IMMUNE RESPONSES TO VSV/HIV/SIV HYBRIDS IN MACAQUES

该项目的目标是产生包膜的重组体， 编码HIV 89.6 Env蛋白的水泡性口炎病毒（VSV） 和SIVmac 239 Gag蛋白，表征对这些的免疫应答。 粘膜和肌内免疫后恒河猴中的病毒， 并评估保护猴子免受 致病性SIV/HIV杂合体，SHIV-89.6P。除了对SIV进行编码之外， 和HIV蛋白质，要构建的病毒将有包膜， 两种VSV血清型之一或来自第二种水疱病毒。使用 不同的信封将允许升压而不中和 矢量本身。罗斯博士的实验室率先开发了 的VSV作为载体，并已表明，VSV重组编码的， 流感血凝素蛋白是有效活鼻内疫苗， 保护小鼠免受流感病毒的致命攻击。VSV 载体可用于表达大量外源基因， 包括编码完整HIV Env和Gag蛋白的基因的信息。 VSV/HIV env病毒诱导CTL对HIV Env蛋白以及HIV 中和抗体。马克思博士和尼克松博士（他们是共同的， 这项申请的调查人员）已经取得了初步结果 显示出对由活的HIV-Env编码的HIV Env的强抗体和CTL应答， VSV载体在恒河猴中的可检测的发病机制的情况下。 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 中和抗体和定量CTL前体频率Env 和Gag以及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 并用于具有足够对照的致病性SHIV攻击实验。 SHIV攻毒后将进行病毒定量分析。 接种动物和对照动物的负荷、健康和免疫状态。

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.