Efficacy of chimeric DNA vaccines encoding Eimeria tenella 5401 and chicken IFN-γ or IL-2 against coccidiosis in chickens
Efficacy of chimeric DNA vaccines encoding Eimeria tenella 5401 and chicken IFN-γ or IL-2 against coccidiosis in chickens
复制标题
编码柔嫩艾美耳球虫 5401 和鸡 IFN-γ 或 IL-2 的嵌合 DNA 疫苗对抗鸡球虫病的功效
DOI：
10.1016/j.exppara.2015.05.003
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发表时间：
2015-09-01
影响因子：
2.1
通讯作者：
Li, Xiangrui
中科院分区：
文献类型：
--
作者：
Song, Xiaokai;Huang, Xinmei;Li, Xiangrui
编码柔嫩艾美耳球虫（Eimeria tenella，E. tenella）表面抗原5401，并对其抗E. tenella攻击。将5401的开放阅读框（ORF）克隆到原核表达载体pGEX-4 T2中进行表达，并对表达的重组蛋白进行Western blot鉴定。将5401的ORF和鸡细胞因子IFN-γ或IL-2基因依次克隆到真核表达载体pVAX 1中，构建成DNA疫苗pVAX-5401-IFN-γ、pVAX-5401-IL-2和pVAX-5401。用逆转录-聚合酶链反应和Western blot检测目的基因在体内的表达。将14日龄的鸡以7天的间隔分别用100 μ g质粒pVAX-5401、pVAX-5401-IFN-γ和pVAX-5401-IL-2或200 μ g重组5401蛋白通过腿部肌肉注射接种两次。第二次接种后7天，除未攻毒对照组外，所有鸡均经口接种5 × 10（4）个E. tenella。攻毒后第7天，所有鸡称重并屠宰以确定免疫效果。结果表明，该重组蛋白分子量约为90 kDa，能与可溶性子孢子抗血清发生特异性反应。动物实验表明，与未免疫攻击对照组和空载体pVAX 1对照组相比，DNA疫苗pVAX-5401、pVAX-5401-IFN-γ和pVAX-5401-1-2以及重组5401蛋白均能明显减轻体重减轻和盲肠病变。此外，pVAX-5401-IFN-γ或pVAX-5401-1-2诱导的抗球虫指数（ACI）为180.01或177.24，其显著高于pVAX-5401。结果表明，5401是一种有效的疫苗候选抗原。这一发现也表明，鸡IFN-γ或IL-2可以有效地提高抗鸡球虫病DNA疫苗的效力。(C)2015 Elsevier Inc. All rights reserved.
Chimeric DNA vaccines encoding Eimeria tenella (E. tenella) surface antigen 5401 were constructed and their efficacies against E. tenella challenge were studied. The open reading frame (ORF) of 5401 was cloned into the prokaryotic expression vector pGEX-4T2 to express the recombinant protein and the expressed recombinant protein was identified by Western blot. The ORF of 5401 and chicken cytokine gene IFN-gamma or IL-2 were cloned into the eukaryotic expression vector pVAX1 consecutively to construct DNA vaccines pVAX-5401-IFN-gamma, pVAX-5401-IL-2 and pVAX-5401. The expression of aim genes in vivo was detected by reverse transcription-polymerase chain reaction and Western blot. Fourteen-day-old chickens were inoculated twice at an interval of 7 days with 100 mu g of plasmids pVAX-5401, pVAX-5401-IFN-gamma and pVAX-5401-IL-2 or 200 fig of recombinant 5401 protein by leg intramuscular injection, respectively. Seven days after the second inoculation, all chickens except the unchallenged control group were challenged orally with 5 x 10(4) sporulated oocysts of E. tenella. Seven days after challenge, all chickens were weighted and slaughtered to determine the effects of immunization. The results showed the recombinant protein was about 90 kDa and reacted with antiserum against soluble sporozoites. The animal experiment showed that all the DNA vaccines pVAX-5401, pVAX-5401-IFN-gamma or pVAX-5401-1-2 and the recombinant 5401 protein could obviously alleviate body weight loss and cecal lesions as compared with non-vaccinated challenged control and empty vector pVAX1control. Furthermore, pVAX-5401-IFN-gamma or pVAX-5401-1-2 induced anti-coccidial index (ACI) of 180.01 or 177.24 which were significantly higher than that of pVAX-5401. The results suggested that 5401 was an effective candidate antigen for vaccine. This finding also suggested that chicken IFN-gamma or IL-2 could effectively improve the efficacies of DNA vaccines against avian coccidiosis. (C) 2015 Elsevier Inc. All rights reserved.