de groot nova se immunology of vaccines2009
TRANSCRIPT
Brown University Brown University University of Rhode IslandUniversity of Rhode Islandand EpiVax, Providence RIand EpiVax, Providence RI
January 2009January 2009
Annie De Groot MDAnnie De Groot MD
Immunology of VaccinesImmunology of Vaccines
OutlineOutline
Vaccine Research and DevelopmentVaccine Research and DevelopmentImmunology of VaccinesImmunology of VaccinesSome Case Studies:Some Case Studies:
tetanustetanussmallpoxsmallpoxpoliopolio
Emerging infectious diseasesEmerging infectious diseasesVaccine technology nowVaccine technology now
Research and DevelopmentResearch and DevelopmentLeads to Vaccine ProductionLeads to Vaccine Production
• It currently costs between 200 and 500 million US dollars to bring a new vaccine from the concept stage to market
• But incentives are present:
The vaccine market has increased fivefold from 1990 to 2000
Annual sales of 8 billion dollars
Less than 2% of the total pharma market but projected to increase
• Major producers (85% of the market)
• GlaxoSmithKline (GSK), Merck, Aventis Pasteur, Wyeth, Novartis
• Main products (>50% of the market)
• Hepatitis B, flu, MMR (measles, mumps, and rubella) and DTP (diphtheria, tetanus, pertussis)
• 40% are produced in the United States and the rest is evenly split between Europe and the rest of the world
Drug Development ProcessDrug Development Process
Laboratory R+DLaboratory R+D
Pre-IND - Safety/Toxicity -> IND filingPre-IND - Safety/Toxicity -> IND filing
Phase I - human safety/toxicityPhase I - human safety/toxicity
Phase II - efficacyPhase II - efficacy
Phase III - extended studies / other drug(s)Phase III - extended studies / other drug(s)
NDA -> FDA ApprovalNDA -> FDA Approval
Post-licensure surveillancePost-licensure surveillance
2008 Success!: 2008 Success!:
New HPV New HPV (Cervical Cancer) Vaccine (Cervical Cancer) Vaccine
almost 100% effective!almost 100% effective!
2007 Failure: 2007 Failure:
• Merck Ad5 HIV VaccineMerck Ad5 HIV Vaccine
Vaccines are Still Big!!Vaccines are Still Big!!Recent Vaccine R and D NewsRecent Vaccine R and D News
November 16, 2007 - - Pfizer buys Coley for $164MNovember 16, 2007 - - Pfizer buys Coley for $164MAugust 15, 2008 - - Pfizer inks vaccine pact with Cytos August 15, 2008 - - Pfizer inks vaccine pact with Cytos October 3, 2008 - - Crucell wins $70M to develop new vaccinesOctober 3, 2008 - - Crucell wins $70M to develop new vaccinesDecember 29, 2008 - - Novartis pays $20M for CMV program (AlphaVax)December 29, 2008 - - Novartis pays $20M for CMV program (AlphaVax)January 7, 2009January 7, 2009 - - Wyeth in talks to buy vaccine maker Crucell- - Wyeth in talks to buy vaccine maker CrucellJanuary 8, 2009 - - GSK unveils $300M vaccines plantJanuary 8, 2009 - - GSK unveils $300M vaccines plant. . . . . .
Why Is Vaccine Research and Why Is Vaccine Research and Development Important?Development Important?
• Immunization saves the lives of Immunization saves the lives of 3 million children3 million children each each year. year.
• 2 million more children could be saved2 million more children could be saved if existing vaccines if existing vaccines were applied on a full-scale worldwidewere applied on a full-scale worldwide
• Vaccines have been made for Vaccines have been made for only 34 of the more than 400 only 34 of the more than 400 known pathogens that are harmful to humans.known pathogens that are harmful to humans.
• What is not said: The small molecule R and D is not What is not said: The small molecule R and D is not producing many new drugs – vaccines are seen as a producing many new drugs – vaccines are seen as a “Pipeline solution”.“Pipeline solution”.
