SIV & HIV 1 RECOMBINANTS SHIVNEF

意义重组SIV克隆是检测HIV-1的一种手段 NEF基因在体内控制病毒载量和进展的功能 猕猴中的猿猴艾滋病。新的抗病毒疗法和 针对HIV-1Nef的疫苗可以在SHIVnef猴子身上进行测试 模特。目的建立一种既能研究HIV-1感染又能研究HIV-1感染的动物模型 目前尚不清楚艾滋病的发病机制。要分析特定的功能 体内HIV-1基因、SIV/HIV-1重组病毒(简称SHIV) 都是通过替换致病克隆中的基因来构建的 SIVmac239与对应的HIV-1基因。我们已经制作了希夫克隆人 携带多种HIV-1nef基因，研究nef在体内的功能。 结果用HIV-1-SF2和HIV-1-SF33nef基因替换 SIVmac239克隆的nef基因。两只幼年恒河猴 在急性期接种SHV-SF2nef的病毒载量较低 在感染和慢性阶段，这些动物是健康的 两年多了。另外两只幼年猕猴被接种了 Shiv-SF33nef。其中一只动物一直保持较低的病毒载量 两年多了。相比之下，第二只动物的病毒含量有所上升 在大约6个月时负载，随后发展为致命的艾滋病样 发病时间为感染后一年。病毒就是从这里恢复的 动物尸检，命名为SIV-SF33nefA。序列分析 该病毒揭示了nef相对于Net的8个氨基酸的变化。 输入病毒克隆中的nef序列。重要的是，SIV-SF33nefA 在幼年猕猴身上产生了一种致命的类似艾滋病的疾病。未来 基因内Shiv-SF33nefA重组体和点突变体方向， 涉及HIV-1-SF33 nef基因，将被构建以确定 哪个序列变化(S)解释了病毒适应和 猕猴的发病机制。关键词HIV-1基因、艾滋病 发病机制，SHIV克隆资助NIH拨款AI38532

Significance Recombinant SHIV clones are a means to examine HIV-1 nef gene function in vivo for control of virus load and progression to simian AIDS in rhesus macaques. Novel antiviral therapies and vaccines against HIV-1 Nef could be tested in the SHIVnef monkey model. 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 various HIV-1 nef genes to study nef function in vivo. Results The HIV-1-SF2 and HIV-1-SF33 nef genes were used to replace the nef gene of the SIVmac239 clone. Two juvenile rhesus macaques inoculated with SHIV-SF2nef showed low virus load in the acute stage of infection and in the chronic stage; these animals were healthy for over two years. Two additional juvenile macaques were inoculated with SHIV-SF33nef. One of these animals has maintained low virus load for over two years. In contrast, the second animal showed a rise in virus load at about 6 months and subsequently developed a fatal AIDS-like disease by one year after infection. Virus was recovered from this animal at necropsy and designated SHIV-SF33nefA. Sequence analysis of this virus revealed about 8 amino acid changes in nef relative to the sequence of nef in the input viral clone. Importantly, SHIV-SF33nefA produced a fatal AIDS-like disease in juvenile macaques. Future Directions Intragenic SHIV-SF33nefA recombinants and point mutants, involving the HIV-1-SF33 nef gene, will be constructed to determine which sequence change(s) accounts for viral adaptation and pathogenesis in rhesus macaques. KEY WORDS HIV-1 genes, AIDS pathogenesis, SHIV clones FUNDING NIH Grant AI38532