COCAINE, BLOOD-BRAIN BARRIER AND DISEASE PROGRESSION

描述：(申请人摘要) 完整的血脑屏障(BBB)保护脑细胞免受多种病毒的侵袭。 可能包括HIV-1在内的感染。假设：可卡因可能会增加 HIV-1对中枢神经系统(CNS)的渗透，可能是通过 肿瘤坏死因子-α(TNF-α)-反应机制，及其改变 T细胞亚群跨血脑屏障和扰动的优先迁移 中枢神经系统的细胞免疫。前期工作：肿瘤坏死因子-α和可卡因 增加无细胞HIV-1在由 人脑微血管内皮细胞(BMVEC)。可卡因被抑制 CD4+CD45RA(记忆)亚群在体外的迁移和WBC及 活体内淋巴细胞计数。方法：HIV-1向下渗透 血脑屏障模型小室(由BMVEC和胎儿星形胶质细胞在 纤维连接蛋白包裹的多孔膜)的p24抗原测定 传染性病毒.T细胞亚群的FACScan分析.ELISA法 细胞因子。 具体目标：在用肿瘤坏死因子-α治疗的血脑屏障模型中，HIV-1的第一穿透性 或可卡因，以及艾滋病毒-1的渗透机制，如跨细胞作用；#2 免疫状态(细胞计数和细胞因子产生)和迁移特性 实验性静脉注射可卡因前后单个核细胞的变化 对可卡因成瘾者的管理；#3肿瘤坏死因子-α， 干扰素-γ、IL-1、IL-4、IL-6、IL-10、IL-12的产生 刺激正常供者单个核细胞的迁移特性 在体外被可卡因。预计成绩；我们将确定(A) 肿瘤坏死因子-α和可卡因是否破坏血脑屏障并增加HIV-1的穿透力 通过跨细胞作用，(B)细胞介导的免疫改变是否 在体内和体外由可卡因诱导，(C)哪些细胞类型分泌 细胞因子对可卡因刺激的反应，以及(D)肿瘤坏死因子-α是否 可卡因成瘾受试者的血浆浓度可能对 对血脑屏障的影响。

DESCRIPTION: (Applicant's Abstract) The intact blood-brain barrier (BBB) protects brain cells from many virus infections possibly including HIV-1. Hypothesis: Cocaine may increase penetration of HIV-1 into the central nervous system (CNS), possibly by a tumor necrosis factor-alpha (TNF-alpha)-responsive mechanism, and alter preferential migration of T cell subsets across the BBB and perturb cell-mediated immunity in CNS. Preliminary work: TNF-alpha and cocaine increased penetration of cell-free HIV-1 across a BBB model constructed with human brain microvascular endothelial cells (BMVEC). Cocaine inhibited transmigration of CD4+CD45RA (memory) subset in vitro and increased WBC and lymphocyte counts in vivo. Methods: HIV-1 penetration into the lower chamber of the BBB model (constructed with BMVEC and fetal astrocytes on a porous membrane coated with fibronectin) measured by p24 antigen assay of infectious virus; FACScan analysis of T cell subsets; Elisa assay of cytokines. Specific Aims: #1 penetration of HIV-1 in BBB models treated with TNF-alpha or cocaine, and the mechanism of HIV-1 penetration, such as transcytosis; #2 immune status (cell counts and cytokine production) and migratory properties of mononuclear cells before and after experimental intravenous cocaine administration in cocaine-addicted individuals; #3 TNF-alpha, interferon-gamma, interleukin (IL)-1, IL-4, IL-6, IL-10, IL-12 production and migratory properties of mononuclear cells from normal donors stimulated by cocaine in vitro. Projected accomplishments; We will determine (a) whether TNF-alpha and cocaine disrupt the BBB and increase HIV-1 penetration by transcytosis, (b) whether alterations of cell-mediated immunity are induced by cocaine in vivo and in vitro, (c) which cell types secrete cytokines in response to cocaine stimulation, and (d) whether TNF-alpha concentrations in the plasma of cocaine-addicted subjects may have adverse effects on the BBB.