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: 5200716
• 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/5200716
• 关键词: AIDS therapy; AIDS vaccines; Macaca nemestrina; antibody titering; disease /disorder model; genetic strain; human immunodeficiency virus 1; model design /development; neutralizing antibody; nonhuman therapy evaluation; polymerase chain reaction; recombinant virus; simian immunodeficiency virus; virus DNA; virus envelope; virus infection mechanism; 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 is sought. We have investigated infection, replication, persistence and clinical changes of HIV-1 in pig-tail macaques. In addition studies are underway to characterize a novel HIV-1/SIV chimeric virus, constructed in our laboratory, for its potential to replicate in pig-tail and rhesus 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 was seen in PT99. We have further analyzed the persistence of HIV-1 sequences in the inoculated monkeys. The results of DNA PCR using HIV-1 gag primers indicated that HIV-1 sequences were detected in the PBMCs of PT86 at all time points tested for up to about a year post-inoculation (last time confirmed) whereas the sequences in PT99 were consistently detected at early time points and then sporadically. In case of PT 87 and PT 89 HIV-1 sequences were detected reproducibly only at an early time point indicating infection of the animals but lack of replication. These results confirm that HIV-1 can infect but not replicate efficiently in monkeys. Furthermore, there are host factors that must be considered in selection of animals for development of an AIDS model.

感染和疾病的动物模型系统的开发， HIV-1对于艾滋病疫苗研究和干预至关重要。 目前， 唯一能被HIV-1感染的动物是黑猩猩; 但不会产生疾病。 由于维护成本巨大， 一只黑猩猩，事实上，这不可能是一个疾病模型， 模式是寻求。 我们研究了感染，复制， HIV-1在猪尾猕猴中的持久性和临床变化。在 另外的研究正在进行中，以表征一种新的HIV-1/SIV嵌合体 病毒，在我们的实验室构建，其复制的潜力， 猪尾猴和恒河猴。 四只猴子接种了两种HIV-1分离株， 它们的env区域：命名为PT86和PT99的两只猴子接受LAV， 其env与实验室适应株HIV-1 IIIB相似; 命名为PT87和PT89的两只猴子接受了初级分离株 （<4代），其env区域与MN菌株相似， 代表了大多数北美HIV-1分离株。 只有PT86和 PT99血清转化。 在PT86的情况下，增加的抗体滴度 免疫印迹分析显示， 在接种后68周（PI）仍然很高。 中和 在PI 6周产生抗体。 从早期的测试时间点来看， 感染性HIV-1在感染后4周从PBMC中分离。 pcr分析 PBMC在早期时间点（PI 4、6、8周）显示RNA表达 并在PI第40周和第52周再次进行。 在PI 84周时，尽管PT86具有 在临床上保持健康，CD4/CD8下降趋势可能是 正在发生。在PT99的情况下，仅针对env蛋白的抗体被检测到。 检测到;然而，没有分离出病毒，CD4/CD8是 稳定 这些结果表明，LAV感染和复制， PT86，而在PT99中没有观察到复制的证据。我们有 进一步分析了HIV-1序列在接种的 猴子用HIV-1 gag引物进行DNA PCR的结果表明， 在所有时间点，PT86的PBMC中均检测到HIV-1序列 接种后长达约一年的检测（最后一次确认） 而PT 99中的序列在早期就被一致地检测到 点，然后零星地。 如果是PT 87和PT 89 HIV-1序列 仅在早期时间点可重复检测到， 动物感染，但缺乏复制。这些结果证实 HIV-1可以感染猴子，但不能在猴子体内有效复制。 此外，在选择时必须考虑宿主因素 用于开发艾滋病模型。