1994 Fiscal Year Annual Research Report

In vitro実験系の開発による自己反応性クローンの選択的分化機構の解析

Research Project

Project/Area Number	04454213
Research Institution	Tokai Univ.School of Med.
Principal Investigator	垣生園子東海大学, 医学部, 教授 (30051618)
Co-Investigator(Kenkyū-buntansha)	佐藤健人東海大学, 医学部, 助手 (50235363) 西村孝司東海大学, 医学部, 助教授 (30143001)
Keywords	細胞抗原レセプター / トランスジェニックマウス / positive selection / negative selection / 胸腺ストローマ細胞 / T細胞クローン
Research Abstract	胸腺内でランダムに発生するT細胞は、分化の段階でpositive及びnegative selectionによって自己・非自己を識別できる個体特有のレパートリーを形成する。本研究では免疫機能の基本となる現象の成立機構を分子レベルで解明することを目指して、理想的実験系開発から開始した。具体的にはT細胞レセプター(TCR)のトランスジェニックマウス(Tg)を作製してその胸腺細胞と既に樹立した胸腺ストローマ細胞クローンを組合わせ、目的とするselection機構誘導に必要な条件や因子の検索を試みた。 (1)OVA特異的MHC class11(I-A^d)拘束性のヘルパーT細胞株より、TCR-α及びβ鎖遺伝子を単離し(VαHDS58-JαBDFL11、Vβ15-Jβ1-Jβ2.2)、受精卵に注入してファウンダーを得た。これらマウスは予想どうり、OVA特異的CD4SP細胞細胞のみが分化した。胸腺に於いてもCD4SP細胞の分化のみが見られた(発表論文7参照)。(2)in vitroに於けるnegative selectionの誘導:対応する抗原に接してapoptosisiが誘導されるのはDPステージで、その誘導には必要なストローマ細胞はI-Adさえ発現していれば胸腺由来でなくても可能であった(発表論文22参照)。(3)positive selectionの引き金:FTOC及びリンパ球の入れ替え実験を行なった結果、、DP細胞が拘束MHCとのinteractionが在る胸腺環境ではTCRの発現は高いが、無い場合は低かった。しかし、後者はCD4からのシグナルを阻止すると高くなった。以上、TCR-Tgを作製し、negative 及びpositive selectionを誘導するin vitroの系を確立し、selection誘導に必要な分子及び条件の一部を明らかにした。さらに、それら結果をもとに、CD4からのシグナルに打ち勝ってTCRの発現低下を阻止することがpositive selectionの引き金であるとのモデルをうちだした。

Research Products
(28 results)

All Other

All Publications (28 results)

