IMMUNE RESPONSES TO VSV/HIV/SIV HYBRIDS IN MACAQUES

这个项目的目标是生成被包膜的重组体 编码HIV 89.6膜蛋白的水疱性口炎病毒(VSV) 和SIVmac239 Gag蛋白，表征了对这些蛋白的免疫反应 粘膜和肌肉免疫后恒河猴体内的病毒， 并评估对猴子免受挑战的保护 致病性SIV/HIV杂交种，SIV-89.6p。除了编码SIV之外 和HIV蛋白，要构建的病毒的包膜将来自 两种VSV血清型中的一种或来自第二种水泡病毒。它的用途 不同的信封将允许助推而不中和 载体本身。罗斯博士的实验室是这项研究的先驱 VSV作为载体，并表明VSV重组体编码一个 流感血凝素蛋白是有效的鼻腔内活疫苗 保护小鼠免受流感病毒的致命挑战。VSV 载体可以用来表达大量的外源基因 信息包括编码完整的HIV、Env和Gag蛋白的基因。 VSV/HIV包膜病毒诱导对HIV包膜蛋白CTL和HIV 小鼠体内的中和抗体。马克思和尼克松博士(他们是 这项申请的调查人员)已获得初步结果 对活体编码的HIV env有较强的抗体和CTL反应 VSV载体在恒河猴中的致病机制尚不明确。 这项拟议的研究将详细检查恒河猴的免疫反应。 猕猴对HSV载体(总抗体、HIV中和抗体、VSV)的反应 中和抗体和针对环境的定量CTL前体频率 GAG和VSV核衣壳蛋白。将使用足够的动物 以便比较短期和长期免疫方案 并在有足够对照的情况下进行致病性希沃克病毒挑战实验。 在SHIV挑战之后，将对病毒进行定量分析 接种疫苗和对照动物的载量、健康和免疫状况。

The goal of this project is to generate recombinants of the enveloped virus, vesicular stomatitis virus (VSV) encoding HIV 89.6 Env protein and SIVmac239 Gag protein, characterize the immune responses to these viruses in rhesus macaques after mucosal and intramuscular immunization, and evaluate protection of the monkeys from a challenge with a pathogenic SIV/HIV hybrid, SHIV-89.6P. In addition to encoding the SIV and HIV proteins, the viruses to be constructed will have envelopes from either of the two VSV serotypes or from a second vesiculovirus. The use of the different envelopes will allow boosting without neutralization of the vector itself. Dr. Rose's laboratory has pioneered the development of VSV as a vector, and has shown that VSV recombinants encoding an influenza hemagglutinin protein are potent live intranasal vaccines that protect mice against a lethal challenge with influenza virus. VSV vectors can be used to express large amounts of foreign genetic information including genes encoding complete HIV Env and Gag proteins. The VSV/HIV env viruses induce CTLs to HIV Env protein as well as HIV neutralizing antibodies in mice. Drs. Marx and Nixon (who are co- investigators on this application) have obtained preliminary results showing strong antibody and CTL responses to HIV Env encoded by a live VSV vector in rhesus macaques in the absence of detectable pathogenesis. The proposed study will examine in detail the immune responses in rhesus macaques to HSV vectors (total antibody, HIV neutralizing antibody, VSV neutralizing antibody, and quantitative CTL precursor frequencies to Env and Gag and to VSV nucleocapsid protein. Sufficient animals will be used to allow comparison of short-term and long-term immunization protocols and for a pathogenic SHIV challenge experiment with sufficient controls. The SHIV challenge will be followed by quantitative analysis of viral load, health, and immune status of the vaccinated and control animals.