OutlineOutline
Vaccine Research and DevelopmentVaccine Research and DevelopmentImmunology of VaccinesImmunology of VaccinesSome Case Studies:Some Case Studies:
tetanustetanussmallpoxsmallpoxpoliopolio
Emerging infectious diseasesEmerging infectious diseasesVaccine technology nowVaccine technology now
What does a vaccine do?What does a vaccine do?
. . . Trains the immune system to recognize . . . Trains the immune system to recognize and fight infectionand fight infection
. . . Without requiring exposure to the . . . Without requiring exposure to the pathogenpathogen
10 Steps to Making A Vaccine 10 Steps to Making A Vaccine
1.1. Define Disease (Chickenpox vs Smallpox)Define Disease (Chickenpox vs Smallpox)
2.2. Define Pathogen (Virus vs Parasite)Define Pathogen (Virus vs Parasite)
3.3. Is there Immunity (If not you are in trouble)Is there Immunity (If not you are in trouble)
4.4. Correlates of Immunity one or many? (Ab? Ag? Correlates of Immunity one or many? (Ab? Ag? CMI?)CMI?)
5.5. Critical Antigens - one or many?Critical Antigens - one or many?
6.6. Animal Model? Does it predict protection?Animal Model? Does it predict protection?
7.7. Prototype Vaccine - Preclinical ProofPrototype Vaccine - Preclinical Proof
8.8. Safety and Toxicity, GMP, StabilitySafety and Toxicity, GMP, Stability
9.9. FDA “IND” ApprovalFDA “IND” Approval
10.10. Post Clinical PhasePost Clinical Phase
• Clinical trials (Phase I, II, III)Clinical trials (Phase I, II, III)
• Approval (and indication)Approval (and indication)
• Distribution / Acceptance / AccessDistribution / Acceptance / Access
3.3. Is there Immunity (If not you are in trouble)Is there Immunity (If not you are in trouble)
4. 4. Correlates of Immunity one or many? (Ab? Ag? Correlates of Immunity one or many? (Ab? Ag? CMI?)CMI?)
Basic Principles of Vaccine Immunology
Innate immunityInnate immunity (e.g., macrophages, (e.g., macrophages, neutrophils, certain molecules) is the first line neutrophils, certain molecules) is the first line of defense. It is fast (usually good-to-go) and of defense. It is fast (usually good-to-go) and usually effective.usually effective.
Adaptive immunityAdaptive immunity (mediated by B and T (mediated by B and T cells) can be slow to respond (several days). cells) can be slow to respond (several days). It is highly effective when the innate immune It is highly effective when the innate immune system cannot fully deal with the threat.system cannot fully deal with the threat.
Primary response (primary immunization) is relatively:Primary response (primary immunization) is relatively:Primary response (primary immunization) is relatively:Primary response (primary immunization) is relatively:
Secondary response (secondary immunization or booster Secondary response (secondary immunization or booster immunization) is relatively:immunization) is relatively:Secondary response (secondary immunization or booster Secondary response (secondary immunization or booster immunization) is relatively:immunization) is relatively:
slow (4-7days)small amount of antibody (low concentration of antibody)low affinity antibodyIgM first, IgG second (equal amounts of IgM and IgG)
slow (4-7days)small amount of antibody (low concentration of antibody)low affinity antibodyIgM first, IgG second (equal amounts of IgM and IgG)
fast (2-4 day)large amounts of antibodyhigh affinity antibodymostly IgG
fast (2-4 day)large amounts of antibodyhigh affinity antibodymostly IgG
Initial SARS Strain
2nd SARS Strain
Initial SARS Strain
Often, a secondary (memory) response is so fast and effective in removing antigens (pathogens), there are few or no symptoms detected by the infected individual (protective immunity).
Secondary responses are the reason we do not get certain infectious diseases more than once.
Secondary responses also explain why vaccinations work. For vaccinations, instead of immunizing with something that makes you sick, a vaccine contains antigens prime the immune response.
