The genetic regulation of natural killer-mediated cell lysis in xenograft

异种移植物中自然杀伤介导的细胞裂解的遗传调控

• 批准号: 10671258
• 负责人: MIYAGAWA Shuji
• 金额: $ 1.98万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671258/
• 关键词: cell-mediated lysis; swine endothelial cell; HLA-G; NK cell; β2-macroglobulin; β2ミクログロプリン

We have investigated the effect of HLA-G and HLA-E to NK-mediated direct cytotoxicity, using hamster ovarian tumor cells (CHO) or swine endothelial cell (SEC) transfectants with these molecules.At first, cDNA of human full-length HLA-G (G1 and G3), HLA-E and β2-macroglobulin were prepared and subcloned into the pCAGGS (β-actin promoter).The surface expression of HLA-G (G1) and β2-macroglobulin were confirmed. In addition, co-transfection of β2-microglobulin with HLA-G1 enhanced the level of expression by several times. On the other hand, HLA-G3 was not expressed on the SEC surface.Concerning HLA-E, the expression was checked by FACS and northern blot. The signal peptide of HLA-E was substituted with that of HLA-G for certain expression.Amelioration of NK cell-mediated lysis by the transfectant molecules on CHO or SEC was then tested as an in vitro delayed-type rejection model of a discordant xenograft. HLA-G1 and HLA-E significantly suppressed the NK cell mediated SEC lysis.The expression of HLA-G and/or HLA-E on the cell surface serves as a new approach in overcoming NK-mediated immunity to xenografts.

本研究利用HLA-G和HLA-E基因转染仓鼠卵巢肿瘤细胞（CHO）和猪内皮细胞（SEC），研究了HLA-G和HLA-E对NK细胞直接杀伤活性的影响（G1和G3），制备HLA-E和β2-巨球蛋白，亚克隆入pCAGGS HLA-G（G1）和β2-巨球蛋白（β 2-macroglobulin，β2-macroglobulin）的表达得到证实。此外，β2-微球蛋白与HLA-G1共转染使表达水平提高了数倍。HLA-E的表达用流式细胞仪和北方印迹法检测。用HLA-G的信号肽取代HLA-E的信号肽进行表达，并在CHO或SEC上检测转染分子对NK细胞介导的溶解的改善作用，作为体外不协调异种移植物的延迟型排斥模型。HLA-G1和HLA-E可显著抑制NK细胞介导的SEC裂解，HLA-G和/或HLA-E在细胞表面的表达为克服NK细胞介导的异种移植免疫提供了新的途径。

项目成果

K.Matsunami,S.Miyagawa et al.: "The regulation of natural tiller-mediated swine endothelial cell lysis by HLA-G(G1 and G3)"Transplantation Proceedings. (in press).

K.Matsunami、S.Miyakawa 等人：“HLA-G（G1 和 G3）对自然分蘖介导的猪内皮细胞裂解的调节”移植论文集。

K.Matsunami,S.Miyagawa et al.: "The protection against natural killer-mediated swine endothelial all lysis by HLA-G(G1 and G3) and HLA-E"Transplantation Proceedings. (in press).

K.Matsunami、S.Miyakawa 等人：“HLA-G（G1 和 G3）和 HLA-E 对自然杀伤介导的猪内皮细胞裂解的保护”移植论文集。

K. Matsunami, S. Miyagawa et al.: "The regulation of natural killer-mediated swine endothelial cell lysis by HLA-G(G1 and G3)"Transplantation Proceedings. (in press).

K. Matsunami、S. Miyakawa 等人：“HLA-G（G1 和 G3）对自然杀伤介导的猪内皮细胞裂解的调节”移植论文集。

K. Matsunami, S. Miyagawa et al.: "The protection against natural killer-mediated swine endothelial cell lysis by HLA-G(G1 and G3) and HLA-E"Transplantation Proceedings. (in press).

K. Matsunami、S. Miyakawa 等人：“HLA-G（G1 和 G3）和 HLA-E 对自然杀伤介导的猪内皮细胞裂解的保护”移植论文集。

K. Matsunami, S. Miyagawa, R. Nakai, M. Yamada, and R. Shirakura.: "The regulation of natural killer-mediated swine endoth elial cell lysis by HLA-G(G1 and G3)."Transplant Proc. (in press).

K. Matsunami、S. Miyakawa、R. Nakai、M. Yamada 和 R. Shirakura.：“HLA-G（G1 和 G3）对自然杀伤剂介导的猪内皮细胞裂解的调节。”移植过程。