NEW MARKER FOR RETROVIRUS-INFECTED LYMPHOCYTES

逆转录病毒感染淋巴细胞的新标记

• 批准号: 3489322
• 负责人: DAVID H BING
• 金额: $ 5万
• 依托单位: CBR LABORATORIES, INC.
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1991-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3489322
• 关键词: AIDS /HIV diagnosis; HIV infections; RNA; Retroviridae; diagnosis design /evaluation; electrophoresis; gene expression; growth factor receptors; human T cell lymphotropic virus type 1; human T cell lymphotropic virus type 2; human immunodeficiency virus; immunofluorescence technique; lymphocyte; messenger RNA; nucleic acid hybridization; platelet derived growth factor; tissue /cell culture; virus cytopathogenic effect; virus infection mechanism; virus protein

The long term objective of this proposal is to develop reagents that will indicate the extent of, or may intervene in the transformation of human lymphocytes infected by retroviruses. Unlike other studies and findings that deal directly with retroviruses and their products, in this Phase I proposal we will study a specific response of lymphocytes to retroviral transformation. The proposed studies are based on our recent findings that human T-cell lines normally express a 5.5 kb MRNA that codes for the receptor for the mitogenic form of the platelet-derived growth factor (PDGF-BB). This receptor (PDGFR-B) is localized on the cell surface. However, HTLV-I-infected T-cell lines express only a 4.8 kb PDGFR-B MRNA that codes for a protein that is localized intracellularly. We have also detected an identical 4.8 kb PDGFR-B MRNA in one HIV-infected T-cell line tested, but only the 5.5 kb MRNA species in B-lymphocytic lines infected with the DNA-virus, Epstein-Barr (EBV). We propose to monitor changes in the size of PDGFR-B MRNA in lymphocytes before and after retrovirus-infection (Aim 1). Further, we will identify and subcellularly localize the PDGFR-B protein (Aim 2). Expression of PDGFR-B mRNAs will be detected by Northern hybridization analysis using a specific PDGFR-B CDNA probe. The presence and molecular size of PDGFR-B protein. in infected cells will be detected by indirect immunofluorescence and radioimmunoprecipitation using a polyclonal antibody raised against a synthetic peptide corresponding to a specific region of the human PDGFR-B. The results of these studies (Phase 1) will indicate whether human lymphocyte transformation by retroviruses correlates with the expression of aberrant PDGFR-B mRNAs of different sizes and consequently different PDGFR- B proteins.

这项提议的长期目标是开发出能够 表明人类转变的程度，或可能干预人类转变 被逆转录病毒感染的淋巴细胞。与其他研究和发现不同 在这个阶段，直接与逆转录病毒及其产品打交道的 我们将研究淋巴细胞对逆转录病毒的特异性反应 转型。建议的研究是基于我们最近的发现，即 人类T细胞株正常表达5.5kb的信使核糖核酸 促有丝分裂形式的血小板衍生生长因子受体 (PDGF-BB)。该受体(PDGFR-B)定位于细胞表面。 然而，HTLV-I感染的T细胞株仅表达4.8kb的PDGFR-B mRNA 这是一种定位于细胞内的蛋白质的编码。我们还有 在一株HIV感染的T细胞中检测到相同的4.8kb PDGFR-B mRNA 检测，但只有5.5kb的mRNA在B淋巴细胞系中感染 与DNA病毒，爱泼斯坦-巴尔(EBV)。我们建议监测以下方面的变化 治疗前后淋巴细胞中PDGFR-B mRNA的大小 逆转录病毒感染(目标1)。此外，我们将识别和亚细胞 定位PDGFR-B蛋白(目标2)。PDGFR-B mRNAs的表达 利用PDGFR-B特异性CDNA进行Northern杂交检测 探测器。PDGFR-B蛋白的存在和分子大小。在受感染的地方 细胞将通过间接免疫荧光和 多克隆抗体的放射免疫沉淀法 与人PDGFR-B特定区域相对应的合成肽。 这些研究(第一阶段)的结果将表明人类 逆转录病毒诱导的淋巴细胞转化与HSP70的表达 不同大小的异常PDGFR-B mRNAs，从而导致不同的PDGFR- B蛋白。