Specificity analysis of cytotoxic T lymphocytes with peptide libraries.

1.基于单个分子的特异性分析。MHC I类分子的定位扫描：使用肽库分析三种MHC I类分子的肽结合特异性（数据库正在准备公开发布）。通过合成和测试> 70个肽的结合能力，揭示了使用文库结合数据预测MHC结合肽的用途和限制。表位肽的鉴定通过上述方法在肿瘤抗原、丙型肝炎病毒和肌营养不良蛋白的序列中预测MHC结合肽作为移植抗原，并通过免疫小鼠或来自人的PBMC鉴定它们的抗原性。可溶性MHC I类分子和可溶性TCR的产生：克隆了MHC I类分子的胞外部分TOR，并在大肠杆菌和杆状病毒中表达.在单细胞基础上进行特异性分析，设计了一系列单氨基酸取代的表位肽。弱活性肽在T细胞识别过程中起拮抗剂的作用。这种活性与它们在识别过程中募集CD8分子的能力密切相关。

1. Specificity analysis on a single molecular basis.a. Positional scanning of MHC class I molecules.Peptide binding specificity was analyzed for three MHC class 1 molecules with the use of peptide libraries (Data Base is in preparation for public release). Use and limit of using the library binding data for prediction of MHC binding peptides were revealed by synthesizing and testing the binding capacity of > 70 peptides.b. Identification of epitope peptides.MHC binding peptides were predicted in the sequence of a tumor antigen, hepatitis C virus and dystrophin, as a transplantation antigen by the above method and their antigenicity was identified by immunizing mice or PBMC from humans.c. Production of soluble MHC class I molecules and a soluble TCR.Extracellular portion of MHC class I molecules, a TOR were cloned and expressed in E.coli and Baculovirus.2. Specificity analysis on a single cell basis.A series of one amino acid substituted epitope peptides were designed. Weakly active peptides acted like antagonists during T cell recognition. Such activity was closely related to their ability to recruit CD8 molecules during recognition.