VIRAL AND CELLULAR DETERMINANTS OF HIV DEMENTIA

仍然没有解决的是为什么艾滋病痴呆症一般不 发展，直到坦率的免疫抑制发作，如果，的确，作为 有证据表明，HIV-1在感染早期进入大脑。虽然 受感染的单核细胞进入大脑的运输经常被 预示着HIV进入中枢神经系统（CNS）的模式， 一些研究已经检查了HIV感染的循环单核细胞内 神经系统疾病的背景下，包括艾滋病痴呆症。同样地， 尽管事实上单核细胞，包括可能的HIV感染者， 其中，从骨髓进入血液，对艾滋病毒宿主知之甚少 细胞间的相互作用这些细胞之间的相互关系 与艾滋病毒感染有关的血液、大脑和骨髓 和艾滋病痴呆症的发展，是这项建议的一个主要重点。 这些相互作用的细胞和病毒方面都将被 研究了来自患有和不患有痴呆症的患者的血液单核细胞， 纵向随访，将检查CD 16细胞表面 增殖细胞核抗原（PCNA）表达。的 在克隆测定中增殖的能力和环状形式的存在 HIV-1 DNA脑和骨髓组织，来自有和没有 将检查痴呆患者是否存在PCNA阳性巨噬细胞 以及环状HIV DNA的存在虽然艾滋病的细胞决定因素 痴呆症可能主要与发生在外界的事件有关。 中枢神经系统，在进入大脑之前，可能许多病毒 这种疾病的决定因素与入境后事件有关。因此，我们认为， 对确定痴呆症的病毒决定因素，补充 对来自脑和血液单核细胞的病毒的HIV表型的研究， 提出了这些来自痴呆症患者和非痴呆症患者的病毒， 评估复制潜力、表达水平、 扩散、引发细胞因子和趋化因子产生能力以及 在体外杀死人类神经元。这些病毒的表型将是 与从相同患者的T细胞中回收的那些相比。 将构建嵌合病毒以鉴定遗传决定簇 与生物特性相关。定点诱变将 用于精确定位关键核苷酸和激活 将检查感染的巨噬细胞中的细胞内信号传导途径。 我们相信对单核细胞运输动力学的理解， 无论是感染的还是未感染的， 艾滋病毒神经系统疾病的成功治疗模式。

Still unresolved is the paradox of why AIDS dementia does not generally develop until the onset of frank immunosuppression, if, indeed, as the evidence indicates, HIV-1 enters the brain early in infection. Although the trafficking of infected monocytes into brain has frequently been heralded a the mode in HIV entry into the central nervous system (CNS), few studies have examined HIV infection of circulating monocytes within the context of neurological disease, including AIDS dementia. Similarly, in spite of the fact that monocytes, including, presumably, HIV-infected ones, enter the blood from the bone marrow, little is known about HIV-host cell interactions in this tissue. The interrelationships of these cellular compartments--blood, brain and bone marrow--with respect to HIV infection and the development of AIDS dementia, is a major focus of this proposal. Both cellular and viral aspects of these interactions will be investigated. Blood monocytes from patients with and without dementia, followed longitudinally, will be examined for CD16 cell surface expression, proliferating cell nuclear antigen (PCNA) expression. the ability to proliferate in clonal assays and the presence of circular forms of HIV-1 DNA. Brain and bone marrow tissues from patients with and without dementia will be examined for the presence of PCNA-positive macrophages and the presence of circular HIV DNA. While cellular determinants of AIDS dementia might be related primarily to events that take place outside of the CNS, prior to entry into brain, it is likely that many of the viral determinants of this disease related to post-entry events. Therefore, towards identification of viral determinants of dementia, complementary studies of HIV phenotype of viruses from brain and blood monocytes are proposed. These viruses, from patients with and without dementia, will be evaluated for replication potential, level of expression, ability to spread, ability to elicit cytokine and chemokine production, and ability to kill human neurons in vitro. The phenotypes of these viruses will be compared with those recovered from T-cells from the same patients. Chimeric viruses will be constructed to identify the genetic determinants associated with the biological properties. Site-directed mutagenesis will be used to pinpoint the critical nucleotide and activation of intracellular signaling pathways in infected macrophages will be examined. We believe an understanding of the dynamics of trafficking of monocytes, both infected and uninfected, is critical for the development of successful treatment modalities for HIV neurological disease.