Welcome to the Clinical Center Grand Rounds,
a weekly series of educational lectures for physicians and
health care professionals broadcast from the Clinical
Center at the National Institutes of Health in
Bethesda, MD.
The NIH Clinical Center is the world's largest hospital totally
dedicated to investigational research and leads the global
effort in training today's investigators and discovering
tomorrow's cures.
Learn more by visiting us online at http://clinicalcenter.nih.gov
I HAVE THE GREAT HONOR TO
PRESENT OUR SPEAKER FOR TODAY,
CHARLES E EGWUAGU, Ph.D.
DR. EGWUAGU IS AN EPIDEMIOLOGIST
AND IMMUNOLOGIST, AND THE CHIEF
OF THE MOLECULAR IMMUNOLOGY
SECTION, NATIONAL EYE INSTITUTE.
HE RECEIVED HIS Ph.D. IN
EPIDEMIOLOGY AND MICROBIOLOGY
AND MASTER'S OF PHILOSOPHY IN
MOLECULAR EPIDEMIOLOGY FROM YALE
UNIVERSITY.
HE ALSO COMPLETED A MASTER'S OF
PUBLIC HEALTH FROM YALE SCHOOL
OF MEDICINE.
FROM 1987 TO 1990, DR. EGWUAGU
WAS A STAFF FELLOW FOR THE
SECTION ON EXPERIMENTAL
IMMUNOLOGY AT NEI.
HE THEN SERVED AS COMMISSIONED
OFFICER OF THE UNITED STATES
PUBLIC HEALTH AS MUCH AS,
ATTAINING RANK OF CAPTAIN.
CURRENTLY HE'S A SENIOR
INVESTIGATOR AND CHIEF OF THE
MOLECULAR IMMUNOLOGY SECTION.
DR. EGWUAGU'S LABORATORY SEEKS
AN INTEGRATED UNDERSTANDING OF
MOLECULAR AND CELL MECHANISMS.
PARTICULAR INTEREST ON THIS
CYTOKINE IS -- IS ON CYTOSINE ,
TO DEVELOP REGULATORY B CELL
IMMUNOTHERAPY FOR AUTOIMMUNE AND
NEURODEGENERATIVE DISEASES.
HE'S A RECIPIENT OF UNITED
STATES PUBLIC HEALTH SERVICE
COME COMMENDATION MEDAL AND
DIRECTOR'S AWARD, AND MENTORING
AWARD FOR HOWARD HUGHES MEDICAL
INSTITUTE IN MONTGOMERY COUNTY
SCHOOL SYSTEM AND NATIONAL
INSTITUTES OF HEALTH INTERNSHIP
PROGRAM.
HE HAS OVER 100 PEER-REVIEWED
PUBLICATIONS, SUPERVISED 60
TRAINEES, FELLOWS, STUDENTS, A
MEMBER OF THE U.S. PUBLIC HEALTH
SERVICE PROFESSIONAL
ASSOCIATION, ASSOCIATION FOR
RESEARCH IN VISION AND
OPHTHALMOLOGY, AND AMERICAN
ASSOCIATION FOR ADVANCEMENT OF
SCIENCE AND AMERICAN ASSOCIATION
OF IMMUNOLOGISTS.
HE IS A STANDING MEMBER OF STUY
SECTION AND NIH CENTRAL TENURE
COMMITTEE.
THE TITLE OF THE PRESENTATION
TODAY IS REGULATORY B CELLS AND
EXOSOMES, UVEITIS, MULTIPLE
SCLEROSIS AND GRAFT-VERSUS-HOST
DISEASE.
JOIN ME IN WELCOMING TODAY'S
GRAND ROUNDS SPEAKER, DR.
EGWUAGU.
>> WELL, I HAVE NO FINANCIAL
DISCLOSURES.
AND I WOULD LIKE TO THANK YOU
FOR THAT WONDERFUL INTRODUCTION.
ALSO TO THANK THE NIH CLINICAL
CENTER GRAND ROUNDS COMMITTEE
FOR THE INVITATION TO SPEAK HERE
THIS AFTERNOON.
RESEARCH IN MY LAB OVER SEVERAL
YEARS FOCUSED ON AUTOIMMUNE
DISEASES.
WE ARE PARTICULARLY INTERESTED
IN UNDERSTANDING WHAT CAUSES AN
ORGAN-SPECIFIC AUTOIMMUNE
DISEASE SUCH AS UVEITIS OR
MULTIPLE SCLEROSIS.
OUR GOAL HAS BEEN TO IDENTIFY
PATHOGENIC LYMPHOCYTES THAT FUEL
THE VICIOUS CYCLES OF RELAPSING
AND REMITTING INFLAMMATION THAT
CHARACTERIZES UVEITIS AND
MULTIPLE SCLEROSIS.
THE ROLE OF T CELLS IN UVEITIS
AND M.S. IS NOW WELL
ESTABLISHED.
WE PUBLISHED A PAPER THAT
PROVIDED EVIDENCE FOR THE ROLE
OF Th17 CELLS IN HUMAN
UVEITIS.
WE AND OTHERS HAVE ALSO
IMPLICATED Th17 CELLS IN MOUSE
MODELS OF MULTIPLE SCLEROSIS.
HOWEVER, THIS AFTERNOON I WILL
TALK ABOUT THE ROLE OF B CELLS
IN SUPPRESSING CNS AUTOIMMUNE
DISEASES.
WITH PARTICULAR FOCUS ON OUR
EFFORTS TO DEVELOP SAFE AND
EFFECTIVE B CELL IMMUNOTHERAPY
FOR UVEITIS.
A GROUP OF POTENTIALLY BLINDING
DISEASES WHICH CAN BE OF
INFECTIOUS OR AUTOIMMUNE
ETIOLOGY.
UVEITIS IS A HETEROGENEOUS
DISEASE DISCUSSING VKH, AND MANY
OTHERS, CAN MANIFEST IN THE
FRONT OF THE EYE AS ANTERIOR
UVEITIS.
IN THE BACK OF THE EYES, AS
POSTERIOR UVEITIS.
OR THROUGHOUT UVEA AS PAN
UVEITIS.
I'VE LISTED CURRENT TREATMENT.
HOWEVER STEROIDS ARE STANDARD F
CARE.
THIS HAS BEEN THE IMPETUS TO
DEVELOP EFFECTIVE NEW DRUGS FOR
THESE POTENTIALLY BLINDING
DISEASES.
WE BELIEVE THAT BIOLOGICS AND
CELL-BASED THERAPIES MAY BE
SAFER ALTERNATIVES TO STEROIDS
FOR TREATMENT OF UVEITIS.
WE'RE THEREFORE FOCUSED ON
MODULATING ACTIVITIES OF IL-12
CYTOKINE FAMILY OF CYTOKINES,
BECAUSE THEY PLAY CRITICAL ROLES
IN HOST IMMUNITY.
THIS INITIATE INNATE AS WELL AS
ADAPTIVE IMMUNE RESPONSES.
