Role of Tetraspan Glycoproteins (CD63) in HIV Entry

超出所提供的空间。HIV感染原代人细胞通常需要与原代受体CD 4和CCRS或CXCR 4的化学动力学受体两者相互作用。我们已经发现了CD 63在HIV感染M0中的潜在作用，因为单克隆抗体（CD 63特异性mAb）可以阻止HIV进入MO，但不能阻止T细胞。CD 63是一种四斯潘膜糖蛋白，最为人所知的是血小板上的a1活化标志物，它几乎普遍存在于人类细胞中，并与整联蛋白密切相关。我们的实验将探讨CD 63在HIV进入M0中的作用。这些研究可能会导致新的抗逆转录病毒疗法的发展，靶向艾滋病毒进入。我们将在五年内实现这些目标：具体目标1。为了检验抗CD 63 mAb可以抑制不同HIV-1原代毒株感染M0的假设，这将包括亚型B和非亚型B毒株，包括一些使用CCRS作为辅助受体（R5）的毒株，以及使用CXCR 4或CCRS（RSX 4）的双嗜性毒株，以及能够在M0中复制的X4毒株。将评估不依赖于CD 4和CCRYCXCR 4相互作用进入的两亲性MLV-Env假型病毒，以确定抗CD 63免疫是否在结合后逆转录病毒进入事件中起作用。具体目标2。检验HIV靶细胞之间的细胞特异性差异影响CD 63在促进HIV进入中的作用的假设。我们将控制和测试不同水平的受体/辅助受体/CD 63表达和活化标志物01对抗CD 63抑制的敏感性的影响。我们还将测试抗CD 63抑制的细胞系模型，这将有助于这些研究中的细胞操作和一致的抑制作用，以及目的3中的机制研究。具体目标3。为了验证CD 63在促进HIV进入巨噬细胞中具有特定作用的假设，我们将确定CD 63介导的HIV进入的潜在机制，包括（a）受体/辅助受体相互作用或与CD 63的共定位，（B）CD 63与gp 120的结合，（c）CD 63在CD 4周转或病毒/受体内吞作用中的作用，以及（d）信号转导。这些实验将根据MLV-Env假型病毒（和X4 HIV-1毒株）的抗CD 63敏感性进行优先排序，如果抑制不依赖于CD 4和CCRS相关事件，则这将指导我们关注晚期事件。这些目标的实现将为HIV-1进入机制提供重要的新见解，并可能为抗逆转录病毒治疗提供重要的新靶点。网站（==

EXCEEDTHE SPACE PROVIDED. HIV infection of primary human cells typically requires interaction with both the primary receptor CD4 and a chemokinecoreceptor either CCRS or CXCR4. We have discovered a potential role for CD63 in HIV infection of M0, as monoclonal antibodies (mAb specific for CD63 can prevent HIV entry into MO, but not T-cells. CD63 is a tetraspan membrane glycoprotein, best known as a1 activation marker on platelets, that is nearly ubiquitous on human cells and is closely associated with integrins. Our experimentswil investigate the role of CD63 in HIV entry into M0. These studies may lead to developmentof novel antiretroviraltherapies targetini HIV entry. We will pursue these Aims over five years: Specific Aim 1. To test the hypothesis that infection of M0by diverse primary HIV-1 strains can be inhibited by anti-CD63 mAb This will include both subtype B and non-subtypeB strains, includingsome that use CCRS as a coreceptor (R5), as well as dual-tropic strains that use either CXCR4 or CCRS (RSX4), and X4 strains able to replicate in M0. Amphotropic MLV-Env-pseudotyped virus which enters independent of CD4 and CCRYCXCR4 interactions, will be assessed to determine if anti-CD63 irhbition acts at post binding retroviral entry events. Specific Aim 2. To test the hypothesis that cell-specific differences between HIV target cells affect the role of CD63 in facilitatini HIV entry. We will control for and test effects of various levels of receptor/coreceptor/CD63 expression and activation markers 01 susceptibility to anti-CD63 inhibition. We will also test cell line models of anti-CD63 inhibition, which would facilitate cel manipulation and the consistencycif lnhibitoryeffects in these studies,and in the mechanistic studiesin Aim 3. Specific Aim 3. To test the hypothesis that CD63 has a specific role in facilitating HIV entry into macrophages, we will identify potential mechanisms of CD63-mediatedHIV entry, including (a) receptor/coreceptor interactions or colocalization with CD63, (b) CD63 binding to gp120, (c) the role of CD63 in CD4 turnover or virus/receptor endocytosis and (d) signal transduction. These experiments will be prioritized based on anti-CD63 susceptibility of MLV-Env pseudotyped virus (and X4 HIV-1 strains), which would direct us to focus on late events if inhibition is not dependent on CD4 and CCRS-associated events. Achieving these Aims will provide important new insights into the mechanism of HIV-1 entry, and may well provide important new targets for antiretroviraltherapy. PERFORMANCESITE( ========================================Section End===========================================