Vaccine strategies: Vaccine strategies: B cells need T helpB cells need T help
Macrophages / APCMacrophages / APC
Two types of T cells: Th and CTLTwo types of T cells: Th and CTL
Th cells provide help to B cellsTh cells provide help to B cells
CTL cells kill virus infected cellsCTL cells kill virus infected cells
Categories of VaccinesCategories of Vaccines
Live attenuatedLive attenuatedWhole KilledWhole KilledSubunitSubunitEpitope-basedEpitope-based
Categories of VaccinesCategories of Vaccines
•Live vaccines
•Are able to replicate in the host
•Attenuated (weakened) so they do not cause disease
•Subunit vaccines
•Part of organism
•Genetic Vaccines
•Part of genes from organism
•Epitope-based vaccines
•Minimal essential information with least cross-reactive material
Live VaccinesLive Vaccines
•Characteristics
•Able to replicate in the host
•Attenuated (weakened) so they do not cause disease
•Advantages
•Induce a broad immune response (cellular and humoral)
•Low doses of vaccine are normally sufficient
•Long-lasting protection are often induced
•Disadvantages
•May cause adverse reactions
•May be transmitted from person to person
Subunit VaccinesSubunit Vaccines
•Relatively easy to produce (not live)
•Induce little anti-viral T cell response (CTL)
•Viral and bacterial proteins are not produced within cells
•Classically produced by inactivating a whole virus or bacterium
•Heat
•Chemicals
•The vaccine may be purified
•Selecting one or a few proteins which confer protection
• Example: HPV Vaccine created from two HPV proteins
• A self-assembling “particle” made of purified protein that is free from whole microorganism cells
Subunit Vaccines: PolysaccharidesSubunit Vaccines: Polysaccharides
•Polysaccharides •Many bacteria have polysaccharides in their outer membrane
•Polysaccharide based vaccines
•Neisseria meningitidis
•Streptococcus pneumoniae
•Generate a T cell-independent response
•Inefficient in children younger than 2 years old
•Overcome by conjugating the polysaccharides to peptides
•This approach used in vaccines against Streptococcus pneumoniae and Haemophilus influenzae.
Subunit Vaccines: ToxoidsSubunit Vaccines: Toxoids•Toxins
•Responsible for the pathogenesis of many bacteria
•Toxoids
•Inactivated toxins
•Toxoid based vaccines
•Bordetella pertussis
•Clostridium tetani
•Corynebacterium diphtheriae
•Inactivation
•Traditionally done by chemical means
•Altering the DNA sequences important to toxicity
Subunit Vaccines: Recombinant Subunit Vaccines: Recombinant •The hepatitis B virus (HBV) vaccine
•Originally based on the surface antigen purified from the blood of chronically infected individuals.
•Due to safety concerns, the HBV vaccine became the first to be produced using recombinant DNA technology (1986)
•Produced in bakers’ yeast (Saccharomyces cerevisiae)
•Virus-like particles (VLPs)
•Viral proteins that self-assemble to particles with the same size as the native virus.
•VLP is the basis of a promising new vaccine against human papilloma virus (HPV)
•Merck
•In phase III
Genetic VaccinesGenetic Vaccines•Introduce DNA or RNA into the host
•Injected (Naked)
•Coated on gold particles
•Carried by viruses
•vaccinia, adenovirus, or alphaviruses
•bacteria such as
•Salmonella typhi, Mycobacterium tuberculosis
•Advantages
•Easy to produce
•Induce cellular response
•Disadvantages
•Low response in 1st generation
Epitope based vaccinesEpitope based vaccines•Advantages (Ishioka et al. [1999]):
•Can be more potent
•Can be controlled better
•Can induce response to a broad range of proteins and subdominant eptiopes (e.g. against tumor antigens where there is tolerance against dominant epitopes)
•Can target multiple conserved epitopes in rapidly mutating pathogens like HIV and Hepatitis C virus (HCV)
•Can be designed to break tolerance
•Can overcome safety concerns associated with entire organisms or proteins
•Epitope-based vaccines have been shown to confer protection in animal models ([Snyder et al., 2004], Rodriguez et al. [1998] and Sette and Sidney [1999]) and De Groot (in Press).
