SIV MOLECULAR CLONES W/ MUTATIONS IN ENV TRANSMEMBRANE

膜蛋白跨膜区突变的意义分析 致病性SIVmac239克隆的（env-TM）结构域可以提供深入了解 该结构域在病毒载量和猿类艾滋病（SAIDS）中的作用 感染猕猴的发病机制。 从此类信息中获得的信息 体内研究将促进减毒活疫苗的开发， 慢病毒疫苗，也可以确定一个新的目标， 抗病毒治疗 目的env-TM域的大部分 SIV在体外组织培养中是病毒复制所必需的 细胞 主要目的是确定这一点的重要性 恒河猴体内用含有 SIVmac239的env-TM细胞内部分的点突变。 结果：在env-TM中， SIVmac239减少了在以下培养物中相应病毒的复制： 猕猴的外周血淋巴细胞。 四只猕猴 接种含有提前终止密码子的病毒克隆， env-TM。 在感染的急性期，这些动物表现出 高病毒载量。 因此，终止密码子的快速逆转 发生，或者env-TM的细胞内部分不需要 高病毒水平。 未来方向病毒将从 感染了突变克隆的猕猴，env基因将被 测序以检测env-TM中的潜在回复。 额外 病毒突变体，含有几个点突变和一个缺失， env-TM将在猕猴中进行病毒载量和（潜在） 疾病进展。 将进行生化实验， 鉴定与细胞内蛋白相互作用的细胞蛋白， 部分env-TM。 关键词SIV，艾滋病，抗病毒治疗

Significance Analysis of mutants in the transmembrane of env (env-TM) domain of the pathogenic SIVmac239 clone may provide insight on the role of this domain in viral load and simian AIDS (SAIDS) pathogenesis in infected macaques. Information obtained from such in vivo studies will facilitate the development of live-attenuated lentivirus vaccines, and may also identify a novel target(s) for antiviral therapy. Objectives A large portion of the env-TM domain of SIV is dispensible for viral replication in vitro in tissue culture cells. The main objective is to determine the importance of this domain in vivo by inoculating rhesus macaques with viruses containing point mutations in intracellular portion of the env-TM of SIVmac239. Results Mutations (missense and stop codons) in the env-TM of SIVmac239 reduced replication of the respective virus in cultures of peripheral blood lymphocytes from macaques. Four macaques were inoculated with a viral clone containing a premature stop codon in env-TM. In the acute phase of infection, these animals are exhibiting high virus load. Thus, either rapid reversion of the stop codon has occurred, or the intracellular portion of env-TM is not required for high virus levels. Future Directions Virus will be recovered from macaques infected with the mutant clone, and the env gene will be sequenced to detect potential reversions in env-TM. An additional viral mutant, containing several point mutations and a deletion in env-TM, will be tested in macaques for virus load and (potential) disease progression. Biochemical experiments will be performed to identify a cellular protein(s) that interacts with the intracellular portion of env-TM. KEYWORDS SIV, AIDS, antiviral therapy