DEVELOPMENT OF MATERIALS WHICH HAVE INHIBITORY ACTIVITY ON GRAFT-VERSUS-HOST DISEASE ACCOPANIED BY BONE MARROW TRANSPLANTATION

对骨髓移植伴随的移植物抗宿主病具有抑制活性的材料的开发

• 批准号: 05557032
• 负责人: ASANO Yoshihiro
• 金额: $ 6.21万
• 依托单位: TOKYO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05557032/
• 关键词: GVHD; CELL DEATH; CLASS I; TH CELL CLONE; MONOCLONAL ANTIBODY; 骨髄移植; 移植片対宿主反応; 活性化リンパ球

The present study was carried out to develop a specific method to inhibit the graft versus host disease which may occur in organ transplantation and blood transfucion. For this purpose, we used a monoclonal antibody (RE2 antibody) which induces cell death specifically in activated lymphocytes but not resting lymphocytes nor somatic cells.The features of the RE2 mAb are following : 1) RE2 directly killed IL2-dependent T cell clones, ConA- or LPS-activated spleen cells and lymph node cells, but not thymocyte, without participation of serum complement. Fab fragments of RE2 had no cytolytic activity, while the cross-linking of Fab fragments reconstituted the cytotoxicity. 2) Giant holes on the cell membrane were formed within 5 minutes after the treatment with RE2, as observed by scanning electron microscopy. There was no indication of DNA fragmentation nor swelling of mitochondria during the cytolysis, suggesting that the cell death is neither apoptosis nor typical necrosis. 3) Expression cloning study revealed that RE2 reacts with a family of MHC class I molecules including H-2K^K.These results provide evidence for a novel pathway of cell death of activated lymphocytes by membrane excitation. It was also speculated that RE2 induces cell death through unknown molecule which associated with MHC class I molecules.

本研究旨在建立一种特异性的方法来抑制器官移植和输血中可能发生的移植物抗宿主病。为此，我们使用了一种能特异性诱导活化淋巴细胞而非静息淋巴细胞和体细胞死亡的单克隆抗体（RE 2抗体），其特点是：1）RE 2可直接杀伤IL 2依赖性T细胞克隆、ConA或LPS活化的脾细胞和淋巴结细胞，而不杀伤胸腺细胞，不需要血清补体的参与。RE 2的Fab片段没有细胞溶解活性，而Fab片段的交联重建了细胞毒性。2)用扫描电镜观察到，RE 2处理后5分钟内细胞膜上形成了巨大的孔。在细胞溶解过程中没有DNA断裂或线粒体肿胀的迹象，表明细胞死亡既不是凋亡也不是典型的坏死。3)表达克隆研究表明，RE 2可与包括H-2K ^K在内的MHC I类分子家族发生反应，这一结果为细胞膜兴奋导致活化淋巴细胞死亡提供了新的证据。推测RE 2可能通过与MHC I类分子相关的未知分子诱导细胞死亡。

项目成果

Matsuoka, S.: "A nevel type of cell deash of cymphocytes induced by a monoclonal antibody without paticipation of coupleneut" J. Exp. Med.181. 2007-2015 (1995)

Matsuoka, S.：“单克隆抗体诱导的一种新类型的淋巴细胞死亡，无需 Coupleneut 的参与”J. Exp。

Kubo, S., T.Nakayama, M.Furutani-Seiki, H.Kishimoto, K.Hashimoto, M -J.Bae, T.Yokochi, N.Takeda, S.Aizawa, Y.Asano, and T.Tada: "A novel form of self tolerance in transgenic mice with autoreactive T cell receptor alpha and beta chein genes." Int.Immunol.

Kubo, S.、T.Nakayama、M.Furutani-Seiki、H.Kishimoto、K.Hashimoto、M-J.Bae、T.Yokochi、N.Takeda、S.Aizawa、Y.Asano 和 T.Tada：

Matsuoka,S.: "A novel type of cell dcath of lymphocytes induced by a monoclonal antibody without participation of complement." Journal of Experimental Medicine.(in press). (1995)

Matsuoka,S.：“一种由单克隆抗体诱导的新型淋巴细胞死亡，无需补体参与。”

Nagasawa,T.: "Negative selection of thymus-dependent CD4^+8^+ intraepithelial lymphocytes by internal superantigens." Cell.Immunol.147. 158-166 (1993)

Nagasawa,T.：“内部超抗原对胸腺依赖性 CD4^8^ 上皮内淋巴细胞的负选择。”

Kubo, S.: "A novel from of self tolerance in transgent wice with autoreactive T cell receptor α and β chaingenes." Int. Immunol.6. 593-602 (1994)

Kubo，S.：“具有自身反应性 T 细胞受体 α 和 β 链基因的转基因小鼠的自我耐受性。”Int. 593-602。