AND THEY ALSO REGULATE THE
INTENSITY AND DURATION OF IMMUNE
RESPONSES.
THEY ALSO SKEW THE
DIFFERENTIATION OF NAIVE
LYMPHOCYTES TOWARDS PATHOGENIC
OR IMMUNOSUPPRESSIVE SUBSETS OF
LYMPHOCYTES.
OUR GOAL HAS BEEN TO MITIGATE
YOU'VE SITES BY TARGETING IL-12
CYTOKINE SIGNALING PATHWAYS THAT
WE CAN USE TO PROMOTE EXPANSION
OF LYMPHOCYTES THAT SECRETE
ANTI-INFLAMMATORY CYTOKINES.
THE IL-12 FAMILY IS COMPRISED OF
FOUR MEMBERS.
IL-12 AND IL-23 ARE PRO INFLAM
TEAR.
IL-27 AND 37 ARE
ANTI-INFLAMMATORY.
EACH MEMBER OF THE IL-12 FAMILY
CYTOKINE IS COMPOSED OF AN ALPHA
CHAIN AND BETA SUBUNIT, BETA
CHAIN.
THE ALPHA SUBUNITS ARE P 35,
P19, AND P 28.
THESE ARE STRUCTURALLY SIMILAR
TO IL-12 SUPER FAMILY CYTOKINES
THAT INCLUDE ONCOSTATIC M,
LEUKEMIA INHIBITORY FACTOR, IL-6
AND MANY OTHERS.
THE BETA SUBUNITS WITH H P 40,
SIMILAR TO INTERFERON TYPE 1
CYTOKINE RECEPTORS.
THE PROFOUND INFLUENCE OF IL-12
FAMILY CYTOKINES ON HOST
IMMUNITY DERIVES FROM THE FACT
THAT EACH OF THE ALPHA AND BETA
SUBUNITS OF THE IL-12 FAMILY
CYTOKINES, THE IMMEDIATE TARGETS
OF TRANSCRIPTION FACTORS THAT
ARE ACTIVATED BY DENDRITIC
CELLS, DURING ANTIGEN PRIME IN
THE LYMPH NODES.
FOR EXAMPLE, PATHOGEN OF
INTEREST IS A GRAM-NEGATIVE
BACTERIA SUCH AS THE ONE THAT
CAUSES MYCOBACTERIA, THAT CAUSES
TUBERCULOSIS, THESE ARE GRAM
POSITIVE BACTERIA, GRAM-NEGATIVE
BACTERIA THAT HAPPEN ON CELL
SURFACE, LIP OH POLYSACCHARIDES
ENGAGE TLR 4 RECEPTORS ON THE
ACTIVITY OF DENDRITIC CELLS
CAUSING THEM TO INDUCE
TRANSCRIPTION FACTOR THAT
MOMENTS SECRETION OF IL-12
CYTOKINES.
THAT PROMOTES THE -- THAT
PROMOTES THE DEVELOPMENT OF TH1
SUBSETS.
HOWEVER, IF THE PATHOGEN IS GRAM
POSITIVE BACTERIA SUCH AS CAUSS
STREP THROATS, THOSE BUGS
CONTAIN GLYCAN ON CELL SURFACES,
THAT INTERACTS CAN TLR2
RECEPTORS IN THE CELL, INDUCING
THEM TO ACTIVATE TRANSCRIPTION
FACTORS THAT PROMOTE PRODUCTION
OF HIGH LEVELS OF IL-23.
IL-23 PROMOTES IMMUNE RESPONSE
TOWARDS TH17 DEVELOPMENTAL
PATHWAY.
ON THAT CHRONIC INFLAMMATION
DENDRITIC CELLS BEGIN TO PRODUCE
IL-27.
FOR THIS SEMINAR I WILL BEGIN BY
PRESENTING OUR DATA, WE SHOW
IL-27 AND IL-35 ARE AFFECTED
BIOLOGICS.
FOR TREATMENT OF UVEITIS OR
ENCEPHALOMYELITIS.
MOST OF OUR STUDIES ARE DONE IN
MICE.
I WOULD ALSO DISCUSS HOW STUDIES
ON THE SUPPRESSION OF UVEITIS BY
THESE CYTOKINES LED TO OUR
DISCOVERY OF TWO NEW REGULATORY
B CELL POPULATION, IN THE
PERITONEAL CAVITY AND HUMAN
UMBILICAL CORD.
THAT SUPPRESSING INFLAMMATION BY
SECRETING IL-27, I-27-BREGS.
THESE CELLS RESIDE MOSTLY IN THE
SPLEEN AND PERIPHERAL LYMPHOID
TISSUES.
THIS SUPPRESSED INFLAMMATION BY
PRODUCING IL-35, AND WE CALL
THESE B REGS I 35 B REGS.
I'LL DISCUSS HOW STUDIES LED TO
A DISCOVERY OF EXOSOMES THAT
CONTAIN IL-27 AND WE'VE NAMED
THEM I27 EXOSOMES, AND B REGS
PRODUCE EXOSOMES THAT CONTAIN
IL-35, I35-EXOSOMES, SUPPRESSING
UVEITIS AND GRAFT-VERSUS-HOST
DISEASE AS STAND-ALONE THERAPY
OR IN COMBINATION WITH THE
CORRESPONDING B REGS THAT
PRODUCE THEM.
I'LL CONCLUDE WITH UNPUBLISHED
DATA PROVIDING NEW INSIGHT ON
IMMUNOBIOLOGY AND BIOSYNTHESIS
OF IL-27 AND IL-35.
AFTER DISCOVERY OF IL-35, THERE
WAS SIGNIFICANT -- HAD BEEN
SIGNIFICANT INTEREST IN USING
THEM THERAPEUTICALLY.
AND THE FIRST ONE OF THE
QUESTIONS WE ASKED WAS IF WE CAN
USE IL-35 TO SUPPRESS UVEITIS.
THE OBSTACLE TO ADDRESSING THIS
ISSUE WAS THAT AT THAT TIME,
BIOLOGIC -- BIOACTIVE IL-35 WAS
NOT AVAILABLE COMMERCIALLY SO WE
HAD TO GENETICALLY ENGINEER
RECOMBINANT IL-35 FOR THESE
STUDIES.
WHAT I'M SHOWING YOU HERE IS
CONSTRUCT THAT WE PRODUCED AND
USED TO EXPRESS THE IL-35 IN
CELLS.
CONSTRUCT CONSISTS OF P 35
cDNA, BOTH JOINED BY IRES,
WHICH IS AN RNA ELEMENT THAT
ALLOWS US TO EXPRESS THIS
PROTEIN AND SECRETE
INDEPENDENTLY.
SO, WE CLONED THIS CONSTRUCT
INTO THE PMI EFFECTOR USE TO
TRANSFECT THESE CELLS TO
GENERATE RECOMBINANT PROTEIN.
THE PURPOSE OF THIS SLIDE HERE
IS JUST TO SHOW YOU THAT WE
PRODUCED HIGHLY PURIFIED IL-35
CYTOKINE.
