NATURAL CYTOTOXIC ACTIVITY TO BACTERIA-INFECTED CELLS

对细菌感染细胞的天然细胞毒活性

• 批准号: 3136067
• 负责人: GARY R. KLIMPEL
• 金额: $ 11.31万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 1991-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3136067
• 关键词: Escherichia coli; HeLa cells; Salmonella typhimurium; Shigella; antibody dependent killer cell; bacillary dysentery; bacterial antigens; bacterial genetics; bacterial proteins; bactericidal immunity; cell population study; gene expression; genetic manipulation; host organism interaction; immunotoxicity; intracellular parasitism; laboratory mouse; lymphocyte; membrane activity; microorganism metabolism; monoclonal antibody; mutant; radioassay; surface antigens; tissue /cell culture; virulence

We have recently demonstrated that human peripheral blood lymphocytes (PBL) contain a population of large granular lymphocytes which preferentially lyse HeLa cells infected with Shigella flexneri. In this proposal we will investigate the effector cells involved in this preferential killing of bacteria- infected cells, and the mechanism by which Shigella pretreatment leads to increased susceptibility to cytotoxic effector cell activity. The surface phenotype of cytotoxic effector cells present in human PBL, and in mouse spleen and gut associated lymphoid tissues (GALT) will be determined using different well characterized monoclonal antisera. The specificity of these cytotoxic effector cells will be addressed by using cold target inhibition and monolayer absorption types of experiments using different tumor cells and virus-infected cells. The susceptibility of bacteria-infected cells to soluble factors produced by natural killer (NK) cells will be investigated and compared to other tumor cells and virus-infected cells. To investigate the mechanism(s) by which Shigella renders a cell susceptible to natural cytotoxic effector cell activity, we will investigate the following: 1) requirement for bacteria invasion, 2) requirement for cell membrane alterations and/or presence of bacteria antigens, and 3) require- ment for cell versus bacteria metabolism. Using recombinant DNA methodology, we will isolate the bacterial gene(s) and define the gene product(s) important for demonstrating natural cytotoxic activity. Finally, we will investigate whether natural cytotoxic activity can be demonstrated against cells infected with other facultative intracellular bacteria. Results from these studies will not only contribute significantly to our understanding of immunity to facultative intracellular bacteria but will also provide the information necessary for establishing in vitro models for immune recognition of bacteria-infected cells.

我们最近证明，人类外周血 淋巴细胞（PBL）含有一群大颗粒 淋巴细胞优先裂解感染的HeLa细胞 福氏志贺氏菌 在本提案中，我们将调查 参与优先杀死细菌的效应细胞 感染的细胞，以及志贺氏菌预处理 导致对细胞毒性效应细胞的敏感性增加 活动 细胞毒性效应细胞的表面表型 存在于人PBL和小鼠脾和肠中， 将使用不同的孔测定淋巴组织（GALT） 特征性单克隆抗血清。 其中的特异性 细胞毒性效应细胞将通过使用冷靶向来解决 抑制和单层吸收类型的实验， 不同的肿瘤细胞和病毒感染的细胞。 易感性 细菌感染的细胞产生的可溶性因子 将研究杀伤（NK）细胞，并与其他肿瘤细胞进行比较。 细胞和病毒感染的细胞。 研究机制 志贺氏菌使细胞对天然的细胞毒素敏感， 效应细胞活性，我们将研究以下内容：1） 细菌侵入的需要，2）细胞膜的需要 细菌抗原的改变和/或存在，以及3）需要- 细胞对细菌代谢的影响。 使用重组DNA 方法学，我们将分离细菌基因并定义 对证明天然细胞毒性很重要的基因产物 活动 最后，我们将研究天然细胞毒性是否 可以证明对感染其他病毒的细胞的活性， 兼性胞内细菌。 这些研究的结果 不仅会大大有助于我们理解 对兼性细胞内细菌的免疫力，但也将 为建立体外模型提供必要的信息 用于细菌感染细胞的免疫识别。