history and evolution of vaccine technologies · yeast dna antigen dna strugnell r et al. chapter 3...
TRANSCRIPT
History and Evolution of Vaccine Technologies Leonard Friedland, MD, FAAPVice President and Director, Scientific Affairs and Public HealthGSK Vaccines
Immunize Nebraska
August 7, 2020
Disclosure
Employed by GSK where I am a vaccine research physician scientist
Presentation at the invitation of Dr. Meera Varman, Professor, Pediatric Infectious Diseases, Creighton University School of Medicine
Presentation is for educational purposes only; this is not a sales, marketing or promotional presentation
Content of presentation will not include unapproved or investigational uses of products or devices
1. CDC. List of Vaccines Used in United States. https://www.cdc.gov/vaccines/vpd/vaccines-list.html. Accessed Jan 2017.2. WHO. Vaccines and Diseases. http://www.who.int/immunization/diseases/en/. Accessed Jan 2017.3. CDC Achievements in Public Health, 1900-1999 Impact of Vaccines Universally Recommended for Children – United States, 1990-1998.https://www.cdc.gov/mmwr/preview/mmwrhtml/00056803.htm. Accessed Jan 2017.
Globally, Many Diseases are Currently Preventable by Vaccination
3
“Vaccines are one of the greatest achievements of biomedical science and public health”3
Global public health1 Regional focus2
Adenovirus AnthraxCholeraDengueEbolaJapanese encephalitisTick-borne encephalitisTyphoid feverRabiesYellow fever
DiphtheriaHaemophilus influenzae type b
Hepatitis AHepatitis BHerpes zosterHuman papillomavirusInfluenzaMeaslesMeningococcal A, B, C, W, YMumps
H5N1 fluPertussisPoliomyelitisPneumococcalRotavirusSmallpox and vacciniaTetanusTuberculosisVaricella
Vaccine Science: Two Centuries of Continuous Research, Improvements, and Achievements
4R. Thom, ‘Jenner: Smallpox is Stemmed, from "The History of Medicine,”’ Collection of the University of Michigan Health System, Gift of Pfizer.Bonanni P, et al. Chapter 1 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
The Science of Immunology Began in the 19th Century
5
Microorganisms were identified as the true cause of infectious diseases
Host cells that ingest and destroy invading microbes were discovered and called ‘phagocytes’
Passive immunotherapy of diphtheria with anti-diphtheria toxin antibodies discovered ‘immune serum’
The antibodies theory was formulated
The concept of ‘natural immunity’ to infection was born
Courtesy of CDC/Janice Haney Carr/Jeff Hageman M.H.S.
Bonanni P, et al. Chapter 1 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
New Technologies Make Vaccines PossibleThat Were Previously Impossible
6
Finco O, Rappuoli R. Front Immunol. 2014;5:1-6.
Reverse Vaccinology
Empirical Approach GlycoconjugationRecombinant
DNA
MenB, GBS, GAS, E. coli, S. aureus,
C. difficile
Diphtheria, Tetanus, Pertussis, Rabies,
Influenza, Smallpox, Polio, BCG
MenACWY, Pneumo,
Hib, GBS, S. aureus
Hepatitis B, Acellular Pertussis,
Lyme, Human papillomavirus
20101930 19901980
In the 1930s, Max Theiler at Rockefeller Foundation used an egg system for the development of an effective yellow fever vaccine, for which he was awarded the
Nobel Prize in Medicine in 1951
Enders, Weller and Robbins at Harvard received the Nobel Prize in Medicine
in 1954 for demonstrating the ability of polio viruses to grow in cell cultures
Cell culture (1950s)Embryonated eggs (1930s)
Courtesy of CDC
Growing Viruses
7
1930s–1950s
Bonanni P, et al. Chapter 1 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
Virus Vaccines
APC, antigen-presenting cell.Callaway E. Nature. 2020; 580(7805): 576–577.
Live-attenuated virusViruses are weakened by being passed through animal or human cells until they gain mutations that limit their ability to cause disease
Cell
Inactivated virusVirus is inactivated using chemicals or heatVirus replicates
APC
Live-attenuated, combined MMR vaccine developed in order to minimize the total number of injections in infantsClinical trial data demonstrate a combined antigen vaccine can be effective and can have an acceptable safety profile
MMR, measles-mumps-rubella
Measles Mumps Rubella
Bonanni P et al. Chapter 1 in: Garçon et al. Understanding Modern Vaccines, Perspectives in Vaccinology, Vol 1, Amsterdam, Elsevier, 2011.
