Regulation of natural killer cell-mediated swine endothelial cell lysis through genetic remodeling of a glycoantigen.
Regulation of natural killer cell-mediated swine endothelial cell lysis through genetic remodeling of a glycoantigen.
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通过糖抗原的遗传重塑来调节自然杀伤细胞介导的猪内皮细胞裂解。
DOI：
10.1093/oxfordjournals.jbchem.a022551
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发表时间：
1999
影响因子：
2.7
通讯作者：
R. Shirakura
中科院分区：
文献类型：
--
作者：
S. Miyagawa;R. Nakai;M. Yamada;M. Tanemura;Y. Ikeda;N. Taniguchi;R. Shirakura
以人外周血单个核细胞(PBMC)或类NK细胞株YT细胞为效应物，以猪内皮细胞(SEC)为靶细胞，研究了糖基转移酶基因对自然杀伤细胞(NK)直接细胞毒作用的影响。将β-1，4-N-乙酰氨基葡萄糖基转移酶III、α2，3-唾液酸基转移酶和α1，2-岩藻糖基转移酶基因导入Sec细胞，获得了几株SEC转基因细胞。这些转基因导致直接和间接NK细胞介导的细胞毒性显著降低，在PBMC中降低了72-94%，在YT细胞中降低了27-72%，同时有效地降低了由α-Gal表位引起的对人类天然抗体的异种抗原性。抗α-Gal表位的单抗或GSI凝集素可显著阻断NK细胞介导的直接细胞毒作用。此外，用α-半乳糖苷酶处理亲本细胞可显著降低细胞毒性。这些结果表明，α-Gal表位不仅参与了超急性排斥反应和急性血管排斥反应，还参与了NK细胞介导的直接细胞毒作用。因此，α-Gal表位的基因重塑以及可能的其他糖抗原的重塑有望成为克服异种移植物间接免疫和直接免疫的新途径。
The effect of remodeling of a glycoantigen such as the alpha-Gal epitope, Galalpha1,3Galbeta1,4GlcNAc-R, by the introduction of glycosyltransferase genes on natural killer (NK) cell-mediated direct cytotoxicity was investigated using human peripheral blood mononuclear cells (PBMC) or an NK-like cell line, YT cells, as an effector, and swine endothelial cells (SEC) as a target. Several SEC transfectants were established by transfection with the genes for beta1,4-N-acetylglucosaminyltransferase III, alpha2, 3-sialyltransferase and alpha1,2-fucosyltransferase. These transfections led to dramatic reductions in both direct and indirect NK cell-mediated cytotoxicity, by 72-94% in the case of PBMC and 27-72% in that of YT cells, in addition to an effective reduction in xenoantigenicity, which is substantially caused by the alpha-Gal epitope, to human natural antibodies. The NK cell-mediated direct cytotoxicity was remarkably blocked by an anti-alpha-Gal epitope monoclonal antibody or GSI lectin which preferentially binds to the epitope. Furthermore, treatment of the parental cells with alpha-galactosidase resulted in a significant reduction in cytotoxicity. These results suggest that the alpha-Gal epitope is involved not only in hyperacute rejection and acute vascular rejection, but also in NK cell-mediated direct cytotoxicity. Thus, the genetic remodeling of the alpha-Gal epitope and probably other glycoantigens as well can be expected to represent a new approach for overcoming not only indirect but also direct immunity to xenografts.