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目的探讨人类巨细胞病毒(HCMV)感染对脐血造血祖细胞(CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk)体外增殖的抑制作用及其机制。方法20例脐血标本收集于正常足月顺产新生儿。实验共分5组:(1)3个HCMV感染组,每个感染组分别加入0.1 mL的103、104及105空斑形成单位(PFU)HCMV-AD169病毒液于培养体系中;(2)灭活对照组,加入同体积灭活HCMV病毒液;(3)空白对照组,不加HCMV病毒液,代之以同体积的IMDM。采用造血祖细胞体外半固体培养技术,培养、观察、计数HCMV-AD169株对脐血CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落数、抑制率和集落维持时间;并用聚合酶链反应(PCR)技术检测集落细胞内HCMV-DNA。结果(1)在造血祖细胞培养体系中加入不同滴度的HCMV-AD169后,104和105PFU滴度感染对CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落形成均有显著的抑制作用,103PFU滴度感染对CFU-Mix及CFU-Mk集落形成有显著的抑制作用,与空白对照组和灭活对照组比较,差异有显著性(P<0.05)。病毒滴度越高,抑制程度越明显(P<0.05)。(2)104和105PFU滴度感染组CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk集落维持时间较对照组明显缩短(P<0.01),103PFU滴度感染组CFU-Mix和CFU-Mk集落维持时间较对照组明显缩短(P<0.01)。(3)PCR显示3个感染组的CFU-GM、CFU-E、CFU-Mix及CFU-Mk集落细胞内均有HCMV-AD169DNA存在。结论HCMV-AD169能直接感染CFU-GM、CFU-E、BFU-E、CFU-Mix及CFU-Mk造血祖细胞,并抑制造血祖细胞的增殖,这可能与HCMV感染患儿出现粒细胞减少、血小板减少和贫血等造血功能紊乱有关。
Objective To investigate the inhibitory effect of human cytomegalovirus (HCMV) infection on the proliferation of cord blood hematopoietic progenitor cells (CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk) in vitro and its mechanism. Methods Twenty cases of umbilical cord blood samples were collected in normal term newborn infants. The experiment was divided into five groups: (1) three HCMV infection groups, each infected group were added with 0.1 mL of 103,104 and 105 plaque-forming units (PFU) of HCMV-AD169 virus solution in the culture system; (2) In the control group, the same volume of inactivated HCMV virus solution was added. (3) The blank control group without HCMV virus solution was replaced by the same volume of IMDM. In vitro semi-solid culture of hematopoietic progenitor cells was used to culture, observe and count the number of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk cells in HCMV- ; HCMV-DNA in colony cells was detected by polymerase chain reaction (PCR). Results (1) After the HCMV-AD169 with different titer was added in the culture system of hematopoietic progenitor cells, the colonies of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU- The inhibition of CFU-Mix and CFU-Mk colony formation was significantly inhibited by 103PFU titer infection, which was significantly different from the blank control group and the inactivated control group (P <0.05). The higher the virus titer, the more obvious the degree of inhibition (P <0.05). (2) CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk colonies in 104 and 105 PFU titer infected groups were significantly shorter than those in control group (P <0.01) Mix and CFU-Mk colony maintenance time was significantly shorter than the control group (P <0.01). (3) PCR showed that there were HCMV-AD169DNA in CFU-GM, CFU-E, CFU-Mix and CFU-Mk cells in three infected groups. Conclusion HCMV-AD169 can directly infect CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk hematopoietic progenitor cells and inhibit the proliferation of hematopoietic progenitor cells. This may be related to the occurrence of neutropenia, Thrombocytopenia and anemia and other hematopoietic disorders related.