AD5/F35腺病毒载体对不同来源造血系统恶性细胞转染效率的研究

Transfection Efficiency of Adenoviral Vector AD5/F35 to Malignant Hematopoietic Cells of Different Origins

本研究比较AD5/F35腺病毒载体对不同来源的血液系统恶性细胞系感染效率的差异和细胞毒性反应。用AD5/F35-EGFP以不同MOI(感染复数)感染不同来源的血液恶性细胞系并用AD5-EGFP作为对照;用流式细胞术检测荧光阳性细胞比例,进行荧光显微镜照相并用MTT法检测病毒对感染靶细胞的杀伤作用。结果表明:对于髓系来源的细胞在MOI为30时,AD5/F35载体的感染效率大于99%,AD5载体在MOI为1000时感染效率为26.4%;对于B系来源的细胞在MOI为1000时,AD5/F35载体感染效率为11.7%,AD5载体为5.7%;AD5/F35和AD5载体都不能有效地感染T系来源细胞,在MOI达到1000也未检测到荧光阳性细胞。在MOI为1000的情况下AD5/F35载体对感染靶细胞也无明显杀伤作用。结论:AD5/F35载体对不同来源的血液恶性细胞的感染具有一定的选择性,对髓系来源细胞感染能力强,对B系来源细胞有一定感染能力,而且对感染靶细胞毒性小。AD5/F35载体优于常用的5型腺病毒载体,在以髓系细胞为靶细胞的基因治疗中有着较好的应用前景。

This study was aimed to investigate the transfection efficiency of adenoviral vector AD5/F35 to hematopoiefic malignant cells lines of various origins and AD5/F35 cytotoxicity. The hematologic malignant cell lines of various origins were transfected by ADS/F35-EGFP at different multiple of infection （ MOI ） and ADS-EGFP was used as control; the proportion of fluorescence positive cells was detected by flow cytometry ; the killing effect of virus on infective target cells was assayed by MTT and observed by fluorescence microscopy. The results showed that the transfection efficiency of AD5/F35 vector to cell line of myeloid origin was 〉 99% at MOI=30, the transfective efficiency of AD5 vector was 26.4% at MOI=1 000 ; the transfection efficiency of ADS/F35 vector and ADS vector to cell line of B cell origin were 11.7% and 5.7%, respectively, at MOI=1 000. AD5/F35 and AD5 vectors could not effectively transfect cells of T cell origin, no fluorescence positive cells were detected at MOI=1 000; no significant killing effect of AD5/ F35 vector on infective target cells was observed at MOI=1 000. It is concluded that ADS/F35 vector infection has difinite selectivity to hematologic malignant cells of varions origin, the infection ability of AD5/F35 vector to cells of myeloid origin is stronger than that to cells of B cell origin, the cytotoxicity of AD5/F35 vector to infective targer cells is small. The ADS/F35 vector is preferable to ADS vector in respect of infection ability and offers good prospects of application in gene therapy for myeloid leukemia cells as target cells.