AND WE DEMONSTRATE HERE THAT THE
IL-35 IS HETERODIMERIC CYTOKINE
BY SEDIMENTATION, EQUILIBRIUM OR
CENTRIFUGATION.
THEY MIGRATE AS HETERODIMER OF
57.7 KILODALTON.
ON THAT REDUCED CONDITION, THEY
MIGRATE AS INDIVIDUAL SUBUNITS.
AND THIS WAS ONE OF THE FIRST
PRODUCTION OF BIOACTIVE
HETERODIMERIC CYTOKINES THAT
COULD BE USED FOR THESE SORT OF
STUDIES.
SO THE FIRST THING WE DID WAS TO
ESTABLISH THE IL-35 IS INDEED
BIOLOGICALLY ACTIVE.
AND WE DID SO BY SHOWING THAT
THE IL-35 CAN SUPPRESS PRIMARY B
CELLS PROLIFERATION, AND DID SO
BY INDUCING B REGS TO PRODUCE
HIGH LEVELS OF IL-10, WHICH IS
AN IMMUNE SUPPRESSIVE CYTOKINE.
INTERESTINGLY, WHY THE IL-35 CAN
SUPPRESS CONVERSIONAL B CELLS,
CAN SUPPRESS PROLIFERATION, IT
INDUCES THE EXPANSION OF VERY
RARE B CELL POPULATION, THAT
PRODUCES IMMUNOSUPPRESSIVE
CYTOKINE IL-10.
AND WE CALL THIS IL-10 B-REGS.
THE LOGICAL EXTENSION OF THIS
WORK IS TO FIND OUT WHETHER WE
COULD USE IL-35 TO SUPPRESS
INFLAMMATIONS IN VIVO AND FOR
THIS WE USED THE EXPERIMENTAL
AUTOIMMUNE UVEITIS MODEL.
EAU IS THE MODEL OF HUMAN
UVEITIS, SHARES
HISTOIMMUNOLOGYIC FEATURES WITH
HUMAN DISEASE, INDUCES MOUSE
STRAINS.
SYMPTOMS OF EAU APPEARS 8, 12,
TO 14 POST IMMUNIZATION, AND
TARGETED DESTRUCTION OF THE
PHOTORECEPTOR LAYER.
IN THE MOUSE WE SEE UNDULATING
CELLS IN THE PHOTORESETTIVE
LAYER, A HALLMARK FEATURE OF
SEVERE UVEITIS IN MICE.
IN THIS STUDY, WE INDUCE EAU IN
MICE, AT THE SAME TIME STARTED
TREATING THEM THE SAME TIME WE
INDUCED EAU STARTED TREATING
THEM WITH 100 NANOGRAMS OF THE
RECOMBINANT IL-35, OR ALSO GAVE
THEM 100 NANOGRAMS OF PMIB,
WHICH IS A CONTROL.
PMIB IS AN EXTRACT DERIVED FROM
CELLS TRANSFECTED WITH MTPMI
DEFECTOR, BY FUNDUSCOPY,
HISTOLOGY, CYTOKINE STAINING,
ANALYSIS.
WHEN YOU LOOK AT FUNDUS IMAGE,
IS THIS WHAT THE FORMAL FUNDUS
LOOKS LIKE, IT HAS THIS
WELL-DEFINED OPTIC DISC.
DURING INFLAMMATION OR IN EAU
THERE'S A SWELLING OF THE OPTIC
DISC, AND WHICH WE CALL PAPULAR
EDEMA.
THESE MICE SHOWN HERE IN THIS
FUNDUS IMAGE IN BLUE, THEY ALSO
DEVELOP RETINAL VASCULITIS, AND
THE WHITE ARROW HERE SHOWS AREAS
OF RETINAL AND CORONAL
INFILTRATES, HALLMARK FEATURES
OF UVEITIS.
AND IN THE HISTOLOGY, STAINING
HERE, THESE AREAS MARKED IN BLUE
ASTERISKS SHOW AREAS OF RETINAL
INFOLDING, WHICH I MENTIONED
EARLIER IS HALLMARK FEATURE OF
SEVERE UVEITIS IN THE MOUSE.
NOW, WHEN YOU LOOK AT THE MICE
THAT WERE TREATED WITH IL-35,
THEY WERE PROTECTED FROM THESE
CARDINAL FEATURES OF UVEITIS
I'VE JUST SHOWN YOU.
WE BELIEVE WE FOUND OUT THAT THE
SUPPRESSION OF UVEITIS DERIVED
FROM THE FACTS THAT IL-35
SUPPRESSED PATHOGENIC CELLS THAT
PRODUCED THE PATHOGENIC
LYMPHOCYTES IL-17 AND INTERFERON
GAMMA.
AT THE SAME TIME THE IL-35
INDUCED EXPANSION OF LYMPHOCYTES
THAT PRODUCED IL-10.
SURPRISINGLY WHAT WE FOUND IS
THAT MOST OF THE IL-10 PRODUCING
B REGS ALSO PRODUCED IL-35, THE
ONES FOUND IN LYMPH NODES
PRODUCED EYE LEVELS OF IL-35.
AS MUCH AS 64% ALSO PRODUCED
IL-35.
THESE ARE THE ONES THAT WE CALL
I35 B-REGS.
IN THIS STUDY, WE DECIDED TO
ISOLATE THOSE B REGS TO EXAMINE
WHETHER THEY CAN BE USED TO
SUPPRESS AND ESTABLISH UVEITIS.
WE ISOLATED CELLS FROM THE
SPLEEN AND LYMPH NODES OF THE
MICE THAT WERE TREATED WITH
IL-35.
AND THEN WE CULTURED THEM,
REACTIVATED THEM WITH IRPP AND
CD40, IL-10 CAPTURE REAGENT TO
SORT THESE CELLS THAT MAKE IL-10
AND ENRICH THEM, WE CAN ENRICH
THEM AND SOME PREPARATION WOULD
GET MORE THAN 90% OF
IL-10-PRODUCING B REG CELLS.
SO, WHAT WE DID IN THIS STUDY
NOW IS TO INDUCE EAU AND GIVE IT
FOUR DAYS LEAD PERIOD TO BUILD
UP CRITICAL MASS OF THE
POTENTIAL PATHOGENIC LYMPHOCYTES
AND THEN HIT THEM WITH THE 10
MILLION B REG CELLS.
WE ALSO IMMUNIZED THEM WITH --
INJECTED THEM 10 MILLION B
CELLS, IL-10, SOME MICE LEFT
THEM UNTREATED WITH ANYTHING.
NOW, WHEN YOU LOOK AT THE FUNDUS
IMAGES, YOU SEE HERE THE
RECAPITULATE ESSENTIAL FEATURES
OF UVEITIS.
LOOKING AT THE HISTOLOGY, IT
GIVES YOU A MUCH MORE COMPLETE
PICTURE.
YOU HAVE SIGNIFICANT
INFILTRATION OF INFLAMMATORY
CELLS IN THE RETINA, IN THE
IRIS, AND IN THE VITREOUS.