1970s Combination Vaccines
Courtesy of CDC/Cynthia S. Goldsmith, William Bellini, PhD
Courtesy of CDC/Dr Fred Murphy Courtesy of CDC/Dr Erskine Palmer
1970s–1980s
Subunit/purified vaccineSplit vaccine
Whole virus vaccine
Pathogen fragmentation
eg, some influenza vaccines
eg, acellular pertussis components: PT, PRN, FHA, FIM; HA in some
influenza vaccines
– Antigen choice: provides immune protection & technologically achievable
– Often reduced immunogenicity versus whole pathogen– Non-infectious, low reactogenicity, acceptable
tolerability– No or limited availability of innate defensive triggers– For subunit vaccines with lower immunogenicity,
adjuvants often needed to compensate– Facilitate supply via synthetic production versus whole
pathogen
Split Pathogen and Subunit Vaccines
10
Strugnell R, et al. Chapter 3 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
New Technologies Make Previously Impossible Vaccines a Reality
11
Reverse Vaccinology
Empirical Approach GlycoconjugationRecombinant
DNA
MenB, GBS, GAS, E. coli, S. aureus,
C. difficile
Diphtheria, Tetanus, Pertussis, Rabies,
Influenza, Smallpox, Polio, BCG
MenACWY, Pneumo,
Hib, GBS, S. aureus
Hepatitis B, Acellular Pertussis,
Lyme, Human papillomavirus
20101930 19901980
Finco O, Rappuoli R. Front Immunol. 2014;5:1-6.
Recombinant Protein Vaccines: HBV
12
Sequence gene encoding HBsAg
Insert gene into yeast expression system
Protein expression
Purification of recombinant protein encoding antigen
(natural assembly into spheres)
Administration as vaccine
Hepatitis B surface antigen
HBsAg= hepatitis B surface antigen; HBV= hepatitis B virus.Strugnell R, et al. Chapter 3 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
1980s
Yeast DNA
Antigen DNA
Strugnell R et al. Chapter 3 in: Garçon et al. Understanding Modern Vaccines, Perspectives in Vaccinology, Vol 1, Amsterdam, Elsevier, 2011.
Elicit immune
response
DNA encoding L1
Transcription Translation
Self-assembly into viral capsids (VLPs)
HPV viral capsid
HPV, human papillomavirus; VLP, virus-like particle
Self-assembly
into pentamers
Purification from
expression system
L1 capsid proteins
1990s Recombinant Protein Vaccines: HPV
Strugnell R et al. Chapter 3 in: Garçon et al. Understanding Modern Vaccines, Perspectives in Vaccinology, Vol 1, Amsterdam, Elsevier, 2011.
B-ce
ll +
antib
ody
quan
tity
Polysaccharide-conjugate vaccine
Polysaccharide vaccine
T-ce
ll qu
antit
y
Polysaccharide-conjugate vaccine
No T-cell response to polysaccharide alone
Antibody, no memory
B-cell
T-cellAntibody
and memory
Polysaccharide-conjugate vaccine
Polysaccharide vaccine
Conjugated protein
1980s-1990s Polysaccharide-conjugate vaccines
Rino Rappuoli, Matthew Bottomley, Ugo D’Oro, Oretta Finco, Ennio De Gregorio J Exp Med 2016:213;469-481
Classical Vaccinology
15
Rappuoli R, et al. J Exp Med. 2016;213:469-481.
Growing Pathogens Reverse Vaccinologydesign from information
Reverse Vaccinology: Human Immunology Instructs Vaccine Antigen Design
16
2000s
Classical Vaccinologygrowing pathogens
Reverse Vaccinologydesign from information
Rappuoli R, et al. J Exp Med. 2016;213:469-481.
B-cell
Express recombinant
proteins
In silico vaccinecandidates
Vaccine Candidates
Reverse Vaccinology: A Genomic Approach for a Meningococcus B Vaccine1,2
171. Ulmer JB, et al. Nat Biotechnol. 2006;24:1377-1383.2. Serruto D, et al. J Biotechnol. 2004;113:15-32.
600 potential vaccine candidates identified
350 proteins successfully expressedin E. coli
91 novel surface-exposedproteins identified
28 novel proteinshave bactericidal
activity
Vaccines Today: An Explosion of New Technologies
18
Reverse Vaccinology
Empirical Approach GlycoconjugationRecombinant
DNA
MenB, GBS, GAS, E. coli, S. aureus,
C. difficile
Diphtheria, Tetanus, Pertussis, Rabies, Influenza, Smallpox, Polio,
BCG
MenACWY, Pneumo, Hib, GBS, S. aureus
Hepatitis B, Acellular Pertussis,
Lyme, Human papillomavirus
Synthetic Biology/RNA/DNA Vaccines
Adjuvants
Structural Vaccinology
Systems Vaccinology
Next Generation Technologies
20101930 19901980 2015
Finco O, Rappuoli R. Front Immunol. 2014;5:1-6.
