PROPAGATION OF THYMUS-DERIVED LYMPHOCYTE LINES

胸腺来源的淋巴细胞系的增殖

• 批准号: 3164552
• 负责人: Nancy H. Ruddle
• 金额: $ 3.14万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3164552
• 关键词: B lymphocyte; T lymphocyte; antibody formation; antibody titering; autoradiography; cancer registry /resource; carbon; cell fusion; cell growth regulation; cellular immunity; centrifugation; chromatography; chromosome complement; clone cells; cytokine receptors; delayed hypersensitivity; flow cytometry; fluorescent dye /probe; gel electrophoresis; gene expression; genetic strain; genetically modified animals; histocompatibility antigens; hybrid cells; immunoglobulins; interleukin 2; iodine; laboratory mouse; leukemia; linkage mapping; lymphokines; macrophage; molecular cloning; neoplasm /cancer immunology; northern blottings; nucleic acid sequence; oncogenic virus; radionuclides; radiotracer; scanning electron microscopy; suppressor T lymphocyte; surface antigens; tissue /cell culture; tritium; tumor necrosis factor beta

The goal of these studies is an in-depth investigation of the thymus derived lymphocyte and its products. Lymphotoxin (LT), a product of antigen or mitogen activated T cells, is being studied as a model lymphokine. The long term goals are to study LT's cell of origin, biochemical nature, mechanism of action, relationship to other lymphokines, regulation and role in vivo. Murine lymphotoxin cDNA has been cloned with mRNA from an interleukin-2 maintained T cell clone. Its gene has been isolated and mapped to chromosome 17. The specific aims of the next 5 years are to analyze the structure of the LT gene, to precisely localize it on chromosome 17, to determine the linkage relationship of the genes for LT and tumor necrosis factor (TNF), to obtain maximal expression of LT cDNA, to analyze regulation of LT and TNF, to study expression of the transfected LT gene and to analyze the 5' regulatory sequence of the LT gene. Goals will be accomplished by sequencing the gene, by identifying LT promoter activity, and by expressing LT DNA in prokaryotic and eurkaryotic cells. Interleukin-2 maintained T cell clones will be exposed to several inducing agents and their LT and TNF mRNA analyzed by Northern blots. The murine LT gene will be ligated to strong promoters and its expression studied in human T cell tumors. Similar vectors will be used to study the effect of LT gene overproduction in transgenic mice. Such mice will be analyzed for autoimmune pathology. If the LT gene is overexpressed in the T cells of these mice, and those cells killed, a murine model for AIDS will be available. LT 5' regulatory sequences will be ligated to a neutral gene, chloramphenical acetyle transferase, in order to study regulation of the LT gene in the absence of effects attributable to the product itself. Studies with transfected T cells, B cells, macrophages and transgenic mice will provide insigt into the nature of T cell specificity of LT expression and eventually lead to an understanding of the molecular basis of LT gene activation by antigen.

这些研究的目的是深入调查 胸腺来源的淋巴细胞及其产物。 淋巴毒素 (LT) 抗原或有丝分裂原激活的 T 细胞的产物，正在研究中 作为模型淋巴因子。 长期目标是研究LT细胞 起源、生化性质、作用机制、关系 对体内其他淋巴因子的调节和作用。 鼠淋巴毒素 cDNA 已与来自小鼠淋巴毒素的 mRNA 一起克隆 IL-2维持T细胞克隆。 其基因已被分离 并映射到17号染色​​体。接下来5个的具体目标 多年的时间来分析 LT 基因的结构，以精确 将其定位在 17 号染色体上，以确定连锁 LT 和肿瘤坏死因子 (TNF) 基因的关系， 获得 LT cDNA 的最大表达，分析调控 LT 和 TNF，以研究转染的 LT 基因的表达 并分析LT基因的5'调控序列。 目标将通过对基因进行测序、识别 LT 启动子活性，并通过在原核生物中表达 LT DNA 和真核细胞。 Interleukin-2 维持的 T 细胞克隆将 暴露于多种诱导剂及其 LT 和 TNF 通过 Northern 印迹分析 mRNA。 小鼠 LT 基因将是 连接到强启动子并在人类 T 细胞中研究其表达 细胞肿瘤。 类似的向量将用于研究效果 转基因小鼠中 LT 基因过量产生。 这样的老鼠将会 分析自身免疫病理学。 如果 LT 基因是 在这些小鼠的 T 细胞中过度表达，并且这些细胞被杀死， 艾滋病小鼠模型将问世。 LT 5'监管 序列将连接到中性基因，氯霉素 乙酰转移酶，以研究 LT 基因的调控 不存在产品本身造成的影响。 研究 转染T细胞、B细胞、巨噬细胞和转基因 小鼠将深入了解 LT 的 T 细胞特异性本质 表达并最终导致理解 抗原激活 LT 基因的分子基础。