AGAIN, YOU SEE THE CARDINAL
FEATURES OF UNDULATING CELLS
WHICH WE CALL RETINAL INFOLDING.
WHAT THIS DATA SUGGESTS IS THAT
B-REGS, THE ONES THAT PRODUCE EX
VIVO, CAN BE USED TO SUPPRESS AN
ESTABLISHED UVEITIS.
WELL, IL-27 IS THE OTHER
IMMUNOSUPPRESSIVE MEMBER OF THE
IL-12 FAMILY.
WE ALSO ASKED WHETHER IT CAN BE
USED AS A BIOLOGIC FOR UVEITIS.
OUR INTEREST IN IL-27 AS A
BIOLOGIC FOR UVEITIS DERIVED
FROM OUR PREVIOUS PUBLICATIONS,
SOME SHOWN HERE, WHETHER WE
SHOWED SUPPRESSION OF UVEITIS IN
MICE CORRELATES WITH
UPREGULATION OF IL-27 IN THE
RETINA AND THIS BY MICROGLIAL
CELLS.
THE FACT THAT IL-27 AND IL-35
ARE IMMUNE SUPPRESSIVE
CYTOKINES, THEY ARE STRUCTURAL
AND FUNCTIONALLY SIMILAR.
WE HAVE HYPOTHESIZED IL-27 MAY
ALSO BE AN EFFECTIVE BIOLOGIC
FOR TREATMENT OF UVEITIS.
WE ALSO POSITED THAT IL-27
PRODUCING B REGS, IL-27 B REGS
MUST ALSO EXIST IN VIVO AS I 35
B REGS.
IN THIS STUDY, WHAT WE DID WAS
TO INDUCE EAU AND EXAMINE
WHETHER IL-35 CAN BE -- CAN
SERVE AS EFFECTIVE BIOLOGIC FOR
MOUSE UVEITIS.
MICE -- EAU WAS INDUCED, AT THE
SAME TIME EAU WAS INDUCED, WE
GAVE THEM IL-27, AT THE TIME OF
EAU INDUCTION.
AND EVERY OTHER DAY TILL DAY 10.
AND WHAT IS SHOWN HERE IN THIS
FUNDUS IMAGE IS THAT INDEED THE
IL-27 CONVERTS PROTECTION
AGAINST SEVERE UVEITIS SHOWN BY
DECREASED DISEASE SCORE,
RECAPITULATED BY HISTOLOGY SHOWN
SIGNIFICANT INFLAMMATIONS IN THE
PBS-TREATED MICE COMPARED TO
IL-TREATED MICE.
WE USE TOMOGRAPHY TO MONITOR EAU
IN MICE.
IT'S A NON-INVASIVE TECHNIQUE
JUST LIKE FUNDUSCOPY, WITH
RECRUITMENT OF INFLAMMATORY
CELLS INTO THE RETINA.
WE ADDRESSED NEXT QUESTION,
WHETHER IL-27 B REGS DO INDEED
EXIST IN VIVO.
BEFORE ADDRESSING THIS ISSUE I
WOULD LIKE TO MAKE -- TO NOTE
THAT THERE ARE TWO TYPES OF B
CELLS IN MAMMALS.
ONE IS B-1 LINEAGE, AND B-2
LINEAGE.
AND B-2 CELLS, THEY DEVELOP IN
THE BONE MARROW, THEY ARE THE
MOST PLENTIFUL B CELLS THAT YOU
FIND IN THE SPLEEN AND IN
PERIPHERAL LYMPHOID TISSUES.
AND THE MOST ALL OF IT, I 35 B
REGS OR B REGS PRODUCING IL-35
DISCUSSED EARLIER, ALL DERIVE
FROM THE B-2 LINEAGE.
WE WERE THEREFORE SURPRISED THAT
THE IL-27 WAS NOT -- THESE B
CELLS DO NOT MAKE IL-27.
INSTEAD, WE FOUND THAT THE IL-27
PRODUCTION WAS RESTRICTED TO THE
B-1 LINEAGE, AND THIS B-1 CELLS,
AND B-1 CELLS AT THE EARLIEST
CELLS, B CELLS, THAT APPEAR IN
DEVELOPMENT.
DERIVED FROM EMBRYONIC YOLK SAC,
RESIDING IN THE PERITONEAL
CAVITY MOSTLY AND UMBILICAL CORD
BLOOD OF HUMANS AND MICE.
WE USE COMBINATION OF
IMMUNOHISTOCHEMISTRY AND
CONFOCAL MICROSCOPY TO
DEMONSTRATE INDEED THE B-1A
CELLS WHEN ACTIVATED, PRODUCE
SIGNIFICANT AMOUNT OF IL-27, AS
SHOWN HERE IN THIS CONFOCAL
IMAGE HERE, AND SHOWN HERE WE
DEMONSTRATE THAT THESE IL-27 MAY
INDEED BE HETERODIMERIC ACTIVE
BIOLOGICAL IL-27.
BUT DEFINITELY PROOF CAME FROM
PROXIMITY LIGATION ASSAY, THIS
IS A VERY, VERY SENSITIVE
TECHNIQUE THAT YOU USE TO DETECT
TWO MOLECULES THAT ARE WITHIN
EACH OTHER, WITHIN 40-NANOMETER
FROM EACH OTHER IN THE MEMBRANE.
WHAT YOU SEE HER IS THIS THESE
RED DOTS, SHOWS THE AREA WHERE
THE 28 AND 3 ARE IN CLOSE
PROXIMITY TO EACH OTHER, LESS
THAN 40 FROM EACH OTHER.
WE ALSO IDENTIFIED THE IL-25,
DETECTED THEM ALSO IN HUMAN
BLOOD, BUT IN MOST ABUNDANCE
AMOUNT OF IL-27 WAS FOUND IN THE
UMBILICAL CORD BLOOD.
WELL, IN THIS STUDY WHAT WE DID
WAS REPEATED THE SAME SORT OF
EXPERIMENT TO ASK HERE NOW
WHETHER GENERATED B REGS CAN BE
USED TO TREAT UVEITIS.
THE ANSWER IS YES.
WE OBTAINED -- GENERATED THE
I27-BREGULARS AND SORT -- B-REGS
AND SORTED THEM.
THESE MICE HERE FROM THE FUNDUS
IMAGES, THEY ARE PROTECTED FROM
UVEITIS WHEN TREATED WITH THE
I27-BREGS INDICATED BY LOWER EAU
SCORES.
AND WE FOUND THE SUPPRESSION OF
THE DISEASE WAS NOT ONLY BY
SUPPRESSING PATHOGENIC
LYMPHOCYTES.
THEY ALSO INDUCED PRODUCTION OF
HIGH LEVELS OF CYTOKINES LIKE
IL-10, IL-35, IL-27.
AND THIS IS REMINISCENT OF WHAT
IS REFERRED TO -- THAT I WOULD
LIKE TO CALL INFECTIOUS
TOLERANCE.
