A NASAL DNA/PROTEIN VACCINE FOR ANTI-HIV ANTIBODY AND CTL

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：探讨Invaplex在非人灵长类动物鼻腔DNA疫苗和蛋白免疫原中的佐剂作用。方法：研究方法。将免疫诊断公司的SIVmac251 rgp130蛋白与编码非感染性SHIV89.6颗粒的pVacc4DNA混合，作为DNA/蛋白质疫苗的试验配方。3组成年MamuA*01阴性雌性恒河猴(每组n=4)分别于0、4、8周鼻腔免疫。免疫前和免疫后每隔一段时间采集外周血、大肠活检和宫颈阴道活检标本，用细胞内细胞因子染色检测SIVmac251 env、HIV89.6 env和SIV Gag肽特异性的CD4和CD8 T细胞应答。用双抗体夹心法检测血清和分泌物(鼻腔、直肠和宫颈阴道)中SIV env和Gag/pol特异性的Ig和Ig抗体。结果：在第3次鼻腔免疫后，第3组猕猴的血清gp130特异性免疫球蛋白和鼻腔、直肠和宫颈阴道分泌物中的gp130特异性IgA水平显著高于未接种佐剂的第1组猕猴，且持续时间更长。第12周时，第3组动物外周血中分泌SIV env特异性干扰素-g的CD4+和CD8+T细胞的百分比也显著增加。结论：目前的数据清楚地表明，Invaplex可以增强对DNA疫苗和蛋白质疫苗的抗体应答。

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. Objective: To determine whether Invaplex could adjuvant nasal DNA vaccines and protein immunogens in nonhuman primates. Methods. The SIVmac251 rgp130 protein (ImmunoDiagnostics, Inc) mixed with pVacc4 DNA encoding noninfectious SHIV89.6 particles was used as a test DNA/protein vaccine formulation. Three groups of adult MamuA*01-negative female rhesus macaques (each n=4) were nasally immunized on weeks 0, 4, and 8. Peripheral blood, colorectal biopsies, and cervicovaginal biopsies were collected before and at intervals after immunization for measurement of SIVmac251 env-, HIV89.6 env-, and SIV gag-peptide specific CD4 and CD8 T cell responses by intracellular cytokine staining for IFN-g, IL-2, TNF-a, and IL-4. ELISA was used to quantitate SIV env- and gag/pol-specific IgG and IgA antibodies in serum and secretions (nasal, rectal and cervicovaginal). Results: After the 3rd nasal vaccination, Group 3 macaques demonstrated significantly higher and more durable levels of gp130-specific IgG in serum and gp130-specific IgA in nasal, rectal, and cervicovaginal secretions when compared to nonadjuvanted Group 1 macaques. Group 3 animals also had significantly greater percentages of SIV env-specific IFN-g-secreting CD4+ and CD8+ T cells in blood on week 12. Conclusions: The current data clearly indicate that Invaplex can enhance antibody responses to DNA vaccines as well as protein-based vaccines.