
WEAKEN IT OR ENACTIVATE IT AND
IMMUNIZE WITH IT AND HOPE IT
PROVIDED A ROBUST IMMUNE
RESPONSE AND YOU WOULD
EFFECTIVELY WEAKEN IT SO IT
DIDN'T CAUSE DISEASE AND IT
WORKED WELL FOR A NUMBER OF
PATHOIENS AND NOW WHAT WE'RE
DOING IS THE REVERSE IS STARTING
WITH PEOPLE, PEOPLE WHO HAVE
MAYBE SURVIVES THE INFECTIONS
THAT WE'RE INTERESTED IN AND
THEY SURVIVED IT IN PART BECAUSE
THEY MOUNTED A ROBUST IMMUNE
RESPONSE.
PARTICULARLY WHAT WE'RE
INTERESTED IN IS PROTECTIVE
ANTIBODIES AND THEN WE LEVERAGED
MAJOR ADVANCES IN ANTIBODY
ISOLATION, SO WE CAN NOW SORT
THESE CELLS AND COME UP WITH
OTHER WAYS FROM ISOLATING
ANTIBODIES FROM PATIENTS AND
FUNCTIONALLY CHARACTERIZE THEM,
IDENTIFY THOSE THAT ARE
NEUTRALIZING, BROADLY
PROTECTIVE, WE CAN DOWN SELECT
AND IDENT WHAT WE CONSIDERED TO
BE THE TOP 5, 10, 20, BEST
ANTIBODIES THAT HUMANS CAN MAKE
AGAINST THAT PATHOGEN.
HOPEFULLY THESE ANTIBODIES ARE
ALSO PROTECTIVE AND THEN THE
GOAL OF THE VACCINE DEVELOPMENT
IS TO CREATE IMMUNOGENS AND
IMMUNIZATION REGIMENS THAT WILL
REENLIST THESE ANTIBODIES IN THE
VACCINE RECIPIENTS.
SO WE COME IN HERE WITH THE
STRUCTURAL BIOLOGY, AND TRY TO
DETERMINE THE HIGH RESOLUTION
STRUCKURES OF THE ANTIBODIES IN
COMPLEX WITH THE ANTIIEN THAT
PRORIDES INSIGHT INTO THE
MECHANISM OF ANTIBODY MEDIATED
NEUTRALIZATION AND IT CAN ALSO
PROVIDE INSIGHT INTO THE ROLE
THE VIRAL GLYCOPROTEINS THAT
IT'S BINDING, TOO, AND
IMPORTANTLY PROVIDES ACCESS TO
THE CHEMISTRY AT THE IRPT FACE
AND THIS ALLOWS FOR STRUCTURE
BASED AND COMPUTATIONAL
APPROACHES TO ANTIGEN DESIGN, SO
HERE WE CAN SEE MAYBE THIS
PARLIAMENT OF THE ANTIBODY IS
BINDING THIS RED PORTION OF THE
ANTIGEN, NOT THE PURPLE PORTION,
MAYBE THE PERIPHERAL PORTION IS
IMMUNO DOMINANT UNLESS IT'S
NONNEUTRALLIZING ANTIBODIES SO
YOU COULD DO THINGS LIKE
SCAFFOLDING AND TRANSPLANT THAT
RED REGION INTO A
SELF-ASSEMBLING NANO PARTICLE
AND TRY TO FOCUS AN IMMUNE
RESPONSE TO THAT PART.
SO A LOT OF THIS IS BASED ON
IMMUNO FOCUS XG ILLICITTING BEST
IN CLASS ANTIBODIES THAT WERE
PREVIOUSLY ISOLATED.
WITH THE STRUCTURAL INFORMATION,
THERE'S SEVERAL DIFFERENT TYPES
OF COMPUTATIONAL DESIGN
APPROACHES AND YOU CAN DID
SEQUENCE BASED DESIGN WHERE
MAYBE YOU'RE TRYING TO PROVIDE
BREDTH AGAINST MANY DIFFERENT
ISOLATES SO WE HAVE DIFFERENT
ISOLATES, SEQUENCES 1-5.
YOU COULD COME UP WITH IDEAS OF
USING LIKE CONSENSUS SEQUENCES
TO DESIGN YOUR ANTIGEN OR COMMON
ANCESTORS THAT WOULD SHARE
RESIDUES FROM BOTH OF THOSE
ISOLATS, YOU DO CONFIRMATIONAL
STABILIZATION WHICH WILL BE THE
FOCUS OF MY TALK, SOME OF THESE
VIRAL GLYCOPROTEINS EXIST IN
MULTIPLE CONFRONTATIONS
PREFUSION AND POST FUSION.
ONE OF THESE IS LIKELY TO BE A
BETTER'MUNE O GEN AND WE
DETERMINE HOW TO STABILIZE THEM
AND IMMUNIZE WITH THE CORRECT
CONFIRMATION AND THEN OTHER
STRUCTURED BASE DESIGN
APPROACHES AS MENTIONED THE
EPITELOMERE SCAFFOLDING WHERE WE
CAN TAKE AN ANTIBODY BINDING
SITE LIKE THIS HELIX FROM THIS
PROTEIN GRAFFED ON TO A SCAFFOLD
AND IMMUNIZE WITH THAT TO FOCUS
THE IMMUNE RESPONSE ON TO JUST 1
PORTION OF THAT PARTICULAR
ANTIGEN.
AND THEN SUBSEQUENTLY THESE CAN
BE DELIVERED IN DIFFERENT WAYS
AND EVEN PRESENTED ON
SELF-ABLING NANO PARTING --
SELF-ASSEMBLING NANO PARTICLES.
WE WILL TALK ABOUT CLASS 1 VIRAL
PROTEINS.
THESE ARE ALL SYNTHESIZED AS
SINGLE CHAIN INACTIVE
PRECURSORS, YOU SEE THE MOST
COMMON 1S, HIV ENVELOPE AND
INFLUENZA HEMATBLIEWTIN AND THEN
THE F PROTEINS FROM PARAMIX O
VIRUSES AND PNEUMOVIRUSES.
THESE ARE GENERALLY EVOLVE
FRIDAY 1 OR 2 ORIGINAL COMMON
ANCESTORS.
THEY NEED TO BE PROTEIN COMPLEX
LYTICALLY ACTIVATED AT A
CONSERVED SITE AND YOU SEE THIS
HAS THE 2 COMPONENTS, THERE'S
MEMBRANE FUSION COMPONENT AND A
FUSION SUPPRESSIVE CAP AND THE
MEMBRANE FUSION MACHINERY,
THAT'S BEEN CONSERVED, AND
EVOLVE FROM A COMMON ANCESTOR,
IT HAS THE 4 ELEMENTS NEEDED TO
PROMOTE MEMBRANE FUSION WHICH IS
A TRANSMEMBRANE DOMAIN, THE
ANCHORS AND THE VIRAL MEMBRANE,
A FUSION PEPTIDE THAT
GERONTOLOGYSTSS RELEASED AFTER
THE PROTEOLYTIC ACTIVATION THAT
GENERATES LITTLE HYDROPHOBIC
STRETCH OF ACIDS THAT WILL
HARPOON INTO THE TARGET CELL
MEMBRANE AND THEN 2 HAVE TO HAVE
REPEATS SO 7 STRETCHES OF IMMUNO
ACIDS, THEY HAVE HYDROPHOBIC
POSITIONS AT WOB AND 4, THEY
WILL INTERACT WITH EACH AND FORM
A HIGHLY STABLED COIL-COIL AND
LATER A 6 HELIX BUNDLE AND THEN
YOU HAVE SOME OF THESE FUSION
SUPPRESSIVE CAPS SO WE LIKE TO
THINK OF THESE AS BEING SPRING
LOADED, THEY WANT TO UPDATER GO
A CONFIRMATIONAL CHANGE FROM
PRETO POSE, AND LIKE A MUSHROOM,
SITS ON TOP AND PREVENTS EARLY
ACTIVATION AND GENERALLY YOU
COUPLE LIKE RECEPTOR BINDING IN
THE CASE OF CORONA VIRUS,
RECEPTOR BINDING TO THE S1
SUBUNIT AND RBD, DESTABILIZES IT
AND CAUSES SHEDDING AND YOU GET
REFOLDING OF S2.
SO THIS IS WHAT WE LIKE TO THINK
FOR HOW THESE WOULD SIT IN THE
MEMBRANE, THEY INITIALLY FOLD
INTO THE METASTABLE STATE, SO
IT'S NOT THE LOWEST ENERGY
STATE, THAT'S THE POST FUSION
ESTATE BUT THEY START OFF IN
THIS METASTABLE PREFUSION
ESTATE, THERE'S SOME ACTIVATION
ENERGY, A TRIGGERING EVENT THAT
CAUSES IT TO REFOLD, YOU THEN
HAVE THE HARPOONING OF THE
HYDROPHOBIC FUSION PEPTIDE INTO
THE HOST CELL MEMBRANE AND AT
THIS POINT IT ADOPTED WHAT WE
CALL AN INTERMEDIATE WHERE THE
PROTEIN IS SPANNING THE BOTH THE
VIRAL MEMBRANE AND THE HOST CELL
MEMBRANE, THIS COLLAPSES IN HALF
AS THE COILED COILS WALK TOWARD
EACH OTHER, THIS BRINGS THE
HOST-CELL MEMBRANE AND VIRAL
MEMBRANE TOGETHER.
