Torabi Azam, Tahmoorespur Mojtaba, Vahedi Fatemeh, Mosavari Nader, Nassiri Mohammadreza
Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran,
Iran J Immunol. 2013 Dec;10(4):205-15.
Tuberculosis is a disease with high morbidity, caused mainly by Mycobaterium tuberculosis (M.tb.). DNA vaccines show a promising future due to their unique advantages over conventional methods. The early-secreted antigen target (ESAT)-6 and culture filtrate protein (CFP)-10 of M.tb. antigens have been identified as vaccine candidates against Mycobacteria and used as subunit vaccines, DNA or protein, in different studies.
To investigate the potential of pcDNA3.1+ plasmid containing CFP-10 and ESAT-6 genes in induction of local immune responses after intramuscular injection in BALB/c mice.
pcDNA 3.1+ CFP-10 and pcDNA3.1+ ESAT-6 plasmids were prepared and defined groups of mice were injected intramuscularly with the plasmids both separately and in combination. The RNA was extracted from muscles after one month and cDNA was made using RT-PCR. The expressions of IL-4, IL-10 and IFN-γ genes cytokines were evaluated using comparative real time PCR.
Expression of IL-4 and IL-10 increased in the injection site of the mice groups which received plasmids encoding ESAT-6 and CFP-10 individually or together. More than 10-fold increase in IFN-γ expression was found in samples taken from mice groups inoculated by plasmids encoding ESAT-6 and CFP-10 individually or together.
pcDNA 3.1+ESAT-6 and pcDNA3.1+CFP-10 plasmids can increase the expression of IFN-γ in mice after immunization.
结核病是一种高发病率的疾病，主要由结核分枝杆菌（M.tb.）引起。DNA疫苗因其相对于传统方法的独特优势而展现出广阔的前景。结核分枝杆菌抗原的早期分泌抗原靶标（ESAT）-6和培养滤液蛋白（CFP）-10已被确定为抗分枝杆菌的候选疫苗，并在不同研究中用作亚单位疫苗、DNA或蛋白质疫苗。
研究含有CFP-10和ESAT-6基因的pcDNA3.1+质粒在BALB/c小鼠肌肉注射后诱导局部免疫反应的潜力。
制备pcDNA 3.1+ CFP-10和pcDNA3.1+ ESAT-6质粒，将特定组的小鼠分别或联合肌肉注射这些质粒。一个月后从肌肉中提取RNA，并使用RT-PCR制备cDNA。使用比较实时PCR评估IL-4、IL-10和IFN-γ基因细胞因子的表达。
单独或共同接受编码ESAT-6和CFP-10质粒的小鼠组注射部位的IL-4和IL-10表达增加。在单独或共同接种编码ESAT-6和CFP-10质粒的小鼠组采集的样本中，IFN-γ表达增加了10倍以上。
pcDNA 3.1+ESAT-6和pcDNA3.1+CFP-10质粒可在免疫后增加小鼠体内IFN-γ的表达。