Vectors
Increasethe level of the immune response
Challenging populationsdue to impaired immune system (eg, elderly, children, immunocompromised)
Current Challenges for Vaccines1
191. Garçon N, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011; Chapter 4: 89-113.2. Petrovsky N, Aguilar JC. Immunol Cell Biol. 2004;82:488-496.
Need for booster vaccinations
Recombinant antigensgenerally less immunogenic than live or attenuated organism vaccine2
Pathogensthat require broad and complex immune response
Need for antigen sparingpotential supply problems (eg, pandemic flu, SARS-CoV-2)
Prolong the duration of the immune response, improve immune memory, and protection
Overcomea weakened immunogenicity
Induce the generation of a high and broad immune response
Reduce the amount of antigen needed (dose-sparing)
Examples of Novel Approaches to Vaccine Design
20
DNA1 Novel adjuvants and adjuvant combinations3
Live vectors1 RNA2Reverse vaccinology1
1. Stanberry L, Strugnell R. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011; Chapter 6: 155-199.2. Geall A, et al. Semin Immunol. 2013;25:152-159. 3. Garçon N, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011; Chapter 3: 61-88.
• Pathogen-derived genetic material coding for the antigens contained in a non-replicating DNA plasmid
• Antigen is expressed by the cells of the vaccine recipient
• Substances included in a vaccine formulation to enhance the quality and strength of the immune response induced by the vaccine antigen(s)
• Targeted antigens encoded by gene(s) incorporated into the vector’s genetic material
• Antigens expressed by a vector (like virus or bacterium) that is non-pathogenic
• Synthetic virus particles include antigen proteins
• Once inside host cell cytoplasm, these self-amplify in large amounts, express antigen proteins and interact with the host immune system
• Computer analysis of the pathogen’s entire genome is conducted to find genes that may be antigenic
• Vaccine candidate identified based on prediction of protein sequences similar to pathogen’s genome sequences
– From Latin, adiuvare: to aid– Substance included in a vaccine to enhance and modulate the quality and/or strength of
the immune response induced by the antigen– Old technology, made new
Adjuvant1,2
21
Adjuvant
1. Bonanni P, et al. Chapter 5 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.2. Garçon N, et al. Chapter 4 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011.
Adjuvants have a long history in the fight against infectious diseases
22Hib=Haemophilus influenzae type b; HPV=human papillomavirus; IPV=inactivated polio vaccine; OPV=oral polio vaccine (live). *IPV is adjuvanted when formulated in combination with diphtheria, tetanus, pertussis-based vaccines but is not adjuvanted when formulated as a stand-alone vaccine.
1. Figure adapted from Di Pasquale A, et al. Vaccines. 2015;3:320-343. 2. CDC Pink Book 2018. https://www.cdc.gov/vaccines/pubs/pinkbook/appendix/appdx-b.html. Accesed September 2018.
No adjuvantAdjuvanted
RabiesTyphoidCholeraPlague
PertussisTetanusTuberculosis
DiphtheriaYellow fever
InfluenzaPneumococcus
Polio, inactivated*Polio, liveMeasles
MumpsRubellaInfluenza
Meningococcus ACWYHib
Pertussis
Hepatitis BHib
Hepatitis ATyphoid
Varicella
TyphoidMeningococcus
Pneumococcus
InfluenzaRotavirus
Meningococcus ACWY
HPVZoster
Smallpox
Adju
vant
dis
cove
ry Pandemic influenza
Influenza
Diphtheria, tetanus, pertussis-based combinations
1750 1800 1900 1910 1920 1940 1990 20101850 20001930 1950 1960 1970 1980
Zoster Hepatitis B
Meningococcus B InfluenzaMalaria
Rationale for adjuvant use in vaccinesWhy do antigens need help?
