人巨细胞病毒体外感染巨核系细胞加重凋亡

为了研究人巨细胞病毒 (HCMV)感染对巨核系细胞加快凋亡的作用机制及HCMV感染引起血小板减少症的机制 ,采用巨核细胞株CHRF 2 88 11和HCMVAD16 9株共同培养 ,用PCR检测HCMVIEA ,用形态学观察、DNALadder形成及AnnexinV/PI流式细胞仪检测分析细胞凋亡情况。结果显示 :HCMVAD16 9株的不同浓度病毒组(10 -3 ,10 -2 ,10 -1)均能显著抑制CHRF细胞的生长。在感染 7天后 ,3组细胞的活率水平分别是 77% ,73%和 6 8% ,而对照组为 98%。用流式细胞仪检测AnnexinV/PI,在感染 7天后 ,10 -3 ,10 -2 和 10 -1病毒组凋亡细胞的百分数是(2 1.3± 2 .4 9) % ,(2 5 .8± 3.6 5 ) %和 (31.4± 3.91) % ,对照组则为 (3.6 8± 1.4 7) %。凋亡率随培养液中病毒浓度的加大和感染后时间的延长而增高 ,二者呈依赖关系。形态学观察和DNALadder的形成进一步证实了凋亡细胞的存在 ,用PCR确定了在CHRF细胞内有HCMVIEA的表达。结论 :HCMV可直接感染巨核系细胞并加重它的凋亡。

Human Cytomegalovirus Aggravates Apoptosis of Human Megakaryocytes via Direct Infection in vitro

The megakaryocyte and platelet lineage may be one of the major sites of human cytomegalovirus (HCMV) infection. However, whether HCMV aggravates apoptosis in normal megakaryocytes was not well investigated. Megakaryocytic cell line CHRF-288-11 and HCMV AD 169 strain were co-cultured in this study. PCR was used to detect the direct infection of the cells by HCMV IEA expression. The apoptotic cells were analyzed by morphologic observation, DNA ladder formation, annexin V/PI and PI assay with flow cytometry. The results showed that HCMV significantly inhibited the growth of CHRF cells in three different concentrations of viral infection groups (10(-3), 10(-2), 10(-1)). The viability levels in each infection groups were 77%, 73% and 68% respectively after incubation for 7 days, compared with 98% in the control group. Using annexin V/PI with flow cytometry, it was shown that the percentages of apoptotic cells viral infection in groups (10(-3), 10(-2), 10(-1)) were (21.3 +/- 2.49)%, (25.8 +/- 3.65)% and (31.4 +/- 3.91)% at 7 days after infection, while the control was (3.68 +/- 1.47)%. The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, PCR detection also showed the direct infection by identification of HCMV IEA expression in CHRF cells. This study suggested that HCMV could directly infect megakaryocytes and aggravated apoptosis in HCMV-infected megakaryocytes.