毒害艾美耳球虫配子体抗原基因DNA疫苗的构建与免疫保护效果评价  

Construction and Evaluation of the Immunoprotection of a DNA Vaccine Targeting Eimeria necatrix Gametocyte Antigen Gene

作　　者：冯永翠 Naing Htet Aung 张馨尹 汪飞燕 王乐乐[1,2,3] 张露 朱玉 许金俊 陶建平[1,2] 刘丹丹 FENG Yongcui;Naing Htet Aung;ZHANG Xinyin;WANG Feiyan;WANG Lele;ZHANG Lu;ZHU Yu;XU Jinjun;TAO Jianping;LIU Dandan(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China;Ganzhou Polytechnic,Ganzhou 341008,China)

机构地区：[1]扬州大学兽医学院,扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,扬州225009 [3]赣州职业技术学院,赣州341008

出　　处：《畜牧兽医学报》2025年第4期1887-1896,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31972698,31602039);江苏省重点研发计划(现代农业,BE2021354);江苏高校优势学科建设四期工程项目(2022-2025);高等学校学科创新引智计划资助(D18007)。

摘　　要：旨在评估毒害艾美耳球虫配子体抗原基因DNA疫苗的抗球虫免疫保护效果。本研究构建了配子体抗原基因的DNA疫苗pVAX-Engam59,并添加CpG ODN2006佐剂免疫雏鸡,通过间接ELISA和qRT-PCR方法分析雏鸡特异性抗体效价和细胞因子(IL-4、IFN-γ和TGF-β4)转录情况;最后,每只雏鸡经嗉囊感染2.5×10^(4)个毒害艾美耳球虫孢子化卵囊,观察各试验组的临床症状、血便排出情况,感染8 d后检测各试验组的卵囊排出量、增重、病变记分等,并计算抗球虫指数。结果表明,与空载体pVAX-1组、未免疫攻虫组、未免疫未攻虫组相比,pVAX-Engam59、pVAX-Engam59+CpG ODN2006免疫雏鸡后血清特异性抗体水平显著升高(P<0.05),细胞因子IFN-γ、IL-4、TGF-β4表达水平显著升高(P<0.05);卵囊减少率分别为64.65%与64.48%,血便数以及病变记分显著降低、相对增重率显著增加(P<0.05);ACI值分别为156.02与159.72。综上,本研究成功构建了毒害艾美耳球虫配子体抗原基因的DNA疫苗,且与pVAX-Engam59相比,pVAX-Engam59+CpG ODN2006佐剂组免疫保护效果较好,二者均接近抗球虫中效水平,有望成为抗球虫病的候选DNA疫苗。The purpose of this experiment was to investigate the immunoprotection of a DNA vaccine targeting Eimeria necatrix gametocyte antigen gene against coccidiosis.In this study,a DNA vaccine of gametocyte antigen gene,pVAX-Engam59,was constructed and combined with CpG ODN2006 adjuvant to immunize the chicks.The level of specific antibody in each group was detected by indirect ELISA.The mRNA transcription of cytokine(IL-4,IFN-γ,and TGF-β4)was analyzed by qRT-PCR.Finally,each chick was infected with 2.5×10^(4) E.necatrix sporula-ted oocysts through the crop,and the clinical symptoms and bloody stools of each group were ob-served.The oocysts output,weight gains and lesion scores of each group were detected after 8 days post-infection.The results showed that the serum specific antibody and cytokine(IL-4,IFN-γ,and TGF-β4)levels of pVAX-Engam59 and pVAX-Engam59+CpG ODN2006 were sig-nificantly increased(P<0.05),and the oocyst reduction ratios were 64.65%and 64.48%,re-spectively.Compared to challenged and unchallenged control group,the number of bloody stools and the lesion scores in the pVAX-Engam59 and pVAX-Engam59+CpG ODN2006 groups were significantly lower(P<0.05),and the relative weight gain rates were significantly higher(P<0.05).The ACI values were 156.02 and 159.72,respectively.In summary,we successfully con-structed DNA vaccine targeting E.necatrix gametocyte antigenic gene,and both pVAX-Engam59+CpG ODN2006 and pVAX-Engam59 showed good immunoprotective effects,which were close to the anti-coccidial intermediate efficiency level,and could be candidates for a DNA vaccine against coccidiosis.

关 键 词：毒害艾美耳球虫 配子体抗原基因 DNA疫苗 免疫保护 

分 类 号：S855.9[农业科学—临床兽医学] S852.723[农业科学—兽医学]