Strugnell R et al. Perspect Vaccinol 2011;1:61–88; Garçon N et al. Perspect Vaccinol 2011;1:89–113 (content on slide from pp. 61, 71, 72, 75, 76, 89, 111)
Illustrative figure based on Strugnell R et al. 2011; Garçon N et al. 2011
Imm
unog
enic
ity
Native pathogen
Replicating(live attenuated
pathogen) Non-replicating(whole inactivated
pathogen)Subunit
(toxoids, split virus,fragments of pathogens) Purified antigens
(recombinant proteins)
High
Low High
Adjuvants(increased immunogenicity and
may have increased reactogenicity)
23
Tolerability
Dose reduction and antigen sparing
e.g. Hepatitis B V, Pandemic flu V
Adjuvants are associated with several potential benefits for vaccine development
24
Increased durationof immune response
e.g. HPV, Herpes zoster V
Improved responses in special populations
e.g. Seasonal flu V, Herpes zoster V
Overcome poor immunogenicity
e.g. Malaria Vaccine
Leroux-Roels I, et al. PLoS One. 2008;3:e1665. RTS, S Clinical Trials Partnership. N Engl J Med. 2011;365:1863-1875. Lal H, et al. N Engl J Med. 2015;372:2087-2096. Roteli-Martins, et al. Hum Vaccin Immunother. 2012;8:390-397. Garçon N, et al. Chapter 4 in: Garçon, et al. Understanding Modern Vaccines: Perspectives in Vaccinology. Vol 1. Amsterdam: Elsevier; 2011. Knuf M, et al. Hum Vaccin Immunother. 2015;11(2):358-76. Denny L, et al. Vaccine. 2013;31:5745-5753. Di Pasquale A et al. Vaccines 2015;3:320–343
Adjuvants–Few Approved, Many in Development
25
Adjuvants in Licensed ProductsAdjuvant Mechanism or
ReceptorLicensedproduct
Aluminum salts Nalp3, ITAM,antigen delivery
Numerous (eg,pertussis, hepatitis,
pneumococcal)
AS04 TLR4 HPV
Emulsions(MF59, AS03)
Immune cellrecruitment,
antigen uptakeInfluenza
AS01 TLR4, inflammasome
Zoster
CpG ODN TLR9 Hepatitis B
Adjuvants in DevelopmentAdjuvant Mechanism or
receptorClinicalphase
ISCOMs (Matrix-M) Unknown 2
dsRNA analogues TLR3 1
Flagellin TLR5 1
C-type lectin ligands Mincle, Nalp3 1
CD1d ligands CD1d 1
GLA-SE TLR4 1
IC31 TLR9 1
CAF01 Mincle, antigendelivery 1
Adapted from Reed SG, et al. Nature Med. 2014;19:1597-1608.
Vaccine Game-changing Technologies
Reverse vaccinology
ATTTATACATGAGACAGACAGACCCCCAGAGACAGACCCCTAGACACAGAGAGAGTATGCAGGACAGGGTTTTTGCCCAGGGTGCAGTATG
Adjuvant Systems
Platform Technologies
Synthetic vaccines
(DNA/RNA) Viral Vectors
Structural vaccinology
Life science revolution driven by insights and data
Potential to improve efficacy; faster, simpler, more efficient development & manufacturing
Reverse Vaccinology and beyond
27
▪ Reverse Vaccinology was first successfully applied to MenB vaccine challenge1
▪ Subsequently Reverse Vaccinolgy was applied to other difficult pathogens, e.g.▪ Group B streptococcus (Maione D et al, Science, 2005)▪ E. coli (Gomes Moriel D et al, PNAS, 2010)
▪ Antigens identified by RV or other methods can potentially be further improved using new technologies e.g. Structural Vaccinology, Reverse Vaccinology 2.02
1. Rappuoli R, et al. Curr Op Microbiol. 2000;3:445-450.2. Rappuoli R, et al. J Exp Med. 2016;213:469-481.
Structural Vaccinology
28
▪ Can now use 3D knowledge of protein structure to design new vaccine antigens with optimized biological and immunogical features1
▪ E.g. design of RSV F antigen engineered as stable pre-fusion conformation2
1. Dormitzer PR et al. Nat Rev Microbiol; 2012;10:807-813.2) McLellan J et al. Science. 2013;342;592-598. 3. Swanson K et al. PNAS. 2011;108:9619-9624.
Example - Respiratory Syncytial Virus (RSV)
Post-fusion FSwanson K et al. (3)
Pre-fusion FMcLellan J et al. (2)
Same protein, different conformation
Concept of nucleic acid-based vaccines
Nucleic acid vaccine
– Express target antigens in the vaccinee
– Transformative,1 simpler technology2
– No requirement for cellculture-based manufacturing
– Scalable and flexible– Types of synthetic nucleic acid-based
vaccines– Plasmid DNA– messenger RNA
Cellular uptake
Immuneresponse
Manufacturein a cell-free system
Injectvaccine
1. Morrison C. Nat Rev Drug Discov 2016;15:521–522; 2. Brito LA et al. Mol Ther 2014;22:2118–2129
Antigen expression
Nucleic Acid Vaccines
APC, antigen-presenting cell; DNA, deoxyribonucleic acid; mRNA, messenger RNA; RNA, ribonucleic acid.Callaway E. Nature. 2020; 580(7805): 576–577.
