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

这个子项目是许多研究子项目中的一个 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， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 粘液传播是艾滋病毒感染的主要途径。已经表明，粘膜早期先天性干扰素应答在抗HIV感染中起重要作用。然而，如果能够增强中和分泌型免疫球蛋白A和G应答，则针对HIV的粘膜免疫应答将更有效。据报道，与HIV血清阳性患者相比，高度暴露、持续血清阴性患者（如冈比亚性工作者和有无保护性关系的HIV感染者的伴侣）在宫颈粘膜中表现出更高的Gag、Pol和Nef特异性T细胞IFN-γ应答[3]。此外，几个研究小组已经表明，Gag在肺和阴道的粘膜部位诱导HIV特异性T细胞和伊加免疫应答[4]，此外Gag [5]、[6]、[7]、[8]、Nef [8]和Pol [8]已经用于许多粘膜免疫方案。因此，选择Gag、Nef和Pol作为抗原与粘膜免疫方法组合预期在HIV特异性免疫应答中具有积极影响。已知基于DNA的疫苗引发细胞介导的和体液介导的免疫应答，然而需要增加它们在人类中的应答幅度。在这个项目中，我们将确定派（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，已被证明具有抗癌和抗HIV活性）在DNA疫苗接种平台上增强HIV特异性免疫应答的免疫调节作用，在接种人源化HLA-A2转基因小鼠后。在这些小鼠中通过ELISPOT和ICC分析测量的细胞和体液介导的免疫应答将与来自假型化牛痘感染小鼠的血清的qPCR分析后的病毒血症控制相关。我们假设，在基于DNA的小鼠粘膜免疫后，先前在我们实验室测试的多抗原免疫调节剂将增强HIV-gag、nef和pol特异性介导的免疫应答。此外，人源化HLA-A2.1小鼠（其具有北美最常见的人类单倍型）将被用作体内模型，并且派由目前用于人类的组分配制的事实将建立该项目临床应用的途径。因此，我们希望这些结果可以轻松安全地转移到诊所，因此可以在人体中进行测试。

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.