Teimourpour Roghayeh, Peeridogaheh Hadi, Teimourpour Amir, Arzanlou Mohsen, Meshkat Zahra
Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.
Department of Epidemiology and Biostatistics, School of Public Health, Tehran University of Medical science, Tehran, Iran.
Iran J Basic Med Sci. 2017 Oct;20(10):1119-1124. doi: 10.22038/IJBMS.2017.9445.
Tuberculosis (TB) has still remained a global health issue. One third of the world's population is infected with tuberculosis and the current BCG vaccine has low efficiency; hence, it is necessary to develop a new vaccine against TB. The aim of the current study was to evaluate the efficiency of a novel DNA vaccine encoding Mtb32C-HBHA antigen in inducing specific immune responses against Mycobacterium tuberculosis.
A DNA plasmid vaccine expressing Mtb32C-HBHA fusion protein was constructed and its ability in protein expression was examined by RT-PCR and Western blot methods. Female BALB/c mice were vaccinated with 100 μg of purified recombinant vector in an attempt to assess its immunogenicity and protective efficacy. Further, the cytokines, IFN-γ, IL-12, IL-4, IL-10, and TGF-β were assessed.
The levels of all the studied cytokines were significantly increased (<0.05) compared with the control group. IFN-γ production in the group receiving DNA vaccine plus BCG was increased compared with those receiving only DNA vaccine or BCG (<0.001).
The immunogenicity of the new chimeric DNA vaccine was confirmed alone and in combination with BCG. Based on the results of the current study, the constructed DNA vaccine induced the expression of Mtb32C-HBHA fusion protein efficiently . Furthermore, high levels of the specific cytokines were induced in mice. By using this DNA vaccine as a booster after BCG, higher amounts of IFN-γ will be produced.
结核病仍然是一个全球健康问题。世界三分之一的人口感染了结核杆菌，而目前的卡介苗效率较低；因此，有必要研发一种新的抗结核疫苗。本研究的目的是评估一种编码Mtb32C - HBHA抗原的新型DNA疫苗在诱导针对结核分枝杆菌的特异性免疫反应方面的效率。
构建了一种表达Mtb32C - HBHA融合蛋白的DNA质粒疫苗，并通过RT - PCR和蛋白质印迹法检测其蛋白质表达能力。用100μg纯化的重组载体对雌性BALB/c小鼠进行免疫接种，以评估其免疫原性和保护效果。此外，还评估了细胞因子IFN -γ、IL - 12、IL - 4、IL - 10和TGF -β。
与对照组相比，所有研究的细胞因子水平均显著升高（<0.05）。接受DNA疫苗加卡介苗组的IFN -γ产生量高于仅接受DNA疫苗或卡介苗组（<0.001）。
新型嵌合DNA疫苗单独以及与卡介苗联合使用时的免疫原性得到了证实。基于本研究结果，构建的DNA疫苗能有效诱导Mtb32C - HBHA融合蛋白的表达。此外，在小鼠中诱导产生了高水平的特异性细胞因子。在卡介苗接种后使用这种DNA疫苗作为加强剂，将产生更高量的IFN -γ。