DNA vaccine
Cell
APC
mRNA
Viral proteins
RNA vaccine(with lipid coat)
The DNA or RNA encoding a viral peptide that promotes an immune response is inserted into human cells, which then
produce large quantities of the specific DNA or RNA
Long lasting, specificEarly, non-specific
Triggering the innate immune response
Triggering the adaptive immune response
• Organized structure and size ideal for pattern recognition receptors ➔efficient triggering of innate immunity1
• Non-pathogenic & no replication in host due to removal of essential genes1
• Infect many cell types, including Antigen Presenting Cells (APC)2
Viral vectors retain immune-stimulating properties of viruses without causing disease
31
• Antigen of interest inserted into viral genome2
• Antigen produced within APC triggering antibodies and cellular immune responses1,2
Image 160235651 from www.shutterstock.com
1. Vittelli A et al. Exp Rev Vaccines 2017; 16:1241-1252; 2.Tatsis N & Ertl H Mol. Ther. 2004; 10:616-629.
Viral Vector Vaccines
APC, antigen-presenting cell.Callaway E. Nature. 2020; 580(7805): 576–577.
Replicating viral vector (such as measles)
Cell
Non-replicating viral vector (such as adenovirus)
Virus replicates
APC
Viruses such as measles or adenovirus are weakened so they
cannot cause disease, and are genetically modified to produce target
antigen proteins
Several vaccine candidates
Repurpose existing tools
Platform technologies can act as ‘on-demand’ solutions to produce vaccines against multiple diseases
331. Strugnell R et al. Perspect Vaccinol 2011;1:61–88; 2. Gilbert SC, Warimwe GM. Vaccine 2017;35:4461–4464
Platform-based vaccine development2Traditional vaccinology1
One technological development
Onetechnological
platform
One vaccine candidate
Multiple rounds of refinement
Accelerated development
How Can These Technologies Help Our Society?
34
Reverse Vaccinology
Empirical Approach GlycoconjugationRecombinant
DNA
MenB, GBS, GAS, E. coli, S. aureus,
C. difficile
Diphtheria, Tetanus, Pertussis, Rabies, Influenza, Smallpox, Polio,
BCG
MenACWY, Pneumo, Hib, GBS, S. aureus
Hepatitis B, Acellular Pertussis,
Lyme, Human papillomavirus
Synthetic Biology
Adjuvants/Human Immune Response
Structural Vaccinology
Next Generation Technologies
Finco O, Rappuoli R. Front Immunol. 2014;5:1-6.
Vectors
ElderlyFluGBSMeningococcalPneumococcalRSVZosterCandidaC. difficileE. coliKlebsiellaP. aeruginosaStaphBreast CancerColorectal CancerProstate Cancer
PregnancyCMVFluGBSHBVMeningococcalPertussisRSVTetanus
Vaccines For Every Age
35Rappuoli R, et al. Nature Rev Immunol. 2011;11:865-872.
Infants & ChildrenDiphtheriaFluGASHAVHBVHibIPVMeningococcalPertussisPneumococcalRotavirusRSVTetanusVaricella Measles
AdolescentsCMVDTaP boostEBVFluHSVHPVMeningococcal
AdultsDiphtheriaFluHBVMeningococcalPertussisRSVTetanus
-3 0 8 18 55 90MONTHS YEARS
Vaccines For Today’s Society
36
Rappuoli R. Vaccine. 2015;33S:B1–B2.
PovertyCholeraDengueETECHAVHBVHEVFluJEVMalariaMenBParasitic infectionsParatyphoidRabiesRotavirusSalmonellaS. entericaS. typhimuriumShigellaTBTyphoid fever
Emerging InfectionsAIDSAnthraxAvian influenzaCholeraCoronavirusesDiphtheriaDengueEbolaEV71MalariaSARSTBSmallpoxWest NileYersinia
TravelersCholeraDengueETECFluHAVHBVJEVMalariaMeningococcalParatyphoidRabiesShigellaTBTyphoid FeverYellow Fever
Patients with Chronic DiseasesCMVFluFungal infectionsP. aeruginosaParainfluenzaRSVStaphTB
Immunotherapy/ Therapeutic Vaccines?CancerAutoimmune diseasesAlzheimer’sChronic infections(HCV, HBV, HPV, HIV…)
Metabolic diseasesAllergyDrug addiction
History and Evolution of Vaccine Technologies Leonard Friedland, MD, FAAPVice President and Director, Scientific Affairs and Public HealthGSK Vaccines
Immunize Nebraska
August 7, 2020
Discussion