EVENTUALLY THE PROTEIN ADOPS THE
POST FUSION CONFIRMATION AND
THEN THE HOST MEMBRANE AND VIRAL
MEMBRANE'S FUSE, YOU NOW HAVE A
FUSION FOR, SO THE GENOME OF THE
VIRUS CAN ENTER THE CELL AND AT
THAT POINT THE CELL IS INFECTED
AND WHAT WE WANT TO DO IS
GENERATE ANTIBODIES AND MAYBE
SMALL MOLL KIEWMS THAT COMBINE
TO THE FUSION STATE AND PREVENT
THE CONFIRMATIONAL CHANGE FROM
OCCURRING, SO IF WE CAN PURIFY
AND ISOLATE ANTIBODIES AND PASS
PERIPHERAL AXIS OR GENERATE
THESE TYPES OF ANTIBODIES
THROUGH VACCINATION, THEY
COMBINED AND PREVENT THE
CONFIRMATIONAL GYMNASTICS AND
THEN THE VIRUS COOPERATE ENTER
AND THAT PROVIDES PROTECTION, A
PROBLEM IS THAT THE PRE-FUSION
STATE AND METASTABLE AND SO
THERE'S A NICE PAPER FROM DAVID
BAKER AND DAVID AGARD DESCRIBING
THIS, SOING IT'S UNSTABLE BUT
NOT TRAPPED.
SO IT'S NOT THE LOWEST FREE
ENERGY STATE, IT'S HIGHER BUT
IT'S KINETICALLY TRAPPED UNTIL
THERE'S AN EVENT THAT GETS IT
OVER THE HILL.
>> SO 1 PROBLEM IS WHEN YOU TRY
TO PURIFY THESE AND LOOK AT
THEM, THEY JUST QUICKLY
TRANSITION TO THE POST FUSION
STATE AND SO MANY TIMES YOU'RE
IMMUNIZING OR WORKING WITH THE
POST FUSION STATE, WHEN THE IDLE
ANTIGEN MAY BE THE PREFUSION
CONFIRMATION, AND SO WE DID A
LOT OF WORK ON THIS BACK WHEN I
WAS A POST DOC HERE AT THE
VACCINE RESEARCH CENTER IN PETER
QUAWNG'S LAB AND WORKING WITH
BARNY GRAHAM TO DETERMINE
STRUCTURES OF THE POST FUSION
AND PREFUSION CONFIRMATIONS OF
THE RESPIRATORY INTERSTITIAL
VIRUS F PROTEIN, THESE ARE TRI
MERRIC SO THERE'S 3 COPIES OF
F-PROTEIN.
AND YOU CAN SEE THAT 1 PROTEIN
COMPLEX MERIS IN RIBBONS, 1
PROTOMERIS IN SURFACE AND 1
PROTOMER IS IN WHITE.
SO WE LOOK AT 1 INDIVIDUAL
PROTOMER THE 1 IN RIBBONS AND WE
CAN APPRECIATE THE CONFIRMATION
WILLA CHANGES THAT OCCUR IN THE
PREAND POST FUSIONITATE.
SO YOU SLEEP APNEA AND OBESITYY
ABOUT HALF THE MOLECULE THAT'S
IN WHITE DOES NOT GO UNDERGO A
CONFIRMATIONAL CHANGE.
SO THESE AMINO ACIDS ARE WITHIN
ABOUT 5
ABOUT 5-ANGSTROM, THERE'S A
LARGE BETA SHEET THAT'S MOSTLY
THE SAME.
BUT THEN THE N-AND C TERMINUS OF
THE F-1 SUBUNIT UNDERGO
REFOLDING AND CONFIRMATIONAL
REARRANGEMENT.
SO THE C-TERMINUS OF F1 PIVOTS
AROUND THIS AMINO ACID HERE AND
THIS FLIPS ALL THE WAY AROUND TO
FORM THE OUTER HELIX OF THIS
HIGHLY STABLE 6 HELIX BUNDLE AND
THEN THE END TERMINUS OF F1
COMPLETELY REFOLDS.
YOU HAVE THE HYDROPHOBIC FUSION
PEPTIDE WHICH IS SEQUESTERED
INSIDE THE CENTER OF THE
PROTEIN, YOU THEN HAVE A HELIX
TURN HELIX, BETA HAIR PIN IN
YELLOW AND GREEN, AND THEN THE
FINAL ALPHA HELIX 4 IN BLUE, ALL
OF THIS FLIPS AROUND AND
POLYMERIZES THE CENTRAL HELIX
AND FORM THIS IS HIGHLY
ELONGATED ALPHA HELIX IN THE
POST FUSION STATE.
SO CAN YOU HAVE THESE ELEMENTS
LIKE THE BETA HAIR PIN IN YELLOW
AND GREEN, ARE REFOLDING AND
TRANSITIONING INTO AN ALPHA
HELIX, SO IT'S A REALLY
SUBSTANTIAL REFOLDING AND
CONFIRMATIONAL CHANGE.
SO WITH THIS INFORMATION, WE'RE
THEN ABLE TO IDENTIFY STABLE
AMINO ACID SUBSTITUTIONS THAT
WOULD LOCK THE PROTEIN IN A
PREPROTEIN STATE SO WE COULD USE
IT AS AN ANTIGEN.
SO TO DO THIS WE IDENTIFY
SEVERAL CLASSES OF STABILIZING
SUBSTITUTIONS, DISULPHIDE BONDS
ARE ALWAYS IDEAL WHEN WE FIND
THEM.
PARTICULARLY IF WEB CONNECTED
CAN GET 1 CYSTINE IN THE
REASONNION THAT MOVES AND
ANOTHER CYSTINE IN A REGION THAT
DOESN'T MOVE, SO THEN THEY COULD
FORM A COVALENT DISULPHIDE BOND,
STAPPLE THOSE 2 REGIONS TOGETHER
AND PREVENT THE CONFIRMATIONAL
CHANGE FROM OCCURRING.
WE CAN ALSO IDENTIFY CAVITY
SUBSTITUTIONS, MANY TIMES IN
THESE PROTEINS THESE EVOLVE TO
BE SOMEWHAT UNSTABLE, SO THERE'S
LITTLE POCKETS AND CAVITIES THAT
THE EVOLUTION THAT IS CREATED
INSTABILITY SO WE CAN FIND THOSE
AND FILL THEM, SO IN THIS CASE,
THERE'S NORMALLY A SMALLER
IMMUNO ACID SIRRINE POINTING TO
THIS CAVITY, AND WE REPLACE IT
WITH A LARGER [INDISCERNIBLE],
AND STABILIZES THE PROTEIN AND
MAKES IT GIFUL TO PULL IT OUT
AND STARVE IT.
THERE'S 6 NEGATIVELY CHARGED
IMMUNO ARK SIDS, CAN YOU MUTATE
THESE TO SPARRAGING OR GLUTAMINE
TO REDUCE THE CHARGE OR
PROPULSION, SOMETIMES WE
ENGINEER IN SALT BRIDGES,
NORMALLY THIS IS AN
R-GENERATEDDINE NEXT TO A
POSITIVELY CHARGED TBLIEWTA MATE
AND NOW YOU GET A
POSITIVE-NEGATIVE CELL BRIDGE TO
HELP STABILIZE IT AND FINALLY
PROLEANS WHICH HAY LOVE AND THEY
WORK REALLY WELL AND THEY'VE
BEEN SHOWN FOR HIV, RSV, AND
LATER FOR COONA VIRUSES, AND
PROTEINS THAT CAP HELIXES OR
THESE RANGES THAT NEED TO
UNDERGO A COIL TO HELIX
TRANSITION, PUTTING IT IN THERE,
RELATIVELY FLEXIBLE AND HELP THE
CHANGE FROM OCCURRING.
WE USED TO DO THIS JUST BY EYE,
NOW THERE'S A LOT OF SERVERS,
TOOLS, AI-ML MODELS TO ASSIST
WITH THIS SO IT'S AN EXCITING
TIME FOR DOING THIS TYPE OF
DESIGN, ULTIMATELY, BACK IN 213,
WE SETTLED ON A SINGLE
DISULPHIDE BONDS THAT LINK THIS
MOVING REGION IN BLUE TO THIS
MOVING REGION IN GRAY AND SO
THAT FORMED THE COVALENT LINKAGE
AND THEN ADDED IN THESE CAVITY
FILLING SUBSIDIARY CONSTITUTIONS
AT THE APEX AND THAT WAS
SUFFICIENT TO KEEP THE MOLECULE
IN THE PREFUSION STATE AND THEN
WHEN BARNY AND COLLEAGUES AT THE
BRC IMMUNIZE MICE AND RECESS MA
CABBINGS WITH A PRIME BOOST AND
LOOK AT THE RSV NEUTRALIZING
ANTIBODY TITERS, WE CAN SEE THAT
HIGHER IS BETTER WITH THE POST
FUSION FORM OF THE FPROTEIN WE
WERE GETTING NEW TITERS IN THE
HUNDREDS AND WITH THE PREFUSION
FORM, WE WERE GETTING TITERS IN
THE HIGH THOUSANDS CLOSE TO
10,000 SO A SUBSTANTIAL INCREASE
USING THE PREFUSION CONFIRMATION
OF THE FPROTEIN AS AN ANTIGEN,
SO LATER WHEN I WAS PROFESSOR AT
DARTMOUTH, WE WORKED WITH A
COMPANY CALLEDAD WHAT AND LAURA
WALKER TRIED TO PROVIDE AN
IMMUNOLOGICAL BASIS FOR WHY
USING THE IMMUNO ILLICITTED SUCH
HIGH ANTIBODY TITERS, SO WORKING
WITH LAUREANO PEELING AWE WERE
ABLE TO ISOLATE 364 RSV F
DIRECTED ANTIBODIES FROM 3 ADULT
DONORS.
ALL OF THESE WERE EXPRESSED AND
PURIFIED AND WE CHARACTERIZED
FOR AFFINITY, EPITOPE,
NEUTRALIZATION POTENCY AND IF WE
LOOK AT THE MOST POTENTLY
NEUTRALIZING ANTIBODIES WE CAN
SEE THAT 60% OF THEM TARGET ONLY
2 EPITOPES, ANTIGENIC SITES THAT
WE CALL ANTIGENIC SITE 5 AND 0.
AND IMPORTANTLY THESE ARE THE
PREFUSION SPECIFIC ANTIGENIC
SITES, SO THOSE SITES ARE ONLY
FOUNDOT PREFUSION FORM OF THE
PROTEIN AND THEY'RE IN A
COMPLETELY DIFFERENT
CONFIRMATION AND POST FUSION.
