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.

该提案的长期目标是开发能够 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 直接处理逆转录病毒及其产物，在第一阶段 建议我们将研究淋巴细胞对逆转录病毒的特异性反应 转变。拟议的研究基于我们最近的发现： 人类 T 细胞系通常表达 5.5 kb 的 mRNA，编码 血小板衍生生长因子促有丝分裂形式的受体 (PDGF-BB)。该受体 (PDGFR-B) 位于细胞表面。 然而，HTLV-I 感染的 T 细胞系仅表达 4.8 kb PDGFR-B mRNA 编码位于细胞内的蛋白质。我们还有 在一个感染 HIV 的 T 细胞系中检测到相同的 4.8 kb PDGFR-B mRNA 进行了测试，但只有 B 淋巴细胞系中的 5.5 kb mRNA 种类被感染 DNA 病毒 Epstein-Barr (EBV)。我们建议监测变化 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，我们将鉴定并亚细胞 定位 PDGFR-B 蛋白（目标 2）。 PDGFR-B mRNA 的表达将是 使用特定的 PDGFR-B CDNA 通过 Northern 杂交分析检测 探测。 PDGFR-B 蛋白的存在和分子大小。在被感染的 细胞将通过间接免疫荧光检测 使用针对 a 的多克隆抗体进行放射免疫沉淀 对应于人 PDGFR-B 特定区域的合成肽。 这些研究（第一阶段）的结果将表明人类是否 逆转录病毒对淋巴细胞的转化与以下物质的表达相关： 不同大小的异常 PDGFR-B mRNA，从而导致不同的 PDGFR- B蛋白。