CTL SPECIFIC FOR ALLO-MHC PLUS SELF AND TUMOR PEPTIDES

ALLO-MHC PLUS 自身肽和肿瘤肽特异性 CTL

• 批准号: 2095025
• 负责人: LINDA A SHERMAN
• 金额: $ 21.81万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2095025
• 关键词: MHC class I antigen; autoantigens; cell free system; clone cells; cytotoxic T lymphocyte; histocompatibility; laboratory mouse; leukocyte activation /transformation; major histocompatibility complex; oncogenes; peptides; tissue /cell culture; transfection; tumor antigens

DESCRIPTION (Adapted from the applicant's abstract): There is an unusually high frequency of CTL clones that recognize allogeneic MHC molecules. While recognition of MHC determinants, per se, has been thought to underlay this phenomenon, recent structural, biochemical, and cellular studies have indicated that the peptide binding groove of the MHC molecule may normally be occupied by endogenously synthesized peptide. This suggests endogenous peptides may contribute to allorecognition. The applicant has recently obtained direct evidence that such peptide-dependent allorecognition is possible for CTL clones specific for H-2Kb. Further, peptides recognized in association with class I appear to be differentially expressed by cells of different origin. This proposal will analyze the generality of these findings and determine their applicability to define self and tumor antigens. CTL raised against Kb expressed on normal or tumor cell lines will be assessed for recognition of Kb expressed on normal cells, tumor cells, or cells of different species that have been transfected with the Kb gene. CTL clones which discriminate among such targets will be further analyzed to define the role of endogenous peptide in Kb recognition. Peptides prepared by cleavage of cytoplasmic proteins from cells recognized by such CTL will be used to sensitize Kb expressing cells which are not recognized. Where recognition appears to be specific for a tumor associated antigen, the capacity of such allogeneic CTL clones to eliminate tumor cells in vivo will be assessed. To learn more about the origin of the peptide antigens defined by allo-CTL, the hypothesis that they are derived from a select group of cellular proteins will be tested by biochemically purifying cytoplasmic proteins prior to cleavage into peptides. In all cases, proof that CTL clones recognize either class I plus a specific peptide or class I, per se, will be obtained using a cell free system for CTL triggering in which stimulatory antigen is provided in the form of purified immobilized class I molecules and specific peptide antigens.

描述（改编自申请人的摘要）：有一个异常 识别同种异体MHC分子的CTL克隆的高频。 虽然对MHC决定因素的识别本身被认为是底层的 这种现象，最近的结构，生化和细胞研究已有 表明MHC分子的肽结合凹槽通常可能 被内源合成的肽占据。 这表明内源性 肽可能有助于同种识别。 申请人最近有 获得了直接证据，表明这种依赖肽的同种识别是 适用于针对H-2KB的CTL克隆。 此外，肽已识别 与级别的I相关联似乎是由细胞差异表达的 不同的起源。 该建议将分析这些建议的一般性 发现并确定其定义自我和肿瘤的适用性 抗原。 针对在正常或肿瘤细胞系上表达的KB提出的CTL 将评估在正常细胞，肿瘤上表达的Kb的识别 用Kb转染的不同物种的细胞或细胞 基因。 区分此类目标的CTL克隆将进一步 分析以定义内源性肽在Kb识别中的作用。 通过识别细胞的细胞质蛋白裂解制备的肽 通过这样的CTL将用于使KB表达细胞的敏感 认可。 识别似乎特定于肿瘤的地方 相关的抗原，这种同种异体CTL克隆消除的能力 将评估体内肿瘤细胞。 了解有关起源的更多信息 由Allo-CTL定义的肽抗原，假设它们是 源自选择的一组细胞蛋白将通过 在裂解之前，生化纯化的细胞质蛋白 肽。 在所有情况下，证明CTL克隆识别任一类 加上特定的肽或I类，本身将使用单元格获得 用于CTL触发的免费系统，其中提供了刺激性抗原 纯化的固定化I类分子和特定肽的形式 抗原。