SIV MOLECULAR CLONES W/ MUTATIONS IN ENV TRANSMEMBRANE

突变体在环境蛋白跨膜中的意义分析 致病SIVmac239克隆的(env-TM)结构域可能提供洞察力 该结构域在病毒载量和猴艾滋病(SAID)中的作用 感染猕猴的致病机制。从该等机构取得的资料 活体研究将促进减毒活组织的发展 慢病毒疫苗，也可能确定一个新的靶点(S) 抗病毒治疗。目标：环境TM领域的很大一部分 SIV对于组织培养中病毒的体外复制是必不可少的 细胞。主要目标是确定这一点的重要性 恒河猴体内接种含不同结构域的病毒 SIVmac239的env-TM胞内部分点突变。 结果肺炎衣原体env-TM基因突变(错义和终止密码子) SIVmac239减少了各自病毒在人的培养物中的复制 猕猴的外周血淋巴细胞。接种了四只猕猴 与一个在env-TM中含有过早终止密码子的病毒克隆 表现出高病毒载量，并显示出免疫缺陷的迹象 疾病。DNA序列分析显示停滞迅速逆转 密码子。接种具有多个突变的病毒克隆的猕猴 在env-TM域中显示低病毒载量，没有迹象表明 疾病。今后生物化学实验的发展方向 确定一种与细胞内蛋白质相互作用的细胞蛋白(S) Env-TM的胞内部分。重要的是，Shiv克隆了 将构建HIV-1 env-TM的突变并对其进行病毒检测 恒河猴的载荷量和致病潜能。关键词猿猴 艾滋病，SIV克隆，环境突变资助NIH赠款AI39415

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 dispensable 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 inoculated with a viral clone containing a premature stop codon in env-TM exhibited high virus load and are showing signs of immunodeficiency disease. DNA sequence analysis revealed rapid reversion of the stop codon. Macaques inoculated with a viral clone with multiple mutations in the env-TM domain are displaying low virus load with no signs of disease. Future Directions Biochemical experiments will be performed to identify a cellular protein(s) that interacts with the intracellular portion of env-TM. Importantly, SHIV clones with mutations in the HIV-1 env-TM will be constructed and tested for virus load and pathogenic potential in rhesus macaques. KEY WORDS simian AIDS, SIV clones, env mutants FUNDING NIH Grant AI39415