ROLE OF NK CYTOTOXIC FACTOR NKCF IN NK CYTOTOXICITY

先前对自然杀伤细胞（NK）溶解机制的研究提供了 刺激分泌模型的证据，并导致假设， 裂解介质从效应细胞转移到靶细胞。 我们实验室的后续工作表明，可溶性天然 杀伤细胞毒性因子（NKCF）可以在上清液中检测到， 用NK靶刺激的效应细胞。 这些因子是NK特异性的 并且仅裂解NK敏感靶。进一步的研究表明，NKCF 符合NK CMC反应的已知特征，我们有 推测NKCF在NK CMC细胞毒性功能中起作用。 的 然后提出了NK CMC的跟随模型， 靶细胞结合时，靶刺激效应子释放NKCF。 然后这些因子结合并裂解靶细胞。初步研究 已经表明NKCF是糖蛋白，并且可以在HPLC上分级分离 凝胶过滤成分子量为20-30的细胞毒性级分 千道尔顿。 此外，我们已经能够生成xeno， 可以抑制NK CMC活性的单克隆抗体， 这些情况会使科索沃解放军的活动失效 在提案中，我们计划：1） 进一步表征和生化纯化NKCF至均匀，2） 表征NKCF特异性抗体（多聚或单克隆），并检查 它们在NK CMC反应中的作用，以及3）与其他 研究者将NKCF与最近描述的溶细胞颗粒进行比较， 细胞毒素和来自NK和CTL系的细胞毒素。 我们将使用 蛋白质纯化和结构分析的常规方法。 将产生针对NKCF的常规和单克隆抗体， 将表征其在阻断NK CMC反应中的作用。 的 所提出的研究在CMC领域是重要的和独特的。 他们是 临床上也很重要，因为它们描述了天然来源的抗肿瘤 可能与免疫治疗相关的细胞溶解分子。 （LB）

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)