THEY DON'T EXIST IN POST.
SO IF YOU IMMUNIZE WITH POST,
YOU WILL NEVER BE ABLE TO
ILLICIT ALL OF THESE PREFUSION
SPECIFIC ANTIBODIES THAT ARE
HIGHLY NEUTRALIZING.
SO THE EXTENT TO WHICH PREFUSION
STABILIZATION WORKS REALLY
DEPENDS ON THE PERCENT OF
ANTIBODIES THAT PEOPLE MAKE THAT
ARE TARGETING EPITOPES
EXCLUSIVELY FOUNDOT PREFUSION
CONFIRMATION.
SO WE HAVE ALL THAT WORK DONE
BACK IN 2013 WITH THE
STABILIZATION OF THE ANTIGEN, IT
WEPT THROUGH THE NORMAL DECADE
OR SO OF CLINICAL TRIALS AND
THEN IN 2022, BOTH GSK AND
PFIZER AND NOW IT'S TOP LINE
PHASE 3 DATA FROM THE RSV
VACCINE CANDIDATES IN OLDER
ADULTS AND THEN, LAST YEAR IN
MAY ISSUES THE FDA APPROVED THE
FIRST RSV VACCINE, AREX BEE FROM
GSK WHICH IS BASED ON THE
MOLECULE THAT WE CREATED SO THIS
IS THE PICTURE BARNY SENT TO ME
IN THE FALL OF LAST YEAR WHERE
HE GETTING VACCINATED MOLECULE
WHILE HOLDING A PREFUSION
MOLECULE THAT HE HELPED CREATE
SO I THINK THAT'S REALLY COOL
AND SOME OF THE POWER OF THE
VACCINE RESEARCH CENTER AND WHAT
CAN BE DONE HERE.
I THEN LEFT THE VRC IN 2000
THREEN TO START MY OWN LAB AT
DARTMOUTH AND BARNY AND I WANTED
TO KEEP WORKING TOGETHER, WE
WERE THINKING WHAT ELSE COULD WE
APPLY STRUCTURE BASED VACCINE TO
WITH OTHER VIRUSES AND THIS WAS
AROUND THE TIME THAT THE MITSD
EAST RESPIRATORY VIRUS HAD
EMERGED IN 2012-2013 IN SAUDI
ARABIA, AND IT WAS THE SECOND
COVID VIRUS, SARS IN 2002, AND
MERS IN 2012, AND WE THOUGHT WE
SHOULD TRY TO FIGURE OUT HOW TO
MAKE OPTIMAL CORONA VIRUSES
ANTICIPATING SPILL OVERS IN THE
FUTURE.
THE OTHER THING IS THEY'RE BOTH
SPIKE PROTEINS, AND WE FELT LIKE
WE COULD USE A SIMILAR PARADIGM,
WHERE WE FIRST NEEDED TO OBTAIN
A 3 DIMENSIONAL TRUCTURE OF THE
PREFUSION STATE OF THE CORONA
VIRUS SPIKE, IDENTIFY
STABILIZING SUBSIDIARY
CONSTITUTIONS AND TEST IT AS AN
IMMUNE VACCINE IMMUNOGEN, BUT
THAT'S WHEN WE SET OUT TO DO
BACK THEN, IT WAS -- TOOK
SEVERAL YEARS AND COMPLICATED,
TRYING TO WORK ON MERS, WHERE WE
COULD NOT GET THE STRUCTURE,
ULTIMATELY WORKED AT THE SCRIPTS
RESEARCH INSTITUTE AND 1 OF THE
OTHER HUMAN CORONA VIRUS, HKU1,
THE COMMON COLD, WAS ABLE TO GET
THIS STRUCTURALLY ARE OF THE
HKU1 SPIKE, AND THIS PROVIDED A
LOT OF INFORMATION ON THE
BIOLOGY AND SPIKE PROTEIN, WE
SAW THE BYPASSING DOMAINS WERE
MEDIATING, WE KNEW THEY WOULD
HAVE TO UPDATER GO A
CONFIRMATIONAL CHANGE TO BIND
RECEPTORS OTHERWISE THERE WOULD
BE SUBSTANTIAL CLASSES, BUT
WHATEVER REASON IN THIS BASE
STATE, ALL THE RBDs WERE IN
THE DOWN CONFIRMATION AND
IDENTIFIED 2 SMALL SUBDOMAINS,
SD1 AND 2, AND MOST IMPORTANTLY
IT PROVIDED STRUCTURE OF THE
SPRING LOADED PROFUSION IMAGERY
WHICH LOOKED A LOT LIKE THE
RSV-F PROTEIN, SINCE THAT TIME,
MY LAB, DAVID'S LAB AND OTHERS
HAVE SOLVED A LOT OF CORONA
VIRUS SPIKE STRUCTURES BOUND TO
ANTIBODIES BOUND TO RECEPTORS
AND NOW WE COULD MAKE A NICE
MOVIE OF HOW THIS WORKS, I THINK
THIS IS BETTER THAN THE
ANIMATION IN THE BEGONING.
SO THE FUSION S-1 DOMAIN, FUSION
MACHINERY IN S2, WE KNOW THE
RBDs HINGE INTO AN UPSTATE,
THOSE BIND RECEPTOR,
DESTABILIZES S1 WHICH FALLS OFF,
AND YOU GET THE FOLDING AND
ALPHA HELIX IN HARPOONING OF THE
HYDROPHOBIC FUSION OF THE TARGET
CELL MEMBRANE AND THEN THE PAIR
INTERMEDIATE COLLAPSES AS THEY
HAVE TO HAVE RESEATS AND COILED
COILS START TO FORM THE 6 HELIX
BUNDLE, IT ZIPPERED UP AND
BRINGS THE VIRAL MEMBRANES AND
HOST CELL MEMBRANES TOGETHER.
SO THESE ARE IMPRESSIVE MACHINES
THAT HAVE EVOLVED TO FUSE THEIR
MEMBRANES WITH OUR MEMBRANES AND
INFECT US WHICH WE'RE CONSTANTLY
TRYING TO STOP THAT FROM
HAPPENING.
SO OUR GOAL AT THIS POINT WAS TO
-- YEAH, BACK IN LIKE 2015
MAYBE, TRYING TO STABILIZE THE
SPIKE PROTEINS.
SO HERE YOU SEE THE COMPAREISON
OF PREFUSION S2, FOR CORONA
VIRUSES, AND THERE'S A LOT OF
SIMILARITY, THEY ALL HAVE --
THEY HAVE A CENTRAL HELIX IN
BLUE THAT DOES NOT UNDERGO MUCH
CHANGE BETWEEN PREAND POST.
IT'S JUST SMALLER AND RSV-F, AND
YOU HAVE THE ELEMENTS IN THE
FIRST TO HAVE REPEAT IN GREEN,
YELLOW, ORANGE, RED, SAME, ALL
THOSE SAME COLORS AND ALL OF
THAT'S GOING TO POLYMERIZE AND
STACK AND FORM THAT REALLY LONG
ALPHA HELIX THAT THEN SHOOTS THE
FUSION PEPTIDE INTO THE TARGET
CELL MEMBRANE, SO WE TRIED TO DO
SOME OF THE SIMILAR TRICKS WITH
DISULPHIDE BONDS, CAVITY FILLING
SUBSIDIARY TUITIONS, BUT WE ARE
HAVING ISSUES, BECAUSE FOR RSV,
WE HAD THE STRUCTURE AND WE WERE
TRYING TO MAKE RSV F VERYIANTS.
HERE WE HAVE THE HK1 STRUCTURE
BUT WE'RE TRYING TO MAKE
VERIANTS OF MERS, AND THOSE ARE
ONLY 35% SEQUENCE IESH DEPTITY
AND THINGS LIKE CAVITY FILLING
AND DISULPHIDE BONDS HAVE TO BE
MORE PRECISE THAN THAT.
SO WE WENT BACK TO USING
PROLEANS WHICH WORKED WELL FOR
RSVF, FROM HANS AND HIS GROUP AT
[INDISCERNIBLE].
THEY ENDED UP PUTTING A PROLEAN
BETWEEN THE CENTRAL HELIX OF
RSVF AND THE FIRST HAD TO HAVE
REPEAT ELEMENT AND THAT DEPENDS
ON CONFIRMATION OF SECONDARY
STRUCTURE AND WHEN WE LOOK AT
THAT SAME REGION IN CORONA VIRUS
SPIKES WE SEE IT'S HIGHLY CON7ED
SO THESE ARE DIFFERENT BETA
CORONA VIRUSES, DIFFERENT
LINEAGES, HK1, MERS, SARS, THEY
ALL HAVE 3 CONSERVED IMMUNO
ACIDS, RLD, 2 NONCONSERVED AMINO
ACIDS, A CONSERVED TBLIEWT MATE
AND MOST OF IT IS CONSERVED, SO
MY POST DOC STARTED PERFORMING
PROLEAN SCANNING OR MUTATING
EACH OF THESE AMINO ACIDS FOR
PROLEAN IN MERS AND THEN WE LOOK
AT THE AMOUNT OF PROTEIN
EXPRESSION, GENERALLY USING THAT
AS A PROXY FOR IMPROVED
STABILITY AND SO THE WILD-TYPE
MERS SPIKE USING WILD-TYPE
SEQUENCE, WE COULD BARELY
PRODUCE ANY, IT'S JUST THE FAINT
BAND BUT A SINGLE PROLEAN
SUBSIDIARY TUITION GAVE US
ALMOST A 20 FOLD BOOST IN
PROTEIN EXPRESSION WHICH IS
AMAZING BECAUSE THESE ARE 1200
PROTEINS THAT TRIEMERRIZE THEIR
METASTABLE, GLYCOSYLATEDDED AND
MAINTAINING A SINGLE ACID TO
PROLEAN COULD GIVE YOU THAT MUCH
OF A BOOST.