Therapeutic vaccinesTherapeutic vaccines• Vaccines to treat the patients that already have a disease
• Targets
• Tumors
• AIDS
• Allergies
• Autoimmune diseases
• Hepatitis B
• Tuberculosis
• Malaria
• Helicobacter pylori
• Concept
• suppress/boost existing immunity or induce immune responses.
Cancer vaccinesCancer vaccines• Break the tolerance of the immune system against tumors
• 3 types
1. Whole tumor cells, peptides derived from tumor cells in vitro, or heat shock proteins prepared from autologous tumor cells
2. Tumor-specific antigen–defined vaccines
3. Vaccines aiming to increase the amount of dendritic cells (DCs) that can initiate a long-lasting T cell response against tumors.
• Therapeutic cancer vaccines can induce antitumor immune responses in humans with cancer
• Antigenic variation is a major problem that therapeutic vaccines against cancer face
• Tools from genomics and bioinformatics may circumvent these problems
Allergy vaccinesAllergy vaccines
• Increasing occurrence of allergies in industrialized countriesIncreasing occurrence of allergies in industrialized countries
• The traditional approach is to vaccinate with small doses of The traditional approach is to vaccinate with small doses of purified allergenpurified allergen
• Second-generation vaccines are under development based on Second-generation vaccines are under development based on recombinant technologyrecombinant technology
• Genetically engineered Bet v 1 vaccine can reduce pollen-Genetically engineered Bet v 1 vaccine can reduce pollen-specific IgE memory response significantlyspecific IgE memory response significantly
• Example of switching a “wrong” immune response to a less Example of switching a “wrong” immune response to a less harmful one.harmful one.
OutlineOutline
Vaccine Research and DevelopmentVaccine Research and DevelopmentImmunology of VaccinesImmunology of VaccinesSome Case Studies:Some Case Studies:
tetanustetanussmallpoxsmallpoxpoliopolio
Emerging infectious diseasesEmerging infectious diseasesVaccine technology nowVaccine technology now
TETANUSTETANUS
SMALLPOXSMALLPOX
POLIOPOLIO
Wild Poliovirus, 1988
Wild Poliovirus, 2004
Progress in Polio EradicationNew Polio Cases linked to Nigerian Boycott, 2005
10 steps to making a vaccine
1.1. PathogenPathogen2.2. Correlates of immunityCorrelates of immunity3.3. Critical antigensCritical antigens4.4. Animal modelAnimal model5.5. Delivery methodDelivery method6.6. Preclinical confirmationPreclinical confirmation7.7. FDA Approval FDA Approval 8.8. Clinical TrialClinical Trial9.9. DistributionDistribution10.10.AcceptanceAcceptance
Wild Poliovirus, 2006Wild Poliovirus, 2006
Wild Poliovirus, 2007Wild Poliovirus, 2007
Can we Eradicate Polio?Can we Eradicate Polio?
