A GENETIC AND BIOLOGICAL ANALYSIS OF HIV-1 AND HIV-2

HIV-1 和 HIV-2 的遗传和生物学分析

• 批准号: 3768914
• 负责人: K PEDEN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/3768914
• 关键词: CD4 molecule; HIV envelope protein gp120; HIV envelope protein gp41; T lymphocyte; clone cells; gene mutation; human immunodeficiency virus 1; human immunodeficiency virus 2; immunoglobulin G; virion; virus genetics; virus infection mechanism; virus replication

Virus derived from an infectious molecular clone of ELI was found to be infectious in PBMC, was found to exhibit a delayed time course of infection on CEM and H9 cells, but was unable to infect U937 cells. The virus that arose from the H9 cells was able to infect CEM and H9 cells without any delay in replication kinetics. In addition, this passaged virus was able to infect U937 cells. Two regions in the envelope gene were found to have mutations, and these changes were shown to confer the enhanced replicative capacity and expanded host range characteristics of the passaged virus. A Val-7 substitution in gp41 conferred the ability to replicate in T cell lines, while the additional Arg-427 change in gp120 was required before the virus could replicate in the promonocytic U937 cell line. The effect of these mutations on several parameters of envelope structure has been investigated. We have determined that the Met-7 to Val substitution in gp41 does not have a significant effect on spontaneous gp120 shedding although it increases the affinity of virions for CD4-IgG and amount of the CD4-induced dissociation of gp120 from virions. With both the gp41 Val-7 and the gp120 Arg-427 substitutions, the affinity of the virions for CD4-IgG, the spontaneous gp120 release, and the CD4-induced gp120 shedding are increased further. Thus, there is a correlation with the ability of a virus to replicate in cell lines and the tendency of its envelope to release gp120 after association with CD4. Mutations were constructed in the nef genes of several strains of HIV-1 and HIV-2 in order to ascertain the effect of Nef on virus replication in vitro. The effect of Nef on virus replication was more variable when tested on various CD4-positive cell lines. In some cases, there was no detectable difference in the replication kinetics between the Nef mutant and wild type viruses. These results demonstrate that the effects of Nef are variable and depend on the particular host-virus system, and that Nef is usually a positive factor for the replication of HIV in vitro.

发现来源于ELI的感染性分子克隆的病毒是 在PBMC中具有感染性，被发现表现出延迟的时间过程， 感染CEM和H9细胞，但不能感染U937细胞。 的 从H9细胞产生的病毒能够感染CEM和H9细胞 而不会延迟复制动力学。 此外，这款通过 病毒能够感染U937细胞。 包膜基因中的两个区域 被发现有突变，这些变化被证明赋予了 增强复制能力和扩大宿主范围的特点 传代病毒。 gp 41中的瓦尔-7取代赋予了以下能力： 在T细胞系中复制，而在gp 120中额外的Arg-427变化 在病毒可以在前单核细胞U937中复制之前， 细胞系 这些突变对几个参数的影响， 包络结构进行了研究。 我们确定 gp 41中Met-7至瓦尔的取代对细胞凋亡没有显著影响。 自发的gp 120脱落，尽管它增加了病毒粒子的亲和力 CD 4-IgG和CD 4诱导的gp 120从 病毒体。 在gp 41瓦尔-7和gp 120 Arg-427置换的情况下， 病毒粒子对CD 4-IgG的亲和力，自发gp 120释放， 并且CD 4诱导的gp 120脱落进一步增加。 由此可见，有 与病毒在细胞系中复制的能力相关， 其包膜与CD 4结合后释放gp 120的趋势。 在几种HIV-1病毒株的nef基因中构建了突变 和HIV-2，以确定Nef对病毒复制的影响。 体外 Nef对病毒复制的影响在以下情况下更易变： 在不同的CD 4阳性细胞系上测试。 在某些情况下， Nef突变体之间复制动力学的可检测差异 和野生型病毒。 这些结果表明，Nef 是可变的，取决于特定的宿主病毒系统，Nef 通常是HIV体外复制的阳性因子。