SEVERAL ON PROLEANS IN THIS
REGION ALSO IMPROVED EXPRESSION
AND WE HAD 2 PRO LINES THAT WE
CALLED 2 P AND THOSE WERE THE 50
FOLD BOOST.
WHEN WE FIEWRIFY THESE SPIKE
PROTEIN AND LOOK AT THEM BY
CHROMEATOGRAPHY, THERE'S THE
WILD STIEP, THE 2 PROLINE SPIKE,
WILD-TYPE MERS SPIKE IS A
MIXTURE OF PREFUSION AND POST
FUSION THAT OVERTIME
IRREVERSIBLY TRIGGERS AND ADOPTS
THE POST FUSION CONFIRMATION,
WHERE THE 2 PROLINE MERS WAS
HOMOGENIUS PREFUSION SPIKE, WE
SEND THIS TO BARNY AT THE LAB IN
HIS TIME AND THEY IMMUNIZED MICE
WITH THE 2 PROLEAN DOSE, THE
WILD-TYPE IN BLUE OR S1 SUBUNIT
IN ORANGE AND THEN LOOKED AT THE
NEUTRALIZING ANTIBODY TITER SO
HIGHER IS BETTER AND AGAIN WE
SAW IMMUNIZING WITH THE 2
PROLEAN FORM OF SPIKE,
ILLICITTED A 5-10 FOLD HIGHER
NEUTRALIZING ANTIBODY TITER THAN
WHILED TYPE OR S1.
IT WASN'T AS BIG AS WHAT WE SAW
FOR RSV, BUT STILL A SIGNIFICANT
DIFFERENCE.
IT WAS ALSO NICE ABOUT THE
STABILIZATION TRIP IS BECAUSE
THE REGION IS SO WELL CONSERVED
WE COULD PUT THE SAME 2 PROLINES
IN ANY 2 BATTA CORONA VIRUS SO
WE DID THIS FOR SARS FROM 2002
AND WE COULD SEE THE WILD-TYPE
IN RED, TO PROLINE IN BLUE,
WILD-TYPE SARS IN PROFUSION 2
FUSION, 2 PROLINE SARS WAS FOR
PREFUSION, THIS WAS SUFFICIENT
FOR THE CRYOEM STUDIES, AND WE
CLONED A BAT CARONA VIRUS, AND
WE HAD 1 AND SHOWED THAT THE 2
PROLINE AND WE PUBLISHED BACK IN
2017 AND PATENTED IT.
WE WERE DISAPPOINTED IT GOT
REJECTED FROM 5 JOURNALS AND SO
THAT WAS TOUGH, NOBODY FOUND
THIS TO BE SIGNIFICANT AND
CORONA VIRUSES ARE REGIONALLY
CONTAINS AND SOME OTHER THINGS
BUT SO THAT WAS DISHEARTENING.
SO THEN I MOVED THE LAB TO HAVE
ACCESS TO ELECTRON MICROSCOPES,
AND THEN IN 2018, WE HEARD ABOUT
THE WUHAN, AND THEY HAD
IEE, AUDIENCE DEBTIFIED IT BY
THE NOVEL CORONA VIRUS, ON
JANUARY 10th THEY RELEASED THE
SEQUENCE ONLINE, AND WITHIN
DAYS, NIANSHUANG WANG STARTED
WORKING ON IT, AND BY AROUND
FEBRUARY 10th OR 11th WE HAD
SUBMITAD I MANUCRYPT TO SCIENCE
AND BIOARCHIVE DESCRIBING THE
STRUCTURE OF THE PROLINE 2
SPIKES.
WE WENT FROM A GENOME SEQUENCE
ONLINE TO SURVEYS SUBMITTED
MANUCRYPT IN 1 MONTH AND WE WILL
NEVER DO THAT AGAIN.
[LAUGHTER]
BUT IT RESULT INDEED A SCIENCE
PAPER, CAME BACK WITH THE
REVIEWS IN 48 HOURS AND NO
REVISIONS, WHICH WAS AMAZING, WE
WERE TRENDING ON REDIT, SO THAT
WAS MY PHONE SCREEN CAPTURE.
THE PAPER'S BEEN CITED OVER
10,000 TIMES SO IT'S ALL DOWN
HILL FOR ME FROM HERE, BUT IT'S
GOOD TO ARE A GOOD 1 AND WHAT
WE'RE MOST EXCITED ABOUT IS ALL
4 OF THE COVID VACCINES
AUTHORIZED FOR USE IN THE UNITED
STATES USED STABILIZED SPIKE
PROTEIN.
SO MODERNA'S CONTAIN 986 AND 7
AS DOES PFIZER AND BION TECH'S
COMMUNITY.
JOHNSON AND JOHNSON HAD THE
ADENOVIRUS, THEY ALSO USED THE
ON 2 PROLINES AND AS DID
NOVAVACS, SO ANOTHER GREAT
COLLABORATION BETWEEN ACADEMIA
AND GOVERNMENT AND WORKING WITH
BARNY THAT'S LED TO ANTIGENS
BEING INCORPORATED INTO HIGHLY
SUCCESSFUL VACCINES.
WE THEN WENT ON TO MAKE AN EVEN
MORE STABILIZED VERSION THAT WE
CALLED HEXAPRO, CONTAINED 4
ADDITIONAL PROLEANS, THAT
SUBSTANTIALLY BOOSTS EXPRESSION
OF 2 P, SO 2 P WE COULD MAKE A
LITTLE BIT OF, MAYBE AROUND, 10
NO PROBABLY MORE -- A COUPLE MIG
DUALAS PER LITER, AND THEN
HEXAPRO WE DID GET CLOSER, WE
DID DETERMINE THE CRYOEM
STRUCTURE, WASN'T MUCH OF A
CHANGE BETWEEN HEXAPRO AND 2 P,
THIS WAS LED BY POST DOC
[INDISCERNIBLE].
RATHER THAN SENDING THE PLASMA
OUT TO A BUNCH OF LABS WE ADD
THEM TO ADD GENE AND THEY
DISTRIBUTED TO 227 LABS AROUND
THE WORLD, SO A LOT OF
RESEARCHERS USING AND WORKING ON
CORONA VIRUS ARE USING THE 6
PIECE SPIKE FOR STRUCTURAL
STUDIES, ANTIBODY ISOLATION AND
IT'S BEEN INCORPORATED INTO
SEVERAL NEW VACCINES FOR LOW AND
MIDDLE INCOME COUNTRIES THAT
WERE REALLY EXCITED ABOUT, AND
THIS IS NOW APPROVED FOR USE IN
THAILAND THAT WAS PARTNERED WITH
PATH, SO IT'S CALLED HXPGPO
VACCINES AND THAT'S HEXAPRO AND
IN MEXICO IT'S CALLED THE PAT
RIA VACCINE, SO THAT'S USED AS A
BOOSTER AS WELL.
WE'VE BEEN WORKING ON VIRAL
PROTEINS BUT WE'VE BEEN TRYING
TO PUSH STRUCTURE BASED ANTIGEN
DESIGN TO OTHER VIRUSES AND EVEN
SOME BACTERIA.
THE VIRAL FUSION PROTEINS, THE
CLASS 1S THAT I MENTIONED, YOU
HAVE CLASS 2s THAT GENERALLY
-- THEY'RE A LITTLE DIFFERENT.
SOMETIMES THEY START OFF AS
DIMERS, TETRAMERS, HIGHER ORDER
AND THEN GO THROUGH AN
INTERMEDIATE AND THEN END UP AS
A FOAOF THE FUSION MOLECULE LIKE
FLAVI VIRUSES AND BUNYA VIRUSES,
AND THEN CLASS 3 LIKE HERPES
VIRUSES, AND I WAS GOING TO TALK
ABOUT WORK IN CLASS 2.
ONE OF THE VIRUSES THAT WE'VE
BEEN FOCUSING ON IS USING THE
CLASS 2 VERSION PROTEINS IS THE
HEMORRHAGIC FEVER FIELD
FUNCTIONS HAVE YOU SEEN, THIS IS
THE MOST WIDE SPREAD TICK-BORNE
INFECTION IN HUMANS, IT USES THE
HARD BODY TICKS AS THE VECTOR.
GENERALLY THE LIFE CYCLE
INVOLVES THE SMALL MAMMALS AND
BIRDS IT CAN EVENTUALLY GET ON
LIVESTOCK AND UNDULATES, THE
INFECTION IN THESE ANIMALS
CAUSES A TRANSIENT VIREMIA, THE
ANIMALS DON'T GET SICK BUT WHEN
HUMANS GET INFECTED YOU GET A
HEMONNAGIC FEVER WITH A CASE
FATALITY RATE OF 10-40%.
SO WE'RE NOT NORMALLY PART OF
ITS LIFE CYCLE, WE'RE MORE OF A
DEAD END HOST BUT IT
PARTICULARLY BAD FOR HUMANS AND
THE INDEMMIC ZONES ARE
EXPANDING.
THERE'S BEEN SOME CASES IN
EUROPE, SPAIN AND ITALY AND IN
PART DUE TO INCREASING
TEMPERATURES, MIEG RATTERY BIRDS
AND LIVESTOCK TRADE.
CCHFV BELONGS TO THE VIRUS AND
THESE ARE A MAJOR FOCUS OF SOME
OF THE INITIATIVES FUNDED BY
NIAID, LIKE THESE REVAMP GRANTS
THAT WE'RE A PART OF AND TRY TO
GENERATE MORE, TOOLS REAGENTS,
ANTIBODY ANDS VACCINES FOR BUNYA
VIRUSES, THESE ARE TRI SEGMENTED
VIRUSES, THESE GENOMES ARE TRI
SEGMENTED THEY HAVE SMALL,
MEDIUM AND LARGE GENOME
SEQUENTS.
THE MEDIUM SEGMENT ENCODES THE
GLYCOPROTEIN PRECURSOR COMPLEX.
