Cationic Liposome-DNA Complexes as Immunomodulators and Vaccine Adjuvants
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1 Cationic Liposome-DNA Complexes as Immunomodulators and Vaccine Adjuvants September 29, 28
2 Platform Technology Proprietary Cationic Lipid-DNA Complexes Cationic/Neutral Lipid + DNA = JVRS-1 JVRS-1 + Antigen (Non-Coding) Delivery System: Proprietary Synthetic Cationic Lipid DNA Plasmid: Containing CpG + Non-CpG motifs Applications: Therapeutic Setting: JVRS-1 (IV dosing) Vaccine Setting: JVRS-1 + Antigen constructs (SC or IM)
3 Mechanism of Action Cationic Lipid-DNA Complexes = Cationic Lipid = DNA Plasmid = Antigen T cells (CTL) B cells (Antibody) JVRS-1 Vaccine DC Endosomal TLR TLR TLR TLR TLR TAP/ER MHC ADAPTIVE IMMUNITY JVRS-1 Therapeutic Mø TLR Pathway DAI Pathway Nucleus INNATE IMMUNITY Cytosolic Cytokines NK cells Extracellular Preferential Facilitated Endosomal TLR & IFN Induction Pathways Protection Targeting Cell Entry Retention Unique Delivery Features Mechanisms of Action
4 CLDC as an Immunomodulator
5 Superior Activation of Innate Immunity Driven by JVRS-1 Complex T H 1 Cytokine Response Indicators NK Cell Expansion IL-12 (pg/ml) Dose = 1ug IV injection Contro l LPS IL-12 Poly I:C Lipid DNA JVRS-1 % NK1.1+ cells Spleen Lung Control JVRS-1 Dow et al, 1999; Journal of Immunology
6 JVRS-1 Therapeutic Dosing of Mice with Punta Toro Virus Infection 1 Doses % Survival ug 3. ug.3 ug.3 ug Placebo Gowen et al, 26; Antiviral Research Days Post Infection IP Treatment 24 hrs post-infection 1-11 mice/group
7 JVRS-1 Hepatitis B Therapeutics Response in HBV Transgenic Mice LIVER HBV DNA (pg/ug of tissue) HB-BS1 Transgenic HBV Mice Control JVRS-1 Days 1, 7, &13 (.25mg/kg) ADV nucleoside Daily x 14 days (1mg/kg) Morrey et al, 28 Antiviral Research TREATMENT ADV-adefovir dipivoxil (nucleoside analog)
8 JVRS-1 Hepatitis B Therapeutics Response in HBV Transgenic Mice 3 25 ug HBV Liver DNA ug.5ug.5ug.5ug Morrey, et al, 28 Antiviral Research
9 CLDC as an Vaccine Adjuvant
10 CD4+ T Cell Responses Following JVRS-1/gp61 Vaccination Comparison with Natural Infection 2 % gp61-specific CD4 T Cells Control LCMV JVRS-1/gp61 I-A b gp61 tetramers x CD44 JVRS-1 = 2X ip immunizations Lymphocytic choriomeningitis virus (LCMV) infection Zaks et al. (26) J. Immunol. 176:7335
11 Superior Adaptive Immune Response CTL Induction by JVRS-1 vs. Other Adjuvants JVRS-1 + OVA DNA (OVA) vaccine Vaccinia virus (OVA) vaccine Dendritic Cell + OVA CpG + OVA CpG + liposome + OVA CFA + OVA OVA protein or (OVA) gene OVA peptide = SIINFEKL Two injections % Ag-specific CD8 + T cells Tetramer Staining Zaks et al. (26) J. Immunol. 176:7335
12 Example Study Recombinant H1 Recombinant H1 (A/New Caledonia/2/99) vaccinated 1x or 2x with or without adjuvant Assessment of antibody and splenocyte response Drifted H1N1 challenge with 2xLD 5 (1x vaccination) or 6xLD 5 (2x vaccination) Supported by 5R41AI Lipid/DNA/Antigen Complexes for Influenza A Viral Vaccination Results demonstrate: Enhancement of antibody response (particularly IgG2a) Enhanced cytokine response of splenocytes from vaccinated mice restimulated with vaccine antigen Increased resistance to drifted H1 challenge
13 Antibody Response Following Vaccination with Suboptimal Amount of Recombinant H1 Antibody Titer (EC5) Antibody Titer (EC5) Anti-H1 IgG Anti-H1 IgG1 Antibody Titer (EC5) Anti-H1 IgG2a Two vaccinations H1 New Caledonia/2/99
14 HAI Antibody Response Following Vaccination with Suboptimal Amount of Recombinant H1 2 New Caledonia HAI 1 PR/8/34 HAI HAI 1 HAI 5 Vaccination Vaccination One or two vaccinations H1 New Caledonia/2/99
15 Splenocyte Response Following Vaccination with Suboptimal Amount of Recombinant H1 Single Vaccination Prime/Boost Vaccination 6 15 IFN-g (ng/ml) 4 2 IFN-g (ng/ml) 1 5 Restimulation Antigens Restimulation Antigens H1 New Caledonia/2/99
16 Body Weight Loss Following Single Vaccination with Recombinant H1 and Challenge with 2xLD 5 H1N % Body weight change 9 8 H1-NC H1-NC/J1 H1/JVRS-1 NT 7 6 day day 5 day 1 day 15 day 2 Days Post Challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
17 Survival Following Single Vaccination with Recombinant H1 and Challenge with 2xLD 5 H1N1 Percent survival H1/JVRS-1 H1 NT day of post-challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
18 Body Weight Loss Following Prime/Boost Vaccination with Recombinant H1 and Challenge with 6xLD 5 H1N % Original Body Weight H1 H1/JVRS-1 NT 6 day day 5 day 1 day 15 day 2 Day of post-challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
19 Survival Following Prime/Boost Vaccination with Recombinant H1 and Challenge with 6xLD 5 H1N1 Percent survival H1/JVRS-1 H1 NT day of post-challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
20 Body Weight Loss Following Adoptive Transfer of Serum from Mice Following Prime/Boost and Challenge with 2xLD 5 H1N1 11 % Original Body Weight H1/JVRS1-serum H1 only-serum NT-serum NT day post challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
21 Survival Following Adoptive Transfer of Serum from Mice Following Prime/Boost and Challenge with 2xLD 5 H1N1 Percent Survival H1/JVRS1-serum H1 only-serum NT-serum NT Day Post Challenge H1 New Caledonia/2/99 PR/8/34 (H1N1) challenge
22 Summary CLDC and in particular the JVRS-1 formulation has been shown to be an effective immunostimulant or adjuvant in mice, rats, rabbits, dogs, cats, chickens, cattle, and non-human primates Particularly effective in enhancement of T H 1 immune response (IgG2a, CD4, and CD8 response) Demonstrated polyfunctional T-cell response in nonhuman primates Human clinical trials Phase I TIV+JVRS-1 (1% enrolled) Phase I HCV Immunotherapeutic (IND activated) Phase I AML Immunotherapeutic (IND activated)
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