【摘要】 目的　探讨人巨细胞病毒 (HCMV)对造血系统的影响及其机制。方法　采用造血祖细胞体外半固体培养技术 ,研究HCMVAD16 9株对粒 巨噬系祖细胞 (CFU GM)、红系祖细胞 (CFU E)、多向造血祖细胞 (CFU Mix)及巨核系祖细胞 (CFU MK)集落生长的影响 ;分别用原位聚合酶链反应 (IS PCR)和聚合酶链反应 (PCR)检测CFU GM、CFU Mix、CFU MK及CFU E集落细胞中的HCMVDNA ;用逆转录聚合酶链反应 (RT PCR)检测CFU E集落细胞中的晚期抗原 (lateantigen ,LA)mRNA及CFU MK集落细胞中的前早期抗原 (immediateearlyantigen ,IEA) )mRNA ;用免疫组化方法检测CFU Mix集落细胞中HCMV早期蛋白P5 2。结果　HCMVAD16 9株在体外能明显抑制CFU GM、CFU E ,CFU Mix及CFU MK的生长 ,抑制程度与病毒感染滴度呈剂量依赖关系 ;经IS PCR或PCR检测发现病毒感染组CFU GM ,CFU E ,CFU Mix及CFU MK细胞中有HCMVDNA存在 ;RT PCR检测发现病毒感染组CFU MK细胞中有IEAmRNA的表达 ,而CFU E细胞中的LAmRNA为阴性 ;免疫组化 (IHC)检测发现病毒感染组CFU Mix集落细胞中有HCMVP5 2的表达。结论　HCMVAD16 9可抑制CFU GM ,CFU E ,CFU Mix及CFU MK的分化和增殖 ;HCMVAD16 9对造血系统有直接损害作用。此作用可能是HCMV感染所致的粒细胞和血小板减少及贫血的更多还原

【Abstract】 Objective To investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system. Methods Semi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique. Results HCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P<0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10 3 plague forming unit (PFU), 10 4 PFU and 10 5 PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10 4 PFU group was also identified by immunohistochemical staining. Conclusion HCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia.更多还原