AND SO, FOR HANTA VIRUSES, IT
HAS THE RELATIVELY SIMPLE
SEGMENT CAN HAS THE M-PROTEIN
AND GC, GC IS THE VIRAL FUSION
MEMBRANE, AND GN IS AN ACCESSORY
PROTEIN THAT HELPS KEEP GC IN
THE PREFUSION STATE AND
SOMETIMES INVOLVED IN RECEPTOR
BINDING BUT FOR CCHFV, THE
SEGMENT IS MORE COMPLICATED.
IT STILL HAS GC WHICH IS THE
CONSERVED FUSION PROTEIN, IT HAS
NSN, SMALLER GN, PROTEIN CALLED
GP38 THAT WAS THOUGHT TO BE A
SECRETED MOLECULE AND THEN A
MUSEON LIKE DOMAIN.
SO WE'VE BEEN WORKING PART OF
THIS LARGE CONSORTIUM, I WILL
SHOW YOU A PICTURE CALLED
PROMETHUOUS, FUNDED BY U19 TO
REALLY TRY TO UNDERSTAND AND GET
MORE STRUCTURES AND ANTIBODIES
IN CREATE ANTIGENS FOR CCHFV, 1
OF THE INTERESTING THINGS ABOUT
BUNYA VIRUSES IS THAT THE GNG C
DIMERS OLIGMERRIZE TO HIGHER
ORDER STATES.
SO WE CAN SEE FOR THE VIRUSES
THESE WILL END UP FORMING THESE
TRI PORTAL COMPLEXES FOR JUNTA
VIRUSES GN AND GC FORM TETRAMERS
AND THOSE TETRAMERS FORM
LATTICES.
FOR FLAVIVIRUSES YOU GET
PENTAMERS AND HEXAMERS, 1 THING
WE DON'T KNOW IS WHAT THE HIGHER
ORDER IS FOR CCHFV, WE'RE STILL
TRYING TO ANALYZE THAT, WE CAN'T
DIRECTLY VISUALIZE IT UNDER THE
MICROSCOPE, THERE WAS A GROUP IN
2018 THAT PUBLISHED USING HAZARA
VIRUS WHICH IS RELATED TO CCHFV,
BUT ONLY EFFECTS ANIMALS BUT IT
WAS SO DIFFICULT TO TELL, IF
THERE WAS A HIGHER ORDER ASSEM
MYELIN, SO THAT'S STILL AN
UNKNOWN QUESTION.
THIS IS THE PROMETHUOUS GROUP,
SO IT WAS A CEDR, LED BY
[INDISCERNIBLE]'S GROUP AT
ALBERT EINSTEIN AND WE'VE BEEN
WORKING TOGETHER FOR 5 YEARS NOW
AND I THINK I'VE REALLY HELPED
PUSH THE FIELD FORWARD AND
CREATED A LOT OF INTERESTING
REAGENTS AND INSIGHT INTOS THE
CCHFV.
SOME OF THE INITIAL WORK WE'RE
EXCITED WITH, MOSTLY WITH CAR
TECH, MAP BIOAND ADAM AB, THEY
WERE ABLE TO ISOLATE 361
MONOCLONAL ANTIBODIES FROM 4
CCHFV UGANDAN DONEARS, MORE THAN
75% OF THEM WERE GC SPECIFIC,
WHEREAS SOME WERE ABLE TO BIND
THIS PROTEIN CALLED GP38, WHICH
IS THOUGHT TO BE SECRETED.
LAURA'S GROUP PERFORMED YEAST
BASED COMPETITION BINDING
STUDIES AND THEY WERE ABLE TO
IDENTIFY 6 ANTIGENIC SITES WITH
ANTIGENIC SITE 1 BEING THE MOST
DOMINANT.
COMPARED TO SMALLER NUMBERS FOR
THE ANTIGENIC SITES.
NUMEROUS SITES WERE POLARIZING
AND OTHER 38 ANTIBODIES WERE
NONNEUTRALLIZING.
THESE WERE THEN TESTED AT USAM
RID, ANDREW HERBERT'S GROUP, AND
THE LEAD NEUTRALIZING
ANTIBODIESS AND COMBINATIONS AND
WERE ABLE TO AFFORD PROPHYLACTIC
PROTECTION BUT NOT THERAPEUTIC
PROTECTION FROM CHALLENGE.
THESE ARE MOUSE EXPERIMENTS AND
YOU HAVE TO DO THESE WITH IFNAR,
YOU CAN SEE IF THESE ARE GIVEN,
IF THE ANTIBODIES WERE GIVEN 1
DAY BEFORE CHALLENGE THEN THESE
NEUTRALIZING ANTIBODIES WERE
ABLE TO PROTECT, BUT IF THEY
WERE GIVEN 1 DAY AFTER THEN
THERE'S VERY LITTLE PROTECTION
THAT WAS AFFORDED.
BUT STILL IN TERMS OF PROVE LAX
EXCITATORY THESE COULD BE
INTERESTING ANTIBODIES TO MAYBE
HELP PROTECT PEOPLE WHO ARE
GOING INTO AND OUTBREAK OF AN
INDEMMIC ZONE, IT WAS REALLY BS
SIGHTING IS JOHN'S GROUP PART OF
THE CONSORTIUM CREATED A BI
SPECIFIC ANTIBODY AND THEY TOOK
2 OF THE BEST ANTIBODIES, 1
AGAINST SITE 3, AND 1 AGAINST
SITE 1 AND THEY MADE THIS DUAL
V-DOMAIN CHIMERA AND
INTERESTINGLY, THIS BY SPECIFIC
WAS PROTECTIVE WHEN GIVEN 1 DAY
AFTER CHALLENGE AND SO, WE'RE
VERY EXCITED BY THE SPECIFIC AND
THEY'RE WORKING UP METHODS OF
PRODUCING IT IN SCALE.
BUT WE WANT TO UNDERSTAND THE
MECHANISM BEHIND THE ANTIBODIES
WERE NEUTRALIZING AND PERHAPS
THE BASIS OF THE SYNERGY, SO A
GRAD STUDENT IN MY GROUP AT THE
TIME [INDISCERNIBLE] WAS ABLE TO
CRYSTALLIZE THE INDIVIDUAL FABS
AS WELL AS COMPLEX OF THE GC
PROTEIN BOUND TO EACH OF THOSE 2
SITE 1 AND SITE 3 ANTIBODIES.
AND SO IN THIS CASE WE SAW A GC,
WE WERE JUST GETTING A MONOMERIC
FORM OF GC.
WE ALSO WERE PUT IN TOUCH WITH
FELIX RAY'S GROUP WHO WE HEARD
HAD A POST FUSION TRI MERRIC
STRUCTURE OF GC SO WE COMBINED
FORCES TO MAKE 1 NICE MANUSCRIPT
TO PROVIDE MORE INSIGHT INTO THE
GC PROTEIN, WE COULD SEE THE
MONOMERIC FORM, THERE'S A DOMAIN
MISSING DOMAIN 3 WE COULD
OBSERVE BUT IN THE POST FUSION
TRIEMER, THIS FLIPS AROUND AND
HELPS STABILIZE THE LOW ENERGY
POST FUSION STATE.
IF WE LOOK AT THE SITE 1
ANTIBODIES WHICH WERE MOST
IMMUNO DOMINANT, THESE RECOGNIZE
THE FUSION LOOPS, SO FOR THE
CLASS 1S, THEY HAVE A FUSION
PEPTIDE, WHICH SHOOTS INTO THE
MEMBRANE, FOR CLASS 2S THEY
CONTAIN 2 OR 3 LOOPS THAT HAVE
HIDE O PHOBIC ACIDS LIKE
TRYPTOPHAN AND [INDISCERNIBLE],
THAT WILL INSERT INTO THE OUTER
LEAF OF THE MEMBRANE.
WHAT'S INTERESTING THAT THE
FUSION LOOPS THEMSELVES CAN
EXIST IN A PREFUSION AND POST
FUSION STATE.
SO THIS WAS -- WE CAN LOOK AT
THE STRUCTURES FROM A PORTAL
VIRUS WHICH IS A JUNTA VIRUS,
THAT THEY DETERMINED STRUCTURES
OF AND YOU CAN SEE THAT THE GC
FUSION LOOP ANDS IN THE POST
FUSION STATE HAD ALL THE
HYDROPHOBIC RESIDUES STICKS OUT,
THE TRYPTOPHANS, READY TO INSERT
IN THE MEMBRANE BUT IN THEIR
PREFUSION STRAWKURE BOUND TO GN,
NOW THE FEIGNAL ALANINE IS IN
THIS POSITION AND THE TRYPTOPHAN
HAS MOVED DOWN SO IT HAS
REPACKED THE LOOPS AND HIDE THE
HYDROPHOBIC RESIDUES.
SO WE SAW IN THE POST FUSION TRI
MERRIC STRUCTURE, THE FUSION
LOOPS WERE IN THE POST
FUSIONITATE, BUT THE ANTIBODY
THAT WAS POTENTLY NEUTRALIZING
IS ACTUALLY RECOGNIZING THE POST
FUSION CONFIRMATION OF THESE
FUSION LOOPS SUGGESTING IT'S
ENGAGING THE FUSION LOOPS AFTER
GN HAS DISASSOCIATED BUT PRIOR
TO FUSION IN THE LOOPS IN THE
TARGET CELL MEMBRANE.
ANTIBODY 36121, WHICH IS AGAINST
ANTIGEN EXCITE 3 AND MOST OF THE
OTHER ANTIGENIC SITES FOR NOW,
THESE BIND THE MONOMERBUT ARE
INCOMPATIBLE WITH THE BOINDING
POST FUSION TRIEMER, SO IF THE
ANTIBODY IS BOUND, THE OTHER
PROTEIN COMPLEX MERS CANNOT
BIND.
