DEVELOPMENT OF A MONKEY MODEL FOR TESTING OF ANTIVIRAL AGENTS AGAINST HIV-1

开发用于测试 HIV-1 抗病毒药物的猴子模型

• 批准号: 3748151
• 负责人: A S KHAN
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3748151
• 关键词: AIDS therapy; Macaca nemestrina; antiviral agents; blood chemistry; cytotoxic T lymphocyte; disease /disorder model; drug screening /evaluation; genetic strain; helper T lymphocyte; human immunodeficiency virus 1; model design /development; nonhuman therapy evaluation; polymerase chain reaction; recombinant DNA; suppressor T lymphocyte; virus DNA; virus envelope; western blottings

The development of an animal model system for infection and disease by HIV-1 is essential for AIDS vaccine studies and intervention. Currently, the only animal species which can be infected by HIV-1 is the chimpanzee; however, no disease is produced. Due to the enormous cost of maintaining a chimpanzee and the fact that this cannot be a disease model, a better model was sought. Based upon a report from the Washington Primate Research Center, studies were undertaken to investigate HIV-1 infection in pig-tail monkeys. Four monkeys were inoculated with two isolates of HIV-1 which differ in their env regions: two monkeys designated as PT86 and PT99 received LAV, which is similar in its env to the laboratory adapted strain, HIV-1 IIIB; and two monkeys designated as PT87 and PT89 received a primary isolate (< 4 passages) which is similar in its env region to the MN strain, which represents the majority of North American HIV-1 isolates. Only PT86 and PT99 seroconverted. In case of PT86, an increasing antibody titer against all the HIV-1 proteins was seen by Western Blot analysis which has remained high at 68 weeks post inoculation (PI). Neutralizing antibodies developed at 6 weeks PI. From the early test time points, infectious HIV-1 was isolated from PBMCs at 4 weeks PI. PCR analysis of PBMCs indicated RNA expression at early time points (4, 6, 8 weeks PI) and again at 40 and 52 weeks PI. At 84 weeks PI, although PT86 has remained clinically healthy, a downward trend in CD4/CD8 may be occurring. In case of PT99, antibodies to only the env proteins were detected; however no virus has been isolated and the CD4/CD8 is stable. These results indicated that LAV infected and replicated in PT86 whereas no evidence of replication has thusfar been seen in PT99. Studies are underway to analyze infection of PT87 and PT89 by the primary HIV-1 isolate. To achieve HIV-1 disease in monkeys, a high level of persistent virus infection must be established. Thus, studies are in progress to analyze various host factors and viral determinants contributing towards the inability of HIV-1 to establish a persistent high level infection in monkeys. One approach has been construction of a novel recombinant HIV-1 DNA in which the nef and LTR genes have been substituted by SIV239, a pathogenic SIV isolate. This novel virus, designated as SHIV-LTR has proven to replicate in monkeys cells at a higher efficiency than the parent HIV-1. Studies are being performed to analyze replication and disease potential of SHIV-LTR virus in monkeys.

一种感染和疾病动物模型系统的研制 HIV-1对于艾滋病疫苗研究和干预是必不可少的。目前， 唯一可能感染HIV-1病毒的动物物种是黑猩猩； 然而，不会产生疾病。由于维护成本巨大 黑猩猩和事实，这不能是一个疾病模型，一个更好的 寻求的是模特。根据华盛顿灵长类动物的一份报告 研究中心，开展了调查HIV-1感染的研究 在猪尾猴身上。给四只猴子接种了两个分离株 不同包膜区域的HIV-1：两只被指定为PT86的猴子 和PT99接收LAV，其环境与实验室相似 HIV-1 IIIB适应株；以及两只被指定为PT87和PT89的猴子 收到与其环境相似的初级分离物(&lt；4代) 地区的MN菌株，它代表了北美的大部分 HIV-1分离株。仅PT86和PT99血清转换。在PT86的情况下， 所有HIV-1蛋白的抗体效价都在增加 Western Blot分析在68周后仍保持在较高水平 接种(PI)。6周龄时出现中和抗体。从… 在早期检测时间点，从PBMC中分离到传染性HIV-1 4周后开始给药。外周血单核细胞的聚合酶链式反应分析显示早期有RNA表达 术后4、6、8周(PI)，40、52周(PI)。在84周时 PI，虽然PT86临床上保持健康，但在 可能正在发生CD4/CD8。在PT99的情况下，仅针对环境的抗体 检测到蛋白质；但没有分离到病毒，而且 CD_4/CD_8稳定。这些结果表明，LAV感染和 在PT86中复制，而到目前为止还没有复制的证据 在PT99中可以看到。分析PT87和PT89感染的研究正在进行中 由主要的HIV-1分离株。为了在猴子身上实现HIV-1疾病，一个 必须建立高水平的持续病毒感染。因此， 分析各种宿主因素和病毒的研究正在进行中 导致艾滋病毒-1无法确定的决定因素 在猴子中持续的高水平感染。一种方法是 含nef和LtR基因的新型重组HIV-1 DNA的构建 基因已经被致病的SIV分离株SIV239取代。这 一种名为SIV-LTR的新型病毒已被证明可在猴子身上复制 细胞的效率比亲本HIV-1更高。研究正在进行中 用来分析SHIV-LTR病毒的复制和致病潜力 在猴子身上。