Host Cell Determinants and Retroviral Infection

宿主细胞决定因素和逆转录病毒感染

基本信息

批准号：
6623084
负责人：
GHALIB ABBAS ALKHATIB
金额：
$ 14.9万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-01 至 2005-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6623084
关键词：
CD4 molecule CD8 molecule T lymphocyte genetic techniques host organism interaction human T cell lymphotropic virus type 1 human tissue tissue /cell culture virus infection mechanism virus receptors

项目摘要

DESCRIPTION (Provided by applicant): The human T cell leukemia virus type 1 (HTLV-1) preferentially infects and transforms CD4+ T lymphocytes in vivo and in vitro. The molecular basis underlying this cell tropism is not understood. A number of candidate molecules have been proposed as potential receptors, however, none of them fulfils the requirements for an obligate viral receptor. The overall goal of this application is to determine the host cell molecules involved in HTLV-1 infection. The first hypothesis to be tested is that HTLV-1 utilizes CD4 for entry into susceptible T lymphocytes. Clinical support for this hypothesis is evident in the preferential infection and transformation of CD4+ T cells in the majority of infected patients. We have recently produced preliminary data to support this hypothesis. By using a novel vaccinia-based reporter gene activation assay developed in our laboratory, we are the first to demonstrate that CD4 expression renders murine cells susceptible for HTLV-1 envelope (Env)mediated cell fusion. The second hypothesis to be tested is that, in addition to CD4, a "coreceptor" is required for the preferential infection of CD4+ T lymphocytes in vivo. This hypothesis is based on preliminary data generated in our laboratory showing that transfection of a HeLa cell cDNA library into NIH-3T3 cells can render them fusogenic with HTLV-1Env and that co-expression of CD4 resulted in ten fold increase in the cell fusion signal. Our preliminary data relied largely on a cell fusion assay that measures the fusogenic potential of Env. The purpose of this application is to obtain further evidence for the role of CD4 in infectivity assays and identify the putative HTLV-1 "coreceptor." The specific aims of this proposal are: 1) Analyze the role of CD4 and CD8 in HTLV-1 infection. 2) Determine the identity of the HTLV- 1 "coreceptor." The first specific aim will utilize HTLV- 1 pseudotyped viruses for infection of a receptor-negative cell line to validate the role of CD4 and correlate cell fusion data with viral infectivity. The second specific aim will utilize a novel functional cDNA screening strategy previously used to isolate the HIV coreceptors. Unlike previously used methods, the proposed screening approach makes no assumptions about the mode of action of the fusion coreceptor, except that it allows a receptor-negative cell type to undergo fusion with cells expressing the HTLV- 1 Env. The experimental methods will utilize procedures and reagents previously utilized by the investigator during his published description of the HIV coreceptors.

描述（由申请人提供）：人类T细胞白血病病毒类型1 （HTLV-1）优先感染并在体内转化CD4+ T淋巴细胞体外。该细胞向乳液的分子基础尚不清楚。一个已经提出了候选分子的数量作为潜在受体，但是，他们都没有符合专性病毒受体的要求。该应用的总体目标是确定宿主细胞分子参与HTLV-1感染。要测试的第一个假设是HTLV-1 利用CD4进入易感T淋巴细胞。临床支持该假设在优先感染和转化中很明显大多数感染患者的CD4+ T细胞。我们最近生产了初步数据以支持这一假设。通过使用新颖的基于疫苗的记者基因激活分析在我们的实验室开发，我们是第一个证明CD4表达使鼠细胞易受HTLV-1 包膜（Env）介导的细胞融合。要检验的第二个假设是，除了CD4之外，优先感染还需要一个“ cocector” 在体内的CD4+ T淋巴细胞。该假设基于初步数据在我们的实验室中产生的，表明转染HeLa细胞cDNA 进入NIH-3T3细胞的库可以使它们与HTLV-1ENV相融合，并且 CD4的共表达导致细胞融合信号增加了十倍。我们的初步数据主要依赖于测量的细胞融合测定法 Env的融合潜力。本申请的目的是获得 CD4在感染性测定中的作用的进一步证据，并确定推定的HTLV-1“ colecector”。该提案的具体目的是： 1）分析CD4和CD8在HTLV-1感染中的作用。 2）确定htlv-1“共感染者”的身份。第一个特定目的将利用HTLV-1伪型病毒进行感染受体阴性细胞系以验证CD4的作用并相关的细胞具有病毒感染性的融合数据。第二个特定目标将利用以前用于隔离HIV的新型功能性cDNA筛选策略受体受体。与以前使用的方法不同，提出的筛选方法除了它允许受体阴性细胞类型与细胞融合表达htlv-1 env。实验方法将使用程序以及调查员以前在其出版期间使用的试剂艾滋病毒感受器的描述。