OutlineOutline
Vaccine Research and DevelopmentVaccine Research and DevelopmentImmunology of VaccinesImmunology of VaccinesSome Case Studies:Some Case Studies:
tetanustetanussmallpoxsmallpoxpoliopolio
Emerging infectious diseasesEmerging infectious diseasesVaccine technology nowVaccine technology now
EMERGING INFECTIOUS EMERGING INFECTIOUS DISEASES SINCE 1990DISEASES SINCE 1990
• 1993 (US) - Hantavirus pulmonary syndrome (Sin nombre virus)1993 (US) - Hantavirus pulmonary syndrome (Sin nombre virus)• 1994 (US) – Human granulocyte ehrlichiosis1994 (US) – Human granulocyte ehrlichiosis• 1995 (Worldwide) - Kaposi sarcoma (HHV-8)1995 (Worldwide) - Kaposi sarcoma (HHV-8)• 1995 (US) – Cyclosporiasis from raspberries1995 (US) – Cyclosporiasis from raspberries• 1996 (England) – Variant Creutzfeld-Jakob disease (vCJD)1996 (England) – Variant Creutzfeld-Jakob disease (vCJD)• 1997 (Japan) – Vancomycin-intermediate 1997 (Japan) – Vancomycin-intermediate S. aureusS. aureus• 1998 (Malaysia) – Nipah virus 1998 (Malaysia) – Nipah virus • 1999 (US) - West Nile encephalitis (West Nile virus)1999 (US) - West Nile encephalitis (West Nile virus)• 2001 (US) - Anthrax attack via letters2001 (US) - Anthrax attack via letters• 2001 (Netherlands) – Human metapneumovirus2001 (Netherlands) – Human metapneumovirus• 2002 (US) – Vancomycin-resistant 2002 (US) – Vancomycin-resistant S. aureusS. aureus• 2003 (China 2003 (China worldwide) - Severe acute respiratory syndrome worldwide) - Severe acute respiratory syndrome
(coronavirus)(coronavirus)• 2003 (US) - Monkeypox2003 (US) - Monkeypox
What’s next? What’s next?
Emerging Diseases Worst Case ScenarioEmerging Diseases Worst Case Scenario What are the critical elementsWhat are the critical elements
• Highly infectious pathogenHighly infectious pathogen• Circumstances that permit transmissionCircumstances that permit transmission
– CrowdingCrowding– TravelTravel– VectorsVectors
• Lack of preparednessLack of preparedness• Lack of treatmentLack of treatment• Lack of vaccineLack of vaccine
EMERGING INFECTIOUS EMERGING INFECTIOUS DISEASES SINCE 1990DISEASES SINCE 1990
• 1993 (US) - Hantavirus pulmonary syndrome (Sin nombre virus)1993 (US) - Hantavirus pulmonary syndrome (Sin nombre virus)• 1994 (US) – Human granulocyte ehrlichiosis1994 (US) – Human granulocyte ehrlichiosis• 1995 (Worldwide) - Kaposi sarcoma (HHV-8)1995 (Worldwide) - Kaposi sarcoma (HHV-8)• 1995 (US) – Cyclosporiasis from raspberries1995 (US) – Cyclosporiasis from raspberries• 1996 (England) – Variant Creutzfeld-Jakob disease (vCJD)1996 (England) – Variant Creutzfeld-Jakob disease (vCJD)• 1997 (Japan) – Vancomycin-intermediate 1997 (Japan) – Vancomycin-intermediate S. aureusS. aureus• 1998 (Malaysia) – Nipah virus 1998 (Malaysia) – Nipah virus • 1999 (US) - West Nile encephalitis (West Nile virus)1999 (US) - West Nile encephalitis (West Nile virus)• 2001 (US) - Anthrax attack via letters2001 (US) - Anthrax attack via letters• 2001 (Netherlands) – Human metapneumovirus2001 (Netherlands) – Human metapneumovirus• 2002 (US) – Vancomycin-resistant 2002 (US) – Vancomycin-resistant S. aureusS. aureus• 2003 (China 2003 (China worldwide) - Severe acute respiratory syndrome worldwide) - Severe acute respiratory syndrome
(coronavirus)(coronavirus)• 2003 (US) - Monkeypox2003 (US) - Monkeypox
• 2004 (Asia) – Avian influenza (H5N1)2004 (Asia) – Avian influenza (H5N1)
The FLUThe FLU
OutlineOutline
Vaccine Research and DevelopmentVaccine Research and DevelopmentImmunology of VaccinesImmunology of VaccinesSome Case Studies:Some Case Studies:
tetanustetanussmallpoxsmallpoxpoliopolio
Emerging infectious diseasesEmerging infectious diseasesVaccine technology nowVaccine technology now
