GP120 INDEPENDENT INFECTION OF CD4(-) CELLS BY HIV-1

GP120 HIV-1 对 CD4(-) 细胞的独立感染

• 批准号: 6495224
• 负责人: Shen Pang
• 金额: $ 25.39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6495224
• 关键词: AIDS therapy; HIV envelope protein gp120; HIV infections; T lymphocyte; cell line; centrifugation; confocal scanning microscopy; enzyme linked immunosorbent assay; flow cytometry; fluorescence microscopy; gel electrophoresis; helper T lymphocyte; human immunodeficiency virus 1; latent virus infection; molecular cloning; polymerase chain reaction; subtraction hybridization; virus infection mechanism; virus integration; virus protein; virus replication

We have developed a new approach to sensitively and quantitatively identify HIV-1-infected cells, and found that HIV-1 significantly infects some types of CD4(-) cells independently of gp120. Further studies have indicated that not only can CD4(-) cells be infected by HIV-1 through this mechanism, but also CD4(+) T-lymphocytes. These results propose two very important possibilities: 1) HIV-1 can exploit some types of CD4(-) cells to serve as viral reservoirs in patients; and 2) HIV-1 can reduce expression of Env in infected cells as a strategy to evade the host immune response and therefore reside in these cells as long-term low-level persistent infection in a latent state. In this application, we will quantitatively characterize gp120- independent HIV-1 infection, and elucidate the molecular mechanism by which this occurs. In Specific Aim 1, CD4(-) cell lines derived from various human tissues will be infected with three strains of HIV-1 Env(-) virus. The susceptibility of CD4(-) cells to HIV-1 infection, the rates of viral replication in infected cells, the life-span of the infected cells, and the expression of viral genes in these cell lines will be studied. The results from Aim 1 studies will be informative to assess the seriousness of overall HIV-1 infection of CD4(-) cells in patients. In Specific Aim 2, we will study the mechanism of gp120-independent infection. We will characterize the infection course of HIV-1 gp120- independent infection, including viral entry, reverse transcription, nuclear transportation, viral DNA integration, and the generation of viral RNA genomes using an HIV-susceptible CD4(-) cell line, LNCaP, infected by HIV-1 Env(-) virus. We will also identify genes that are involved in gp120-independent HIV-1 infection using subtractive hybridization. The results from these studies will be very informative for assessing the seriousness of CD4(-) cell hosted HIV-1 reservoirs in patients and for developing an approach to interrupt the infection of CD4(-) cells by HIV-1.

我们已经开发了一种新的方法来敏感和定量鉴定HIV-1感染的细胞，并发现HIV-1显着感染某些类型的CD4（ - ）细胞，独立于GP120。进一步的研究表明，CD4（ - ）细胞不仅可以通过这种机制感染HIV-1，还可以通过CD4（+）T淋巴细胞感染。这些结果提出了两种非常重要的可能性：1）HIV-1可以利用某些类型的CD4（ - ）细胞作为患者的病毒储层； 2）HIV-1可以减少感染细胞中ENV的表达，作为逃避宿主免疫反应的策略，因此将其作为潜在状态下的长期低水平持续感染驻留在这些细胞中。在此应用中，我们将定量地表征GP120-独立的HIV-1感染，并阐明发生这种情况的分子机制。在特定的目标1中，衍生自各种人体组织的CD4（ - ）细胞系将感染三种HIV-1 Env（ - ）病毒。 CD4（ - ）细胞对HIV-1感染的敏感性，感染细胞中病毒复制的速率，感染细胞的寿命以及这些细胞系中病毒基因的表达。 AIM 1研究的结果将有用，以评估患者CD4（ - ）细胞总体HIV-1感染的严重性。在特定目标2中，我们将研究与GP120独立感染的机制。我们将表征HIV-1 GP120-独立感染的感染过程，包括病毒式进入，逆转录，核转运，病毒DNA整合以及使用HIV敏感的CD4（ - ）细胞系，LNCAP，HIV-1 Enk感染的病毒RNA基因组的产生。我们还将使用减法杂交确定与GP120无关HIV-1感染有关的基因。这些研究的结果将非常有用，以评估患者中CD4（ - ）托管HIV-1储层的严重性以及开发一种通过HIV-1中断CD4（ - ）细胞感染的方法。