Cell-mediated immune responses generated after DNA delivered by
April 10, 2018 | Author: Anonymous | Category: N/A
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Cell-mediated immune responses generated after DNA delivered by either Biojector or Electroporation and boosted with a heterologous insert recombinant poxvirus September 13, 2011 Jeffrey R. Currier, C PhD Military HIV R Milit Research hP Program Division of Retrovirology, WRAIR
Background • RV262: Phase I study of the safety and immunogenicity of g) administered by y either PENNVAX™-G DNA ((Env and Gag) Biojector® 2000 or by CELLECTRA® Intramuscular Electroporation device followed by administration of MVACMDR (HIV-1 (HIV 1 CM235 env/CM240 en /CM240 gag/pol boost in healthy, health HIV uninfected adults • Part A: A Open-Label Open Label safet safety study st d conducted cond cted at the MHRP in Rockville, MD in 12 healthy, HIV uninfected adults • Part B: B Randomized Randomi ed placebo placebo-controlled controlled st study d opening in Kenya, Tanzania and Uganda (Oct’ 2011) with an n=80, HIV uninfected adults
PENNVAX™-G DNA (Prime) • Consists of 4 DNA plasmids:
3 plasmids each e expressing pressing an HIV Env En consensus consens s modified gp140 proteins for subtypes A, C and D respectively
1 plasmid encoding a Gag consensus consens s protein deri derived ed from subtypes A, B, C and D
• Administered in 4 mg doses, doses 1 ml volume (days 0 0, 28) 28):
2mg of the Gag expressing plasmid
0.7 mg of each Env expressing plasmid
• Two delivery y methods using g either:
BIOJECTOR® device: needle-free injection intra-muscularly
CELLECTRA® device: de ice in vivo i o intra-muscular intra m sc lar electroporation
MVA-CMDR (Boost) • Live, non-replicating, double recombinant, MVA vector
Expressing CRF01_AE Env (gp140) with a truncated cytoplasmic t l i tail t il
Expressing CRF01_AE Gag/Pol native open-reading frame (i t (integrase d l t d and deleted d reverse ttranscriptase i t functionally f ti ll inactivated)
108 pfu f administered d i i t d iintra-muscularly t l l on study t d days d 84, 84 168
Proven safety and immunogenicity as a stand-alone product
RV262 DNA/MVA Study Schedule PENNVAX™- G DNA ((EP or BioJect))
6-month vaccination schedule
MVA-CMDR
0
1
3
6
12
27
(time in months) V2
V7
V10
V13
V15
Immune monitoring schedule IFN- Elispot and Multi-functional ICS Pre-vaccination and 2 weeks post DNA and each MVA vaccination
RV262 Preliminary Safety – Part A • First-in-man safety evaluation of PENNVAX™-G: Consensus Env A, C, D, and CG4 plasmids
• Testing in vivo electroporation versus Biojector for DNA delivery • Total n=13 subjects j enrolled, 2 studyy unrelated withdrawals • No related SAEs, no pregnancies, no HIV infections • To date, DNA prime and boosting with MVA has been well tolerated
RV262 Cellular Immunogenicity (CMI) OBJECTIVES • Compare the relative immunogenicity of electroporation versus Biojector delivered DNA followed by an MVA boost • Use a standard IFN- Elispot assay and prime/boost matched peptides y using g flow cytometry y y for function and p phenotype yp ((14 color,, • Real-time analysis 16 parameter assay) Functions: IFN-, , IL-2,, MIP-1, , TNF-,, and CD107 Phenotyping: Naïve, Central memory, Effector memory, Effector cells ICS performed only with boost (MVA-CMDR insert) matched peptides • PBMC were tested Pre-vaccination,, 2 weeks post p 2x DNA prime p and 2 weeks posts each MVA boost
HIV-specific Response Rate by IFN- Elispot
HIV Env-specific Response by IFN- Elispot 193 146 ((62-555)) ((12-367))
MVA-CMDR ENV-PP (CM235)
76 98 (72-342) (15-215) PENNVAX-G ENV C PP ENV-C-PP
12 32 ((3-65)) ((23-72))
Point Prevalence for T Cell Responses (ICS)
• Positive response >0.05% of all antigen-specific T cells (and 3x BG) • Functional assessment has the caveat of boost (MVA) matched peptides
HIV antigen-specific CD4 T cells (any function)
MVA-CMDR ENV-PP (CM235)
MVA-CMDR GAG-PP (CM240)
HIV antigen-specific CD8 T cells (any function)
MVA-CMDR ENV-PP (CM235)
MVA-CMDR GAG-PP (CM240)
HIV-specific T Cell Responses by Function
Gating Strategy: Effector/EM/CM Discrimination CD8 T Cells
EFFECTOR MEMORY CD45RO+/CD45RA+/ CCR7-/CD28+/-
CENTRAL MEMORY CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL CD45RA C 5 +/C /CD45RO 5 OCCR7-/CD28+/-
NAIVE T CELL CD45RA+/CD45ROCCR7+/CD28+
HIV-specific T Cell Responses by Phenotype
Effector memory CD4+ T cells are generated CD4+ T cells
EFFECTOR MEMORY CD45RO+/CD45RACCR7-/CD28+/-
CENTRAL MEMORY CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL CD45RA+/CD45ROCCR7-/CD28+/-
NAIVE T CELL CD45RA+/CD45ROCCR7+/CD28+
Eff / Effector memory CD8+ T cells are generated CD8+ T cells
EFFECTOR MEMORY CD45RO+/CD45RACCR7-/CD28+/-
CENTRAL MEMORY CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL CD45RA+/CD45ROCCR7-/CD28+/-
NAIVE T CELL CD45RA+/CD45ROCCR7+/CD28+
Preliminary Conclusions • Few responses detected post-DNA using either MVA-matched peptides (ICS) or DNA-matched peptides (Elispot) • After the 1st MVA boost: 11/11 subjects and 5/11 subjects respectively have Env-specific CD4 and d CD8 T cellll responses iin th the 0.05 0 05 – 0.57% 0 57% range Gag responses seen in 6/11 and 2/11 subjects for CD4 and CD8 T cells ll respectively ti l after ft th the 1st MVA vaccination i ti • CD8 responses are predominantly EM after the MVA boosts • CD4 responses are a balance of EM and CM (2:1) after the MVA boosts • Elispot Eli t responses iin di directt agreementt with ith th the ICS d data t • Diminution of response rate and magnitude after 2nd MVA vaccination
Acknowledgements MHRP Rockville Mary Marovich, Protocol Chair/PI Tina Tong, Manager, Bioproduction Tyesha Jackson, GLP/QC Lab Silvia Ratto-Kim, Co-Investigator Merlin Robb Jerome Kim Nelson Michael
AFRIMS, Bangkok Mark de Souza, Laboratory Director Viseth Ngauy, Protocol co co-PI PI
MHRP Cellular Immunology Laboratory Doris Thelian Erick Herrera Vicky Lo
Commercial Partners N. Sardesai, Innovio A. Khan, Innovio J. Yan, Innovio M. Bagarazzi, Innovio
MHRP Vaccine Clinical Research Clinic Julie Ake Cheun Yen Lau Lei Zheng
Laboratory of Viral Diseases, NIAID Patricia Earl Leigh Anne Eller Bernard Moss
University of Pennsylvania D Weiner D. J. Boyer
NIH Division of AIDS and DoD: Funding, Funding Support, Support Sponsorship, Sponsorship Oversight
Supplemental Slides
Vector-specific CD8 T cell Response Boosting
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