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

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。本研究旨在评价由聚丙交酯-乙交酯(PLG)微粒组成的临床级DNA疫苗和临床级重组包膜蛋白疫苗(DNA/PLG Prime‘Protein Boost)。这项研究的第二个目标是比较穆拉塔支原体和束支支原体的免疫反应。分别于0、4、8周用表达HIV-1 Gag和env的DNA疫苗免疫猕猴。以阳离子PLG微粒为载体构建表达载体。在第24、36和48周，用重组HIV gp140env蛋白和等量的MF-59佐剂混合进行加强免疫。定期给动物放血，并进行免疫功能分析。用双抗体夹心法测定血清中Env和Gag特异性免疫球蛋白的滴度。2例束状病毒血清转化为GAG，但无1例阳性。所有动物在一次蛋白质强化后都出现了阳性的环境滴度，并在59周的研究中一直保持阳性。这两个物种的滴度相似。第二次包膜蛋白免疫后获得的所有血清均为HIV中和抗体阳性，毛滴度和束状滴度相同。用细胞因子流式细胞术(CFC)对Env和Gag特异性的CD4+和CD8+T细胞进行定量。仅在一个时间点检测到1个乳头状肌和1个束状肌的CD4+T细胞GAG特异性反应。在DNA/PLG预备期未检测到包膜特异性的CD4+T细胞反应。然而，在第三次蛋白质增强后，除了1个束状肌外，所有6个伞状肌都有阳性的CD4+反应。CD8+T细胞反应在两个物种中都很低，GAG特异性反应在第三次DNA/PLG免疫后两周达到峰值，在3次蛋白质强化后出现环境特异性反应。来调查观察到的物种之间的差异？关于CD4+T细胞反应的研究，这项研究扩展到检测CD4+CD25+调节性T细胞是否抑制束支原体的抗原特异性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.