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用表达柔嫩艾美耳球虫3-1E蛋白的枯草芽孢杆菌对鸡进行口服免疫后,鸡对柔嫩艾美耳球虫感染的保护性免疫。

Protective immunity against Eimeria tenella infection in chickens following oral immunization with Bacillus subtilis expressing Eimeria tenella 3-1E protein.

作者信息

Lin Zhiwei, Shi Yanyun, Deng Bin, Mao Xiangfei, Yu Dongyou, Li Weifen

机构信息

Key Laboratory of Molecular Animal Nutrition, Ministry of Education, College of Animal Sciences, Zhejiang University, 388 Yuhangtang Road, West Lake District, Hangzhou, 310058, People's Republic of China.

出版信息

Parasitol Res. 2015 Sep;114(9):3229-36. doi: 10.1007/s00436-015-4539-3. Epub 2015 May 22.

Abstract

The current experiment was conducted to construct recombinant Bacillus subtilis WB600 expressing Eimeria tenella 3-1E protein to investigate the oral immunization protective effects against E. tenella. The merozoite surface antigen 3-1E gene of E. tenella was introduced into the pBS-H1 expression vector with a novel signal peptide sequence. After the electro-transformation, the expression of objective protein in B. subtilis WB600 was detected by Western blot. The results showed that the recombinant B. subtilis strain with the ability of high-level secretion of 3-1E was constructed successfully. Seven-day-old broiler chickens were orally vaccinated with B. subtilis WB600 harboring 3-1E (B.S-pBS-H1-3-1E) or B. subtilis WB600 with empty plasmid (B.S-pBS-H1) 10 days prior to challenge with sporulated E. tenella oocysts. The results showed the recombinant B. subtilis strain with the ability of high-level secretion of 3-1E was constructed successfully. Vaccination with B.S-pBS-H1-3-1E strain significantly increased the anti-coccidial index and reduced cecal lesion scores compared with the positive control group (chickens were challenged with sporulated E. tenella oocysts without oral administration of B.S-pBS-H1-3-1E strain) and B.S-pBS-H1 group. Ceca mucosal sIgA, secretion, and IL-2, IL-12, IFN-γ, and IL-10 level after challenge were greater in the B.S-pBS-H1-3-1E group than in the positive control group. Taken together, these results indicated that B. subtilis WB600 harboring 3-1E protein induces protective immunity against E. tenella.

摘要

进行当前实验以构建表达柔嫩艾美耳球虫3-1E蛋白的重组枯草芽孢杆菌WB600,以研究其对柔嫩艾美耳球虫的口服免疫保护作用。将柔嫩艾美耳球虫的裂殖子表面抗原3-1E基因导入带有新信号肽序列的pBS-H1表达载体。电转化后,通过蛋白质免疫印迹法检测枯草芽孢杆菌WB600中目的蛋白的表达。结果表明成功构建了具有高水平分泌3-1E能力的重组枯草芽孢杆菌菌株。在经孢子化的柔嫩艾美耳球虫卵囊攻毒前10天,用携带3-1E的枯草芽孢杆菌WB600(B.S-pBS-H1-3-1E)或携带空质粒的枯草芽孢杆菌WB600(B.S-pBS-H1)对7日龄肉鸡进行口服免疫。结果表明成功构建了具有高水平分泌3-1E能力的重组枯草芽孢杆菌菌株。与阳性对照组(用经孢子化的柔嫩艾美耳球虫卵囊攻毒但未口服B.S-pBS-H1-3-1E菌株的鸡)和B.S-pBS-H1组相比,用B.S-pBS-H1-3-1E菌株免疫显著提高了抗球虫指数并降低了盲肠病变评分。攻毒后,B.S-pBS-H1-3-1E组盲肠黏膜sIgA、分泌水平以及IL-2、IL-12、IFN-γ和IL-10水平均高于阳性对照组。综上所述,这些结果表明携带3-1E蛋白的枯草芽孢杆菌WB600可诱导对柔嫩艾美耳球虫的保护性免疫。

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