HOWEVER, BECAUSE WE HAD A
PRODUCTION OF IL-10 AND IL-35,
TWO OF OUR REVIEWERS OF OUR
PAPERS INSISTED WE PROVIDE
EVIDENCE THIS DISEASE
SUPPRESSION HERE IS NOT CAUSED
BY THESE OTHER CYTOKINES.
AND FOR THIS WE DECIDED TO USE A
CRISPR TECHNOLOGY TO GENETICALLY
ENGINEER MICE WITH TARGETED
DELETION OF THE IL-27 RECEPTOR
FROM THE B CELLS.
NOW, WHEN WE REPEATED THIS
EXPERIMENT WE FOUND THE BREGS
CAN NO LONGER SUPPRESS DISEASE.
SUGGESTING IN ADDITION TO ONE OF
THE MECHANISMS USED IS BY
INDUCING INFECTIOUS TOLERANCE
WHICH IS A MECHANISM BY WHICH
INFLAMMATORY CYTOKINES CAN
CONVERT B CELLS TO REGULATORY
CELLS, AND THAT'S WHAT WE'RE
SEEING WITH INCREASE IN CELLS
THAT COULD MAKE OTHER CYTOKINES,
ALSO CAUSES EXPANSION OF IL-27.
AND THIS IS WHAT'S RESPONSIBLE
FOR THE SUPPRESSION OF THIS
DISEASE.
AT THE BEGINNING I TOLD YOU THAT
WE ALSO ARE INTERESTED IN
DEVELOPING BIOLOGICS, THAT CAN
BE USED TO TREAT MULTIPLE
SCLEROSIS.
FOR THESE STUDIES WE USED
EXPERIMENTAL AUTOIMMUNE
ENCEPHALOMYELITIS MODEL, EAE.
WHICH IS A MODEL OF MULTIPLE
SCLEROSIS.
WE INDUCED DISEASE BY IMMUNIZING
MICE WITH MOG PEPTIDES, IN
ADJUVANTS, FOLLOWED MICE FOR
ABOUT 30 DAYS.
EXAMINED ALL THE SYMPTOMS THAT
CHARACTERIZED EAE IN MICE.
AND AS YOU CAN SEE, THESE MICE
HERE WERE PROTECTED FROM SEVERE
EAE.
THE SAME DATA SHOWN HERE BY
HISTOLOGY, THESE BLACK ARROWS
INDICATE AREAS OF INFLAMMATION,
AND AS YOU CAN SEE MICE THAT
WERE THE CONTROL MICE HERE
DEVELOPED -- HAD SIGNIFICANT
INFILTRATION OF CELLS INTO THE
BRAIN AND SPINAL CORD.
AND WE ALSO USED BLUE DYE TO
STAIN THE SECTIONS, AND THIS
ALLOWS US TO DETECT AREAS OF
DEMYELINATION, WHICH IS AN
IMPORTANT ASPECT OF THE DISEASE
IN MULTIPLE SCLEROSIS.
AS YOU CAN SEE, MICE THAT
RECEIVED IL-27 WERE PROTECTED
FROM THESE HALLMARK FEATURES OF
EAE IN MICE.
IN THIS STUDY WHAT WE DID THEN
WAS TO USE WHAT WE CALL TWO
CONGENIC MOUSE STRAINS, ONE IS
CD45.2, ONE IS CD45.1.
WE ISOLATED THE BREGULARS FROM
THE CD45.2 MICE, INDUCED EAE, IN
THESE MICE.
AND THEN TREATED THEM WITH THESE
BREGS, THIS SYSTEM ALLOWS US TO
MONITOR AND TRACE AND FIND OUT
THE SOURCE OF THE BREGS THAT
CONTRIBUTED IN THE SUPPRESSION
OF THE DISEASE.
AND WHAT THIS DATA IS SHOWING
HERE IS THAT BOTH THE DONOR
CELLS THAT WERE THE SOURCE OF
THE BREGS, THEY THEY ALSO EXPAND
IN VIVO, IN THE ANIMALS,
RECIPIENTS, IN ADDITION THE
ANIMALS THAT THE RECIPIENT MICE
ALSO -- NORMAL CELLS WERE ALSO
CONVERTED, TO REGULATORY CELLS
THAT MAKE IL-27 AND IL-10.
AND THE MECHANISM BY WHICH THEY
CONTRIBUTED TO SUPPRESS EAE HERE
IN THIS MODEL.
I WILL NOW SWITCH GEARS AND TELL
YOU ABOUT AN EXOSOME
IMMUNOTHERAPY.
DURING CHARACTERIZATION OF THE
IL-27 BREGS AND IL-35 BREGS WE
DISCOVERED THAT THEY SECRETE
EXOSOMES THAT CONTAIN IL-27 AND
IL-35, WE CALL THESE EXOSOMES
IL-27 AND IL-35 EXOSOMES, THEY
RANGE FROM 50 TO 150 NANOMETERS,
AND THIS ENABLES THEM TO BE ABLE
TO CROSS THE BLOOD-BRAIN BARRIER
AND BLOOD-RETINAL BARRIER MAKING
THEM IDEAL VEHICLES FOR
THERAPEUTIC DELIVERY OF IMMUNE
SUPPRESSIVE CYTOKINES LIKE IL-35
AND IL-27 INTO THE BRAIN OR THE
RETINA.
WE BELIEVE THAT EXOSOMES ARE
MAJOR THERAPIES FOR INFLAMMATORY
DISEASES.
AND I WILL PROVIDE DATA, OUR
CONCLUSION DERIVES FROM THE FACT
THAT WE'RE ABLE TO USE IL-35
EXOSOMES AND IL-27 TO SUPPRESS
UVEITIS AND GRAFT-VERSUS-HOST
DISEASE.
SO IN THIS STUDY WE ISOLATED I
35 EXOSOMES FROM CULTURES FROM
THE SUPER NATES OF CULTURES OF I
35 BREGS, EXTRACELLULAR
VESICLES, USING TO ISOLATE THE
EXOSOMES, AND EXOSOMES RANGE AS
SHOWN FROM 50 TO 150 NANOMETERS.
WE INDUCED EAU IN THESE MICE.
AFTER NINE DAYS STARTED TREATING
THEM WITH -- WE GAVE THEM 20
MILLION OF THE I 35 EXOSOMES
RETRO ORBITALLY, INJECTED MORE
OR LESS IN THE EYE.
AND THEN EVALUATED THE EFFECT OF
THE EXOSOMES BY FUNDUSCOPY,
HISTOLOGY, OCT AND ERG.
AGAIN, IT GAVE RESOUNDING
ANSWER, THAT THE EXOSOMES CAN
SUPPRESS THE UVEITIS IN THESE
MICE.
AND SUPPRESSION DERIVES FROM
INHIBITING PRO-INFLAMMATORY
CYTOKINES AND UPREGULATING
IMMUNOSUPPRESSIVE CYTOKINES AS
DEMONSTRATED BY ELISA.
WE REPEATED THE SAME EXPERIMENT
FOR IL-27 EXOSOMES, WHICH WE SAW
ISOLATED FROM CULTURES OF I 27
BREGS, RECAPITULATED SAME
RESULTS WITH IL-35 EXOSOMES.
