IMMUNE RESPONSE TO HIV-1 ENCODED ANTIGENS IN MICE

小鼠对 HIV-1 编码抗原的免疫反应

• 批准号: 3141967
• 负责人: MAN-SUN M SY
• 金额: $ 30.24万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3141967
• 关键词: CD4 molecule; HIV infections; antibody; antibody formation; antigen presenting cell; cellular immunity; cytotoxic T lymphocyte; enzyme linked immunosorbent assay; gene expression; genetic manipulation; human immunodeficiency virus 1; humoral immunity; hybridomas; immunofluorescence technique; laboratory mouse; lymphocyte; lymphokines; monoclonal antibody; monocyte; neoplastic cell; surface antigens; tissue /cell culture; transfection; virus antigen

The objectives of this proposal are to study: 1. The relationship between CD-4 Internalization and susceptibility to HIV-1 Infection. We transfected the human CD-4 gene into murine T cell hybridomas, one of these hybridomas can be infected by HIV-1. The cell biology of the CD-4 molecules will be studied with particular emphasis on the mechanism of CD-4 internalization. The relationship between the capacity of the T cell hybridomas to internalize CD-4 molecules and their sensitivity to HIV-1 infection will be investigated. We will investigate the effects of HIV-1 infection on the activation of CD-4 bearing T cell hybridomas. 2. Effects of HIV-1 gene expression on the physiology of lymphocytes and monocytes. We transfected the tatIII gene of HIV- l into a murine B cell lymphoma (A20), a murine antigen specific T cell hybridoma (7E2) and a murine monocyte tumor line (P388D1). These cell lines were chosen because their surface antigens, antigen specificities, and antibody and lymphokine production have been well characterized. In addition, these cell types are similar to the cell types known to be infected by HIV-l, T cells, monocytes and some B cells. We will continue to study the effects of the tatIII gene on the expression of surface antigens and the physiology of these transfected cell lines. 3. Immune responses against HIV-l encoded antigens in mice. We have immunized normal mice with tatIII transfected cell lines and generated antigen specific CTL and polyclonal antisera specific for tatIII transfected cell lines. The epitopes that are essential for the elicitation of CTL will be identified. We plan to characterize the new proteins induced by the tatIII gene. The results from these experiments should provide useful insights into the mode of action of the tatIII gene and the immune responses against tatIII gene products. We plan to immunize mice with HIV- l infected murine cell lines and study T cell mediated and humoral immune responses to HIV-l infected cells in vivo. These investigations utilize in vivo and in vitro models of HIV-l infection in the murine system. These model systems are designed to facilitate discovery of the mechanisms of HIV-l entry into host cells and the effects of HIV-l gene expression on cell physiology and function. By studying the immune response to murine cell lines either transfected with individual genes or infected with HIV-l, we will be studying the mode of action of individual viral genes and the host immune responses against HIV-l. These experiments will provide fundamental insights into the pathogenesis of HIV-l infection.

本提案的目标是研究： 1. CD-4内化与 易感染HIV-1。 我们转染了人类CD-4 基因导入鼠T细胞杂交瘤，这些杂交瘤之一可以是 感染HIV-1。 CD-4分子的细胞生物学将是 特别强调CD-4的机制 内化 T的容量之间的关系 细胞杂交瘤内化CD-4分子及其敏感性 将对HIV-1感染进行调查。 我们将调查 HIV-1感染对携带CD-4的T细胞活化的影响 杂交瘤。 2. HIV-1基因表达对小鼠生理功能的影响 淋巴细胞和单核细胞。 我们把艾滋病病毒的tatIII基因- 1转化为鼠B细胞淋巴瘤（A20），鼠抗原特异性 T细胞杂交瘤（7 E2）和鼠单核细胞肿瘤系（P388 D1）。 选择这些细胞系是因为它们的表面抗原， 抗原特异性，抗体和淋巴因子的产生 被很好地描述了。 此外，这些细胞类型是相似的 已知被HIV-1感染的细胞类型，T细胞，单核细胞， 和一些B细胞。 我们会继续研究 tatIII基因对表面抗原表达的影响， 这些转染细胞系的生理学。 3. 小鼠中针对HIV-1编码抗原的免疫应答。 我们用tatIII转染的细胞系免疫正常小鼠， 并产生了抗原特异性CTL和多克隆抗血清， 对于tatIII转染的细胞系。 这些重要的表位 将确定诱导CTL的条件。 我们计划 表征由tatIII基因诱导的新蛋白质。 的 这些实验的结果应该提供有用的见解， tatIII基因的作用方式和免疫应答 针对tatIII基因产物。 我们计划用艾滋病病毒免疫老鼠- l感染小鼠细胞系并研究T细胞介导和体液免疫 体内对HIV-1感染细胞的免疫应答。 这些研究利用HIV-1的体内和体外模型 感染小鼠系统。 这些模型系统被设计成 为了便于发现HIV-1进入宿主的机制， 细胞和HIV-1基因表达对细胞生理的影响 和功能 通过研究对小鼠细胞系的免疫反应， 无论是用单个基因转染还是感染HIV-1， 我们将研究单个病毒基因的作用方式 以及宿主对HIV-1的免疫应答。 这些实验 将为HIV-1的发病机制提供基本的见解 感染