The Old Way of Making VaccinesThe Old Way of Making Vaccines
shake and bakeshake and bake
The New Way of Making VaccinesThe New Way of Making Vaccines
The Even Newer WayThe Even Newer Way
In vitro screeningIn vitro screening
epitopeepitope
BioinformaticsBioinformatics
Less than the entire pathogen is requiredLess than the entire pathogen is required
Hepatitis VirusHepatitis Virusor or
VaccineVaccine
Epitope Subset = ImmunomeEpitope Subset = Immunome
Immune systemImmune system‘‘filter’filter’
T cell epitope T cell epitope At Intersection of Immune ResponseAt Intersection of Immune Response
EpiVax: Accelerating EpiVax: Accelerating Vaccines and Biologics Vaccines and Biologics
Research and DevelopmentResearch and Development
EpiVax: Accelerating EpiVax: Accelerating Vaccines and Biologics Vaccines and Biologics
Research and DevelopmentResearch and Development
Anne S. De GrootAnne S. De Groot1, 2,3,1, 2,3, L.Moise L.Moise1, 31, 3, J.A. McMurry, J.A. McMurry11, W. Yang, W. Yang11, William Martin, William Martin11
11EpiVax, Inc. EpiVax, Inc. 22Brown Medical School and Brown Medical School and 33University of Rhode IslandUniversity of Rhode Island
[email protected]@EpiVax.com http://www.EpiVax.com http://www.EpiVax.com
We think about what a vaccine does. . . We think about what a vaccine does. . .
. . . Trains the immune system to recognize . . . Trains the immune system to recognize and fight infectionand fight infection
. . . Without requiring exposure to the . . . Without requiring exposure to the pathogenpathogen
using “epitopes” = chains of amino acidsusing “epitopes” = chains of amino acids
Current Vaccine–Related NIH Funding Current Vaccine–Related NIH Funding
1R43AI058376 "A novel Smallpox Vaccine Derived from the VV/VAR Immunome“
1R43AI065036 "A Genome-Derived, Epitope-Driven H pylori Vaccine“
1R43AI058326 "A Genome-Derived, Epitope-Driven Tularemia Vaccine"
1R43AI075830-01“Optimization of a Multivalent Tuberculosis Vaccine”
7R01AI050528 (new R21: Optimization of HIV Vaccine Delivery) Epitope Driven HIV Vaccine Development
Unfunded: Influenza, HPV, EBV
In Silico EpiMatrix / ClustiMer / OptiMatrix [class I and class II alleles]
Conservatrix / BlastiMer/. EpiAssembler/ VaccineCAD
In Vitro HLA binding assay
ELISpot - ELISA - Multiplex ELISA - FACS - T regulatory T cell profiling
In Vector DNA prime/peptide (pseudoprotein boost) vaccines
Vaccine delivery / formulation optimization / detolerizing delivery agents
In Vivo HLA DR3, DR4 transgenic mice
HLA class I transgenic mice
Vaccination, Comparative studies
EpiVax Genome-derived, epitope-driven vaccine approach:
Prime-boost Smallpox Vaccine
Immunization Sacrifice Birth
1. epitope DNA vaccine prime2. epitope peptide boost
1. control DNA prime2. control peptide boost
Week 0 Week 8-14
IFN
-gam
ma
and
mu
ltip
lex
EL
ISA
Challenge
Let
hal
In
trat
ran
asal
Ch
alle
ng
e
3 mice week 16 Week 18
Results: 100% survival of Vaccinated mice vs. 17% of placebo
100%
0 20 40 60 80 100
100%
No significant weight loss in vaccinated mice – surviving mice in placebo arm are regaining weight
HIV Vaccine DevelopmentHIV Vaccine Development
The GAIA HIV Vaccine
• In Development since 1998 - More than 300 epitopes mapped• Highly Variable Pathogen – Conserved epitopes• HLA Diversity -- 6 HLA supertypes• T cell help -- Immunogenic consensus sequence epitopes• Validation in HLA transgenic mice -- Good progress.
Better Vaccines and Health for AllBetter Vaccines and Health for All
Our Hope for the FutureOur Hope for the Future
Fearless ScienceFearless Science
QUESTIONSQUESTIONS
EpiVax: Science without Fear/ Fearless EpiVax: Science without Fear/ Fearless ScienceScience