The strategy for inhibiting NK cell activity by gene technology

利用基因技术抑制NK细胞活性的策略

• 批准号: 12470273
• 负责人: SHIRAKURA Ryota
• 金额: $ 10.3万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470273/
• 关键词: NK cell; HLA-E; HLA-G1; pig eridothelial cell; hlycosyltransferase; 糖転位酵素; β_2microglobulin; β2 microglobulin

Natural killer (NK) cell mediated response plays an important role in xenograft rejection. The strategy for inhibiting NK cell activity to PEC was investigated using HLA-G and HLA-E on pig endothelial cell (PEC).1. The transfected HLA-G1 was easily expressed on PEC. The expressed HLA-G1 significantly suppressed NK-mediated PEC cell lysis.The HLA-G3 was not expressed on PEC. Therefore, the gene introduction of HLA-G3 in PEC showed no protective effect from human NK cells.2. The PEC transfectant with HLA-E genes did not expressed the HLA molecule. However, the modification of the leader sequence of the HLA-E gene successfully induced the expression of the HLA-E molecule. Furthermore, the transfectant expressed both HLA-G1 and HLA-E molecules, thus efficiently enhancing the inhibition of NK-mediated PEC lysis.3. The co-effect of HLA-G1 expression and the remodeling of glycoantigens such as the α-Gal epitope by the introduction of glycosyltransferase genes, was evaluated.The effect of HLA-G1 on NK cell-mediated PEC lysis was lower than that by the glycosyltransferases. In the case of the co-transfectants, such as HLA-G1 + N-acetylglucosaminyltransferase-III and HLA-G1 + α1,2fucosyltransferase, showed significant reductions in direct NK cell-mediated cytotoxicity, compared with the single HLA-G1 transfectant.

自然杀伤（NK）细胞介导的反应在异种移植排斥反应中起重要作用。用HLA-G和HLA-E对猪内皮细胞（PEC）进行抑制NK细胞对PEC活性的策略研究。转染后的HLA-G1易在PEC上表达。表达的HLA-G1显著抑制nk介导的PEC细胞裂解。HLA-G3未在PEC上表达。因此，在PEC中引入HLA-G3基因对人NK细胞没有保护作用。转染HLA- e基因的PEC不表达HLA分子。然而，修饰HLA-E基因的先导序列成功诱导了HLA-E分子的表达。此外，该转染物同时表达HLA-G1和HLA-E分子，从而有效增强了对nk介导的PEC裂解的抑制作用。通过糖基转移酶基因的引入，评估了HLA-G1表达与α-Gal表位等糖抗原重塑的共同作用。HLA-G1对NK细胞介导的PEC裂解的作用低于糖基转移酶。在共转染的情况下，如HLA-G1 + n -乙酰氨基葡萄糖转移酶- iii和HLA-G1 + α1,2聚焦转移酶，与单一HLA-G1转染相比，显示出直接NK细胞介导的细胞毒性显著降低。

项目成果

Miyagawa S, Nakai R, Matsunami K, Kusama T, Shirakura R, et al.: "Co-effect of HLA-G1 and Glycosyltransfeses in Reducing NK cell-Mediated Pig Endothelial Cell Lysis"Transplant Immunol. (In press).

Miyakawa S、Nakai R、Matsunami K、Kusama T、Shirakura R 等人：“HLA-G1 和糖基转移在减少 NK 细胞介导的猪内皮细胞裂解中的共同作用”移植免疫学。

K. Matsunami, S. Miyagawa, et al.: "The possible use of HLA-G1 and G3 in the inhibition of NK cell mediated swine endothelial cell lysis"Clin. Exp. Immunol.. 126. 165-172 (2001)

K. Matsunami、S. Miyakawa 等人：“HLA-G1 和 G3 在抑制 NK 细胞介导的猪内皮细胞裂解中的可能用途”Clin。

K.Matsunami, S.Miyagawa, R.Nakai, A.Murase, R.Shirakura et al.: "The possible use of HLA-G1 and G3 in the inhibition of NK cell mediated swine endothelial cell lysis"Clin.Exp.Immunol. 126. 165-172 (2001)

K.Matsunami、S.Miyakawa、R.Nakai、A.Murase、R.Shirakura 等人：“HLA-G1 和 G3 在抑制 NK 细胞介导的猪内皮细胞裂解中的可能用途”Clin.Exp.Immunol

K.Matsunami, S.Miyagawa, et al.: "The Regulation of Natural Killer-Mediated Swine Endothelial Cell Lysis by HLA-G(G1 and G3)"Transplant Proc. 32. 2087-2088 (2000)

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”移植程序。

Matsunami k, Miyagawa S, Nakai R, Shirakura R, et al.: "Regulates Its Expression and Downregulates NK Cell-Mediated Swine Endothelial Cell Lysis"Transplantation. 27;73. 1582-1589 (2002)

Matsunami k、Miyakawa S、Nakai R、Shirakura R 等人：“调节其表达并下调 NK 细胞介导的猪内皮细胞裂解”移植。