DEVELOPMENT OF ANTI-HIV VECTORS IN MONOCYTES/MACROPHAGES

单核细胞/巨噬细胞中抗 HIV 载体的开发

• 批准号: 3144772
• 负责人: DANIEL G TENEN
• 金额: $ 17.54万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-04-01 至 1993-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3144772
• 关键词: AIDS; CD antigens; CD4 molecule; HIV infections; antisense nucleic acid; bone marrow; cell differentiation; chronic myelogenous leukemia; cytokine; disease vectors; gene expression; gene therapy; genetic markers; genetic promoter element; genetic transcription; human immunodeficiency virus 1; human immunodeficiency virus 2; immunological substance; leukocyte adhesion molecules; macrophage; model design /development; monocyte; nucleic acid sequence; oncogenes; phorbols; regulatory gene; tissue /cell culture; tissue donors; transposon /insertion element; virus RNA; virus genetics; virus protein; virus replication

While many of the devastating effects of the Acquired Immune Deficiency Syndrome (AIDS) appear to result from the infection of CD4+ lymphocytes, it is clear that human immunodeficiency virus (HIV), the causative agent, can also infect monocytes and macrophages. Infection of monocytic cells may be important int he initiation and propagation of persistent, low-level infection; in AIDS encephalopathy; in the release of inflammatory mediators, like tumor necrosis factor and interleukin 1 (IL-1); in suppression of hematopoiesis, and in the immune defects seen in AIDS. The objective of this proposal is to develop vectors which are capable of blocking HIV expression in monocytic cells, using cell-type specific promoters driving the expression of anti-HIV constructs. The specific aims are: (1) To study the effects of HIV infection of monocytic cells, particularly with respect to the effects on expression of monocytic cell surface markers and differentiation of these cells. We shall measure the expression of CD14, CD18, and other monocytic markers in myeloid cell lines, in monocytes and macrophages, and in bone marrow myeloid cells susceptible to different isolates of HIV-1 and HIV-2; and investigate the effects of differentiating agents like phorbol esters and cytokines on the expression of these markers in HIV infected versus uninfected cells. (2) To characterize in detail the promoters and regulatory elements of monocyte specific genes, particularly CD14 and CD18, and to isolate those elements of the promoters which are capable of driving the expression of heterologous genes in monocytes and macrophages. (3) To use these characterized promoters to develop vectors encoding of blocking the expression of specific HIV gene products essential for HIV viral replication in monocytic lines. (4) To develop vectors containing these monocytic promoter/anti-HIV constructs which can be efficiently introduced into bone marrow cells and block HIV infection of these cells. These studies should provide a focused but comprehensive approach to development of a model of gene therapy for AIDS.

虽然获得性免疫缺陷造成的许多破坏性影响 综合症（艾滋病）似乎是由 CD4+ 淋巴细胞感染引起的，它 很明显，人类免疫缺陷病毒（HIV）作为病原体，可以 也感染单核细胞和巨噬细胞。 单核细胞的感染可能是 重要的是持久性、低水平的启动和传播 感染;艾滋病脑病；在释放炎症的过程中 介质，如肿瘤坏死因子和白细胞介素 1 (IL-1)；在 造血抑制以及艾滋病中出现的免疫缺陷。 这 该提案的目标是开发能够 使用细胞类型特异性阻断单核细胞中的 HIV 表达 驱动抗HIV构建体表达的启动子。 具体目标 是：（1）研究HIV感染对单核细胞的影响， 特别是对于单核细胞表达的影响 这些细胞的表面标记和分化。 我们将测量 骨髓细胞中 CD14、CD18 和其他单核细胞标志物的表达 单核细胞和巨噬细胞以及骨髓髓样细胞中 对不同的 HIV-1 和 HIV-2 分离株易感；并调查 佛波酯和细胞因子等分化剂对 这些标记物在 HIV 感染细胞与未感染细胞中的表达。 (2) 详细描述单核细胞的启动子和调控元件 特定基因，特别是 CD14 和 CD18，并分离这些元件 能够驱动表达的启动子 单核细胞和巨噬细胞中的异源基因。 (3) 使用这些 表征启动子以开发编码阻断 HIV病毒所必需的特定HIV基因产物的表达 单核细胞系中的复制。 (4) 开发含有这些的载体 可有效导入的单核细胞启动子/抗 HIV 构建体 进入骨髓细胞并阻止这些细胞的艾滋病毒感染。 这些研究应该提供一个有重点但全面的方法 开发艾滋病基因治疗模型。