SIV & HIV 1 RECOMBINANTS SHIVSUBTYPE B ENV

意义重组SHIVenv克隆是一种分析 HIV-1 env基因在体内跨粘膜传播中的作用 膜和SAIDS发病机制。 此外，基于 HIV-1免疫原可以通过用这样的抗体进行攻击来测试功效。 重组病毒克隆。 目的建立一种动物模型， HIV-1感染与艾滋病发病机制尚不清楚。 分析 HIV-1特异性基因在体内的功能，SIV/HIV-1重组体 病毒（命名为SHIV）已经通过替换 具有对应HIV-1基因的致病性克隆SIVmac 239。 我们有 制备了含有各种HIV-1包膜基因的SHIV克隆 亚型B分离株，并分析了这些重组病毒， 在幼年和新生猕猴体内。 结果一株致病性SHIV （命名为SHIV-33 A），其含有HIV-1-SF 33的env基因， 通过在幼年恒河猴中一次传代获得。 SIV-33A 在青少年猕猴中产生了猿类艾滋病， 静脉内或粘膜（口腔和阴道粘膜）途径。 在 此外，新生猕猴（2日龄）也出现猴艾滋病 静脉注射后。 中国对虾囊膜蛋白基因的序列分析 SHIV-33 A相对于输入物显示了约15个氨基酸的变化， SHIV-33克隆。 所有显示免疫缺陷疾病的动物也 在两种情况下，CD 4 T细胞均表现出快速和持续的下降， 外周血和淋巴结。 SHIV-33 A的体外分析 显示细胞病变增加和组织中复制增加 培养细胞。 基因内SHIV-33重组体和 正在构建涉及HIV-1 SF 33 env基因的点突变体 为了确定哪种序列变化导致病毒适应， 和发病机制。 还将进行研究，以确定 急性期CD 4 T细胞快速耗竭的机制 感染致病性SHIV-33 A株。 功能域 SHIV-33 A将通过构建具有点的env基因进行分析， 缺失突变;这些突变病毒克隆将在 组织培养细胞和恒河猴。 关键词HIV-1基因， 艾滋病发病机制，SHIV克隆

Significance Recombinant SHIVenv clones are a means to analyze the role of HIV-1 env genes in vivo for viral transmission across mucosal membranes and for SAIDS pathogenesis. Additionally, vaccines based on HIV-1 immunogens can be tested for efficacy by challenge with such recombinant viral clones. Objectives An animal model to study both HIV-1 infection and AIDS pathogenesis is not available. To analyze function of specific HIV-1 genes in vivo, SIV/HIV-1 recombinant viruses (designated SHIV) have been constructed by replacing genes in the pathogenic clone SIVmac239 with counterpart HIV-1 genes. We have made SHIV clones containing the envelope (env) gene of various HIV-1 subtype-B isolates, and have analyzed these recombinant viruses in vivo in juvenile and newborn macaques. Results A pathogenic SHIV (designated SHIV-33A), containing the env gene of HIV-1-SF33, was obtained by one passage in a juvenile rhesus macaque. SHIV-33A produced simian AIDS in juvenile macaques when given by either the intravenous or mucosal membrane (oral and vaginal mucosa) routes. In addition, newborn macaques (2 days of age) also developed simian AIDS after intravenous inoculation. Sequence analysis of the env gene of SHIV-33A revealed about 15 amino acid changes, relative to the input SHIV-33 clone. All animals showing immunodeficiency disease also exhibited a rapid and sustained decline in CD4 T-cells in both peripheral blood and lymph nodes. In vitro analysis of SHIV-33A revealed increased cytopathicity and increased replication in tissue culture cells. Future Directions Intragenic SHIV-33 recombinants and point mutants, involving the HIV-1SF33 env gene, are being constructed to determine which sequence change(s) accounts for viral adaptation and pathogenesis. Studies will also be performed to determine the mechanism of rapid depletion of CD4 T-cells in the acute stage of infection with the pathogenic SHIV-33A strain. Functional domains of SHIV-33A will be analyzed by constructing env genes with point and deletion mutations; these mutant viral clones will be examined in tissue culture cells and in rhesus macaques. KEY WORDS HIV-1 genes, AIDS pathogenesis, SHIV clones FUNDING NIH Grant AI41907