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目的 探讨减少异体干细胞移植中移植物抗宿主病(GVHD) ,同时最大限度提高移植物抗白血病 (GVL )效应的有效方法 .方法 采用逆转录病毒介导的基因转移方法将 型单纯疱疹病毒胸苷激酶 (HSV- TK)基因、绿色荧光蛋白(GFP)基因及抗新霉素 (N eo R)基因插入人外周血单个核细胞 (PBMC) ,MTT法测定丙氧鸟苷 (GCV)对转化细胞抑制率 .结果 转化细胞表达 GFP,且主要为 T淋巴细胞 ,占11.4% ,而且被转化的 T淋巴细胞中 ,CD4阳性细胞转化率较高 ,占 7.6 % ,CD8,CD19,CD33阳性细胞转化率分别为2 .9% ,2 .1%和 4.7% .PCR鉴定表明 ,转染的人外周血单个核细胞 DNA中整合有 Neo R基因 . MTT法测定丙氧鸟苷(GCV)对转化细胞与未转化细胞抑制率 ,显示转化细胞生长明显受抑 .结论 异体干细胞移植后 ,如产生严重 GVHD应用 GCV选择性清除 HSV- TK基因转导的 T淋巴细胞 ,使控制 GVHD已成可能 .
Objective To explore an effective method to reduce graft-versus-host disease (GVHD) in allogeneic stem cell transplantation and to maximize the effect of graft-versus-leukemia (GVL) .Methods The retroviral-mediated gene transfer method was used to transfer herpes simplex virus thymidine (HSV-TK) gene, green fluorescent protein (GFP) gene and neo-renin (N eo R) gene were inserted into human peripheral blood mononuclear cells (PBMCs). The effects of gonadoside (GCV) Results The transfected cells expressed GFP, mainly T lymphocytes, accounting for 11.4%. Among the T lymphocytes transformed, the percentages of CD4 positive cells were higher, accounting for 7.6%. The conversion rate of CD8, CD19 and CD33 positive cells Were 2.9%, 2.1% and 4.7% respectively.PCR identification showed that the Neo R gene was integrated into the DNA of the peripheral blood mononuclear cells transfected by human gastric cancer cells.MTT assay was used to determine the effect of gonadoside (GCV) The inhibition rate of untransformed cells showed that the growth of transformed cells was significantly inhibited.Conclusion After allogeneic stem cell transplantation, it is possible to control GVHD by GCV selective removal of T lymphocytes transduced by HSV-TK gene in severe GVHD.