Liu Wen-Jun, Jin Run-Ming, Fu Xiao-Dong, Liu Bin, Guo Qu-Lian, Deng Zheng-Hua
Department of Pediatrics, Union Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430022, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2006 Apr;8(2):85-9.
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.
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-AD169 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-AD169 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.
HCMV-AD169 (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-AD169 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-dependent. 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-AD169 (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-AD169 DNA in the colony cells of the three Infection groups.
HCMV-AD169 strain can infect hematopoietic progenitors of cord blood and inhibit the proliferation of hematopoietic progenitors, associated with anemia, neutropenia and thrombocytopenia in HCMV patients.
本研究旨在探讨人巨细胞病毒(HCMV)对脐血中粒-巨噬细胞集落形成单位(CFU-GM)、红系集落形成单位(CFU-E)、爆式红系集落形成单位(BFU-E)、多能集落形成单位(CFU-Mix)和巨核细胞集落形成单位(CFU-Mk)祖细胞体外增殖的影响及其可能机制。
从自然分娩的正常足月新生儿脐带静脉采集20份脐血标本。本研究分为五组:3个感染组,分别向培养体系中加入0.1 mL 10³、10⁴和10⁵空斑形成单位(PFU)的HCMV-AD169病毒溶液;1个灭活对照组,加入等体积的灭活病毒溶液;1个空白对照组(无HCMV病毒感染的正常祖细胞培养体系)。采用集落形成单位分析法检测HCMV-AD169株对脐血CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的集落形成、抑制率及集落维持时间的影响。应用PCR技术检测培养的CFU-GM、CFU-E、CFU-Mix和CFU-Mk集落细胞中HCMV-DNA的存在情况。
低浓度的HCMV-AD169(10³PFU)对CFU-Mix和CFU-Mk的集落形成有抑制作用(P<0.05),而10⁵PFU和10⁴PFU的HCMV-AD169与空白对照组和灭活对照组相比,导致CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的集落减少(P<0.05)。HCMV对集落形成的抑制作用呈剂量依赖性。10⁵PFU和10⁴PFU HCMV感染组中CFU-GM、CFU-E、BFU-E、CFU-Mix和CFU-Mk的集落维持时间明显短于两个对照组(P<0.01)。低浓度的HCMV-AD169(10³PFU)感染导致CFU-Mix和CFU-Mk的集落维持时间缩短(P<0.01),但对CFU-GM、CFU-E和BFU-E的集落维持时间无影响。PCR扩增显示三个感染组的集落细胞中存在HCMV-AD169 DNA。
HCMV-AD169株可感染脐血造血祖细胞并抑制造血祖细胞的增殖,这与HCMV感染患者的贫血、中性粒细胞减少和血小板减少有关。