THE ADDITIONAL DATA THAT WE GOT
FROM THIS DATA, FROM THIS STUDY,
IS THAT ONE OF THE WAYS THAT THE
EXOSOMES INHIBIT INFLAMMATIONS
IS INDUCING CHECKPOINT
INHIBITORS LIKE PD-1, CTLA-4,
INDUCES INCREASED EXPRESSION IN
T CELLS AND THIS CHECKPOINT
INHIBITORS ARE KNOWN TO INDUCE T
CELL EXHAUSTION AND SO ONE OF
THE WAYS THAT THEY USE
SUPPRESSING THE PATHOGENIC
LYMPHOCYTES IS BY INDUCING T
CELL EXHAUSTION.
I WOULD NOW TURN TO
GRAFT-VERSUS-HOST DISEASE WHICH
IS ANOTHER INFLAMMATORY DISEASE.
GRAFT-VERSUS-HOST DISEASE IS
CHRONIC AUTOIMMUNE DISORDER THAT
CAN AFFECT MOST TISSUES OF THE
BODY.
WE THERE EXAMINED WHETHER WE
COULD USE I 35 BREGS OR EXOSOMES
TO TREAT GRAFT-VERSUS-HOST
DISEASE.
THIS IS A MODEL OF THE GvHD
MODEL WE USED IN THIS STUDY,
CONSISTS OF USING TWO MICE THAT
DIFFER IN THEIR H2K LOCUS.
THE RECIPIENT MICE WE GAVE IT
10.4 GRAY UNIT OF RADIATION, FOR
15 MINUTES.
AND SIX TO EIGHT HOURS BEFORE
TRANSPLANTATION.
FOR THE TRANSPLANTATION, WE USED
DONOR MICE HERE, AND WE
OBTAINED -- WE TRANSPLANTED 10
MILLION T CELL DEPLETED BONE
MARROW CELLS.
FOR THE ANIMAL THAT WOULD HAVE
GRAFT-VERSUS-HOST DISEASE WE
GAVE THEM THE BONE MARROW CELL
AS WELL AS 50 MILLION ALLOGENEIC
SPLEEN CELLS.
FOR THE TREATMENT GROUP, WE GAVE
THEM THE SAME REGIMENT AS
GRAFT-VERSUS-HOST DISEASE BUT
ALSO INCLUDED 1.5 MILLION I35
BREGS.
HISTOLOGICAL EXAMINATION WERE
RESTRICTED TO LUNGS, SPLEEN,
LIVER, COLON.
THIS IS SCORING METHOD TO SCORE
GRAFT-VERSUS-HOST DISEASE,
LOOKED AT BODY WEIGHT CHANGE,
POSTURE, ACTIVITY OF THE MICE,
SKIN INTEGRITY.
AND THE GRAFT-VERSUS-HOST
DISEASE SCORE IS SUBMISSION OF
THESE FIVE PARAMETERS.
AND I WOULD LIKE TO MENTION HERE
THIS WAS A VERY LABOR INTENSIVE
STUDY BECAUSE A LOT OF POSTDOCS
AND STUDENTS HAD TO GO TO THE
ANIMAL ROOM EVERY DAY TO EXAMINE
THESE MICE.
AND IT WAS PARTICULARLY
CHALLENGING DURING THE COVID
PANDEMIC.
NOW, IN THIS MODEL THAT WE USE,
MICE THAT DID NOT RECEIVE ANY
TREATMENT THAT HAD
GRAFT-VERSUS-HOST DISEASE, THEY
DIED WITHIN 45 DAYS.
ALL OF THEM.
100%.
AS SHOWN HERE MICE THAT RECEIVED
IL-35 TREATMENT, THEY SURVIVED
TILL 90 DAYS.
AND HOWEVER IT'S ONLY 60% OF
THEM THAT SURVIVED TO 90 DAYS
BUT IT'S A VERY, VERY
SIGNIFICANT PROGRESS IN THIS
PARTICULAR THERAPEUTIC APPROACH.
AND ALSO, YOU SEE HERE THAT IT
ALSO SUPPRESSED MICE THAT HAS
SUPPRESSION OF CLINICAL SYMPTOMS
OF GRAFT-VERSUS-HOST DISEASE IN
MICE, I 35 BREGS, STUDY TISSUES
FROM THE LIVER, THE COLON, THE
SPLEEN, AND THE BRAIN.
AND WE SENT THIS OUT
COMMERCIALLY FOR UNBIASED
PATHOLOGIST TO SCORE THE
DISEASE.
AND THIS IS THE RESULT THAT THEY
CAME UP WITH.
AND AGAIN, ACROSS THE BOARD,
WHETHER LIVER OR COLON OR SPLEEN
OR LUNG, THE DISEASE IS
SIGNIFICANTLY REDUCED.
AND DISEASE REDUCTION DERIVED
FROM THE FACT THAT THE IL-35
BREGS SUPPRESS CYTOKINES THAT
CAUSE WHAT WE CALL CYTOKINE
STORM, WHICH IS RESPONSIBLE TO A
GREAT EXTENT TO THE PATHOLOGY
THAT'S SEEN IN GRAFT-VERSUS-HOST
DISEASE.
WE LOOKED AT THE CELLS IN
DIFFERENT TISSUES, AND, AGAIN,
IN THE LIVER, WE SEE SIGNIFICANT
EXPANSION OF IL-10-PRODUCING B
CELLS AND IL-35-PRODUCING B
CELLS IN THIS MODEL.
WE REPEATED THIS EXPERIMENT
USING THE I35 EXOSOMES, AGAIN
SHOWING WE -- ALL OF OUR
EXOSOMES, SOME WE VERIFIED USING
ELECTROMICROSCOPY TO SHOW HERE
THEY ALL RANGE IN SIZE BETWEEN
50 AND 150 NANOMETERS.
AGAIN, I 35 EXOSOME PROLONGED
SURVIVAL OF GRAFT-VERSUS-HOST
DISEASE MICE SHOWN IN THIS, 80%
OF THEM SURVIVED, BY DAY 40 MOST
OF THE 45, MOST ALL OF THE
GRAFT-VERSUS-HOST DISEASE THAT
WERE NOT MICE WHO WERE NOT
TREATED DIED.
AGAIN, THE I35 EXOSOMES
EXPRESSED GRAFT-VERSUS-HOST
DISEASE IN THIS MODEL.
AND, AGAIN, DISEASE SUPPRESSION
DERIVED FROM SUPPRESSION OF
CYTOKINES THAT MEDIATE CYTOKINE
STORM, WHICH CONTRIBUTES HEAVILY
FOR THE DISEASE PATHOLOGY.
AGAIN, WHEN WE LOOKED AT THE
CELLS, WE SEE THAT THE T CELLS
IN THESE ANIMALS, THEY EXPRESS
HIGH LEVELS OF CTLA-4, PD-1 AND
CHECKPOINT INHIBITORS PLAY A
ROLE ON THE INTEGRITY OF CELLS,
T CELL EXHAUSTION, REDUCED
EFFECTIVE NUMBER IN VIVO.
