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: 8098853
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8098853
• 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; Glycoproteins; 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与主受体CD4和趋化因子受体CCR5或CXCR4的相互作用，然后是ENV GP41诱导的融合。其他因素和过程的参与很可能是可能的，其重要性可能在原理细胞，巨噬细胞（M？）和T细胞之间有所不同。在筛选髓样特异性单克隆抗体（MAB）库后，发现抗CD63 MAB阻止HIV进入主要M？，但不能阻断T细胞。 CD63是四跨质膜糖蛋白家族的成员，该家族与HIV感染有关（MS＃1），而其他四跨质蛋白已与HTLV-I（CD82），FIV（CD9）和肝炎病毒（CD81）感染有关。最近的研究表明，GST融合蛋白的抑制作用，其中包括CD63第二个细胞外环（EC2），该蛋白进一步支持CD63在HIV感染中的特定作用。自上次提交以来的新数据还暗示了CD63在HIV的内体处理中的作用，因此我们将研究这两个早期事件以及在HIV生命周期后期发生的事件，以更充分地描述CD63的潜在作用在HIV感染中。这些研究可能导致新型抗逆转录病毒疗法的发展。我们将在五年内追求这些目标：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中的病毒摄取和启动M中的逆转录？和T细胞/细胞系。目的2。为了测试CD63通常在T细胞/细胞系中感染CD63的假设，我们将评估病毒蛋白的运输和组装，尤其是GAG（以及ENV），以及没有CD63沉默带有小的抑制RNA（siRNA）。我们还将使Rab蛋白保持沉默，这些变异是细胞GTP酶，通过内体途径流动细胞（和病毒）蛋白，以识别受CD63影响的HIV运输步骤。目的3。检验CD63在M的感染中起作用的假设？ （和其他细胞）通过多种原发性HIV 1菌株，我们将评估亚型B和非囊型B菌株的抗CD63和CD63-EC2易感性，包括专门使用CCR5的抗CCR5（R5，CXCR4） - 热带菌株（R5x4）。

项目成果

Impact of human immunodeficiency virus type 1 reverse transcriptase inhibitor drug resistance mutation interactions on phenotypic susceptibility.

人类免疫缺陷病毒1型逆转录酶抑制剂耐药突变相互作用对表型易感性的影响。

• DOI: 10.1089/aid.2007.0244
• 发表时间: 2008
• 期刊: AIDS research and human retroviruses
• 影响因子: 1.5
• 作者: Trivedi,Vinod;VonLindern,Jana;Montes-Walters,Miguel;Rojo,DanielR;Shell,ElisabethJ;Parkin,Neil;O'Brien,WilliamA;Ferguson,MoniqueR
• 通讯作者: Ferguson,MoniqueR

Quantitative PCR used to assess HIV-1 integration and 2-LTR circle formation in human macrophages, peripheral blood lymphocytes and a CD4+ cell line.

• DOI: 10.1186/1743-422x-7-354
• 发表时间: 2010-12-03
• 期刊: Virology journal
• 影响因子: 4.8
• 作者: Friedrich B;Li G;Dziuba N;Ferguson MR
• 通讯作者: Ferguson MR