PROJECT 1: DNA AND MVA IMMUNOGENS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定是研究者机构。大量的努力已经放在比较体外剂量反应曲线的表达与体内剂量反应曲线的免疫原性。已经在293 T细胞中进行了体外表达测试，并使用FACS分析对表达Gag和Env的细胞进行评分。已在BALB/c小鼠中进行了免疫原性试验，并使用ICS对应答细胞进行评分。这些研究的结果表明，体外表达试验对产品浓度差异的评分比体内免疫原性试验准确得多。在MVA剂量的1000倍范围内，Gag和Env表达细胞的数量增加了接近1000倍。相比之下，在体内MVA引发和加强后，MVA剂量的100倍差异仅导致响应Gag的CD 8细胞增加4倍，响应Gag的CD 4细胞增加2倍。在DNA初免/MVA加强方案中，MVA剂量增加100倍导致对Gag应答的CD 8细胞增加6.5倍，而对Gag应答的CD 4 T细胞没有增加。对Env的反应显示MVA剂量对CD 8或CD 4反应没有可检测的影响。 已经在恒河猴中开始了关于我们的进化枝B和进化枝C HIV-1疫苗中ADA和IN 3 Env分别引发中和Ab应答的能力的研究。

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.