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目的:研究体内α(1,3)半乳糖转移酶α(1,3)GT反义抑制降低Galα(1,3)Gal(Gal表位)异种抗原的作用。方法:原核显微注射法产生转基因小鼠,PCR和Southern-blot鉴定转基因的整合,RT-PCR检测小鼠α(1,3)GT的表达,组织学染色检查脾脏形态,免疫荧光检测Gal表位,流式细胞仪分析人血清中异种天然抗体(IgM和IgG)和补体(C3c)对鼠源细胞的结合。结果:猪反义α(1,3)半乳糖转移酶转基因小鼠发育正常,但部分杂合体小鼠脾脏有细胞坏死特征。转基因小鼠细胞表面Gal表位数量明显减少。与野生型对照相比,转基因小鼠多种组织的细胞对人血清异种天然抗体的结合降低30％-60％,对补体的结合降低40％-50％。结论:通过反义基因抑制α(1,3)半乳糖转移酶的表达能有效降低人异种反应性。
AIM: To investigate the antisense inhibition of α (1,3) galactosyltransferase α (1,3) GT in vivo by reducing Galα (1,3) Gal (Gal epitope) heterologous antigen. Methods: Transgenic mice were produced by pronuclear microinjection. The integration of transgene was confirmed by PCR and Southern-blot. The expression of α (1,3) GT was detected by RT-PCR. The morphology of spleen was examined by histological staining. The binding of murine cells to different types of human antibodies (IgM and IgG) and complement (C3c) in human serum was analyzed by flow cytometry. Results: The pig antisense α (1,3) galactosyltransferase transgenic mice developed normally, but some heterozygous mice had the characteristics of cell necrosis. The number of Gal epitopes on the surface of transgenic mice was significantly reduced. Compared with the wild-type control, cells from various tissues of transgenic mice showed a 30% -60% reduction in binding to human serum-xenoantibodies and a 40% -50% reduction in complement fixation. CONCLUSION: Inhibition of α (1,3) galactosyltransferase expression by antisense gene is effective in reducing human xenoreactivity.