THERE WILL BE SUBSTANTIAL
CLASSES AND SO ULTIMATELY YOU
THINK THAT YOU GET GN'S
ASSOCIATION, GC REFOLDING,
INSERTION OF THE FUSION LOOPS AT
THE TARGET CELL MEMBRANE,
COLLAPSE OF THE PREINTERMEDIATE
AND ADOPTION OF THE POST FUSION
STATE AND THESE ANTIBODIES ARE
TARGETING 2 DIFFERENT STEPS OF
THE FUSION PROCESS, THE SITE 1
ANTIBODIES COMBINED THE RELEASED
LOOPS AFTER THEY REFOLD INTO THE
POST FUSION CONFIRMATION, THIS
WILL PREVENT MEMBRANE INSERTION
AND THE SITE 3 WOULD PREVENT THE
ASSEMBLY OF THE HIGHLY STABLE
POST FUSION CONFIRMATION, SO
YOU'RE TARGETING DIFFERENT STEPS
AND PERHAPS THAT'S WHY WE'RE
GETTING SYNERGY.
WE STILL HAVE BEEN TRYING TO DO
MORE WORK ON ASSEMBLING
STRUCTURES OF THE LARGER COMPLEX
FOR CCHFV, BACK THIS 2020 WE
ALSO SAW THE STRUCTURE OF GP38
BUT THAT DIDN'T PROVIDE A LOT OF
INSIGHTS.
WE WERE HOPING IT WOULD LOOK
LIKE SOMETHING AND PROVIDE
FUNCTION BUT IT DID NOT.
AGAIN, IT'S THOUGHT TO BE
SECRETED AND IS THE TARGET OF
PROTECTED BUT NONNEUTRALLIZING
ANTIBODYINGS AND SO WE TRIED TO
GET STRUCTURES OF GN AND A
COMPLEX OF ALL OF THESE TOGETHER
AND A HIGHER ORDER OF OLIGMERRIC
STATE, ALPHA FOLD 2 COMING OUT
WAS A REALLY BIG ADVANCE AND
FELIX PUBLISHED A NICE REVIEW ON
THIS WHERE THEY USE ALPHA FOLD
AND SHOWED THAT THE STRUCTURE OF
JUNTA VIRUS GN WOULD LOOK VERY
SIMILAR TO THE ALPHA 2 FOLD 2
MODEL, OF CCHFV GN AND PLUS
GP38, SO GP38 ISN'T JUST A
SECRETED PROTEIN, IT'S ACTUALLY
SORT OF GN LIKE, BUT ULTIMATELY
GETS CUT AND DISASSOCIATES.
SO WE WANTED TO TEST THIS MODEL,
SO ELIZABETH MCFADDEN A GRAD
STUDENT IN MY LAB, SHE USED THE
AF2 MODEL TO IDENTIFY AND DESIGN
A STABILIZING DISULPHIDE BOND
THAT WOULD LINK GN IN ORANGE TO
GP38 IN BLUE.
ALL RIGHT, WE MADE THIS
CONSTRUCT, SO WE WOULD EXPRESS
THIS PROTEIN, THE SELECT CRYPTS
AND SINGLE BAN FOR GP38 AND GN
WHICH WE COULD SEE THERE.
IF WE ADDED IN PREDICTED
DISULPHIDE BOND THEN WE GOT A
VERY LARGE BOOST IN PROTEIN
EXPRESSION INDICATING THAT THE
DISULPHIDE IS FORMING AND THAT
THE MODEL IN THAT REGION IS
LIKELY TO BE CORRECT.
WE WOULD THEN ENGINEER AN IRPT
FERON SITE THAT WOULD ALLOW
CLEAVAGE BETWEEN GP38 AND GN, SO
ON A NONREDUCING GEL, WE COULD
SEE 2 BANDS FOR GP38, AND THEN
ON THE NONREDUCING GEL, WE
SHOWED THAT IT WAS FORMING AND
WITHOUT THE DISULPHIDE BOND IN
ALLOWING CLEAVAGE TO, OCCUR WE
GET NO EXPRESSION SO IT'S VERY
UNSTABLE, JUST GP38 AND GN
WITHOUT THE STABILIZING BOND.
INTERESTINGLY THE PROTEIN IS
QUITE STABLE, THE GP38
DISULPHIDE BOND HAD A MELTING
TIP OF AROUND 60-DEGREES.
ENTIOUS LIZ BETH WAS ABLE TO
CRYSTALLIZE AND SOLVE THE
STRUCTURE OF GP38 N AND THEN WE
WANT OFFICE OF DIVERSITY TO GET
THE FULL COMPLEX WITH GC AND WE
TOOK THE APPROACH THAT FELIX'S
GROUP USED FOR JUNTA VIRUS WHERE
THEY LINK WITH A GLIESER LINKER
AND WHICH COULD BE CLEAVED AND
THEN WE ADD A DISULPHIDE BOND
AND ADD GC, AND HAD A MELTING
TEMP OF AROUND 65-DEGREES AND
ALLOWED ELIZABETH TO SHOW THE
STRUCTURES OF GM, CPC, 38 AND
IMPORTANTLY SOME OF THE
ANTIBODIES WE HAD ISOLATED
PREVIOUSLY, WERE USED HERE TO
ADD SOME MASS AND HELP IMPROVE
SIGNAL IN CONTRAST SO WE COULD
GET THE STRUCTURE.
WHAT IT SHOW SYSTEM THAT GN IS
WEDGED IN BETWEEN GP38 AND GC,
AND OVERALL THE GP38, GN FOR
CCHFV LOOKS LIKE JUNTA VIRUS GN,
INTERESTINGLY WHERE IN JUNTA
VIRUS THEY USE THIS LOOP TO HELP
COVER THE FUSION LOOPS FROM GC,
HERE AT AN NLIFE LINK GLYCAN
THAT'S EMANATING OUT FROM GN
THAT INTERACTING WITH AND
STABILIZES THE HYDROPHOBIC
FUSION RESIDUES OF GC WHICH WE
THOUGHT WAS REALLY INTERESTING
AND THEN ELIZABETH MADE A MOVIE
TO SHOW THE PREFUSION AND POST
FUSION DYNAMICS AND STATE.
SO CAN YOU START TO APPRECIATE
HOW EVEN JUST THESE LOOPS CAN
STILL UNDERGO CONFIRMATIONAL
CHANGE IN BOTH HIGH AND THEN
EXPOSE THE FUSION RESIDUES AND
HYDROPHOBIC RESIDUES FOR
INSERTION INTO THE TARGET CELL
MEMBRANE.
WE THEN WORKED WITH SIDE WHAT
AND SHELLY'S GROUP TO IMMUNIZE
THESE PROTEIN ANDS SEE HOW THEY
WORK.
SO HERE WE'RE USING C57 MICE
WITH THE ANTIBODY AGAINST AN
INTERFERON RECEPTOR.
THERE'S A PRIME BOOST FOLLOWED
BY A CHALLENGE. AND WHAT WE SAW
WAS THAT ALL THE PROTEINS WITH
GP38, PROVIDED SOME PARTIAL
PROTECTION, LIKE 40%, GC ALONE,
DESPITE ILLICITTING NEUTRALIZED
ANTIBODIES DOES NOT PROVIDE A
LOT OF PROTECTION, WHEN WE LOOK
MORE CLOSELY, AND WE LOOK AT
THOSE GROUPS, AND THEIR ABILITY
TO ILLICIT GP38 AND THE BINDING
ANTIBODIES ARE ENGINEERED
CONSTRUCT IN PINK THE
HETEROTRIEMER THAT WE GOT THE
STRUCTURE ON, WORKED REALLY WELL
ON ELICITING GP38 ANTIBODIES AND
4 OUT OF 5 MICE THAT ILLICITTED
STRONG ANTIBODY RESPONSES
SURVIVED, WHEREAS 4 DIDN'T.
SO WE THINK THIS IMMUNOGEN IS
DOING SOMETHING AND IF WE CAN
IMPROVE AND GET A BETTER
RESPONSE FROM ALL THE MICE WE
CAN GET UP TO 80 OR NIEBT%
PRODUCTION EMPLOY SO TO
SUMMARIZE THE WORK ON CCHFV,
THIS BIG CONSORTIUM THAT'S
ENDING ALTHOUGH WILL BE
REINCARNATED UNDER A NEW U19
CALLED REVAMP, WE'VE BEEN ABLE
TO ISOLATE GN, GC, NO GN
ISOLATED TO DATE BECAUSE THAT'S
BECAUSE GN IS SMALL RELATIVE, -
IT'S WEDGED BETWEEN GP38 AND GC,
WE DETERMINED STRUCTURES OF
GP38, GC, HETEROTRIEMER, WE
THINK THIS SUGGESTS THAT GP 38
IS NOT JUST A SECRETED PROTEIN,
IT ACTUALLY IS A STRUCTURAL
PROTEIN INITIALLY AND THEN AFTER
PROTEOLYSIS, GP 38, DISSOCIATES
FROM GN AND THEN GP 38 MAY HAVE
AN ADDITIONAL ROLE WORKING WITH
HARRIS' GROUP, SHE SHOWED IT IS
SIMILAR TO DENGUE NS1 AND
PRODUCES VASCULAR LEAK AND MAYBE
THAT'S WHY ANTIBODIES TARGETING
GP38 ARE NONNEUTRALLIZING BUT
PROTECTIVE.
WEAVER CONTINUING TO ENGINEER
THE HETEROTRIEMER AS A VACCINE
ASPECT GEN, DIFFERENT WAYS OF
THE DISPLAYING IT, WE'RE STILL
TRYING TO CHARACTERIZE THE
HIGHER ORDER OLIGMERRIZATION
STATE FOR KRRK CHFV, CLIEK O
PROTEINS AND IMPORTANTLY WE NOW
HAVE THESE ANTIGENS AS NEW
PROBES TO GO BACK AND TRY TO
ISOLATE ANTIBODIES THAT MAYBE
BRIDGE ACROSS GC, GN AND GP38
THAT COULD USEFUL
PROPHYLACTICALLY.
