Wang Wenhuan, Feng Fangfang, Lv Jinhui, Xie Zixin, Chen Jun, Zhang Lifang, Li Wenshu
Department of Microbiology and Immunology, Wenzhou Medical University , Wenzhou, China .
Viral Immunol. 2017 Sep;30(7):508-515. doi: 10.1089/vim.2016.0135. Epub 2017 Apr 24.
To prepare the dominant multiepitope fusion antigen ROP2-SAG1 (RSmultiepitope) from Toxoplasma gondii in a prokaryotic system, the major immunodominant region (MIR) of the human hepatitis B virus core antigen (HBcAg(MIR)) was used as a delivery vector. The gene encoding the RSmultiepitope was inserted into HBcAg(MIR), and rHBcAg(MIR)-RSmultiepitope was prepared, purified, and administered to BALB/c mice through intradermal injection. An indirect enzyme-linked immunosorbent assay analysis based on a multiepitope peptide facilitated the specific differentiation of sera obtained from mice immunized with the rHBcAg(MIR)-RSmultiepitope protein, and high titers (greater than 1:6,400) of specific anti-RSmultiepitope antibodies were obtained. Immunized splenocytes demonstrated enhanced IFN-γ production. Based on these results, the HBcAg(MIR) vector is easily applied in vitro for targeting the RSmultiepitope and efficiently presents this target epitope for the induction of significant humoral and cellular immune responses. This study offers a novel strategy for the design of a target epitope delivery system for a toxoplasmosis vaccine.
为了在原核系统中制备来自刚地弓形虫的优势多表位融合抗原ROP2-SAG1(RS多表位),将人乙型肝炎病毒核心抗原的主要免疫优势区(HBcAg(MIR))用作递送载体。将编码RS多表位的基因插入HBcAg(MIR)中,制备、纯化rHBcAg(MIR)-RS多表位,并通过皮内注射给予BALB/c小鼠。基于多表位肽的间接酶联免疫吸附测定分析有助于特异性区分用rHBcAg(MIR)-RS多表位蛋白免疫的小鼠获得的血清,并获得了高滴度(大于1:6400)的特异性抗RS多表位抗体。免疫的脾细胞显示出增强的IFN-γ产生。基于这些结果,HBcAg(MIR)载体易于在体外用于靶向RS多表位,并有效呈递该靶表位以诱导显著的体液和细胞免疫反应。本研究为弓形虫病疫苗的靶表位递送系统设计提供了一种新策略。