I WILL NOW SUMMARIZE.
WHAT THE TAKEHOME MESSAGE FROM
ALL THE STUDIES I TOLD YOU WILL
REGULATORY B CELLS AND THEIR
EXOSOMES, WELL, THE TAKEHOME
MESSAGE IS THAT THE BREGS AND
EXOSOMES THEY PRODUCE, POTENTIAL
EMERGING IMMUNOTHERAPIES FOR
UVEITIS, MULTIPLE SCLEROSIS, AND
GRAFT-VERSUS-HOST DISEASE.
I WOULD END BY SHARING WITH YOU
GAPS IN KNOWLEDGE BETWEEN THESE
AND CYTOKINES IL-35 AND IL-27.
THEY ARE COMPRISED OF TWO
SUBUNITS.
WE KNOW THE GENES THAT CODE, WE
KNOW CELLS THAT PRODUCE THEM,
MOSTLY PRELIMINARY -- LYMPHOCYD
MYELOID CELLS, WE KNOW SOME
REGULATORY PROPERTIES.
EACH OF THE SUBUNIT PROTEINS
ENCODE SIGNAL PEPTIDES, TWO
SUBUNITS THAT MAKE IL-35 AND
IL-27 EVENTUALLY ASSOCIATE VERVE
TO FORM FUNCTIONAL ON IL-27
CYTOKINE.
HOWEVER, NOBODY KNOWS THE
MECHANISM BY WHICH THESE ARE
CAUSED.
AND THE DATA THAT WE HAVE IS
THAT SO CONTRARY TO PREVAILING
VIEW THAT IL-35 AND IL-27 ARE
SECRETED CYTOKINES, I WILL SHOW
YOU DATA THAT THE P 35 IS
TRANSMEMBRANE PROTEIN AND NOT A
SOLUBLE PROTEIN, AND PROVIDE
EVIDENCE THAT THE EBI3 IS THE
ONLY ONE CYTOPLASMIC OR SECRETED
PROTEIN AND SHOW EARLIER RECENT
DATA THAT IN RESPONSE TO INTENSE
INFLAMMATION, IL-35 -- EBI 3,
GOES TO THE MEMBRANE, CYCLES
BACK, ASSOCIATES WITH P 35 TO
FORM MEMBRANE BOUND IL-35
CYTOKINES.
SO IL-35 WE DON'T BELIEVE IS
SOLUBLE PROTEIN.
WE BELIEVE IT'S MEMBRANE BOUND
CYTOKINE.
THIS DATA HERE, WE USE
COMBINATION OF
IMMUNOHISTOCHEMISTRY AND
CONFOCAL MICROSCOPY, AND THESE
ANALYSIS WE ACTIVATED THESE
CELLS AND AS YOU CAN SEE HERE IN
THE PLASMA MEMBRANE HERE IN
GREEN, YOU SEE THE P 35.
IT'S NOT IN THE CYTOPLASM.
AND THIS BLUE AREA HERE IS JUST
DEPICTS THE NUCLEUS, THE DYE.
WHEN YOU LOOK AT EPI 3 AS YOU
SEE IT'S NOT ON THE MEMBRANE,
IT'S MOSTLY IN THE CYTOPLASM.
AND I'M SHOWING YOU THIS PROTEIN
HERE.
IT'S A CD8 CD81 PROTEIN, SIGNAG
PROTEINS TO PLASMA MEMBRANE
PARTICULARLY CD19 PROTEIN.
INTO THE PLASMA MEMBRANE.
WHAT WE SHOW HERE IS THAT THE P
35 CO-LOCALIZES WITH CD81, MAYBE
CD81 PLAYS A ROLE IN TRAFFICKING
THE P 35 TO THE PLASMA MEMBRANE
AND AGAIN WHEN YOU LOOK AT THE
IMAGE HERE EBI 3 STAYS IN THE
CYTOPLASM, RETICULUM, SHOWING
THE INTERACTION BETWEEN 81 AND P
35 HERE BUT PROXIMITY LIGATION
ASSAY THAT INTERACT.
HOWEVER, DURING PROLONGED
ACTIVATION OF THESE CELLS, AND
THIS DATA WAS GENERATED BY A
GRADUATE STUDENT WHO JUST
RECEIVED A Ph.D. THIS PAST
YEAR, AND WHAT WE SEE IS THAT
THE EBI3 THEN EITHER MIGRATES
FROM THE CYTOPLASM OR
TRANSPORTED FROM THE CYTOPLASM
TO THE PLASMA MEMBRANE OR FROM
THE CYTOSOL, AND FORMS
TRIMOLECULAR COMPLEX WITH CD81,
P35, AND EBI3 TO FORM MEMBRANE
BOUND IL-35.
WE DID THE SIMILAR THING WITH
THE IL-27 BREGS, AS YOU CAN
APPRECIATE HERE THE EBI - 28 IS
FOUND IN THE PLASMA MEMBRANE, TO
SOME EXTENT IN CYTO CYTOPLASM.
EBI 3 IS MOSTLY CYTOPLASMIC.
WHEN YOU DO THE IMAGE HERE YOU
SEE CO-LOCALIZATION OF CD81 AND
P28.
MEANWHILE EBI3 IS MOSTLY SO
CYTOPLASMIC, SHOWN BETTER HERE.
IN THIS STUDY HERE WE DID ONE
ADDITIONAL THING, LABELED ALSO
DETECT CALRETICULIN IN CYTOPLASM
AND ENDOPLASMIC RETICULUM.
I WILL STOP HERE AND INTRODUCE
YOU TO SOME OF THE VERY HAPPY
PEOPLE IN MY LAB WHO DID ALL
THIS WORK THAT I DESCRIBED HERE
THIS AFTERNOON.
I WOULD LIKE TO ACKNOWLEDGE MY
COLLEAGUES AT THE EYE INSTITUTE,
PARTICULARLY THE ONE WHO
RECRUITED ME AS A NEWLY MINTED
Ph.D. TO WORK WITH HIM.
I'VE BEEN HERE IN THE NEI ON THE
NIH SINCE THEN.
THIS IS THE ONLY JOB I'VE HAD AS
AN ADULT.
AND I LOVE WORKING AT NIH.
AND MY COLLEAGUES, RACHEL AND
CHI-CHAO-CHAN, COLLABORATORS AND
FRIENDS, WE PUBLISHED SEVERAL
PAPERS TOGETHER, AND HAVE
ENJOYED WORKING WITH THEM ALL
THESE YEARS AT NIH.
ROBERT NUSSENBLATT WAS AT BRANCH
LAB CHIEF WHO DIED A FEW YEARS
AGO, WE STILL MISS HIM.
YINGYOU AND MARY HAVE BEEN --
WE'VE COLLABORATED ON MANY
MANUSCRIPTS, THEY PLAY IMPORTANT
ROLE IN SOME OF OUR ANIMAL MODEL
MAL STUDIES.
