不同基因型别乙型脑炎病毒在不同细胞系上的生长特点比较

Comparison of growth characteristics of different genotypes of Japanese encephalitis virus in different cell lines

目的研究不同基因型别乙型脑炎病毒在不同细胞系上的生长特点, 为乙脑病毒的研究中细胞系的选择提供科学依据。方法选择BHK-21、Vero、C6/36、PK-15、DF-1、N2a、SH-sy5y和MDCK细胞系, 通过空斑测定法和RT-qPCR法评价基因1型(G1, NX1889株)、基因3型(G3, P3株)和基因5型(G5, XZ0934株)乙脑病毒在上述细胞系中的增殖能力。结果 3种基因型别乙脑病毒在BHK-21、Vero、C6/36、DF-1、N2a和PK-15细胞系中存在明显的细胞病变效应(CPE), 在同一种细胞系中引起CPE的特点并未观察到明显不同。病毒感染SH-sy5y和MDCK细胞系后没有明显的CPE出现, 但是在SH-sy5y细胞系中存在病毒的增殖, 而MDCK细胞系为非敏感细胞系。G1、G3和G5型乙脑病毒在BHK-21、Vero和SH-sy5y细胞系中的增殖曲线没有明显差异;在C6/36和PK-15细胞系中, G1型乙脑病毒的滴度高于其他两型乙脑病毒;在DF-1细胞系中, G5型高于其他两者, 在N2a细胞系中, G5型低于其他两者。结论 3种不同基因型别的乙脑病毒在不同细胞系中的增殖存在差异, 可为针对乙脑病毒不同方向的研究提供选择参考。

Objective To analyze the growth characteristics of different genotypes of Japanese encephalitis virus(JEV)in different cell lines,and to provide scientific basis for the selection of cell lines in the study of JEV.Methods BHK-21,Vero,C6/36,PK-15,DF-1,N2a,SH-sy5y and MDCK cell lines were selected.The proliferation ability of genotype 1(NX1889 strain),genotype 3(P3 strain)and genotype 5(XZ0934 strain)JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results Significant cytopathogenic effects(CPE)were observed in BHK-21,Vero,C6/36,DF-1,N2a and PK-15 cell lines across all three JEV genotypes.However,no significant differences in CPE characteristics were observed within the same cell line.SH-sy5y and MDCK cell lines did not show significant CPE,but virus proliferation was detected in SH-sy5y cell line,while MDCK cell line were found to be insensitive to JEV.No significant difference was observed in the proliferation curves of G1,G3 and G5 JEV in BHK-21,Vero and SH-sy5y cell lines.In C6/36 and PK-15 cell lines,the titer of G1 JEV was higher than that of G3 and G5.In DF-1 cell line,G5 demonstrated a higher titer than the other two genotypes,whereas in N2a cell line,G5 showed a lower titer than the other two.Conclusions There are differences in the proliferation of three different genotypes of JEV in different cell lines,which can provide reference for the study of JEV in different directions.