IMMUNOGENICITY IN MACAQUES OF A DNA/OLIGOMERIC ENVELOPE PROTEIN HIV VACCINE

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。本研究旨在评估用聚乳酸-乙醇酸酯（PLG）微粒配制的临床级DNA和使用DNA/PLG prime '蛋白增强方案的临床级重组Env蛋白HIV疫苗。该研究的第二个目标是比较mulatta和束状分枝杆菌的免疫反应。在第0、4和8周给猕猴接种表达HIV-1 gag和env的质粒dna。质粒采用阳离子PLG微粒配制。重组HIV gp140env蛋白与等体积的MF-59佐剂混合，在第24、36和48周肌肉注射加强疫苗。定期放血，进行免疫功能测定。ELISA法检测Env和gag特异性血浆IgG滴度。两个束状肌血清转化为gag，但没有一个mulatta阳性滴度。在一次蛋白质增加后，所有动物的env滴度都呈阳性，并在整个59周的研究中保持阳性。这两个物种的滴度相似。第二次env蛋白疫苗接种后获得的所有血清HIV中和抗体均呈阳性，mulatta和fascularis的滴度相当。细胞因子流式细胞术（CFC）定量检测Env和gag特异性CD4+和CD8+ T细胞。CD4+ t细胞gag特异性反应仅在1个mulatta和1个束状肌的一个时间点检测到。在DNA/PLG启动阶段未检测到env特异性CD4+ t细胞反应。然而，在第三次蛋白质增加后，所有6只mulatta只有1只束状体有阳性的CD4+反应。两种动物的CD8+ t细胞反应均较低，gag特异性反应在第三次DNA/PLG免疫后两周达到峰值，而在3次蛋白增强后出现特异性反应。为了调查物种之间的差异？CD4+ T细胞反应，本研究扩展到检测CD4+CD25+调节性T细胞是否抑制束状支原体抗原特异性T细胞反应。我们的结果表明，情况可能并非如此。

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. This study was designed to evaluate clinical-grade DNA formulated with poly(lactide-coglycolide) (PLG) microparticles and clinical-grade, recombinant Env protein vaccines for HIV using a DNA/PLG prime ' protein boost regimen. A secondary goal of the study was to compare the immune responses of M. mulatta and M. fascicularis. Macaques were vaccinated at weeks 0, 4, and 8 with plasmid DNAs expressing HIV-1 gag and env. Plasmids were formulated with cationic PLG microparticles. Booster vaccinations with recombinant HIV gp140env protein mixed with an equal volume of MF-59 adjuvant were given intramuscularly at weeks 24, 36 and 48. Animals were bled periodically and immune function assays performed. Env- and gag-specific plasma IgG titers were measured by ELISA. Two fascicularis seroconverted to gag, but none of the mulatta had positive titers. All animals developed positive env titers after one protein boost and remained positive throughout the 59-week study. Titers for the two species were similar. All sera obtained after the second env protein vaccination were positive for HIV neutralizing antibodies with mulatta and fascicularis having comparable titers. Env- and gag-specific CD4+ and CD8+ T cells were quantified by cytokine flow cytometry (CFC). CD4+ T-cell gag-specific responses were detected only at one time point in 1 mulatta and 1 fascicularis. No env-specific CD4+ T-cell responses were detected during the DNA/PLG priming phase. After the third protein boost, however, all 6 mulatta but only 1 fascicularis had positive CD4+ responses. CD8+ T-cell responses were low in both species with gag-specific responses peaking at two weeks after the third DNA/PLG immunization env-specific responses after 3 protein boosts. To investigate the differences observed between the species? CD4+ T-cell responses, this study was extended to examine whether CD4+CD25+ regulatory T cells suppressed the antigen-specific T-cell responses in M. fascicularis. Our results indicated that probably this was not the case.