Department of Pathobiology, Faculty of Veterinary Medicine, University of Tehran, PO Box 14155-6453, Tehran, Iran.
Virus Res. 2012 Aug;167(2):219-25. doi: 10.1016/j.virusres.2012.05.002. Epub 2012 May 16.
There is a growing concern regarding continuous risk of emerging a new influenza pandemic. It is highlighted the need for novel vaccination techniques that quickly and effectively employed to respond to such threats. Although, DNA vaccine is a simple and effective approach to induce antigen specific immune responses, their potency requires further improvement. DNA vaccine encoding conserved antigen of influenza virus could provide protection in various animal models. Therefore, we constructed a plasmid vector encoding M2e-HSP70c sequences, pcDNA/MHc, as a candidate for universal influenza vaccine. The expression of newly constructed vectors was verified by transient transfection of mammalian cells (HEK293T cell line) and western blot analysis using commercial antibodies. Mice were injected subcutaneously (s.c.) by the help of electroporation (IEP) in the footpad area and boosted without IEP with 100 μg of constructed vector. Furthermore, the potency of this construct to provoke humoral immune responses and its protectivity against lethal dose of viral challenge were evaluated. Based on our study, the fusion construct was immunogenic in mice and was able to confer both protection against lethal challenge of H1N1 virus and reduce viral load in lung homogenates of the infected mice.
人们越来越担心新的流感大流行的持续风险。这凸显了需要新的疫苗接种技术,以便快速有效地应对这些威胁。虽然 DNA 疫苗是一种简单有效的诱导抗原特异性免疫应答的方法,但它们的效力需要进一步提高。针对流感病毒保守抗原的 DNA 疫苗可以在各种动物模型中提供保护。因此,我们构建了一个编码流感病毒 M2e-HSP70c 序列的质粒载体 pcDNA/MHc,作为通用流感疫苗的候选物。使用商业抗体通过瞬时转染哺乳动物细胞(HEK293T 细胞系)和 Western blot 分析来验证新构建载体的表达。通过足底区域的电穿孔(IEP)帮助,将构建的载体以 100 μg 的剂量皮下(s.c.)注射到小鼠体内,并在没有 IEP 的情况下加强注射。此外,还评估了该构建体引发体液免疫应答的效力及其对致死剂量病毒攻击的保护作用。根据我们的研究,融合构建体在小鼠中具有免疫原性,能够提供针对 H1N1 病毒致死性攻击的保护,并降低感染小鼠肺匀浆中的病毒载量。