人巨细胞病毒对多向造血祖细胞的影响

Effect of human Cytomegalovirus on multipotential hematopoietic progenitors

目的 探讨人巨细胞病毒 (HCMV)对多向造血祖细胞 (CFU Mix)的抑制作用及其机制。方法 采用造血祖细胞体外半固体培养技术 ,研究HCMVAD 16 9株对脐血CFU Mix集落生长的影响 ;用原位聚合酶链反应 (IS PCR)检测集落细胞中的HCMVDNA ;用免疫组化方法检测集落细胞中HCMV早期蛋白P 5 2。结果 HCMVAD 16 9在体外能明显抑制CFU Mix集落生长 ,集落数随病毒感染滴度的增高而减少 ,抑制幅度亦加大的趋势 ;经IS PCR检测发现病毒感染组CFU Mix集落细胞中有HCMVDNA存在 ;免疫组化检测发现病毒感染组CFU Mix集落细胞中有HCMVP 5 2的表达。结论 HCMVAD 16 9可抑制CFU Mix的分化、增殖 ;HCMVAD 16 9对CFU Mix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、贫血的主要原因 ;并证实CFU

Objective To investigate the mechanism and the suppression effect of human Cytomegalovirus (HCMV) on multipotential hematopoietic progenitors(CFU Mix).Methods Colony forming unit assay was applied to observe the effect of HCMV AD169 strain on CFU Mix of cord blood. The technique of IS PCR was also used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU Mix. HCMV early protein P52 was detected with immunohistochemical technique.Results (1)The numbers of CFU Mix colonies were 140±26 in blank group and 136±23 in mock group, and showed a decrease tendency with the increase of HCMV infection, which were 106±14 colonies in 10 3 PFU HCMV group, 89±14 in 10 4 PFU HCMV group and 60±10 in 10 5 PFU HCMV group. The CFU Mix ( P <0.05, n =18) formation was inhibited significantly after HCMV AD 169 strain infection compared with the control group. (2) HCMV AD 169 containing fluids (10 3, 10 4 and 10 5 PFU/ml) were added to the culture system respectively. The suppression effect showed a dose dependent fashion: 21.8% inhibition with 10 3 PFU of HCMV, 34.7% with 10 4 PFU and 55.5% with 10 5 PFU in the CFU Mix assay. (3) HCMV DNA was positively detected in the colony cell of viral infection group by IS PCR, while negative in the control group. (4) HCMV early protein P52 was also identified in the cells of CFU Mix colonies of viral group by immunocytochemical staining.Conclusion HCMV AD169 strain inhibited the differentiation and proliferation of CFU Mix by infecting the hematopoietic progenitors. HCMV may cause the suppression of hematopoiesis by direct infection, which may be the reason of HCMV infection associating with thrombocytopenia, neutropenia, and anemia. There were early expressions of HCMV protein in colony cells infected by virus.