人巨细胞病毒体外感染巨核系细胞的实验研究

Experimental study of in vitro infection of human megakaryocytes by human cytomegalovirus

目的探讨人巨细胞病毒(HCMV)对人巨核系细胞的抑制作用及其机制。方法采用体外悬浮培养巨核系细胞技术,观察HCMV AD169株对巨核系祖细胞生长的影响;用聚合酶链反应(PCR)检测HCMV感染巨核系细胞中HCMV即刻早期蛋白(IE)DNA的表达;用流式细胞术检测HCMV感染巨核系细胞上HCMVpp65抗原的表达。结果HCMV AD169株在体外能明显抑制巨核系细胞的增殖。悬浮培养第12天,与对照组相比,HCMV感染组的巨核细胞数减少了82.19%(P<0.05)。HCMV感染巨核系细胞有IE蛋白DNA和pp65抗原的表达,其感染率为(24.5±3.1)%。结论HCMV AD169株在体外能抑制巨核系细胞的增殖,提示HCMV感染引起的血小板减少与其抑制巨核系细胞增殖有关。

Objective To investigate the suppression effect and the mechanism of human cytomegalovirus （HCMV） on human megakaryocytes. Methods Megakaryocytes of 18 cord blood samples in a liquid culture system were infected by HCMV AD169 strain. HCMV immediate early （IE） protein DNA was detected by PCR. Whether there was expression of HCMV pp65 protein in infected megakaryocytes was detected by FACS. Results HCMV AD169 suppressed the formation of megakaryocytes in vitro significantly. Compared with control, the number of megakaryocytes infected by HCMV AD169 decreased 82. 19% （ P 〈 0. 05）. HCMV IE DNA was positively detected in the cultured cells. FACS detected 2-color fluorescent staining with antibodies directed against CD41a and HCMV pp65 antigen on megakaryocytes infected by HCMV, and the infection rate was （24.5±3.1 ） %. Conclusion HCMV AD169 strain inhibited the proliferation and differentiation of megakaryocytes and their precursors by directly infecting them.