[Publications] 1.Kuge,S.: "Superantigen-induced human CD4^+ helper/killer T cell phenomenon:Selective induction of Th1 type of helper/killer T cells and application to tumore immunotherapy." J.Immunol. (in press.). (1995)
[Publications] 2.Makino,K.: "In vitro expansion of human CD4^+ helper/killer T cells reactive to biological response modifier,OK-432." J.Jpn.Soc.Cancer Ther.30. 540-547 (1995)
[Publications] 3.Hattori,K.: "Bispecific antibody-mediated cytotoxicity by CD4^+ and CD8^+ -activated T cells generated from leukemia patients after allogeneic bonemarrow transplantation." Bone Marrow Transplant.(in press). (1995)
[Publications] 4.Ogata,T.: "In vitro targeting of CD8^+ T cells infiltrated into human head and neck squamous cell carcinoma using anti-CD3 x anti-c-erbB-2 bispecific antibody." Biomedical J.2. 15-20 (1995)
[Publications] 5.Kuge,S.: "IL-12 auguments the generation of autologous tumor-reactive CD8^+ cytotoxic T lymphocytes from tumore-infiltrating lymphocytes." Jpn.J.Cancer Res.(in press). (1995)
[Publications] 6.Ohta,S.: "Tumore-associated glycoantigen, syalyl Lewis^a as a target for bispecific antibody-derected adoptive tumor immunotherapy." Immunol.Letters. (in press.). (1995)
[Publications] 7.Sato,T.: "Naive T cells can mediate delayed-type hyper sensitivety (DTH)response in transgenic mice." Eur.J.Immunol. 24. 1512-1516 (1994)
[Publications] 8.Hozumi,K.: "Pro-T cells in fetal thymus express c-kit and RAG-2 but lack the rearangemente of TCR β." Eur.J.Immunol.24. 1339-1344 (1994)
[Publications] 9.Hozumi,K.: "Implication of the common γ chain of the IL-7 receptor in intrathymic development of pro-T cells." Int.Immunol.6. 1451-1454 (1994)
[Publications] 10.Shimada,A.: "Acceleration of diabetes in young NOD mice with peritoneal macrophages." Diabetes Reseach and Clinical Practice. 24. 69-76 (1994)
[Publications] 11.Makino,K.: "Expression of tumore-associated glycoantigen,sialyl Lewis^a,in human head and neck squamous cell carcinoma and its application to tumore immunotherapy." Jpn.J.Cancer Res.85. 887-891 (1994)
[Publications] 12.Tanaka,Y.: "Inhibition of inflammatory liver injury by a monoclonal antibody against lymphocyte function-associated antigen-1." J.mmunol.151. 5088-5095 (1993)
[Publications] 13.Kasuga,A.: "The role of cytotoxic macrophage in non-obese diabetic mice:cytotoxicity against murine mastocytoma and beta-cell lines." Diabetologia. 36. 1252-1257 (1993)
[Publications] 14.Ohta,S.: "Expression of sialyl lewis^X antigen on human T cells." Cellular Immunol.151. 491-497 (1993)
[Publications] 15.Tamauchi,H.: "CD4^+CD8^+cells lacking self-Mls reactiove T cells are induced in mesenteric lymph nodes of Salmonella enteritidis-infected mice." Immunol.Letters.37. 123-130 (1993)
[Publications] 16.Sydora,B.C.: "Intestinal intraepithelial lymphocytes preferetially repopulate the intestial epithelium." Int.Immunol.5. 743-751 (1993)
[Publications] 17.Hozumi,K.: "Characterization of the T cells in aged rat bone marrow." Immunol.Letters.36. 137-144 (1993)
[Publications] 18.Urano,K.: "Potentiation of therapeutic effect of recombinant tumour necrosis factor against B16 mouse melanoma by combination with recombinant interleukin 2." Cytokine. 5. 224-229 (1993)
[Publications] 19.Akiyama,M.: "Lymphocyte infiltration of the skin in transgenic mice carrying the human interlerkin-2 gene." Dermatological Research. 285. 379-384 (1993)
[Publications] 20.Gao,X.: "Thymic nurse cell clone supports the differentiation of CD4^-8^- thymocytes into CD4^+8^+ thymocytes in vitro." Immunol.Letters. 35. 169-176 (1993)
[Publications] 21.Tsukamoto,H.: "Specific targeting of in vitro-activated human antitumour efector cells using anti-CD3 X anti-c-crbB-2 bispecific antibody." Immunol.& Cell Biol.71. 109-115 (1993)
[Publications] 22.Iwabuchi,K.: "Cellular and peptide requirements for in vitro clonal deletion of immature thymocytes." Proc.Nat.Acad.Sci.USA. 89. 9000-9004 (1992)
[Publications] 23.Nishimura,T.: "Generation,propagation and targeting of human CD4^+ helper/killer T cells induced by anti-CD3 monoclonal antibody plus recombinant IL-2:an efficient atrategy for adaptive tumor immunotherapy." J.Immunol. 148. 285-291 (1992)
[Publications] 24.Nishimura,T.: "Human c-cebB-2 Proto-oncogene product as a arget for bispecific-antibody-directed adoptive tumor immunotherapy." Int.J.Cancer. 50. 800-804 (1992)
[Publications] 25.Yanagita,Y.: "Natural cytotoxic T cells responsible for anti-CD3-induced cytotoxicity in mice." Immunol.Letters. 31. 137-142 (1992)
[Publications] 26.Takeuchi,Y.: "Perforin is expressed in CTL populations generated in vivo." Immunol.Letters. 31. 183-188 (1992)
[Publications] 27.Nakamura,Y.: "Large-scale culture system of human CD4^+6 helper/killer T cells for the application to adoptive tumour immunotherapy." Br.J.Cancer. 66. 20-26 (1992)
[Publications] 穂積勝人、垣生園子: "現代病理学大系:6T細胞の発生と分化(胸腺内分化)" 中山書店, 13 (1994)

1994 Fiscal Year Annual Research Report

In vitro実験系の開発による自己反応性クローンの選択的分化機構の解析

Principal Investigator

垣生 園子 東海大学, 医学部, 教授 (30051618)

Research Products

[Publications] 1.Kuge,S.: "Superantigen-induced human CD4^+ helper/killer T cell phenomenon:Selective induction of Th1 type of helper/killer T cells and application to tumore immunotherapy." J.Immunol. (in press.). (1995)

[Publications] 2.Makino,K.: "In vitro expansion of human CD4^+ helper/killer T cells reactive to biological response modifier,OK-432." J.Jpn.Soc.Cancer Ther.30. 540-547 (1995)

[Publications] 3.Hattori,K.: "Bispecific antibody-mediated cytotoxicity by CD4^+ and CD8^+ -activated T cells generated from leukemia patients after allogeneic bonemarrow transplantation." Bone Marrow Transplant.(in press). (1995)

[Publications] 4.Ogata,T.: "In vitro targeting of CD8^+ T cells infiltrated into human head and neck squamous cell carcinoma using anti-CD3 x anti-c-erbB-2 bispecific antibody." Biomedical J.2. 15-20 (1995)

