Molecular analysis for TCR-MHC-peptide interaction in thymic selection in vitro and in vivo

体外和体内胸腺选择中TCR-MHC-肽相互作用的分子分析

• 批准号: 08839017
• 负责人: FUKUI Yoshinori
• 金额: $ 1.34万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08839017/
• 关键词: a single MHC; peptide complex; T cell receptor; GPI; transgenic mice; thymic selection; MHC; in vitro; T細胞分化

The T cell repertoire is shaped by positive and negative selection through the interaction of alphabeta T cell receptor with self-peptide bound to self-major histocompatibility complex (MHC). To elucidate TCR/MHC/peptide interaction determining this reciprocal selection process, we have tried to develop in vitro and in vivo system where a single MHC/peptide ligand is expressed.First, we developed a soluble MHC molecule of which cytoplasimic domain has been replaced with glycosyl-phophatidylinositol (GPI). This molecule complexed with a single peptide was found to be expressed on the cell-surface of any cells, by just incubation the complex with cells. However, the GPI anchored MHC/peptide complex expressed on thymic epithelial cells could not support T cell differentiation.Second, we developed transgenic mouse lines expressing MHC molecules covalently bound to a single peptide at different levels in the thymus. By comparing the fate of T cells in these lines, we found that a single MHC/peptide ligand, depending on the expression level in thymus, can serve as both positively and negatively selecting ligand.

T细胞库是通过α T细胞受体与自身主要组织相容性复合体（MHC）结合的自身肽相互作用进行阳性和阴性选择而形成的。为了阐明TCR/MHC/肽相互作用决定这一相互选择过程，我们尝试在体外和体内建立表达单个MHC/肽配体的系统。发现这种与单个肽复合的分子在任何细胞的细胞表面上表达，只要将复合物与细胞孵育即可。然而，在胸腺上皮细胞上表达的GPI锚定的MHC/肽复合物不能支持T细胞分化。第二，我们开发了在胸腺中不同水平表达与单个肽共价结合的MHC分子的转基因小鼠系。通过比较这些细胞系中T细胞的命运，我们发现单个MHC/肽配体，取决于胸腺中的表达水平，可以作为阳性和阴性选择配体。

项目成果

Hori T.: "Japanese cedar pollinosis and HLA-DP5" Tissue Antigens. 47. 485-491 (1996)

Hori T.：“日本雪松花粉病和 HLA-DP5”组织抗原。

Savoie CJ.: "MHC class I bound peptides of a colon carcinoma cell line,a ki-ras gene-trageted progeny cell line and a B cell line." Cancer Letters. (in press).

Savoie CJ.：“结肠癌细胞系、ki-ras 基因靶向子代细胞系和 B 细胞系的 MHC I 类结合肽。”

Tana T.: "A HLA binding motif-aided peptide epitope library: A novel library design for the screening of HLA-DR4-restricted antigenic peptides recognized by CD4_+ T cells." J.Human Genet.(in press).

Tana T.：“HLA 结合基序辅助肽表位文库：一种用于筛选 CD4_T 细胞识别的 HLA-DR4 限制性抗原肽的新颖文库设计。”

Mori K.: "Persistent elevation of immunoglobulin G titer against the C region of recombinant group A streptococcal M protein in patients with rheumatic fever" Pediatric Research. 39. 336-342 (1996)

Mori K.：“风湿热患者中针对重组 A 组链球菌 M 蛋白 C 区的免疫球蛋白 G 滴度持续升高”儿科研究。

Fukui,Y.,et al.: "Positive and negative CD4+ thymocyte selection by a single MHC class II/peptide ligand affected by its expression level in the thymus." Immunity. 6. 401-410 (1997)

Fukui,Y.,et al.：“单个 MHC II 类/肽配体的阳性和阴性 CD4 胸腺细胞选择受其在胸腺中表达水平的影响。”