吉林大学学报（医学版）2016，Vol.42Issue(6)：1071-1075,5.DOI:10.13481/j.1671-587x.20160606
最低限度免疫定义基因表达的HCV多表位DNA疫苗对小鼠细胞免疫的诱导作用
Induction of cellular immunity by HCV multi-epitope DNA vaccine prepared with minimalistic immunologically defined gene expression method in mice
摘要
Abstract
Objective:To study the feasibility of hepatitis C virus (HCV) multi-epitope DNA vaccines prepared by minimalistic immunologically defined gene expression,and to clarify its potential application value in the field of DNA vaccines.Methods:A minimalistic immunologically defined gene expression DNA vaccine (1 346 bp) containing CMV promoter, HCV 1b subtype multi-epitope,and bovine growth hormone (BCG)polyA sequence was prepared by artificial synthesis and PCR methods,then it was named M-HCV-epi.A control DNA fragment, which structure gene was replaced by a DNA fragment without homologous to HCV,was prepared and named V-pcDNA3.1. A total of 12 ICQ mice were randomly divided into experimental group (n=6)and control group (n=6 );the mice were subcutaneously injected with M-HCV-epi and V-pcDNA3.1 with the dosage of 20 μg, respectively.The levels of interferon-γ (IFN-γ)mRNA in spleen cells of the immunized mice were detected by QRT-PCR method.A total of 3 HCV 1b subtype epitope polypeptides and a control polypeptide were synthetically prepared.The levels of IFN-γin culture supernatant of spleen cells were detected by ELISA method after the cells were stimulated with the polypeptides prepared above.Results:Compared with control group,the level of IFN-γmRNA in spleen cells of the mice in experiment group was increased to (1.50±0.18)times (P<0.05);compared with control group,the level of IFN-γin the culture supernatant of spleen cells of the mice in experiment group was increased (P<0.05).Conclusion:The HCV multi-epitope DNA vaccine with minimalistic immunologically defined gene expression can induce the cellular immune response, and may have application values in the field of DNA vaccines.关键词
最低限度免疫定义基因表达/丙型肝炎病毒/多表位/DNA疫苗/干扰素γKey words
minimalistic immunologically defined gene expression/hepatitis C virus/muti-epitope/DNA vaccine/interferonγ分类
生物科学引用本文复制引用
辛桂杰,朱海超,李昊,温剑平..最低限度免疫定义基因表达的HCV多表位DNA疫苗对小鼠细胞免疫的诱导作用[J].吉林大学学报（医学版）,2016,42(6):1071-1075,5.基金项目
吉林省科技厅科研基金资助课题 （）
