人巨细胞病毒感染致造血祖细胞增殖抑制与更昔洛韦的影响(英文)  

Inhibitory effect of ganciclovir on proliferation of cord blood hematopoietic progenitor cells after infection of human cytomegalovirus in vitro

作　　者：刘文君[1] 刘斌[1] 郭渠莲[1] 付晓冬[1] 邓正华[1] 

机构地区：[1]泸州医学院附属医院儿科,四川省泸州市646000

出　　处：《中国组织工程研究与临床康复》2008年第3期593-597,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：四川省教育厅课题资助项目(2004A058)~~

摘　　要：背景:临床研究显示,骨髓移植后人巨细胞病毒感染可阻碍血小板植入、出现表型异常的外周血白细胞,导致骨髓移植失败,这可能是由于人巨细胞病毒感染直接影响了造血祖细胞所致。目的:观察更昔洛韦对人巨细胞病毒感染所致脐血粒-巨噬系祖细胞、红系祖细胞、淋巴系祖细胞、多向造血祖细胞及巨核系祖细胞体外增殖抑制的影响及更昔洛韦对此的保护作用。设计:对比观察性实验。单位:泸州医学院附属医院分子生物学实验室。对象:正常足月顺产新生儿脐血标本20例,每份10mL,由泸州医学院附属医院妇产科提供,产妇对本实验知情同意。实验经医院伦理委员会批准。方法:实验于2004-06/2006-12在泸州医学院附属医院分子生物学实验室完成。采用脐血标本造血祖细胞培养技术,观察、计数100TCID50人巨细胞病毒-AD169感染粒-巨噬系祖细胞、红系祖细胞、淋巴系祖细胞、多向造血祖细胞及巨核系祖细胞后各祖细胞集落数,记录集落维持时间。用PCR和荧光定量PCR技术检测集落细胞内人巨细胞病毒-AD169DNA。将7.5mg/L更昔洛韦作用于人巨细胞病毒感染的造血祖细胞,再分别计集落数、集落维持时间及集落细胞内人巨细胞病毒-AD169DNA。设正常培养的造血祖细胞为空白对照组,设与含灭活性人巨细胞病毒的正常造血祖细胞培养系统为灭活对照组。主要观察指标:①祖细胞集落数、祖细胞集落维持时间。②祖细胞集落细胞内人巨细胞病毒-AD169DNA。结果:①集落数和集落维持时间:人巨细胞病毒感染的祖细胞集落数较对照组明显减少(P<0.01),集落维持时间较对照组明显缩短(P<0.01)。在人巨细胞病毒感染的集落细胞内检测到人巨细胞病毒-DNA的存在;更昔洛韦作用于人巨细胞病毒感染的祖细胞后,与人巨细胞病毒感染的祖细胞比较集落数明显提高,差异有非常显著性意义(P<0.01),集�BACKGROUND： Clinically, in patients undergoing hematopoietic stem cell transplantation （HSCT）, human cytomegalovirus （HCMV） can be associated with delayed platelet engraftment, phenotypically abnormal peripheral blood leukocytes, and graft rejection, possibly through a direct viral effect on hematopoietic progenitor cells after HCMV infection. OBJECTIVE： To investigate the inhibitory effect of ganciclovir （GCV） on proliferation of colony forming unit （CFU） granulocyte-macrophage （CFU-GM）, CFU-erythroid （CFU-E）, CFU T-lymphocyte （CFU-TL）, CFU-multipotential （CFU-Mix） and CFU-megakaryocyte （CFU-Mk） progenitor cells of cord blood （CB） and the protective effects on them. DESIGN： Contrast observational study. SETTING： Department of Molecular Biology, Affiliated Hospital of Luzhou Medical College. PARTICIPANTS： A total of 20 cord blood （CB） samples （with 10 mL for each sample） from fetal umbilical vein of normal term spontaneous delivery neonates were provided by the Department of Gynaecology and Obstetrics, Affiliated Hospital of Luzhou Medical College. All the patients were informed and agreed with the experiment, METHODS： The experiment was carried out in the Department of Molecular Biology, Affiliated Hospital of Luzhou Medical College from June 2004 to December 2006. Colony forming unit-assay was applied to observe the suppression effect of HCMV-AD169 strain on CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk of CB with the presence of GCV. The techniques of polymerase chain reaction （PCR） and fluorescence quantification PCR were used to demonstrate the existence of HCMV-AD169 DNA in the colony cells of cultured CFU-GM, CFU-E, CFU-TL, CFU-Mix and CFU-Mk. Normal progenitor cells culture system was regarded as blank control group; normal progenitor cells culture system with inactivated HCMV fluid as inactivated （IV） control group. MAIN OUTCOME MEASURES： （1） The number and maintaining duration of colonies of cultured progenitor cells were counted by u

关 键 词：巨细胞病毒属 造血干细胞 更昔洛韦 脐血 

分 类 号：R511[医药卫生—内科学] R285.5[医药卫生—临床医学]