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

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 中心，但不一定是研究者所在的机构。 目的：探讨Invaplex在非人灵长类动物鼻用DNA疫苗和蛋白免疫原中的佐剂作用。 方法.将SIVmac 251 rgp 130蛋白（ImmunoDiagnostics，Inc）与编码非感染性SHIV89.6颗粒的pVacc 4 DNA混合用作测试DNA/蛋白疫苗制剂。在第0、4和8周对三组成年MamuA*01阴性雌性恒河猴（每组n=4）进行鼻免疫。 在免疫之前和免疫之后间隔收集外周血、结肠直肠活检和宫颈阴道活检，以通过IFN-γ、IL-2、TNF-α和IL-4的细胞内细胞因子染色测量SIVmac 251 env-、HIV 89.6 env-和SIV gag-肽特异性CD 4和CD 8 T细胞应答。ELISA用于定量血清和分泌物（鼻、直肠和宫颈阴道）中的SIV env和gag/pol特异性IgG和伊加抗体。结果如下：第3次鼻内接种后，与未佐剂化的第1组猕猴相比，第3组猕猴血清中gp 130特异性IgG水平和鼻、直肠和宫颈阴道分泌物中gp 130特异性伊加水平显著更高且更持久。 在第12周，第3组动物的血液中SIV env特异性分泌IFN-γ的CD 4+和CD 8 + T细胞的百分比也显著更高。 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.