Role of Tetraspan Glycoproteins (CD63) in HIV Replication

四跨糖蛋白 (CD63) 在 HIV 复制中的作用

• 批准号: 7633282
• 负责人: Monique Regail Ferguson
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7633282
• 关键词: Affect; CCR5 gene; CD4 Antigens; CD81 gene; CXCR4 gene; Cell Line; Cell Nucleus; Cell surface; Cells; Chimeric Proteins; Data; Development; Down-Regulation; Endosomes; Event; Family; Feline Immunodeficiency Virus; Flow Cytometry; Gagging; Guanosine Triphosphate Phosphohydrolases; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Hepatitis C virus; Human; Human T-lymphotropic virus 1; Infection; KAI1 gene; Lead; Libraries; Life Cycle Stages; Membrane Glycoproteins; Membrane Microdomains; Messenger RNA; Monoclonal Antibodies; Myelogenous; Pathway interactions; Platelet Activation; Play; Predisposition; Process; Production; Proteins; RNA Interference; Recycling; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Role; Screening procedure; Small Interfering RNA; T-Lymphocyte; Testing; Time; Viral; Viral Proteins; Virus; Virus Diseases; Western Blotting; Work; antiretroviral therapy; chemokine; chemokine receptor; extracellular; human PHEMX protein; macrophage; melanoma; member; novel; receptor; research study; trafficking; uptake

DESCRIPTION (provided by applicant): HIV infection of primary human cells typically requires interaction of Env gp120 with both the primary receptor CD4 and a chemokine receptor, either CCR5 or CXCR4, followed by Env gp41-induced fusion. Involvement of other factors and processes are likely, and their importance may differ between the principle target cells, macrophages (M?) and T-cells. Upon screening of a myeloid-specific monoclonal antibody (mAb) library, anti-CD63 mAb was found to block HIV entry into primary M?, but not T-cells. CD63 is a member of the tetraspanin membrane glycoprotein family, now implicated in HIV infection (MS #1), and other tetraspanins have been implicated in infection with HTLV-I (CD82), FIV (CD9) and Hepatitis C virus (CD81). Recent studies have shown inhibition with a GST fusion protein that includes the CD63 second extracellular loop (EC2), which further supports a specific role for CD63 in HIV infection. New data since the last submission also suggests a role for CD63 in endosomal processing of HIV, and so we will look at both early events, as well as events that occur later in the HIV life cycle, to more fully delineate the potential role of CD63 in HIV infection. These studies may lead to development of novel antiretroviral therapies. We will pursue these aims over five years: Aim 1. To test the hypothesis that CD63 is involved with early HIV replication events, we will investigate virus uptake and initiation of reverse transcription in M? and T cells/cell lines. Aim 2. To test the hypothesis that CD63 is generally required for infection of primary M?, as well at T cells/cell lines, we will assess trafficking and assembly of viral proteins, particularly Gag (and also Env) with and without CD63 silencing with small inhibiting RNAs (siRNA). We will also silence varions Rab proteins, which are cellular GTPases that traffic cellular (and viral) proteins through endosomal pathways in order to identify steps of HIV trafficking affected by CD63. Aim 3. To test the hypothesis that CD63 plays a role in infection of M? (and other cells) by diverse primary HIV 1 strains, we will assess anti-CD63 and CD63-EC2 susceptibility of both subtype B and non-subtype B strains, including those that use CCR5 exclusively (R5, CXCR4 exclusively (X4) or dual-tropic strains (R5X4).

描述（由申请人提供）：原代人类细胞的 HIV 感染通常需要 Env gp120 与初级受体 CD4 和趋化因子受体（CCR5 或 CXCR4）相互作用，然后是 Env gp41 诱导的融合。可能还涉及其他因素和过程，并且它们的重要性在主要靶细胞、巨噬细胞 (M?) 和 T 细胞之间可能有所不同。在筛选骨髓特异性单克隆抗体 (mAb) 文库后，发现抗 CD63 mAb 可以阻止 HIV 进入原代 M？细胞，但不能阻止 T 细胞。 CD63 是四跨膜蛋白膜糖蛋白家族的成员，目前与 HIV 感染 (MS #1) 有关，其他四跨膜蛋白与 HTLV-I (CD82)、FIV (CD9) 和丙型肝炎病毒 (CD81) 感染有关。最近的研究表明，包含 CD63 第二胞外环 (EC2) 的 GST 融合蛋白具有抑制作用，这进一步支持了 CD63 在 HIV 感染中的特定作用。自上次提交以来的新数据也表明 CD63 在 HIV 内体加工中发挥作用，因此我们将研究早期事件以及 HIV 生命周期后期发生的事件，以更全面地描述 CD63 在 HIV 感染中的潜在作用。这些研究可能会导致新型抗逆转录病毒疗法的开发。我们将在五年内实现这些目标： 目标 1. 为了检验 CD63 参与早期 HIV 复制事件的假设，我们将研究 M? 中病毒的摄取和逆转录的启动。和 T 细胞/细胞系。目标 2. 为了检验原代 M? 以及 T 细胞/细胞系的感染通常需要 CD63 的假设，我们将评估病毒蛋白的运输和组装，特别是使用或不使用小抑制 RNA (siRNA) 沉默 CD63 的 Gag（以及 Env）。我们还将沉默 Rab 蛋白变种，Rab 蛋白是细胞 GTP 酶，通过内体途径运输细胞（和病毒）蛋白，以确定受 CD63 影响的 HIV 运输步骤。目标 3. 检验 CD63 在 M? 感染中发挥作用的假设。通过不同的原发性 HIV 1 毒株（和其他细胞），我们将评估 B 亚型和非 B 亚型毒株的抗 CD63 和 CD63-EC2 敏感性，包括那些仅使用 CCR5（R5、仅使用 CXCR4 (X4) 或双嗜毒株 (R5X4)）的毒株。