ROLE OF NK CYTOTOXIC FACTOR NKCF IN NK CYTOTOXICITY

NK 细胞毒性因子 NKCF 在 NK 细胞毒性中的作用

• 批准号: 3173359
• 负责人: BENJAMIN BONAVIDA
• 金额: $ 11.92万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 1989-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3173359
• 关键词: antibody formation; cell mediated cytotoxicity; gel electrophoresis; high performance liquid chromatography; immunochemistry; immunotoxicity; leukocyte activation /transformation; monoclonal antibody; neoplasm /cancer immunology; tissue /cell culture

Previous studies of the natural killer (NK) lytic mechanism have provided evidence for the stimulus secretion model and led to the postulation that lytic mediators are transferred from the effector to the target cell. Subsequent work in our laboratory has demonstrated that soluble natural killer cytotoxic factors (NKCF) can be detected in the supernatant of effector cells stimulated with NK targets. These factors are NK specific and lyse only NK sensitive targets. Further studies have shown that NKCF conform with known characteristics of the NK CMC reaction and we have postulated that NKCF plays a role in NK CMC cytotoxicity function. The following model for NK CMC was then proposed by which, following effector target cell binding, the target stimulates the effector to release NKCF. These factors then bind to and lyse the target cells. Preliminary studies have shown that NKCF are glycoproteins and can be fractionated on HPLC gel-filtration into cytotoxic fractions of molecular weight 20-30 kilodaltons. Furthermore, we have been able to generate xeno and monoclonal antibodies that can inhibit NK CMC sctivity and in a few instances neutralize NKCF activity. In the proposal we plan to: 1) further characterize and biochemically purify NKCF to homogeneity, 2) characterize antibodies (poly or monoclonal) specific for NKCF, and examine their role in NK CMC reactions, and 3) in collaboration with other investigators, compare NKCF with recently described cytolytic granules or cytotoxins and lymphotoxins derived from NK and CTL lines. We will use conventional methods of protein purification and structure analysis. Conventional and monoclonal antibodies will be generated against NKCF and will be characterized for their role in blocking NK CMC reactions. The studies proposed are significant and unique in the field of CMC. They are also clinically important since they describe naturally derived antitumor cytolytic molecules that may be relevant in immunotherapy. (LB)

先前关于自然杀手（NK）裂解机制的研究已提供 刺激性分泌模型的证据，并导致了这样的假设 溶血介质从效应子转移到靶细胞。 随后在我们实验室的工作表明可溶性自然 可以在上清液中检测到杀手细胞毒性因子（NKCF） 用NK靶标刺激的效应细胞。 这些因素是NK特定的 只裂解NK敏感目标。进一步的研究表明NKCF 符合NK CMC反应的已知特征，我们有 假定NKCF在NK CMC细胞毒性函数中起作用。 这 然后提出了以下NK CMC模型 靶细胞结合，目标刺激效应子释放NKCF。 这些因素然后结合并裂解目标细胞。初步研究 已经表明NKCF是糖蛋白，可以在HPLC上分离 分子量的细胞毒性部分20-30 千瓦尔顿。 此外，我们能够生成Xeno和 可以抑制NK CMC sctitive的单克隆抗体，其中一些抗体 实例中和NKCF活性。 在提案中，我们计划：1） 进一步的特征和生化纯化的NKCF为同质性，2） 表征特异性NKCF的抗体（聚或单克隆），并检查 它们在NK CMC反应中的作用，以及3）与其他合作 研究人员，将NKCF与最近描述的细胞溶解颗粒或 衍生自NK和CTL系的细胞毒素和淋巴毒素。 我们将使用 蛋白质纯化和结构分析的常规方法。 将对NKCF产生常规和单克隆抗体 其在阻断NK CMC反应中的作用将以特征为特征。 这 提出的研究在CMC领域很重要，并且是独一无二的。 他们是 也很重要，因为它们描述了自然得出的抗肿瘤 可能与免疫疗法相关的细胞溶解分子。 （磅）