ROLE OF NK CYTOTOXIC FACTOR NKCF IN NK CYTOTOXICITY

以前对自然杀伤(NK)溶解机制的研究已经提供了 刺激分泌模型的证据，并导致假设 裂解介质从效应器转移到靶细胞。 我们实验室的后续工作证明，天然可溶物质 细胞培养上清中可检测到杀伤细胞毒因子(NKCF) NK靶点刺激的效应细胞。这些因素是NK特有的 而Lyse只对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 推测NKCF在NK-CMC细胞毒作用中起一定作用。这个 在此基础上，提出了NK CMC的跟随式模型，并给出了跟随器 靶细胞结合，靶刺激效应器释放NKCF。 然后这些因子结合并裂解靶细胞。初步研究 已经证明NKCF是糖蛋白，可以在高效液相色谱上进行分离 凝胶过滤成相对分子质量为20-30的细胞毒级分 千道尔顿。此外，我们已经能够产生Xeno和 能抑制NK CMC活性的单抗和在少数 实例中和了NKCF活动。在提案中，我们计划：1) 进一步表征和生化提纯NKCF，使其达到均一，2) 鉴定NKCF特异性抗体(多克隆或单抗)，并检测 它们在NK CMC反应中的作用，以及3)与其他 研究人员将NKCF与最近描述的细胞溶解颗粒或 NK和CTL来源的细胞毒素和淋巴毒素。我们将使用 传统的蛋白质纯化和结构分析方法。 将产生针对NKCF和NKCF的常规和克隆抗体 将以其在阻断NK CMC反应中的作用为特征。这个 所提出的研究在CMC领域具有重要而独特的意义。他们是 临床上也很重要，因为他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。(磅)

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)