[Publications] 5.Kuge,S.: "IL-12 auguments the generation of autologous tumor-reactive CD8^+ cytotoxic T lymphocytes from tumore-infiltrating lymphocytes." Jpn.J.Cancer Res.(in press). (1995)

[Publications] 6.Ohta,S.: "Tumore-associated glycoantigen, syalyl Lewis^a as a target for bispecific antibody-derected adoptive tumor immunotherapy." Immunol.Letters. (in press.). (1995)

[Publications] 7.Sato,T.: "Naive T cells can mediate delayed-type hyper sensitivety (DTH)response in transgenic mice." Eur.J.Immunol. 24. 1512-1516 (1994)

[Publications] 8.Hozumi,K.: "Pro-T cells in fetal thymus express c-kit and RAG-2 but lack the rearangemente of TCR β." Eur.J.Immunol.24. 1339-1344 (1994)

[Publications] 9.Hozumi,K.: "Implication of the common γ chain of the IL-7 receptor in intrathymic development of pro-T cells." Int.Immunol.6. 1451-1454 (1994)

[Publications] 10.Shimada,A.: "Acceleration of diabetes in young NOD mice with peritoneal macrophages." Diabetes Reseach and Clinical Practice. 24. 69-76 (1994)

[Publications] 11.Makino,K.: "Expression of tumore-associated glycoantigen,sialyl Lewis^a,in human head and neck squamous cell carcinoma and its application to tumore immunotherapy." Jpn.J.Cancer Res.85. 887-891 (1994)

[Publications] 12.Tanaka,Y.: "Inhibition of inflammatory liver injury by a monoclonal antibody against lymphocyte function-associated antigen-1." J.mmunol.151. 5088-5095 (1993)

[Publications] 13.Kasuga,A.: "The role of cytotoxic macrophage in non-obese diabetic mice:cytotoxicity against murine mastocytoma and beta-cell lines." Diabetologia. 36. 1252-1257 (1993)

[Publications] 14.Ohta,S.: "Expression of sialyl lewis^X antigen on human T cells." Cellular Immunol.151. 491-497 (1993)

[Publications] 15.Tamauchi,H.: "CD4^+CD8^+cells lacking self-Mls reactiove T cells are induced in mesenteric lymph nodes of Salmonella enteritidis-infected mice." Immunol.Letters.37. 123-130 (1993)

[Publications] 16.Sydora,B.C.: "Intestinal intraepithelial lymphocytes preferetially repopulate the intestial epithelium." Int.Immunol.5. 743-751 (1993)

[Publications] 17.Hozumi,K.: "Characterization of the T cells in aged rat bone marrow." Immunol.Letters.36. 137-144 (1993)

[Publications] 18.Urano,K.: "Potentiation of therapeutic effect of recombinant tumour necrosis factor against B16 mouse melanoma by combination with recombinant interleukin 2." Cytokine. 5. 224-229 (1993)

[Publications] 19.Akiyama,M.: "Lymphocyte infiltration of the skin in transgenic mice carrying the human interlerkin-2 gene." Dermatological Research. 285. 379-384 (1993)

[Publications] 20.Gao,X.: "Thymic nurse cell clone supports the differentiation of CD4^-8^- thymocytes into CD4^+8^+ thymocytes in vitro." Immunol.Letters. 35. 169-176 (1993)

[Publications] 21.Tsukamoto,H.: "Specific targeting of in vitro-activated human antitumour efector cells using anti-CD3 X anti-c-crbB-2 bispecific antibody." Immunol.& Cell Biol.71. 109-115 (1993)

[Publications] 22.Iwabuchi,K.: "Cellular and peptide requirements for in vitro clonal deletion of immature thymocytes." Proc.Nat.Acad.Sci.USA. 89. 9000-9004 (1992)

[Publications] 23.Nishimura,T.: "Generation,propagation and targeting of human CD4^+ helper/killer T cells induced by anti-CD3 monoclonal antibody plus recombinant IL-2:an efficient atrategy for adaptive tumor immunotherapy." J.Immunol. 148. 285-291 (1992)

[Publications] 24.Nishimura,T.: "Human c-cebB-2 Proto-oncogene product as a arget for bispecific-antibody-directed adoptive tumor immunotherapy." Int.J.Cancer. 50. 800-804 (1992)

[Publications] 25.Yanagita,Y.: "Natural cytotoxic T cells responsible for anti-CD3-induced cytotoxicity in mice." Immunol.Letters. 31. 137-142 (1992)

[Publications] 26.Takeuchi,Y.: "Perforin is expressed in CTL populations generated in vivo." Immunol.Letters. 31. 183-188 (1992)

[Publications] 27.Nakamura,Y.: "Large-scale culture system of human CD4^+6 helper/killer T cells for the application to adoptive tumour immunotherapy." Br.J.Cancer. 66. 20-26 (1992)

[Publications] 穂積勝人、垣生園子: "現代病理学大系:6T細胞の発生と分化(胸腺内分化)" 中山書店, 13 (1994)

垣生園子東海大学, 医学部, 教授 (30051618)