NOVEL STRATEGIES FOR IMMUNIZATION AGAINST PERTUSSIS AND RSV INFECTIONS

Srinivas Garlapati

Vaccine and Infectious Disease OrganizationSaskatoon, Canada

A novel adjuvant platform for neonates

Polyphos-phazenes

Delivery of Antigen

HostDefense Peptides

Recruitment

CpG ODN

Immune stimulation

Antigen

Platform technology: Pertussis, RSV, others….

Courtesy: Parallel solutions Inc

Generation of microparticles

Andrianov et al, 1998

Generation of microparticles

1000X 5000X

OVA CpG Merge

Garlapati et al 2008

Garlapati et al 2010

Release Kinetics

Garlapati et al 2010

Local microenvironment

Alum SOL

MP

MP

Triple Combo Soluble Formulation

Activation of IRF by cytosolic pattern recognition receptors

Antigen Presentation PathwaySOL

Triple Combo Microparticle

IL-1β Pathway

MPs induce STAT and IRF transcription genes

MP induce genes in the NFkβ complex

PCEP particle formulation by Ion-bridging with poly-amines

Single step process by simple mixing PCEP with natural poly-amine Chitosan or synthetic poly-ally-amine hydrochloride (PAH) – FDA approved

NaCl-CaCl2 PAH

Chitosan

PCEP- CaCl2 MPs

PCEP- PAH MPs

PCEP- CHIT MPs

PCEP particle uptake by DCsPCEP particle uptake by DCs

Garlapati et al manuscript in submission

Garlapati et al manuscript in submission

RSV Vaccine Development

RSV MARKET POTENTIAL

Non-Replicating RSV vaccines need TLR Adjuvant

Immune response FI-RSV Reason Read outHumoral response Low Avidity Non-neutralizing Ab-

Ag complexes,Complement Activation,Immune pathology

ELISA-Urea Wash reduces AbsorbanceHigh virus titersHigh Ab/VNT RatioLow IgG2a/IgG1 Ratio

Secretory Antibodies Low IgA Parenteral route Low IgA AbsCytokines Pro-inflammatory

IL1β & IL-15Th2 IL-4, IL-5, IL-13ChemokinesMIP1β, MCP-1, IL-8

Eosinophila,Peri-vasculitisPeri-bronchiolitisBroncho-constrictionMucus

CD4+ T cells High Prolonged innate immune responses,Suppress CD8+ T cells

CD8+ T-cells Low Defective viral clearancd

High virus shedding

RSV VACCINE PIPELINE

Discovery Pre-Clinical Phase-I Phase-II Phase-III2020

MEDI-559MEDI-534 (PIV3)

Novavax VLP F & G

PEV4 (virosome)

Protollin- RSV

RSV287Nanobio RSV

Sendai Virus vectorAlpha Virus vector

Virosomes P3CSK4rRSVΔG

DNA electoporation

Adenovirus vector

0 21 44 48 50

Immunize Boost Challenge

Kill 1

Kill 2

Spleens - ELISPOTLung-Fragment culturesBAL

Groups

1.Saline/RSV challenge2.Ag IN3.Ag Triple Combo SOL IN4.Ag Triple Combo MP IN5.Ag IM6.Ag Triple Combo SOL IM7.Ag Triple Combo MP IM

Triple Combo formulation with ΔF protein

Viral SheddingLung Cytokines

RSV-specific humoral responses in serum.

Serum Virus Neutralization TitersVirus Titers

Lung Fragment Culture

Cytokine ELISPOT of Splenocytes

Cytokines in Lung Homogenates Post-RSV Challenge

Acknowledgements….Acknowledgements….

Nelson EngGael AurayTadele KirosRachelle BuchananStacy StromMonika PolewiczAleksandra GraciaLaura LatimerMarlene SniderRavinder Garg

Volker GerdtsLorne BabiukAndrew PotterSylvia v.d. HurkHeather Wilson George MutwiriJim HolmstromHugh Townsend

VIDO

Funding: - B. & M. Gates Foundation - Canadian Institute of Health Research - Krembil Foundation - National Science and Engineering Research Council - Saskatchewan Health Research Foundation

Animal CareBarry WalkerSherryOthers…