腺病毒介导CTLA4Ig基因转染骨髓基质细胞及其对CD34^+细胞粘附力的影响

Transfection of CTLA4Ig-gene in BMSCs and its effects on adherence of the cells to CD34^+ cells

目的 通过腺病毒介导CTLA4Ig在骨髓基质细胞 (Bonemarrowstromalcells ,BMSCs)中的表达 ,探讨CTLA4Ig基因修饰的BMSCs对CD3 4+ 细胞的粘附力的变化。方法 以CTLA4Ig 重组腺病毒按感染复数 (Mutiplicityofinfection ,MOI) 5 0转染BMSCs ,免疫组织细胞化学确定CTLA4Ig的表达 ,免疫磁珠 (MACS)阳性选择分选骨髓CD3 4+ 细胞 ,流式细胞仪检测其纯度 ,通过测定粘附于BMSCs的3 H TdR标记CD3 4+ 细胞的cpm值观察CTLA4Ig基因修饰的骨髓基质细胞的粘附力变化。结果 免疫细胞化学染色示CTLA4Ig表达阳性率达 85 % (MOI =5 0 ) ,弥漫性定位于细胞质 ;免疫磁珠阳性选择获取的骨髓CD3 4+ 细胞纯度可高达 97 8% ;转染组与对照组BMSCs对CD3 4+ 细胞的粘附力无显著差异 (P >0 0 5 )。结论 CTLA4Ig 重组腺病毒能有效介导CTLA4Ig基因在BMSCs中表达 。

Objective To study the changes of adhesion ability to CD34 + cells of CTLA4Ig modified bone marrow stromal cells(BMSCs) mediated by adenovirus vector. Methods The expression and location of CTLA4Ig in the transfected BMSCs were confirmed by immunocytochemical techniques. Values of cpm from the CD34 + cell selected by MACS and marked by 3H TdR after the adhesion to BMSCs were measured to evaluate the changes of adhesion ability due to transfection. Results The rate of CTLA4Ig expression in the transfected BMSCs (MOT=50) was as high as 85%, and the expression was located disseminatedly in the cytoplasm of BMSCs. But the adhesion ability of the transfected BMSCs to CD34 + cells was not significantly different ( P >0.05) from the untransfected BMSCs. Conclusion CTLA4Ig recombinant adenovirus can effect the mediation of CTLA4Ig gene expression in BMSCs but can not effect on the adhesion ability.