SIV MOLECULAR CLONES W/ MUTATIONS IN ENV TRANSMEMBRANE

ENV 跨膜突变的 SIV 分子克隆

• 批准号: 6116678
• 负责人: PAUL A LUCIW
• 金额: $ 7.98万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6116678
• 关键词: AIDS; Mammalia; communicable diseases; immunology; inflammation; lymphatic system; virus

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

膜蛋白跨膜区突变的意义分析 致病性SIVmac 239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在体外组织培养中不适于病毒复制 细胞 主要目的是确定这一点的重要性 恒河猴体内用含有 SIVmac 239的env-TM细胞内部分的点突变。 结果：在env-TM中， SIVmac 239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 用在env-TM中含有提前终止密码子的病毒克隆 显示出高病毒载量，并显示出免疫缺陷的迹象 疾病 DNA序列分析显示，停止的快速逆转 密码子 用具有多个突变的病毒克隆接种的猕猴 在env-TM域中显示低病毒负载， 疾病 未来方向生化实验将进行 以鉴定与所述蛋白质相互作用的细胞蛋白质， env-TM细胞内部分。 重要的是，SHIV克隆与 将构建HIV-1 env-TM中的突变， 在恒河猴中的负载和致病潜力。 关键词猿猴 AIDS，SIV clones，env mutants FUNDING NIH Grant AI39415