Specificity analysis of cytotoxic T lymphocytes with peptide libraries.

1.单分子基础上的特异性分析。MHC-I类分子的位置扫描：利用多肽库(数据库正在准备公开发布)分析了三个MHC-1类分子的多肽结合特异性。通过合成和测试&gt；70多肽的结合能力，揭示了利用文库结合数据预测MHC结合多肽的用途和局限性。表位多肽的鉴定：用上述方法预测肿瘤抗原、丙型肝炎病毒和抗肌萎缩蛋白的MHC结合多肽作为移植抗原，并用免疫小鼠或人的PBMC鉴定其抗原性。目的：1.克隆MHC-I类分子胞外部分TOR基因，并在大肠杆菌和杆状病毒中表达。单细胞特异性分析：设计了一系列单氨基酸取代表位多肽。弱活性多肽在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招募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.