RAFAEL IS THE HEAD OF THE FLOW
CYTOMETRY SCORE AND HE'S BEEN AN
INVALUABLE MEMBER AND HELPED US
IN MANY OF OUR LYMPHOCYTE
STUDIES.
LIJIN AND GUANPUA PLAYED
IMPORTANT ROLE TO GENERATE MICE
WITH TARGETED DELETION OF IL-27
RECEPTOR FROM B CELLS.
AND MONIKA AND YURI, THEY PLAYED
SEMINAL IMPORTANT ROLE IN HPLC
PURIFICATION OF THE IL-35
CYTOKINE.
ROBERT AND MONES, THEY HELPED US
A LOT WITH THE CONFOCAL
MICROSCOPY AND THE E.M. IMAGE
WAS GENERATED BY MONIS.
PAUL WINGFIELD HELPED US WITH
CHARACTERIZING THE HETERODIMERIC
IL-35 TO DEMONSTRATE IT'S
HETERODIMERIC PROTEIN.
LAST BUT NOT LEAST, MICHELE K
EVANS, SENIOR INVESTIGATOR AT
NATIONAL AGING INSTITUTE, WHO
HAPPENED TO BE MY WIFE.
SHE'S THE ONE WHO INTRODUCED ME
TO THE WONDERFUL FIELD OF
EXOSOMES.
AND THIS OCCURRED ONE EVENING
DURING DINNER, COMPLAINING ABOUT
THE IL-27, IL-35 BREGS WITH
SUBUNITS, THERAPEUTICALLY
DIFFICULT TO BE ABLE TO PREDICT
HOW MUCH OF THEM FORM
HETERODIMER.
AND IN VIVO.
THE THERAPY.
SHE ASKED HAVE YOU TRIED TO MAKE
EXOSOMES?
THEY MAY SIMPLIFY YOUR LIFE.
JUST DOING THAT EXPERIMENT
THAT'S HOW COME WE CAME TO FIND
OUT THEY MAKE EXOSOMES AND IT'S
OPENED A WONDERFUL AREA OF
RESEARCH IN MY LAB.
THANK YOU ALL VERY, VERY MUCH
FOR LISTENING.
AND I'LL BE HAPPY TO TAKE ANY
QUESTIONS IF WE HAVE TIME.
>> THANK YOU, DR. EGWUAGU, FOR
SUCH A WONDERFUL TALK ON THE
STORY OF THE IL-35 AND IL-27 AND
EXOSOMES.
THIS IS FANTASTIC.
WE DO HAVE SOME QUESTIONS.
LET ME START FIRST.
I GUESS THE IL-35 STORY THAT YOU
STARTED OUT WITH, YOU KNOW, VERY
INTRIGUING.
DO YOU THINK THAT THERAPIES
COULD BE USED, USING IL-35 AND
TOWARDS OTHER DISEASES LIKE
MACULAR DEGENERATION OR DIABETIC
RETINOPATHY, OTHER TYPES OF
DISEASES?
>> YES, I DO.
THEY CAN BE APPLIED TO MOST
INFLAMMATORY DISEASES.
HOWEVER, MY OUR PREDICTION IS
THAT WE MAY BE BETTER OFF USING
EXOSOMES BECAUSE THEY HAVE MORE
ACCESS.
THEY CAN EASILY ENTER DIFFERENT
TISSUES, ESPECIALLY FOR LIKE
MACULAR DEGENERATION, YOU
COULD -- THEY HAVE ACCESS TO THE
CNS AND MUCH MORE USEFUL.
BASICALLY IS THE ACTIVE CYTOKINE
ITSELF, SO THE BREGS OR EXOSOMS
ON VECTORS.
I BELIEVE THEY CAN BE APPLICABLE
TO MANY INFLAMMATORY DISEASE.
HOWEVER, THEY SHOULD BE
INHIBITED IN CANCER, BECAUSE
THEY WOULD SUPPRESS CANCER
IMMUNOTHERAPY.
>> VERY INTERESTING.
THANK YOU.
WE HAVE A QUESTION THAT WAS
E-MAILED FROM ONE OF OUR NHLBI
COLLEAGUES.
COULD I 27, I 35 B CELL
TREATMENTS BE HELPFUL IN
TREATING DANGEROUS CYTOKINE
STORMS WITH CAR T CELL TREATMENT
IN THE FUTURE?
>> I DON'T SEE WHY NOT.
BASICALLY WHAT IT DOES IS THAT
IT'S ABLE TO INHIBIT LYMPHOCYTES
THAT MAKE THOSE CYTOKINES THAT
INDUCE, AND IT'S ACROSS THE
BOARD.
SO THAT IT'S NOT JUST LIKE ONE
CYTOKINE INHIBITS A REPERTOIRE
OF CYTOKINES, WHETHER IT'S IL-1,
INTERFERON GAMMA, LOOKED AT TYPE
I INTERFERONS BUT INTERFERON
GAMMA AND TNF AND ALL THE OTHER
CYTOKINES, YES, THEY CAN INHIBIT
THEM.
YES.
>> GREAT.
THANK YOU.
I HAVE ANOTHER QUESTION.
YOU DESCRIBED A LOT TO US TODAY
RELATED TO THE I 35 SORT OF
STORY, IF YOU WILL.
IS THERE MORE FOR THE IL-27
STORY THAT MAY BE FORTHCOMING?
OR DID ONE PATH JUST LEAD MORE
TOWARDS THE IL-35?
>> AS A MATTER OF FACT, THE
IL-27 IS MORE INTRIGUING BECAUSE
IT'S AN INNATE-LIKE LYMPHOCYTE,
AND WHILE IL-27 CAN INDUCE IL-35
AND IL-10, IL-35 CANNOT INDUCE
IL-27.
OKAY.
SO IT HAS A MUCH MORE ROBUST
ACTIVITY.
THE OTHER THING I DIDN'T POINT
OUT, A LOT OF BREGS, THEY ARE
SPECIFIC TO PARTICULAR DISEASES.
IL-27 CAN BE ELICITED JUST USING
LPS.
SO THE IL-27 WE GENERATE TO
INHIBIT EAE WE COULD USE IT TO
INHIBIT EAU.
SO, IT HAS A MUCH MORE GLOBAL
IMPACT.
AND PART OF IT IS BECAUSE IT'S
REALLY AN INNATE-LIKE CELL,
MEANWHILE THE IL-35 BREGS IS
ADAPTED BRREG REALIZE MORE RICH,
YOU COULD USE IT TO INHIBIT
DIVERSE AUTOIMMUNE DISEASES.
>>THANK YOU FOR SUCH A
WONDERFUL TALK AND TAKING TIME
TO ANSWER QUESTIONS FROM THE
AUDIENCE.
WE'RE AT 1:00.
THIS WILL CONCLUDE OUR SESSION
FOR GRAND ROUNDS.
THANK YOU VERY MUCH.
AND THANK YOU TO THE AUDIENCE
FOR YOUR LISTENING AND YOUR
PARTICIPATION.