1995 Fiscal Year Annual Research Report

移植腎浸潤細胞におけるT細胞抗原レセプターの解析

Research Project

Project/Area Number	06671613
Research Institution	Kitasato University
Principal Investigator	小幡文弥北里大学, 医療衛生学部, 助教授 (60129236)
Keywords	T細胞レセプター / 移植腎拒絶反応 / クローナリティー
Research Abstract	昨年度解析した移植拒絶腎浸潤T細胞のうち1例は、極めて著しいオリゴクローナリティーを示した。すなわち、無作為に選んだ35個のTCRβ鎖cDNAのうち,Vβ17.1-Jβ2.2でCDR3配列も同一のcDNAが17個(全体の約半数)検出された。今年度は、この1例についてさらに確認のためsingle cell解析を行った。まず浸潤細胞に抗CD25抗体と免疫ビーズを作用させて、顕微鏡下でCD25^<+-> single cellsを25個拾った。single cellsからmRNAを分離し、random primerにてcDNAを合成後、Vβ17特異的primerを用いたPCRを行って、各single cellsがVβ17を表現しているか否かを検証した。その結果,6個のsingle cellsがVβ17表現T細胞であることがわかった。さらにその増殖産物をゲルから回収し、塩基配列を決定したところ,6個のうちの5個が同一のCDR3配列を有することが確認された。また,CD4/8特異的primerを用いてPCR解析を行った結果、上述の同一CDR3配列を有する5個の細胞中3個がCD8陽性細胞と考えられた。(残りの2個がCD4-/CD8-を示したのは検出感度の限界のためと考えられる。)これらの結果から、この臨床検体の場合,effector phaseのCD8^+T細胞のうち,アロHLA抗原に対する親和性がより高いT細胞がin vivoでクローン拡大をおこしたものと考えられる。

Research Products
(7 results)

All Other

All Publications (7 results)

[Publications] Obata F. et al.: "T-cell oligoclonality in renal allograft-infiltrating lymphocytes demonstrated by restriced T-cell receptor diversity." Transpl. Proc.27. 471-474 (1995)
[Publications] Watanabe K. et al.: "Factors influencing long-term operational tolerance in dogs treated with fractionated lymphoid irradiation, donor bone marrow cell infusion and fk506." Transpl. Proc.27. 221-224 (1995)
[Publications] Ohnuma H. et al.: "Endognous peptides bound to HLA-A2 molecules expressed by a stomach cancer cell line." J. Exp. Cancer Res.14. 163-170 (1995)
[Publications] Obata F. et al.: "Association of the HLA-DPB1^*0901-DRB1^*1502 haplotype with gastirc cancer in the Japanese." Ann. Cancer Res. Ther.4. 99-102 (1995)
[Publications] Onda K. et al.: "T-cell receptor (TCR) diversity expressed by CD4^+ T cells activated by primary allogeneic HLA-DR stimulation : Estimation of the degree of CDR3 diversity." Scand. J. Immunol.(in press).
[Publications] Kaneko T and Obata F: "Allogeneic recognition of HLA-DRB1^*0406 by T cells with HLA-DRB1^*0403 : Role of amino acid residue 37 of the β sheet in T cell response." Immunobiology. (in press).
[Publications] 小幡文弥: "今日の移植(特集分子免疫学からみた移植免疫)" 日本医学館, (1995)

1995 Fiscal Year Annual Research Report

移植腎浸潤細胞におけるT細胞抗原レセプターの解析

Principal Investigator

小幡文弥北里大学, 医療衛生学部, 助教授 (60129236)

Research Products

[Publications] Obata F. et al.: "T-cell oligoclonality in renal allograft-infiltrating lymphocytes demonstrated by restriced T-cell receptor diversity." Transpl. Proc.27. 471-474 (1995)

[Publications] Watanabe K. et al.: "Factors influencing long-term operational tolerance in dogs treated with fractionated lymphoid irradiation, donor bone marrow cell infusion and fk506." Transpl. Proc.27. 221-224 (1995)

[Publications] Ohnuma H. et al.: "Endognous peptides bound to HLA-A2 molecules expressed by a stomach cancer cell line." J. Exp. Cancer Res.14. 163-170 (1995)

[Publications] Obata F. et al.: "Association of the HLA-DPB1^0901-DRB1^1502 haplotype with gastirc cancer in the Japanese." Ann. Cancer Res. Ther.4. 99-102 (1995)

[Publications] Onda K. et al.: "T-cell receptor (TCR) diversity expressed by CD4^+ T cells activated by primary allogeneic HLA-DR stimulation : Estimation of the degree of CDR3 diversity." Scand. J. Immunol.(in press).

[Publications] Kaneko T and Obata F: "Allogeneic recognition of HLA-DRB1^0406 by T cells with HLA-DRB1^0403 : Role of amino acid residue 37 of the β sheet in T cell response." Immunobiology. (in press).

[Publications] 小幡文弥: "今日の移植(特集分子免疫学からみた移植免疫)" 日本医学館, (1995)

1995 Fiscal Year Annual Research Report

移植腎浸潤細胞におけるT細胞抗原レセプターの解析

Principal Investigator

小幡 文弥 北里大学, 医療衛生学部, 助教授 (60129236)

Research Products

[Publications] Obata F. et al.: "T-cell oligoclonality in renal allograft-infiltrating lymphocytes demonstrated by restriced T-cell receptor diversity." Transpl. Proc.27. 471-474 (1995)

[Publications] Watanabe K. et al.: "Factors influencing long-term operational tolerance in dogs treated with fractionated lymphoid irradiation, donor bone marrow cell infusion and fk506." Transpl. Proc.27. 221-224 (1995)

[Publications] Ohnuma H. et al.: "Endognous peptides bound to HLA-A2 molecules expressed by a stomach cancer cell line." J. Exp. Cancer Res.14. 163-170 (1995)

[Publications] Obata F. et al.: "Association of the HLA-DPB1^*0901-DRB1^*1502 haplotype with gastirc cancer in the Japanese." Ann. Cancer Res. Ther.4. 99-102 (1995)

[Publications] Onda K. et al.: "T-cell receptor (TCR) diversity expressed by CD4^+ T cells activated by primary allogeneic HLA-DR stimulation : Estimation of the degree of CDR3 diversity." Scand. J. Immunol.(in press).

[Publications] Kaneko T and Obata F: "Allogeneic recognition of HLA-DRB1^*0406 by T cells with HLA-DRB1^*0403 : Role of amino acid residue 37 of the β sheet in T cell response." Immunobiology. (in press).

[Publications] 小幡文弥: "今日の移植(特集 分子免疫学からみた移植免疫)" 日本医学館, (1995)

小幡文弥北里大学, 医療衛生学部, 助教授 (60129236)

[Publications] Obata F. et al.: "Association of the HLA-DPB1^0901-DRB1^1502 haplotype with gastirc cancer in the Japanese." Ann. Cancer Res. Ther.4. 99-102 (1995)

[Publications] Kaneko T and Obata F: "Allogeneic recognition of HLA-DRB1^0406 by T cells with HLA-DRB1^0403 : Role of amino acid residue 37 of the β sheet in T cell response." Immunobiology. (in press).

[Publications] 小幡文弥: "今日の移植(特集分子免疫学からみた移植免疫)" 日本医学館, (1995)