ADDITIONALLY WE DOING A LOT OF
WORK WITH FUNDING FROM SEPE, NEW
ARP A-H GRANT AND TRYING TO
DEVELOP THESE ANTIGEN LIBRARIES
FOR DISEASE X, ARP A-H IS MORE
FOR HE WERIES VIRUSES, CEPI IS
WORKING TO MAKE MORE OF THESE,
AND CREATING THESE ANTIGENS,
CREATING THEM TESTING THEM AND
HAVING THEM READY.
MUCH LIKE WHAT WE DID WITH MERS
AND HAVING THE 2 P READY FOR
SARS-COV-2, WE WANT TO IDENTIFY
STABILIZING SUBSIDIARY
CONSTITUTIONS THAT COULD BE USED
WHEN THE NEXT VIRUS SPILLS OVER.
WORKING WITH JIMMY GOLLIHAR, WE
MADE STABILIZING PROTEINS FOR
ALL THESE DIFFERENT VIRUSES AND
VIRAL ANTIIENS AND SO IN LESS
THAN 1 YEAR WE MADE 8 VIRAL
GLYCOPROTEINS AND OVER 32
VARIANT VS BEEN ARK ASSESSESSED
AND WE HAVE MANY MORE IN THE CUE
NOW.
SO I HIGHLIGHTED A LOT OF PEOPLE
IN MY LAB, AGAIN THIS HAS BEEN
REALLY COLLABORATIVE, WITH GREAT
COLLABORATION WITH BARNY GRAHAM
AND COLLEAGUES AT THE VACCINE
RESEARCH CENTER, ANDREW WARD'S
GROUP FOR THE INITIAL WORK ON
CRYOEM OF HK1.
ALTHOUGH THE PROMETHUOUS,
COLLEAGUES, IN THE CCHFV GROUP
EMPLOY GENEROUS NIH FUNDING
RO-1S AND U19S.
THANK YOU VERY MUCH FOR YOUR
ATTENTION AND HAPPY TO TAKE ANY
QUESTIONS.
[ APPLAUSE ]
>> THANK YOU.
TOUR DEFORCE INDEED.
ANY QUESTIONS FROM THE AUDIENCE,
NOTHING HAS WOMAN IN ONLINE YET.
>> THAT MAY HAVE BEEN MORE
STRUCTURED --
>> [LAUGHTER]
>> HELLO, GREAT TALK, SO I HAVE
A QUESTION FOR YOUR ANTIBODIES
THAT TARGET POST FUSION?
>> ONLY POST FUSION OR ONLY --
>> POST FUSION IS THAT HAPPENING
OUTSIDE THE CELL, WHEN THE CELL
IS INFECTED, ENDO ZONE, RECEPTOR
BINDING WHEN THOSE ARE POSED?
>> GOOD QUESTION.
ANTIBODIES THAT ALSO BIND POST,
THEY BIND PREAS WELL, THAT'S WHY
THEY'RE NEUTRALIZING IS BECAUSE
OF THE ACTIVITY TO BIND PRE, AND
WE THINK THOSE ARE BINDING
PREFUSIONOT SURFACE OF THE
VARIANTS.
SOME OF THE OTHER 1S MAY BE MORE
INTERESTING IF THEY'RE BINDING
-- LIKE EPITOPES THAT BECOME
TRANSIENTLY EXPOSED AND IN SOME
CASES THERE ARE SOME VIRUSES
WHERE MAYBE THEY NEED TO BE
TAKEN UP IN THE ENDOSOME, AND
CLEAVAGE OCCURS AND SO IN THAT
CASE YOU NEED THE ANTIBODY TO
GET TAKEN UP INTO THE ENDOSOME,
SO I'M ALSO THINK OF EBOLA, AND
[INDISCERNIBLE] GROUP HAS DONE
SOME SPECIFICS WHERE 1 ARM BINDS
AND THE OTHER ARM IS READY ONCE
IT'S TAKEN UP IN THE ENDOSOME,
WHICH IS KIND OF INTERESTING, SO
I THINK A LOT OF COOL ANTIBODY
ENGINEERING COULD BE APPLIED
HERE.
>> YEAH, REALLY GOOD TALK.
AND YOU SHOWED IN CCHFV, PREDICT
1 STATE CLOSE TO THE REAL
STRUCTURE SO I WONDER BECAUSE
YOU HAVE ALREADY HAD PREFUSION,
POST FUSION, CAN ALPHA FOLD
PREDICT BOTH IN KNOWN VIRUSES?
>> GREAT QUESTION.
IT DEPENDS A LITTLE BIT.
PEOPLE HAVE CREATED SOME MODELS
WHERE YOU USE TEMPLATES TO HELP
BIAS IT.
BUT THEN YOU'RE KIND OF BIASING
IT, YEAH, SO ON ITS OWN, IT MAY
PICK A PREOR A POST OR WE'VE
SEEN IT CAN PROVIDE LIKE A
CHIMERA, WHICH SOME OF THE
SECONDARY STRUCTURES IS MORE IN
PREAND OTHERS HAVE FLIPPED OUT
AND ARE MORE IN POST.
YEAH, THAT HAS BEEN A LIMITATION
BUY-IN THAT WILL PROBABLY
IMPROVE AS MORE STRUCTURES ARE
AVAILABLE AND THE TEMPLATES DO
HELP BIAS IT IN THAT DIRECTION
EMPLOY THE GOAL FOR CEPI IS TO
GO FROM A NOVEL VIRUS SPILLING
OVER, THE SEQUENCE BEING
ICE'SALATED, ALPHA FOLD
STRUCTURALLY STRUCTURES OF ALL
THE DEFINITE STATES, AI ML BASED
SUBSTITUTION IDEBTICTION AND MR
NA MANUFACTURING WITHOUT EVER
GOING INTO A WET LAB.
>> FUTURE.
>> YEAH, MEMBRANES, MEMBRANES,
THE MEMBRANES OF THE CELLULAR
MEMBRANES AND THE CELL AND THE
ENDO PLASMA, MEMBRANE THEY HAVE
DIFFERENT [INDISCERNIBLE]
>> THE VIRAL MEMBRANES COME FROM
THE CELL MEMBRANES, THEY'RE
PULLED, THAT'S WHERE THE VIRUS
GETS THE MEMBRANE.
>> AND THE PEPTIDE EMBEDDED INTO
THESE MEMBRANES, NOW HAS A
LENGTH OF ABOUT 16, 17 OR
SOMETHING LIKE THAT BUT IF YOU
HAVE GO INTO OTHER MEMBRANE, AND
IN THE [INDISCERNIBLE], THERE'S
SMALL LENGTH IF YOU EXTENDED
LENGTH THE PROTEIN WILL GO TO
THE CELL SURFACE MEMBRANE SO
THAT IS IMPORTANT TO BE TAKEN
INTO CONSIDERATION, WHAT IS THE
MEMBRANE [INDISCERNIBLE] OR THE
THICKNESS OF THE MEMBRANE?
>> YEAH, I WOULD SAY, MOST OF
ALL OF THESE STRUCTURES I'VE
PRESENTED HAVE BEEN FROM SOLUBLE
ECTODOMAINS, WE'RE NOW DOING
MORE STRUCTURESOT SURFACE OF
VESICLES OR VIRUSES THEMSELVES
AND WE'RE STARTING TO SEE THE
TMs AND MAYBE EVEN SOME BOUND
LIPIDS AND THINGS INTO THE
PROTEIN SO WE'RE EXCITED TO
MOVE,A WAY FROM PURIFIED PROTEIN
ANDS INTO MORE OF THE NATIVE
CONTEXT.
>> OKAY.
>> QUESTION FROM THE SOUTH SIDE
HERE.
>> SO IT MAKES PERFECT SENSE
THAT LOCKING THE PREFUSION
DOMAIN FROM INTERACTING WITH THE
RECEPTOR ON THE INFECTED CELL
PROFESSIONAL INTEGRITY PREVENTS
INFECTION BUT WHAT ACTUALLY
HAPPENS TO THE VIRUS THAT IS
CODED WITH THE ANTIPREFUSION
ANTIBODIES.
DOES THAT GET FAIG O
SIGNIFYITOSED, REPRESENTED AS
CROSS PRESENTED ANTIIENS AS
WELL?
>> YEAH, WE KNOW THEY CAN GET
TAKEN UP BY MACROPHAGES, IT CAN
ALSO BE ENDOSIGNIFYITOSED AND
DESTROYED.
THERE MAY BE OTHER OUTCOMES.
>> DO THEY CAUSE RENAL PROBLEMS?
>> THAT I DO NOT KNOW.
>> I DO HAVE A QUESTION ONLINE.
I NOTICE THAD YOUR GROUP DIDN'T
TRY TO TACKLE THE HIV 1 ENV
PROTEIN, ENVELOPE, WHICH IS ALSO
A CLASS 1 FUSION PEPTIDE, IS
THERE A REASON FOR THAT?
>> BECAUSE THERE'S ALREADY GREAT
INVESTIGATORS WORKING ON IT HERE
AT THE VACCINE RESEARCH CENTER
AND ELSEWHERE, SO WE TRY TO
MAYBE GO TO SOME FIELDS THAT
PEOPLE AREN'T WORKING ON OR ARE
A LITTLE UNDERSTUDIED RATHER
THAN COMPETING WITH THE HEAVY
WEIGHTS FOR HIV AND UNIVERSAL
FLU AND SOME OF MY COLLEAGUES
HERE.
>> OKAY, VERY WELL.
>> DIPLOMATIC ANSWER.
>> DR. SHORE?
>> YEAH, WONDERFUL TALK.
