DEVELOPMENT OF ANTI-HIV VECTORS IN MONOCYTES/MACROPHAGES

虽然获得性免疫缺陷的许多破坏性影响 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)，即病原体，可以 也会感染单核细胞和巨噬细胞。单核细胞感染可能是 重要的是启动和传播持久的、低水平的 感染；在艾滋病脑病中；在炎症的释放中 介质，如肿瘤坏死因子和白介素1(IL-1)； 抑制造血，并在艾滋病的免疫缺陷中出现。这个 这项提议的目标是开发能够 阻断单核细胞中HIV的表达，使用细胞类型特异性 驱动抗HIV结构表达的启动子。具体目标 (1)研究HIV感染对单核细胞的影响， 尤其是对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 CD14、CD18等单核细胞标志物在髓系细胞中的表达 系，单核细胞和巨噬细胞，以及骨髓髓系细胞 对不同的HIV-1和HIV-2分离株敏感；并调查 佛波酯、细胞因子等分化因子对细胞生长的影响 这些标记物在HIV感染的细胞和未感染的细胞中的表达。(2) 详细描述单核细胞的启动子和调控元件 特异性基因，特别是CD14和CD18，并分离这些元件 能够驱动基因表达的启动子 单核细胞和巨噬细胞中的异源基因。(3)使用这些工具 特征化启动子以开发编码阻断 HIV病毒必需的特异性HIV基因产物的表达 在单核细胞系中复制。(4)开发含有这些基因的载体 可有效导入的单核细胞启动子/抗HIV载体 进入骨髓细胞并阻断这些细胞的艾滋病毒感染。 这些研究应该提供一种有重点但全面的方法来 开发一种艾滋病基因治疗模式。

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.