TCR--PEPTIDE--MHC INTERACTION IN T CELL CROSSREACTIVITY

TCR--肽--T 细胞交叉反应中的 MHC 相互作用

• 批准号: 6350450
• 负责人: JANKO Z. NIKOLICH
• 金额: $ 13.47万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6350450
• 关键词: MHC class I antigen; T cell receptor; X ray crystallography; cellular immunity; cross immunity; crystallization; cytotoxic T lymphocyte; genetically modified animals; laboratory mouse; laboratory rat; protein structure function; site directed mutagenesis; transfection

The aim of this proposal is to elucidate T cell receptor (TCR) crossreactivity at the molecular level. The unique model developed involves crossreactive TCR recognition of the immunodominant viral peptide coupled to two very closely related class I MHC variants. TCRs to be studied share the same TCRbeta chains, but some see only one of the complexes (monospecific recognition) while others see both (crossreactive recognition). This indicates that the TCRalpha chain must play a decisive role in crossreactivity. TCR sequencing, generation and analysis of single TCR chain transgenic (scTCRTg mice), Crystal structure determination and TCR mutagenesis will be used to dissect the molecular contacts that permit crossreactivity, a phenomenon underlying alloreactivity, tolerance induction, transplant rejection and autoimmunity. The aims are: 1. To analyze functional reactivity and TCR sequence of crossreactive and monospecific CTL clones from normal and scTCRTg mice and establish the preliminary contacts that define crossreactive and monospecific TCR:peptide:MHC interactions for closely related TCRs. 2. To determine the x-ray structures of the monospecific and crossreactive TCRs with the appropriate pep:MHC complexes, and use site-directed mutagenesis to definitively dissect crossreactive recognition.

该提案的目的是在分子水平上阐明 T 细胞受体 (TCR) 交叉反应性。开发的独特模型涉及对与两个非常密切相关的 I 类 MHC 变体偶联的免疫显性病毒肽进行交叉反应 TCR 识别。待研究的 TCR 共享相同的 TCRbeta 链，但有些只看到其中一种复合物（单特异性识别），而另一些则看到两种复合物（交叉反应识别）。这表明TCRα链必定在交叉反应中起决定性作用。 TCR 测序、单 TCR 链转基因（scTCRTg 小鼠）的生成和分析、晶体结构测定和 TCR 诱变将用于剖析允许交叉反应的分子接触，交叉反应是同种异体反应、耐受诱导、移植排斥和自身免疫的潜在现象。目的是： 1. 分析来自正常小鼠和 scTCRTg 小鼠的交叉反应性和单特异性 CTL 克隆的功能反应性和 TCR 序列，并建立定义密切相关 TCR 的交叉反应性和单特异性 TCR:肽:MHC 相互作用的初步联系。 2. 使用适当的 pep:MHC 复合物确定单特异性和交叉反应性 TCR 的 X 射线结构，并使用定点诱变来明确剖析交叉反应性识别。