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， 感染可能包括HIV-1。 假设：CO2可能增加 HIV-1渗透到中枢神经系统（CNS），可能是通过 肿瘤坏死因子-α（TNF-α）反应机制，并改变 T细胞亚群优先迁移通过血脑屏障， CNS中的细胞介导免疫。 前期工作：TNF-α和可卡因 无细胞HIV-1穿过BBB模型的渗透增加， 人脑微血管内皮细胞（BMVEC）。 辅酶抑制 体外CD 4 + CD 45 RA（记忆）亚群的迁移和WBC增加， 体内淋巴细胞计数。 方法：HIV-1渗透入下 BBB模型的腔室（用BMVEC和胎儿星形胶质细胞构建在 用纤连蛋白包被的多孔膜）的p24抗原测定法测量。 传染性病毒; T细胞亚群FACScan分析; ELISA测定 细胞因子 特定目的：在TNF-α治疗的BBB模型中HIV-1的渗透率为1 或可卡因，以及HIV-1渗透的机制，如转胞吞作用; #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核细胞 可卡因成瘾个体的给药; #3 TNF-α， 干扰素-γ、白细胞介素（IL）-1、IL-4、IL-6、IL-10、IL-12产生 和来自正常供体的单核细胞的迁移特性 可卡因体外培养。 预期成绩;我们将确定（a） TNF-α和可卡因是否会破坏血脑屏障并增加HIV-1的渗透 （B）细胞介导的免疫的改变是否 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响应，以及（d）TNF-α是否 可卡因成瘾受试者血浆中的浓度可能具有不利的 对BBB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.