ANTI HIV MOLECULAR THERAPEUTICS TARGETING CHEMOKINE RECEPTORS

人类免疫缺陷病毒I型（HIV-1）诱发的脑病是一种神秘的综合征，发生在相当数量的艾滋病患者中，以认知、运动和感觉缺陷为特征。该综合征的潜在神经发病机制尚不清楚。最近，已经证明α和β趋化因子受体是HIV-1的主要共受体，并决定细胞特异性病毒株的趋向性。初步研究表明趋化因子和趋化因子受体在中枢神经系统（CNS）细胞上表达。同样，在HIV-1感染期间，中枢神经系统中趋化因子和趋化因子受体表达的改变也有报道。因此，趋化因子和趋化因子受体可能直接参与hiv -1诱导的脑病。我们建议利用细胞内免疫方法和单链可变片段（SFvs）抑制特定cns细胞中的趋化因子受体。初步研究将开发人类和小鼠的抗趋化因子受体SFvs。CXCR-4、CCR3趋化因子受体将是相关的初始分子靶点。在人类和小鼠SFVS发展到相关趋化因子受体后，这些构建物将被测试其抑制HIV-1在原代人类中枢神经系统细胞类型中的复制的能力。此外，在HIV-1感染期间改变趋化因子与基于中枢神经系统的细胞相互作用的影响将被表征。小鼠白血病病毒载体和新设计的慢病毒载体，转导非增殖细胞类型，将被利用。基于趋化因子受体胞内抑制，将分析其对中枢神经系统细胞（如凋亡）的分子机制。这些研究将直接与该计划中的几个项目相互作用，并承诺有助于了解HIV-1感染的神经发病机制，并朝着我们为HIV-1感染患者设计相关和互补的中枢神经系统治疗的长期目标迈进。

Human immunodeficiency virus type I (HIV-1)-induced encephalopathy is an enigmatic syndrome which occurs in a sizable group of patients with AIDS, characterized by cognitive, motor and sensory deficits. The underlying neuropathogenesis of this syndrome remains unclear. Recently, it has been demonstrated that alpha- and beta-chemokine receptors are the major co-receptors for HIV-1, and determine cell- specific viral strain tropism. Initial studies have now demonstrated that chemokine and chemokine receptors are expressed on central nervous system, (CNS)-based cells. As well, alterations in chemokine and chemokine receptor expression in the CNS during HIV-1 infection have also now been reported. As such, chemokine and chemokine receptors may b directly involved with HIV-1induced encephalopathy. We propose to inhibit chemokine receptors in specific CNS-based cells utilizing intracellular immunization approaches and single chain variable fragments (SFvs). Initial studies will develop anti-chemokine receptor SFvs, both human and murine. The CXCR-4, CCR3 chemokine receptors will be the relevant initial molecular targets. After development of human and murine SFVS to the relevant chemokine receptors, these constructs will be tested for their ability to inhibit HIV-1 replication in primary human CNS cell-types. In addition, the effects of altering chemokine interactions with CNS-based cells during HIV-1 infection will be characterized. Both murine leukemia virus vectors and newly designed lentiviral vectors, which transduce non-proliferating cell-types, will be utilized. The molecular mechanisms effects on CNS-based cells (e.g., apoptosis) will be analyzed, based on chemokine receptor intracellular inhibition. These studies will directly interact with several projects within this program and promise to both assist in understanding the neuropathogenesis of HIV-1 infection and move towards our long-term goal of designing relevant and complementary CNS therapeutics in patients with HIV-1 infection.