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首页> 外文期刊>Journal of Virological Methods >A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice
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A suicidal DNA vaccine expressing the fusion protein of peste des petits ruminants virus induces both humoral and cell-mediated immune responses in mice

机译:表达小反刍动物病毒融合蛋白的自杀性DNA疫苗在小鼠中诱导体液和细胞介导的免疫反应

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摘要

Peste des petits ruminants (PPR), a highly contagious disease induced by PPR virus (PPRV), affects sheep and goats. PPRV fusion (F) protein is important for the induction of immune responses against PPRV. We constructed a Semliki Forest virus (SFV) replicon-vectored DNA vaccine ("suicidal DNA vaccine") and evaluated its immunogenicity in BALB/c mice. The F gene of PPRV was cloned and inserted into the SFV replicon-based vector pSCA1. The antigenicity of the resultant plasmid pSCA1/F was identified by indirect immunofluorescence and western blotting. BALB/c mice were then intramuscularly injected with pSCA1/F three times at 14-d intervals. Specific antibodies and virus-neutralizing antibodies against PPRV were quantified by indirect ELISA and microneutralization tests, respectively. Cell-mediated immune responses were examined by cytokine and lymphocyte proliferation assays. The pSCA1/F expressed F protein in vitro and induced specific and neutralizing antibody production, and lymphocyte proliferation in mice. Mice vaccinated with pSCA1/F had increased IL-2 and IL-10 levels after 24-h post first immunization. IFN-gamma and TNF-alpha levels increased from that time point and gradually decreased thereafter. Thus, the Semliki Forest virus replicon-vectored DNA vaccine expressing the F protein of PPRV induced both humoral and cell-mediated immune responses in mice. This could be considered as a novel strategy for vaccine development against PPR. (C) 2015 Elsevier B.V. All rights reserved.
机译:小反刍兽疫(PPR)是一种由PPR病毒(PPRV)引起的高度传染性疾病,会影响绵羊和山羊。 PPRV融合蛋白(F)对于诱导针对PPRV的免疫反应非常重要。我们构建了Semliki森林病毒(SFV)复制子载体DNA疫苗(“自杀性DNA疫苗”),并评估了其在BALB / c小鼠中的免疫原性。克隆了PPRV的F基因,并将其插入基于SFV复制子的载体pSCA1中。通过间接免疫荧光和蛋白质印迹鉴定所得质粒pSCA1 / F的抗原性。然后以14天的间隔向BALB / c小鼠肌肉内注射pSCA1 / F 3次。分别通过间接ELISA和微中和试验对针对PPRV的特异性抗体和病毒中和抗体进行定量。通过细胞因子和淋巴细胞增殖测定法检查细胞介导的免疫应答。 pSCA1 / F在体外表达F蛋白,并诱导特异性和中和抗体的产生以及小鼠淋巴细胞的增殖。首次免疫后24小时,接种pSCA1 / F的小鼠的IL-2和IL-10水平升高。从那个时间点开始,IFN-γ和TNF-α水平升高,此后逐渐降低。因此,表达PPRV F蛋白的Semliki Forest病毒复制子载体DNA疫苗在小鼠中诱导了体液和细胞介导的免疫反应。这可以被认为是针对PPR疫苗开发的新策略。 (C)2015 Elsevier B.V.保留所有权利。

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