IMMUNOMODULATOR-MEDIATED ENHANCEMENT OF ANTI-HIV-SPECIFIC IMMUNE RESPONSE

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。 粘膜传播是人类免疫缺陷病毒感染的主要途径。已有研究表明，黏膜早期先天干扰素反应在抗HIV感染中起着重要作用。然而，如果中和分泌型免疫球蛋白A和G的反应能够增强，针对HIV的粘膜免疫反应将更加有效。据报道，与HIV血清阳性患者相比，暴露于高危环境、血清持续阴性的患者，如冈比亚性工作者和有无保护性关系的HIV感染者的伴侣，宫颈黏膜中的Gag、POL和Nef特异性T细胞干扰素-γ反应较高[3]。此外，几个小组已经证明，GAG在肺和阴道的粘膜部位诱导HIV特异性T细胞和IgA免疫反应[4]，此外，GAG[5]、[6]、[7]、[8]、Nef[8]和POL[8]已被用于许多粘膜免疫方案。因此，选择Gag、Nef和Pol作为抗原，结合粘膜免疫方法，有望对HIV特异性免疫应答产生积极影响。众所周知，基于DNA的疫苗既可以诱导细胞免疫反应，也可以诱导体液免疫反应，但有必要增加它们在人类身上的反应幅度。在这个项目中，我们将确定PAI(一种多抗原免疫增强剂，由流感和呼吸道疫苗的混合物组成，被证明具有抗癌和抗HIV活性)在DNA疫苗平台上增强HIV特异性免疫反应的免疫调节作用。通过ELISPOT和ICC分析在这些小鼠中测量的细胞和体液介导的免疫反应将与假型痘苗病毒感染小鼠血清的qPCR分析后病毒血症的控制相关。我们推测，先前在我们实验室测试的多抗原免疫调节剂，在基于DNA的小鼠粘膜免疫后，将增强HIV-Gag、nef和pol特异性介导的免疫反应。此外，拥有北美最常见的人类单倍型的人源化HL A-A2.1小鼠将被用作体内模型，并且PAI是由目前人类使用的成分配制的；这将为该项目的临床应用铺平道路。因此，我们希望这些结果可以轻松和安全地转移到临床上，因此可以在人体上进行测试。

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. Mucosal transmission is the principal route of HIV infection. It has been shown that the mucosal early innate interferon response plays an important role against HIV infection. However, the mucosal immune response against HIV would be more effective if the neutralizing secretory immunoglobulins A and G response could be enhanced. It has been reported that highly exposed, persistently seronegative patients such as Gambian sex workers, and partners of HIV infected persons who have unprotected sexual relations exhibit higher Gag, Pol and Nef-specific T cell IFN-gamma responses in cervical mucosa, as compared to HIV-seropositive patients [3]. Moreover, several groups have shown that Gag induces an HIV-specific T-cell and IgA immune responses at mucosal sites of lung and vaginal tract [4], besides that Gag [5], [6], [7], [8], Nef [8] and Pol [8] have been used in numerous mucosal immunization protocols. Therefore, the selection of Gag, Nef and Pol as antigens in combination with a mucosal immunization approach is expected to have a positive impact in the HIV-specific immune responses. DNA based vaccines are known to elicit both: cell- and humoral-mediated immune responses, however there is a need to increase the amplitude of their response in humans. On this project, we will determine the immunomodulatory effect of PAI (a polyantigenic immunopotentiator consisting of a mixture of influenza and respiratory vaccines that was proven to have anti-cancer and anti-HIV activities) in the enhancement of the HIV-specific immune response on a DNA vaccination platform, after vaccination of humanized HLA-A2.transgenic mice. Cell- and humoral-mediated immune responses, as measured by ELISPOT and ICC analysis in these mice, will be correlated with control of viremia after qPCR analysis of serum from pseudotyped vaccinia infected mice. We hypothesize that the Polyantigenic Immunomodulator, previously tested in our laboratory, will enhance the HIV-gag, nef and pol specific mediated immune responses, after a DNA based mucosal immunization in mice. Moreover, the facts that humanized HLA-A2.1 mice, which possess the most common human haplotype in North America, will be used as the in vivo model, and that PAI is formulated from components currently used in humans; will build a pathway towards a clinical application of this project. We therefore expect that these results could be moved easily and safely into the clinics, and therefore, could be tested in humans.