人巨细胞病毒感染对脐血淋巴系祖细胞增殖的影响

目的探讨人巨细胞病毒(HCMV)感染对脐血淋巴系祖细胞(CFU-TL)集落增殖能力的影响。方法采用甲基纤维素半固体培养CFU-TL集落,不同浓度HCMV-AD169持续攻击CFU-TL,观察计数CFU-TL集落数、抑制率、集落峰值时间及集落维持时间;用MTT法测量HCMV对宿主细胞增殖活性的影响;用流式细胞仪测定相应表面抗原阳性T-淋巴细胞百分率;并用聚合酶链反应(PCR)技术检测集落细胞内HCMV-AD169DNA。结果HCMV感染组CFU-TL集落数明显低于对照组(P<0.01),抑制程度与病毒感染滴度呈剂量依赖关系;HCMV感染组CFU-TL集落维持时间明显缩短,而各组集落出现时间及集落维持时间无明显差异;HCMV感染组48h、72h宿主细胞增殖指数分别1.59、1.28,感染96h、120h的增殖指数分别为0.58、0.32.;CD3+细胞较培养前明显提高,占集落组成细胞的(90.4±5.6)%,表明集落组成细胞中绝大部分细胞为T淋巴细胞;通过PCR,在HCMV感染组CFU-TL集落细胞内检测到HCMV-AD169存在。结论HCMV在体外试验中能抑制CFU-TL的分化和增殖;淋巴系祖细胞是HCMV的宿主细胞之一,HCMV-AD169能直接感染淋巴系祖细胞。此作用可能是HCMV感染所致的免疫功能低下的主要原因之一。提示HCMV感染引起的细胞免疫功能紊乱与CFU-TL受该病毒抑制有关。

Effect of human cytomegalovirus on colony unit formation of lymphocyte progenitor

Objective] To investigate the effect of human cytomegalovirus (HCMV) on the differentiation and proliferation of colony forming unit T-lymphocyte (CFU-TL). Methods] 20 samples of cord blood were obtained. By the methylcellulose semi-solid colony culture in vitro, the impact of 3 concentrations of HCMVAD169 on the CFU-TL colony formation were surveyed; we measured effect of HCMV on CFU-TL proliferation by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide); the percent of T-lymphocyte was surveyed. HCMV-DNA in CFU-TL was detected by PCR. Result] Compared with the control groups, the CFU-TL colonies in 3 HCMV infection group decreased significantly and with diversity in all the 3 infected groups (P <0.05). It suggested that the suppression and the titer of virus was dose-dependent. The keeping time of CFU-TL colonies infecting HCMV groups was shorter than that of control groups, but the formation time and the peaking time in each group was not different signifficantly. The infected CFU-TL proliferation index after 48 h, 72 h was 1.59, 1.28. The index after 96 h, 120 h was 0.58, 0.32. The colonies consist of CD3+ cell. HCMV-AD169DNA in CFU-TL was detected by PCR in nuclear of CFU-TL. Conclusion] HCMV can inhibit the differentiation and proliferation of lymphocyte progenitors in vitro, which relates to the damage of CFU-TL after HCMV infection.