HSV-1糖蛋白B和糖蛋白D的T细胞表位重组核酸疫苗的构建及其免疫原性  

Construction of a T cell inducing multi-epitope recombinant DNA vaccine consisting of glycoproteins B and D of HSV-1 and assessment of its immunogenicity

作　　者：王丹凤 马小力[1] WANG Danfeng;MA Xiaoli(Department of Ophthalmology,The First Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院眼科,沈阳110001

出　　处：《中国医科大学学报》2021年第2期141-144,167,共5页Journal of China Medical University

基　　金：辽宁省自然科学基金(201602844)。

摘　　要：目的构建Ⅰ型单纯疱疹病毒(HSV-1)糖蛋白B(gB)和糖蛋白D(gD)的T细胞表位重组核酸疫苗,并研究其作为核酸疫苗的免疫原性。方法登录免疫表位数据库(IEDB),查阅已发表的无症状表位,联合运用SYFPEITHI、IEDB等生物信息学软件,预测潜在的HSV-1 gB和gD的T细胞表位。将已知表位和预测表位串联,设计并合成HSV-1 gB、gD的T细胞多表位基因盒(Tg)。将Tg克隆入真核表达载体pVAX,构建真核表达质粒pVAX-Tg。pVAX-Tg转染293T细胞,鉴定其蛋白表达。将pVAX-Tg免疫小鼠,检测小鼠血清γ干扰素(IFN-γ)水平的变化。结果成功构建真核表达质粒pVAX-Tg,且该质粒能够成功转染细胞并表达目的蛋白。将pVAX-Tg免疫小鼠后,血清IFN-γ水平明显升高(P<0.001)。结论本研究成功设计构建HSV-1 gB和gD的T细胞表位核酸疫苗,其能够在体外真核细胞中表达,并能够有效诱导细胞免疫,为HSV-1多表位核酸疫苗的研究奠定了坚实基础。Objective To design and construct a T cell inducing multi-epitope recombinant DNA vaccine consisting of glycoproteins B(gB)and D(gD)of herpes simplex virus type 1(HSV-1),and assess its immunogenicity.Methods Asymptomatic epitopes were obtained by retrieving published literature from IEDB.Bioinformatics packages including SYFPEITHI and IEDB were used to predict T-cell activating epitopes of HSV-1 gB and gD.We combined known epitopes with predicted epitopes to design and synthesize a T-cell activating gene cassette(Tg)expressing multiple epitopes from HSV-1 gB and gD.Tg was cloned into the eukaryotic expression vector pVAX to express pVAX-Tg.293T cells were transfected with pVAX-Tg to assess protein expression,and the serum level of interferon-γin mice vaccinated with pVAX-Tg was measured.Results pVAX-Tg was successfully constructed.pVAX-Tg transfected cells can express the protein,and serum levels of interferon-γincreased significantly after vaccination with pVAX-Tg(P<0.001).Conclusion This study successfully constructed a multi-epitope T cell inducing DNA vaccine against HSV-1 gB and gD.The vaccine can be expressed in eukaryotic cells in vitro and induce strong cellular immunity in mice.This study provides a foundation for further research into HSV-1 DNA vaccines.

关 键 词：Ⅰ型单纯疱疹病毒 糖蛋白B 糖蛋白D T细胞表位 核酸疫苗 

分 类 号：R772.2[医药卫生—眼科]