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

• Project/Area Number: 08839017
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 免疫の制御機構
• Research Institution: Kyushu University
• Principal Investigator: FUKUI Yoshinori Medical Institute of Bioregulation, Kyushu University, Research Associate, 生体防御医学研究所, 助手 (60243961)
• Co-Investigator(Kenkyū-buntansha): KAMIKAWAJI Nobuhiro Medical Institute of Bioregulation, Kyushu University, Associate Professor, 生体防御医学研究所, 助教授 (90224659)
• Project Period (FY): 1996 – 1997
• Project Status: Completed (Fiscal Year 1997)
• Budget Amount: ¥2,100,000 (Direct Cost: ¥2,100,000); Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
• Keywords: a single MHC / peptide complex / T cell receptor / GPI / transgenic mice / thymic selection / MHC / in vitro / T細胞分化

Research Abstract

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.

Research Products

• [Publications] Savoie CJ.: "MHC class I bound peptides of a colon carcinoma cell line,a Ki-ras gene-targeted progeny cell line and a B cell line." Cancer Letters. (in press).
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 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).
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fukui Y.: "Differential requirement of MHC class II molecules expressed on haematopoietic cells for positive selection of CD4+thymocytes in TCRab and TCRb transgeni mice." Int.Immunol.vol.9. 1385-1391 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ishimoto T.: "In Vitro and Vivo evidence for high frequency of I-Ab-reactive CD4+T cells in HLA-DQ or HLA-DRA transgenic mice lacking endogenou MHC class I and/or class II expression." J.Immunol.vol.159. 3717-3722 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Fukui Y.: "Positive and negative CD4+thymocyte selection by a single MHC class II/peptide ligand affected by its expression level in the thymus." Immunity. vol.6. 401-410 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Takeshita T.: "Identification of HLA-DQ6-derived peptide recognized by mouse class I H-2Db-restricted CD8+T cells in HLA-DQ6 transgenic mid." Jpn.J.Human Genet.vol.42. 225-232 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Savoie CJ,et al.: "MHC class I bound peptides of a colon carcinoma cell line, a Ki-ras gene-targeted progeny cell line and a B cell line." Cancer Letters. (in press).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Tana T., et al.: "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).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fukui Y,et al.: "Differential requirement of MHC class II molecules expressed on haematopoietic cells for positive selection of CD4^+ thymocytes in TCRalphabeta and TCRbeta transgenic mice." Int.Immunol.9. 1385-1391 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ishimoto T,et al.: "In Vitro and In Vivo evidence for high frequency of I-A^b-reactive CD4^+ T cells in HLA-DQ or HLA-DRA transgenic mice lacking endogenous MHC class I and/or class II expression." J.Immunol.159. 3717-3722 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Takeshita T,et al.: "Identification of HLA-DQ6-derived peptide recognized by mouse MHC class I H-2D^b-restricted CD8^+ T cells in HLA-DQ6 transgenic mice." Jpn.J.Human Genet.42. 225-232 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 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).
• [Publications] 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).
• [Publications] Fukui Y.: "Differental requirement of MHC class II molecules expressed on haematopoietic cells for positive selection of CD4+ thymocytes in TCRab and TCRb transgenic mice." Int.Immunol.vol.9. 1385-1391 (1997)
• [Publications] Ishimoto T.: "In Vitro and Vivo evidence for high frequency of I-Ab-reactve CD4+ T cells in HLA-DQ or HLA-DRA transgenic mice lacking endogenous MHC class I and/or class II expression." J.Immunol.vol.159. 3717-3722 (1997)
• [Publications] Fukui Y.: "Positive and negative CD4+ thymocyte selection by a single MHC class II/peptide ligand affected by its expression level in the thymus." Immunity. vol.6. 401-410 (1997)
• [Publications] Takeshita T.: "Identification of HLA-DO6-derived peptide recognized by mouse MHC class I H-2Db-restricted CD8+ T cells in HLA-DQ6 transgenic mice." Jpn.J.Human Genet.vol.42. 225-232 (1997)
• [Publications] Fukui Y.: "Positive and negative CD4^+ themocyte selection by a single MHC class II/peptide ligand in vivo affected by its expression level" Immunity. (in press).
• [Publications] Takeshita T.: "Identification of HLA-DQ6-derived peptide recognized by mouse MHC class I H-2D^b-restricted CD8^+ T cells in HLA-DO6 transgenic mice" Jpn.J.Human Genet.(in press).
• [Publications] Hori T.: "Japanese cedar pollinosis and HLA-DP5." Tissue Antigens. 47. 485-491 (1996)
• [Publications] 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)
• [Publications] Min W.: "Identification of an epitope for T cells correlated with antibody response to hepatitis B surface antigen in vaccinated humans." Human Immunol.46. 93-99 (1996)