丙氧鸟苷对转TK基因的人外周血单个核细胞的毒性作用

Ganciclovir toxicity on herpes simplex thymidine ki-nase transduced human peripheral blood mononuc-lear cells

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 .

AIM To explore the way for decreasing graft versus host disease (GVHD) while preserving graft versus leukemia (GVL) effect maximally in allogeneic hematopoietic stem cell transplantation.METHODS Viral thymidine kinase(TK) gene, green fluorescent protein(GFP) gene and the bacterial neomycin resistance(NeoR )gene were introduced into human peripheral blood mononuclear cells(PBMC) with retroviral vector. The transduced cells growth ability were assayed by MTT.RESULTS Transduced PBMNC showed that major transduced cells were T cells and CD4 positive ones took the lead. CD3 expression was 11.4%, CD4, CD8, CD19 and CD33 expression were 7.6%,2.9%,2.1% and 4.7%,respectively. In the genome of the infected target cells, integrated NeoR gene was identified by PCR analysis. The transduced cells' growth ability was inhibited by gancicloir (GCV).CONCLUSION Selectively eliminated TK transduced T lymphocyte in case GVHD develops is possible.