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Construction and immune efficacy of recombinant pseudorabies virus expressing PrM-E proteins of Japanese encephalitis virus genotype ?

机译:表达日本脑炎病毒基因型PrM-E蛋白的重组伪狂犬病病毒的构建和免疫效果

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Japanese encephalitis (JE) is an arboviral disease with high case fatality rates and neurologic or psychiatric sequelae among survivors in Asia, western Pacific countries and northern Australia. Japanese encephalitis virus (JEV) is the cause of JE and the emergence of genotype ? (GI) JEV has displaced genotype III (GIII) as the dominant strains circulating in some Asian regions. The currently available JE vaccines are safe and effective in preventing this disease, but they are developed based on the GIII JEV strains. The recombinant virus PRV TK?/gE?/PrM-E+ which expressed the premembrane (prM) and envelope (E) proteins of JEV SX09S-01 strain (genotype I, GI) was constructed by homologous recombination between the genome of PRV TK?/gE?/LacZ+ digested with EcoRI and plasmid pIE-CAG-PrM-E-BGH. Expression of JEV PrM and E proteins was analyzed by Western blot analysis. Immune efficacy of PRV TK?/gE?/PrM-E+ was further evaluated in mouse model. A recombinant pseudorabies virus (PRV TK?/gE?/PrM-E+) was successfully constructed. Mice experiments showed that PRV TK?/gE?/PrM-E+ could induce a high level of ELISA antibodies against PRV and JEV, as well as high titer of PRV neutralizing antibodies. After challenge with 1?×?107 PFU virulent JEV SX09S-01 strain, the time of death was delayed and the survival rate was improved in PRV TK?/gE?/PrM-E+ vaccinated mice. PRV TK?/gE?/PrM-E+ is a potential vaccine candidate against PRV and JEV GI infection in the future.
机译:日本脑炎(JE)是一种人畜共患病毒疾病,在亚洲,西太平洋国家和澳大利亚北部的幸存者中,病死率很高,并且有神经系统或精神科后遗症。日本脑炎病毒(JEV)是JE的病因和基因型的出现? (GI)JEV已取代基因型III(GIII),成为在亚洲某些地区传播的主要菌株。当前可用的JE疫苗在预防该疾病方面是安全有效的,但它们是基于GIII JEV株开发的。通过PRVTKα基因组之间的同源重组,构建了表达JEV SX09S-01株(基因型I,GI)的前膜(prM)和包膜(E)蛋白的重组病毒PRVTKα/gEα/ PrM-E +。用EcoRI和质粒pIE-CAG-PrM-E-BGH消化的/gEα/ LacZ +。通过蛋白质印迹分析来分析JEV PrM和E蛋白的表达。在小鼠模型中进一步评估了PRVTKα/gEα/ PrM-E +的免疫效力。成功构建了重组伪狂犬病病毒(PRVTKα/gEα/ PrM-E +)。小鼠实验表明,PRVTKα/gEα/ PrM-E +可以诱导高水平的针对PRV和JEV的ELISA抗体,以及高滴度的PRV中和抗体。用1××107 PFU强力JEV SX09S-01毒株攻击后,PRVTKα/gEα/ PrM-E +接种小鼠的死亡时间被延迟,存活率提高。 PRVTKα/gEα/ PrM-E +可能是将来对抗PRV和JEV GI感染的潜在疫苗。

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