CHO-K1细胞糖基化修饰对登革病毒感染的影响  

Effect of glycosylated modification of CHO-K1 cells on dengue virus infection

作　　者：刁莹 王娜 潘文 DIAO Ying;WANG Na;PAN Wen(Institutes of Biology and Medical Sciences,Soochow University,Suzhou 215123,China)

机构地区：[1]苏州大学生物医学研究院,江苏苏州215123

出　　处：《扬州大学学报（农业与生命科学版）》2021年第5期33-38,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金青年基金项目(31400737);江苏省基础医学优势学科资助项目(202101);苏州市科协青年科技人才托举工程项目(202105)。

摘　　要：登革病毒引起的登革热是全球热带亚热带地区主要公共卫生问题。蚊虫细胞来源的高甘露糖型登革病毒更易于感染树突状细胞,但不同糖基化修饰水平是否影响登革病毒感染尚不清楚。以中国仓鼠卵巢CHO-K1细胞及其N-乙酰葡萄糖基转移酶缺陷型Lec1细胞(高甘露型糖基化)作为模型,研究糖基化修饰对登革病毒感染的影响。结果表明:使用相同量登革病毒感染CHO-K1和Lec1细胞,48 h后免疫荧光法检测到Lec1细胞中表达更多的登革病毒E蛋白,qPCR检测到在Lec1细胞中登革病毒E基因复制水平显著高于CHO-K1细胞。Lec1细胞在吸附病毒和病毒入胞水平上也高于CHO-K1细胞。对于含荧光素酶的登革病毒复制子,用酶标仪进行荧光检测显示复制子在CHO-K1和Lec1细胞中的复制水平无明显差异。使用Lec1细胞培养的子代病毒对Vero细胞具有更高的感染性。综上所述,CHO-K1细胞糖基化修饰为Lec1细胞后,更易于感染登革病毒。Dengue fever caused by the dengue virus(DENV)is a major public health problem in tropical and subtropical regions around the world.Mosquito cell-derived high-mannose-type dengue viruses were reported more likely to infect dendritic cells,but whether glycosylation affects virus infection remains unclear.In this study,Chinese hamster ovarian CHO-K1 cells and N-acetylglucosyl transferase-deficient Lec1 cells(high mannose glycosylation)were used as models to study the effects of glycosylated modification on dengue virus infection.The results showed that more dengue virus E protein in Lec1 cells was detected by immunofluorescence and the replication level of dengue virus E gene detected by qPCR in Lec1 cells was significantly higher than that in CHO-K1 cells.They were detected 48 hours after infection with the same amount of dengue virus.Lec1 cells were also higher than CHO-K1 cells in the levels of virus absorption and virus entry.Luciferase reporter assay with microplate reader showed that there was no significant difference in replication levels between CHO-K1 and Lec1 cells in dengue replicons containing luciferase.The progeny virus cultured with Lec1 cells showed higher infectivity to Vero cells.In conclusion,Lec1 cells were more susceptible to dengue virus infection than CHO-K1 cells.

关 键 词：登革病毒 糖基化修饰 中国仓鼠卵巢细胞 酶标仪 

分 类 号：R512.8[医药卫生—内科学]