I'M JUST WONDERING, SO YOU'VE
SHOWN LOTS OF DATA DERIVED FROM
SEQUENTIAL EXPERIMENTS WITH
DIFFERENT CONFIGURURATIONS OF
THE VARIOUS PROTEINS, CAN YOU
NOW USE THAT INFORMATION THAT
YOU'VE GAINED TO GO BACK AND A
PRIORI LOOK AT A GIVEN PROTEIN
AND PREDICT WHICH
CONFIGURURATION WILL BE MOST
IMMUNOGENIC IN A DIFFERENT
VIRUS, OR DO YOU -- EACH TIME
YOU WORK WITH A VIRUS, DO YOU
HAVE TO GO THROUGH THE SAME
SEQUENCE OF EXPERIMENTS?
>> YEAH, I THINK IT STILL HAS TO
BE DONE IM PERICALLY, THE
IMMUNE SYSTEM IS TOO MUCHAVE
BLACK BOX, PEOPLE ARE TRYING TO
COME UP WITH AI BASED IMMUNE
SYSTEM AND THINGS BUT EVEN THEN
-- YOU KNOW DIFFERENT ANIMALS IN
THEIR REPERTOIRES COULD BE
DIFFERENT.
SO I THINK YOU REALLY HAVE TO
TEST IT OUT.
IDEALLY WE WOULD LIKE TO USE
ANTIBODIES AS THE GUIDE, RIGHT?
AND SO, THAT WORKS VERY WELL
WITH RSV IN PARTICULAR, GROUPS
ALREADY ISOLATED PREFUSION
SPECIFIC ANTIBODIES, WE KNEW
THEY WERE SUPER POTENT AND THE
GOAL IS TRYING TO ILLICIT THEM.
FOR THESE OTHER PATHOIENS IT'S
HARD TO GENERATE ANTIBODIES FROM
OR GET CONVALESCENT DONOR SO WE
HAVE TO START OVER, I THINK TEST
IM PERICALLY.
AND THEN SOMETIMES YOU DON'T
KNOW -- MAYBE 1 FORM TESTSAs A
SUBPROTEIN UNIT COULD GIVE A
DEFINITE RESULT AS AN MRNA
DELIVERED SO I THINK YOU NEED TO
TEST IT WITH DIFFERENT
MODALITIES, DIFFERENT DELIVERY
ADJUVANTS AND TEASE IT APART.
WE ALSO THINK THAT REGARDLESS OF
WHAT THE OPTIMAL ANTIGEN IS,
IT'S PROBABLY NOT WHAT NATURE
EVOLVED, SO NATURE'S EVOLVING IT
TO FUNCTION OR NOT EVOLVING IT
TO BE AN OPTIMAL ANTIGEN SO WE
THINK WE SHOULD BE ABLE TO DO
BETTER THAN NATURE, WE SHOULD BE
ABLE TO BRING ANTIGEN
ENGINEERING AND PROVIDE BENEFIT,
AND IT MAY NOT ALWAYS BE PRE,
BUT WE NEED A LOT OF IMMUNOLOGY
AND VIROLOGY AND THAT'S WHY THE
WORK IS SO COLLABORATIVE.
>> THANK YOU.
>> THANKS.
>> HI, JASON, THE FUSION PEPTIDE
IS SO IMPORTANT YOU KNOW BASED
ON ALL YOUR STRUCTURE ANALYSIS,
AND IT'S ALSO QUITE CONSERVED,
SO WHY HAVEN'T WE SEEN THE
FUSION PEPTIDE FOR THE ANTIBODY
IN ALL KINDS OF SAY CLASS 1
VIRUSES.
SHOULD WE SEE MORE OFTEN OF THIS
TYPE OF ANTIBODY?
>> YEAH, GOOD QUESTION, YOU KNOW
-- I THINK PART OF IT DEPENDS
PERHAPS ON THE LOCATION OF THE
FUSION PEPTIDE AND THE PREFUSION
STATE.
SO FOR HIV ENVELOPE AS YOU KNOW,
IT'S MORE ON THE LATERAL SIDE,
SOMEWHAT EXPOSED, THAT'S TRUE
FOR CORONA VIRUSES, PARAMIX O
VIRUSES FOR RSVF, IT'S VARIED
INSIDE SO IT'S REALLY ONLY
TRANSIENTLY EXPOSED.
I WOULD SAY THE B-CELLS HAVE A
VERY SMALL WINDOW BETWEEN AFTER
THE VIRUS HAS ATTACHED AND THEN
YOU HAVE TRIGGERING SO I THINK
FOR RSV, YOU MAY NOT SEE A LOT
OF THEM.
IT'S ALSO POSSIBLE THEY EXIST
AND WE JUST HAVEN'T FOUND THEM,
RIGHT?
MANY TIMES YOU FIND WHAT YOU GO
LOOKING FOR AND A LOT OF IS
BASED DISCOVERY SO I'M UNAWARE
OF PEOPLE THAT HAVE TRIED TO
LIKE USE FUSION PEPTIDE AS BAIT
TO ISOLATE RSV ANTIBODIES BUT
MAYBE PEOPLE HAVE, YOU GO YEAH,
A LOT OF IT IS BAIT DERIFFED
ANTIBODY ISOLATION.
>> [INDISCERNIBLE] SO IT SHOULD
BE ACCESSIBLE?
>> IT SHOULD BE ACCESSIBLE BUT I
THINK IN SOME CASES IN THE CASE
OF RSVF, THE VIRUS IS ATTACHED
SO YOU WOULD NEED B-CELLS TO GET
IN THERE SO I THINK
THERE'SSTERRIC AS WELL AS
TEMPERRAL THINGS TO CONSIDER.
>> HEY, JASON.
>> HEY CHRIS.
SO YOU SHOWED THIS VERY NICE
PIPELINE THAT YOU USUALLY START
WITH ANTIBODIES DERIVED FROM
PATIENTS.
>> IF POSSIBLE.
>> IF POSSIBLE, SO WHAT ARE
OTHER MEANS OF HAVING GOOD
STARTING [INDISCERNIBLE].
DO YOU START BY IMMUNIZING FOR
EXAMPLE, LLAMAS OR ALPACAS OR
OTHER SOURCES OF ANTIBODIES,
LIKE RABBITS?
>> SO THAT'S A GREAT WAY, WE'VE
DONE ANTIBODY INDEPENDENT AS
WELL, WHICH IS REALLY PAIN
STAKING BECAUSE WE CAN'T REALLY
ASSESS THE CONFIRMATION OF ANY
OF THE VARIANTS AND WE
ULTIMATELY NEED TO LOOK BY EM
WHICH IS VERY SLOW AND TEDIOUS,
IF YOU DON'T HAVE THE HUMAN
DERIVED ANTIBODIES YOU SHOULD BE
ABLE TO DERIVE THESE BY
IMMUNIZATION OF SMALL ANIMALS
AND BEST IS TO DO SOME SORT OF
MRNA BASED IMMUNIZATION OF THE
FULL WILD-TYPE PROTEIN WHICH WE
WOULD ASSUME INITIALLY FOLDS IN
THE PREFUSION CONFIRMATION AND
RELATIVELY STABLE AND YOU COULD
IMMUNIZE WITH POST FUSION AS
WELL, HAVING ANTIBODIES, IDEALLY
YOU WANT AN ANTIBODY THAT
RECOGNIZES ALL FORMS AND THEN A
ANTIBODY THAT RECOGNIZES
PREFUSION AND YOU COULD ALSO
HAVE AN ANTIABOUT THAT
RECOGNIZES POST FUSION AND THEN
THE RATIO GIVES YOU THE
PREFUSION PERCENTAGE SO YOU
COULD -- PEOPLE OF IMMUNIZE WITH
PEPTIDES THAT FORM THE HELIX 6
BUNDLE SO SOMETIMES WE HAVE TO
START OFF AND FOR HAD OF THESE
REVAMPS WE HAVE TO -- WE START
OFF WITH REAGENT IMENERATION AND
TRY TO GET THOSE JUST FROM
ANIMALS AND DO YOU HAVE AN
ESTIMATE IF IT IS -- IF YOU GET
BETTER NEUTRALIZING ANTIBODIES
FROM PATIENTS COMPARED TO OTHER
SOURCES?
>> GOOD QUESTION.
I FEEL LIKE MANY OF THE BEST 1S
IN THE DIFFERENT FIELDS ARE
USUALLY FROM HUMANS.
YEAH, THAT'S A CASE WHERE
NEARING THE ANTIBODY WAS REALLY
GOOD COMPARED TO A HUMAN.
AGAIN, SOME OF THAT'S JUST BIAS
MAYBE PEOPLE HAVEN'T LOOKED AS
HARD IN THE 1 ANIMAL AS THE
OTHER.
>> MAYBE 1 LAST QUESTION FROM
ONLINE, IF YOU'RE HOLDING UP.
>> I WILL MAKE IT.
>> BEAUTIFUL TALK.
THANK YOU VERY MUCH.
REGARDING THE STUDIES LOOKING AT
GC STABILIZATION VIA THE END
LINKED GLYCON, COULD THE
MACHINERY BETWEEN HUMANS AND
TICKS OR WHATEVER MODEL USED TO
PROBE THIS INTERACTION GAVE
DIFFERENT RESULTS THAN WHAT WAS
OBSERVED?
NI --
>> I DON'T THINK SO, MOST
GLYCOSYLATION WE'RE SEEING
LAYING ON TO THE GC FUSION LOOPS
IS JUST THE BASE DPLI
COSALATION, THAT'S BETA NANOS
AND THAT'S GENERALLY CONSERVED
IN ALL EUKARYOTES, SO TO THAT'S
DIFFERENTS ON THE BRANCHING AND
TERMINAL RESIDUES, ABOUT YOU
THAT IS THE MORE CONSERVED CORE
GLYCANS THAT ARE RESPONSIBLE SO
I DON'T THINK THAT WOULD HAVE AS
MUCH OF AN IMPACT.
>> I WILL REMIND EVERYONE, NO
MOW QUESTIONS, THERE'S'
RECEPTION OUTSIDE, READY TO GO.
CME NUMBER CODE 57666.
57666.
AND LASTLY LET'S THANK OWEVER
SPEAKER.
[ APPLAUSE ]