MOLECULAR ANALYSIS OF APOPTOSIS IN T-CELLS

T 细胞凋亡的分子分析

• 批准号: 2185350
• 负责人: BARBARA A OSBORNE
• 金额: $ 18.32万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2185350
• 关键词: T cell receptor; T lymphocyte; animal genetic material tag; antibody; antisense nucleic acid; apoptosis; gene expression; genetic library; genetic recombination; genetically modified animals; laboratory mouse; molecular cloning; molecular genetics; nucleic acid sequence; subtraction hybridization; transfection

Programmed cell death (PCD) is a differentiative process that is required for the selective loss of cells during development. The predominant mechanism used for PCD in vertebrates is apoptosis, which is characterized by DNA degradation, chromatin margination along the nuclear envelope and membrane blebbing. While much is known about the triggers which initiate apoptosis, there is little information about the molecular mechanisms which mediate the process. This proposal takes advantage of a transgenic mouse where the majority of the T cells express the same T cell receptor. When this receptor is stimulated by the appropriate antigen in vivo, massive synchronized apoptotic cell death is induced in the T cells. Using these naturally dying thymocytes, we have generated a cDNA library. The library was screened by a plus/minus hybridization protocol, allowing us to isolate several putative cell death genes. It is proposed that cell death recombinants, which will be sequenced, expressed and used to generate antibodies which can aid in the analysis of the putative protein product.Experiments also are presented to examine how the expression of these genes are regulated during cell death in both T cells and non-immune cells.

程序性细胞死亡（PCD）是一个区分过程 在发育过程中选择性损失细胞所必需的。这 脊椎动物中PCD使用的主要机制是凋亡， 以DNA降解为特征，染色质边缘 沿着核包膜和膜爆炸。虽然很多 知道引发凋亡的触发因素，几乎没有 有关介导的分子机制的信息 过程。该建议利用了转基因鼠标 大多数T细胞表达相同的T细胞受体。什么时候 适当的抗原在体内刺激该受体， 在T中诱导大量同步凋亡细胞死亡 细胞。使用这些天然垂死的胸腺细胞，我们产生了一个 cDNA库。图书馆由加号/减 杂交协议，使我们能够隔离几个推定 细胞死亡基因。有人提出细胞死亡重组剂， 将测序，表达和用于生成抗体 可以帮助分析推定的蛋白 也提出了product。 这些基因的表达在两个细胞死亡期间都受到调节 T细胞和非免疫细胞。