Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095, China.
Virol J. 2012 Sep 17;9:204. doi: 10.1186/1743-422X-9-204.
Epitope-based vaccination might play an important role in the protective immunity against Japanese encephalitis virus (JEV) infection. The purpose of the study is to evaluate the immune characteristics of recombinant MVA carrying multi-epitope gene of JEV (rMVA-mep). The synthetic gene containing critical epitopes (B-cell, CTL and Th) of JEV was cloned into the eukaryotic expression vector pGEM-K1L, and the rMVA-mep was prepared. BALB/c mice were immunized with different dosages of purified rMVA-mep and the immune responses were determined in the form of protective response against JEV, antibodies titers (IgG1 and IgG2a), spleen cell lymphocyte proliferation, and the levels of interferon-γ and interleukin-4 cytokines. The results showed that live rMVA-mep elicited strongly immune responses in dose-dependent manner, and the highest level of immune responses was observed from the groups immunized with 107 TCID50 rMVA-mep among the experimental three concentrations. There were almost no difference of cytokines and neutralizing antibody titers among 107 TCID50 rMVA-mep, recombinant ED3 and inactivated JEV vaccine. It was noteworthy that rMVA-mep vaccination potentiates the Th1 and Th2-type immune responses in dose-dependent manner, and was sufficient to protect the mice survival against lethal JEV challenge. These findings demonstrated that rMVA-mep can produce adequate humoral and cellular immune responses, and protection in mice, which suggested that rMVA-mep might be an attractive candidate vaccine for preventing JEV infection.
基于表位的疫苗接种可能在预防日本脑炎病毒(JEV)感染的保护性免疫中发挥重要作用。本研究旨在评估携带 JEV 多表位基因的重组 MVA(rMVA-mep)的免疫特征。含有 JEV 关键表位(B 细胞、CTL 和 Th)的合成基因被克隆到真核表达载体 pGEM-K1L 中,并制备了 rMVA-mep。用不同剂量的纯化 rMVA-mep 免疫 BALB/c 小鼠,并以抗 JEV 保护性反应、抗体滴度(IgG1 和 IgG2a)、脾细胞淋巴细胞增殖以及干扰素-γ和白细胞介素-4 细胞因子水平的形式测定免疫反应。结果表明,活的 rMVA-mep 以剂量依赖性方式引起强烈的免疫反应,在实验的三个浓度中,用 107TCID50 rMVA-mep 免疫的组观察到最高水平的免疫反应。在 107TCID50 rMVA-mep、重组 ED3 和灭活 JEV 疫苗之间,细胞因子和中和抗体滴度几乎没有差异。值得注意的是,rMVA-mep 接种以剂量依赖性方式增强了 Th1 和 Th2 型免疫反应,足以保护小鼠免受致死性 JEV 攻击。这些发现表明 rMVA-mep 可以在小鼠中产生足够的体液和细胞免疫反应和保护,这表明 rMVA-mep 可能是预防 JEV 感染的有吸引力的候选疫苗。