SIV & HIV 1 RECOMBINANTS SHIVNEF

意义重组SHIV克隆是检测HIV-1的一种手段 nef基因在体内用于控制病毒载量和进展的功能 恒河猴的艾滋病 新的抗病毒疗法和 抗HIV-1 Nef的疫苗可以在SHIVnef猴中进行测试 模型 目的建立一种研究HIV-1感染和 艾滋病的发病机制尚不清楚。 具体分析功能 体内HIV-1基因，SIV/HIV-1重组病毒（命名为SHIV） 通过替换致病克隆中的基因 SIVmac 239与对应的HIV-1基因。 我们克隆了SIV病毒 含有多种HIV-1 nef基因，以研究nef在体内的功能。 结果用HIV-1-SF 2和HIV-1-SF 33 nef基因分别替换 SIVmac 239克隆的nef基因。 两只幼年恒河猴 接种SHIV-SF 2nef的小鼠急性期病毒载量较低 感染和慢性阶段;这些动物健康， 超过两年。 另外两只幼年猕猴接种 SHIV-SF33nef. 这些动物中有一只保持了低病毒载量， 超过两年。 相比之下，第二只动物的病毒感染率上升， 在大约6个月的负荷，随后开发了致命的艾滋病样 感染后一年内发病。 病毒是从这个 尸检时的动物，并命名为SHIV-SF 33 nefA。 序列分析 该病毒揭示了nef相对于 输入病毒克隆中的nef序列。 重要的是，SHIV-SF 33 nefA 在幼年猕猴身上产生了致命的艾滋病样疾病。 未来 基因内SHIV-SF 33 nefA重组体和点突变体， 涉及HIV-1-SF 33 nef基因，将被构建以确定 该序列变化解释了病毒适应， 在恒河猴中的发病机制。 关键词HIV-1基因，艾滋病 发病机制，SHIV克隆基金NIH Grant AI 38532

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