人类疱疹病毒6型包膜胆固醇去除及糖蛋白表达

Depeletion of cholesterol content and expression of glycoproteins in envelope of human herpesvirus-6A

目的探讨人类疱疹病毒(HHV)-6型包膜胆固醇去除后,对其包膜糖蛋白表达水平的影响。方法用0,1,2,3,5,10 mmol/L不同浓度的甲基-β-环糊精作用于HHV-6A GS株,经20%蔗糖缓冲液梯度离心,去除甲基-β-环糊精,并纯化病毒,采用胆固醇含量检测试剂盒测定病毒胆固醇含量;纯化后的病毒感染HSB-2细胞后,用免疫蛋白杂交方法检测包膜糖蛋白的表达。结果不同浓度的甲基-β-环糊精作用HHV-6A后,其包膜胆固醇含量与甲基-β-环糊精的浓度呈负相关,从35μg/ml下降到10μg/ml以下;与未经处理的病毒比较,其包膜糖蛋白的表达水平未发生改变,糖蛋白gQ1和gL的表达均处在同一水平。结论甲基-β-环糊精可去除HHV-6A包膜胆固醇,且包膜胆固醇的缺失并未影响包膜糖蛋白的表达。

Objective To study the effect of methyl-β-cyclodextrin on the cholesterol content and the expression of glycoprotein in the envelope of human herpesvirus-6 （HHV-6）. Methods HHV-6A GS strains were treated with 0, 1, 2, 3, 5, 10 mmol/L different concentrations of MβCD. The treated virus was subjected to ultracentrifugation through a 20 96 sucrose cushion to remove MβCD and purified virons, then the cholesterol content of treated HHV-6A was measured with cholesterol assay kit. HSB-2 cells were infected by treated HHV-6A, the expression of viral glycoprotein in the envelope of HHV-6A by using Western blotting post HHV-6A cholesterol depeletion. Results Cholesterol was removed from HHV-6 virions with MβCD, the content of cholesterol decreased from35μg/ml to 10μg/ml below, and was negative relate to the concentration of MβCD, the expression of envelope glycoprotein was not changed post HHV-6A cholesterol depeletion compared with untreated with MβCD. Conclusion MβCD can remove the cholesterol in the HHV-6 envelope, but the expression of envelope glycoprotein was not affected when cholesterol absent.