Immunogenicity of HSV Amplicons in Rhesus Macaques

描述(由申请人提供)：预防人类免疫缺陷病毒(HIV)感染持续传播的最佳战略包括开发有效的疫苗。单纯疱疹病毒(HSV)扩增片段代表了一种新的、有前途的疫苗开发策略。单纯疱疹病毒扩增子作为疫苗策略保持了几个优势。首先，单纯疱疹病毒的扩增不会引起任何传染病。第二，单纯疱疹病毒扩增片段具有广泛的宿主细胞趋向性，能够有效地进入包括粘膜细胞在内的多种细胞类型。第三，单纯疱疹病毒扩增子具有很大的编码能力，这允许开发能够编码多种抗原和/或免疫刺激基因产物的载体。最后，单纯疱疹病毒扩增片段不包含免疫调节基因，而免疫调节基因是复制缺陷或减毒单纯疱疹病毒传递系统的一部分。以前对HSV扩增的研究是在小动物模型系统中进行的。然而，这些扩增子的免疫原性从未在非人类灵长类动物模型系统中进行过测试。这项建议中描述的研究将确定HSV扩增片段是否能在恒河猴中引发显著的免疫反应。具体地说，猕猴将通过肌肉内接种表达SIVmac239 Gag基因产物的HSV扩增片段，或通过不表达任何SIV基因产物的对照扩增片段进行接种。对GAG的细胞免疫反应将通过分析CD4+和CD8+T细胞反应、四聚体染色和抗原特异性细胞因子产生来追踪。体液反应将通过血清和粘膜免疫分析来确定。最后，将通过对自然杀伤细胞的分析和对淋巴细胞增殖和激活变化的纵向研究来确定先天免疫反应。这些研究的成功完成将允许在SIV模型系统中启动疗效试验。未来的研究可能会扩展到利用HSV作为疫苗递送系统来预防人类感染艾滋病毒。此外，这些研究的成功完成将对HSV扩增作为基因和癌症治疗载体的继续发展产生重大影响。

DESCRIPTION (provided by applicant): The best strategy for preventing the continuing spread of infection with the human immunodeficiency virus (HIV) involves the development of an effective vaccine. Herpes simplex virus (HSV) amplicons represent a novel and promising strategy for vaccine development. HSV amplicons maintain several advantages as a vaccine strategy. First, HSV amplicons cannot cause any infectious disease. Second, HSV amplicons have a wide host cell tropism and are able to efficiently enter multiple cell types including mucosal cells. Third, HSV amplicons have a large coding capacity, which allows for the development of vectors that could encode multiple antigens and/or immunostimulatory gene products. Finally, HSV amplicons do not contain the immunomodulatory genes that are a part of replication-defective or attenuated HSV delivery systems. Previous studies with HSV amplicons have been performed in small animal model systems. However, the immunogenicity of these amplicons has never been tested in a non-human primate model system. The studies described in this proposal will determine if HSV amplicons can elicit a significant immune response in the rhesus macaque. Specifically, rhesus macaques will be inoculated via an intramuscular route with HSV amplicons that express the SIVmac239 gag gene product or by control amplicons that do not express any SIV gene product. The cellular immune response to Gag will be followed through analysis of CD4+ and CD8+ T cell responses, tetramer staining, and antigen-specific cytokine production. The humoral response will be determined by analysis of both serum and mucosal immunity. Finally, the innate immune response will be determined through analysis of both natural killer cells and by performing longitudinal studies of changes in lymphocyte proliferation and activation. Successful completion of these studies will allow for efficacy trials to be initiated in the SIV model system. Future studies may be extended to utilize HSV as a vaccine delivery system to prevent HIV infection of humans. In addition, the successful completion of these studies will have a significant impact on the continued development of HSV amplicons as gene and cancer therapy vectors.