人类巨细胞病毒感染对脐血造血祖细胞增殖的影响(英文)

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。

Effect of human cytomegalovirus on proliferation of hematopoietic progenitor cells of cord blood

Objective This study was designed to investigate the effect of human cytomegalovirus(HCMV) on the proliferation of colony forming unit granulocyte-macrophage(CFU-GM),CFU-erythroid(CFU-E),burst forming unit-erythroid(BFU-E),CFU-multipotential(CFU-Mix) and CFU-megakaryocytic(CFU-Mk) progenitor cells of cord blood in vitro as well as the possible mechanism.Methods Twenty cord blood specimens were collected from the umbilical vein of normal full-term neonates delivered spontaneously.This study consisted of five groups: 3 Infection groups in which 0.1 mL 10~3,10~4 and 10~5 plague forming unit(PFU) HCMV-AD_(169) virus solution was added to the culture system,an Inactivated control group in which the equal volume of inactivated virus solution was added,and a Blank control group(normal progenitor cells culture system without HCMV virus infection).Colony forming unit-assay was applied to detect the effects of HCMV-AD_(169) strain on the colony formation,inhibition rate and colony-maintaining duration of CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-Mk of cord blood.PCR technique was used to demonstrate the existence of HCMV-DNA in the colony cells of cultured CFU-GM,CFU-E,CFU-Mix and CFU-Mk.Results HCMV-AD_(169)(10~3 PFU) in low concentration had inhibition effects on colony formation of the CFU-Mix and CFU-MK(P<0.05),whereas 10~5 PFU and 10~4 PFU HCMV-AD_(169) lead to decreased colonies in CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-MK compared with the Blank control and the Inactivated control groups(P<0.05).The suppression effect of HCMV on the colony formation was dose-dependant.The colony-maintaining duration of the CFU-GM,CFU-E,BFU-E,CFU-Mix and CFU-Mk in the 10~5 PFU and 10~4 PFU HCMV infection groups was significantly shorter than that in the two control groups(P<0.01).The low concentration of HCMV-AD_(169)(10~3 PFU) infection resulted in a shortened colony-maintaining duration of the CFU-Mix and CFU-Mk(P<0.01),but had no effects on the colony-maintaining duration of CFU-GM,CFU-E and BFU-E.PCR amplification demonstrated the existence of HCMV-AD_(169) DNA in the colony cells of the three Infection groups.Conclusions HCMV-AD_(169) strain can infect hematopoietic progenitors of cord blood and inhibit the proliferation of hematopoietic progenitors,associated with anemia,neutropenia and thrombocytopenia in HCMV patients.