HEV感染血小板模型的建立及其机制的初步研究  

Establishment of a platelet model of HEV infection and preliminary study on its mechanism

作　　者：程丽姗 付萍[1] 吴秉婷[1] 柯玲[1] 刘鱼[1] CHENG Lishan;FU Ping;WU Bingting;KE Ling;LIU Yu(Institute of Blood Transfusion,Chinese Academy of Medical Sciences,Peking Union Medical College,Chengdu 610052,China)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,成都四川610052

出　　处：《中国输血杂志》2019年第1期11-15,共5页Chinese Journal of Blood Transfusion

基　　金：医科院重大协同创新工程(CIFMS2016-I2M-1-018);中央高校基本科研业务费专项资金(3332018125)

摘　　要：目的研究戊型肝炎病毒(hepatitis E virus, HEV)是否感染血小板并进一步在其内复制,以及血小板与HEV的相互作用情况。方法将纯化的血小板与病毒共培养1 h后在PBS中洗涤2次,22℃下继续震荡培养5 d,每天观察病毒的复制情况。使用荧光定量PCR方法检测HEV核酸浓度的变化;免疫荧光染色观察HEV是否感染血小板;流式细胞术分析HEV是否促进血小板活化。结果随着HEV感染量的增加,血小板感染病毒量逐渐增加并趋于平稳,并且感染率最高约为1.22%,免疫荧光结果可同时观察到HEV结合或进入血小板;培养上清中的病毒浓度与d0相比,d4、d5有明显的增长(P<0.01),且沉淀中的病毒浓度在d2有明显的增加(P<0.01);HEV导致CD62P,CD41双阳性血小板比例增加约8%(P<0.01)。结论 HEV在体外实验中可以感染血小板并促进血小板活化,HEV在血小板内的复制特征需进一步研究验证。Objective To investigate whether hepatitis E virus ( HEV) infects platelets and further replicates in them, and the interaction between platelets and HEV. Methods The purified platelets were co-cultured with the virus for 1 hour, washed twice in PBS,and further incubated at 22℃ for 5 days,and the virus replication was observed every day. Fluorescence quantitative PCR was used to detect changes in HEV nucleic acid concentration;immunofluorescence staining was used to observe whether HEV infected platelets;flow cytometry was used to analyze whether HEV promoted platelet activation. Results With the increase of HEV infection,the amount of platelet infection virus increased gradually and stabilized, and the highest infection rate was about 1. 22%. HEV binding or entry into platelets can be observed simultaneously by immunofluorescence;the virus concentration in the culture supernatant was observed. Compared with day 0,there was a significant increase on days 4 and 5( P<0. 01),and the virus concentration in the sediment increased significantly on the 2nd day ( P<0. 01);the proportion of CD62P and CD41 double positive platelets increased by about 8%,and the difference was statistically significant ( P<0. 01). Conclusion HEV can infect platelets and promote platelet activation in vitro. The replication characteristics of HEV in platelets need further study.

关 键 词：血小板 戊型肝炎病毒 输血传播感染 血小板活化 

分 类 号：R373.2[医药卫生—病原生物学] R331.143[医药卫生—基础医学]