HIV ENVELOPE PEPTIDE BASED VACCINE IN SHIV RHESUS MODEL

描述：(改编自申请者摘要)我们早先假设 针对艾滋病毒引起的艾滋病的有效疫苗接种战略应侧重于 通过使用高度的鸡尾酒启动细胞介导的免疫(CMI) 与多种MHC单倍型反应的保守表位。我们的 长期目标是研制一种以合成肽为基础的艾滋病毒疫苗 因为它提供了定义、安全和经济的优势。至 为了实现这一目标，我们提出了一种创新的方法，包括使用 自体树突状细胞(DC)呈递肽鸡尾酒和 猪瘟病毒诱导有效的CMI应答控制感染和病理 在恒河猴模型中。该疫苗由六种合成疫苗组成的混合物 HIV包膜蛋白中高度保守区域对应的多肽 在我们之前的研究中，我们在一系列动物模型中发现了gp60 (小鼠、恒河猴和黑猩猩)和艾滋病毒感染者的样本(包括 长期无进展者)，能够诱导HIV特异性CMI 回应。SHIV-恒河猴模型最适合测试保护性 多肽-鸡尾酒的疗效是因为，Shiv，一种由以下成分组成的嵌合病毒 HIV包膜和SIV核心，在猕猴中诱导类似艾滋病的疾病，因此 为测试基于HIV env的疫苗提供最佳替代方案 治疗学。我们的研究设计为创新的HIV疫苗方案所涉及 弗氏佐剂多肽免疫恒河猴的初步研究 随后是多肽脉冲自体DC的输注，导致 有效诱导疫苗接种后的增殖性和CTL反应 动物。重要的是，在与Shiv KU-2进行挑战时，有效地清除 循环中的病毒感染细胞和血浆传染性的降低 在所有接种疫苗的动物中都能观察到，但在对照组中没有观察到，尽管穿着制服 最初在所有猴子中都有感染。在其中一只对照猴子身上 恰逢CD4+细胞在三周内急剧下降至50以下， 34周前有消瘦的迹象，这是艾滋病的典型症状。这些结果证明了 针对HIV的多肽疫苗的原理。现在，我们建议使用 同样的六肽鸡尾酒疫苗与自体DC相结合， 作为唯一佐剂，用于在希夫-恒河猴模型中启动保护性免疫。 此外，我们建议采用Shiv-Rheesus模型来检验 六种保守HIV包膜多肽鸡尾酒的免疫原性和免疫效力 基于新型细菌的佐剂黏膜疫苗接种策略 经过改造以消除毒性但保留佐剂能力的毒素。我们 获得的试点数据显示突变形式的霍乱毒素的有效性 和一种迄今未检测到的嗜水气单胞菌细胞毒素，AS 黏膜佐剂诱导HIV包膜特异性Th和CTL应答的模型研究 老鼠。最后，我们建议研制一种DNA疫苗，由一种鸡尾酒组成 含有编码六个保守的HIV包膜的微小基因结构的质粒 多肽，并测试其免疫原性和保护效果 模特。

DESCRIPTION: (Adapted from Applicant's Abstract) We hypothesized earlier that an effective vaccination strategy against HIV-induced AIDS should focus on priming cell-mediated immunity (CMI) by employing a cocktail of highly conserved epitopes identified to be reactive with multiple MHC haplotypes. Our long-term goal is to formulate a synthetic peptide-based vaccine against HIV because it offers the advantage of being defined, safe and economical. To achieve this goal, we proposed an innovative approach that included employing autologous dendritic cells (DC) for presenting the peptide cocktail and inducing efficient CMI responses for control of infection and pathology by SHIV in a rhesus monkey model. The vaccine consisted of a mixture of six synthetic peptides corresponding to highly conserved regions in the HIV envelope protein gpl60 that we identified in our previous studies, in a series of animal models (murine, rhesus and chimpanzee) and samples from HIV infected people (including long-term nonprogressors), to be capable of inducing HIV-specific CMI responses. The SHIV-rhesus model is best suited for testing the protective efficacy of the peptide-cocktail because, SHIV, a chimeric virus comprised of HIV envelope and SIV core, induces AIDS-like disease in macaques, and thus provides the best alternative for testing HIV env-based vaccines and therapeutics. Our study design for the innovation HIV vaccine proposal involved immunizing rhesus monkeys initially with the peptides in Freund's adjuvant followed later by infusions of peptide-pulsed autologous DC that resulted in efficient induction of proliferative and CTL responses in the vaccinated animals. Importantly, upon challenge with SHIV KU -2, efficient clearance of virus infected cells in circulation and reduction in plasma infectivity were observed in all the vaccinated animals but not in the controls, despite uniform infection in all the monkeys initially. In one of the control monkeys this coincided with a precipitous drop in CD4+ cells to below 50 in three weeks, and signs of wasting by week 34, typical of AIDS. These results serve as proof of the principle for a peptide-based vaccine against HIV. Now, we propose to use the same six-peptide cocktail as a vaccine in combination with autologous DC, as sole adjuvant, for priming protective immunity in the SHIV-rhesus model. Additionally, we propose to adopt the SHIV-rhesus model to test the immunogenicity and efficacy of the six-conserved HIV env peptide cocktail for mucosal vaccination strategies employing adjuvants based on novel bacterial toxins that are modified to eliminate toxicity but retain adjuvant capacity. We obtained pilot data showing the effectiveness of mutated forms of cholera toxin and a hitherto untested cytotoxic enterotoxin from Aeromonos hydrophila, as model mucosal adjuvants for inducing HIV env-specific Th and CTL responses in mice. Finally, we propose to formulate a DNA vaccine, consisting of a cocktail of plasmids with mini-gene constructs encoding the six conserved HIV env peptides, and test immunogenicity and protective efficacy in the SHIV-rhesus model.