PROJECT 1: DNA AND MVA IMMUNOGENS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Substantial effort has been placed on comparing in vitro dose response curves for expression with in vivo dose response curves for immunogenicity. In vitro expression tests have been done in 293T cells and used FACS analyses to score cells expressing Gag and Env. Immunogenicity tests have been done in BALB/c mice and used ICS to score responding cells. The results of these studies reveal in vitro expression tests being much more accurate for scoring differences in product concentration than the in vivo immunogenicity tests. The numbers of Gag and Env expressing cells increased by close to 1000-fold over a 1000-fold range of MVA dose. In contrast, following in vivo MVA priming and boosting, a 100- fold difference in MVA dose resulted in only a four-fold increase in CD8 cells responding to Gag and 2-fold difference in CD4 cells responding to Gag. In a DNA prime/MVA boost regimen, a 100-fold increase in MVA dose resulted in a 6.5 fold increase in responding CD8 cells to Gag and no increase in responding CD4 T cells to Gag. Responses to Env revealed no detectable effects of MVA dose on the CD8 or CD4 response. Studies have been initiated in rhesus macaques on the ability of the ADA and IN3 Envs in our clade B and clade C HIV-1 vaccines respectively to elicit neutralizing Ab responses.