College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu 210095, PR China.
Res Vet Sci. 2011 Feb;90(1):72-7. doi: 10.1016/j.rvsc.2010.05.003.
The study describes vaccination experiments with highly immunogenic sporozoite E. acervulina cSZ-2 co-administered with chicken IL-2 (chIL-2) and interferon-γ (chIFN-γ) to determine their efficacies against homologue challenge. The entire coding sequence of cSZ2, chIL-2 and chIFN-γ were cloned into eukaryotic expression vector pVAX1, constructing DNA vaccines pVAX1-cSZ2, pVAX1-chIL-2, pVAX1-chIFN-γ, pVAX1-cSZ2-chIL-2 and pVAX1-cSZ2-chIFN-γ. 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 μg intramuscularly. At 28 days of age, all chickens were inoculated orally with 1×10(5) 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.
本研究描述了用高度免疫原性的疟原虫孢子虫 E. acervulina cSZ-2 与鸡白细胞介素 2（chIL-2）和干扰素-γ（chIFN-γ）共同进行的疫苗接种实验，以确定它们对同源性挑战的功效。cSZ2、chIL-2 和 chIFN-γ 的整个编码序列被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。通过以 100μg 的剂量肌内接种两次对鸡进行实验。在 28 天龄时，除未挑战对照组外，所有鸡均口服接种 1×10(5)个孢子化的柔嫩艾美耳球虫卵囊。攻毒后 7 天，所有鸡称重并屠宰收集十二指肠。结果表明，这些 DNA 疫苗构建成功，抗原基因可在体内有效表达。研究结果还表明，IL-2 与该蛋白具有最佳协同作用，这表明细胞因子与该抗原共同给药是一种通过减轻肠道病变、体重减轻和卵囊计数来增强免疫力的有效方法，对同源性挑战具有部分保护作用。
