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Construction of DNA vaccines encoding Eimeria acervulina cSZ-2 with chicken IL-2 and IFN-gamma and their efficacy against poultry coccidiosis

机译:带有鸡IL-2和IFN-γ的编码小肠艾美球虫cSZ-2的DNA疫苗的构建及其抗禽球虫病的功效

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The study describes vaccination experiments with highly immunogenic sporozoite E. acervulina cSZ-2 coadministered with chicken IL-2 (chIL-2) and interferon-gamma (chIFN-gamma) to determine their efficacies against homologue challenge. The entire coding sequence of cSZ2, chIL-2 and chIFN-gamma were cloned into eukaryotic expression vector pVAX1, constructing DNA vaccines pVAX1-cSZ2, pVAX1-chIL-2, pVAX1-chIFN-gamma, pVAX1-cSZ2-chIL-2 and pVAX1-cSZ2-chIFN-gamma. The expression of target genes in vivo was detected by RT-PCR and Western blot. Chicken experiments were carried out by vaccinating chickens two times at dose rate of 100 jig intramuscularly. At 28 days of age, all chickens were inoculated orally with 1 x 105 sporulated oocysts of E. acervulina except the unchallenged control group. Seven days after challenge, all chickens were weighted and slaughtered for duodenum collection. The results indicated that these DNA vaccines were successfully constructed and the antigen genes could be expressed effectively in vivo. The findings also demonstrated best synergistic effect of IL-2 with this protein which suggested that co-administration of cytokines with this antigen was a powerful method to enhance immunity by alleviating intestinal lesions, body weight loss and oocyst count imparting partial protection against homologous challenge
机译:该研究描述了将高度免疫原性子孢小球藻大肠杆菌cSZ-2与鸡IL-2(chIL-2)和干扰素-γ(chIFN-γ)共同施用的疫苗接种实验,以确定它们对抗同源物攻击的功效。将cSZ2,chIL-2和chIFN-γ的整个编码序列克隆到真核表达载体pVAX1中,构建了DNA疫苗pVAX1-cSZ2,pVAX1-chIL-2,pVAX1-chIFN-γ,pVAX1-cSZ2-chIL-2和pVAX1 -cSZ2-chIFN-γ。通过RT-PCR和Western blot检测体内靶基因的表达。鸡实验是通过肌肉注射100夹具的两次疫苗对鸡进行的。在28天大时,除无挑战性的对照组外,所有鸡均口服1 x 105个带孢子的大肠埃希氏菌卵囊。攻击后7天,将所有鸡称重并屠宰以收集十二指肠。结果表明,这些DNA疫苗已成功构建,抗原基因可在体内有效表达。这些发现还证明了IL-2与该蛋白的最佳协同作用,这表明与该抗原共同施用细胞因子是一种有效的方法,可通过减轻肠道损伤,体重减轻和卵囊计数来增强免疫力,从而部分抵御同源攻击

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