DEC205单链抗体增强巨型艾美耳球虫核酸疫苗免疫原性的评价  

Evaluation of DEC205 single-chain antibody enhancing immunogenicity ofEimeria maxima DNA vaccine

作　　者：陈育琪 黄春芳 李丽娜 魏坤亚 黄志坚[1,2] 黄瑜 殷光文[1,2] CHEN Yu-qi;HUANG Chun-fang;LI Li-na;WEI Kun-ya;HUANG Zhi-jian;HUANG Yu;YIN Guang-wen(College of Animal Sciences(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Animal Medicine Engineering Laboratory,Fuzhou 350002,China;Fujian Provincial Key Laboratory for Avian Diseases Control and Prevention/Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福建福州350002 [2]福建省动物药物工程实验室,福建福州350002 [3]福建省农业科学院畜牧兽医研究所福建省禽病防治重点实验室,福建福州350013

出　　处：《中国兽医科学》2021年第6期705-714,共10页Chinese Veterinary Science

基　　金：国家自然科学基金面上项目(31872466);福建省科技厅引导性项目(2019N0005);福建省禽病防治重点实验室资助课题(FKADL-2019-02)。

摘　　要：为了探究scFvDEC205(DEC205单链抗体)能否增强巨型艾美耳球虫优势抗原片段EmIMP1C的免疫原性,构建重组质粒pVAX1-scFvDEC205-EmIMP1C,转染HEK-293T细胞检验其体外表达情况,于试验鸡大腿肌肉处定期3次注射该重组质粒。结果显示:成功构建重组质粒pVAX1-scFvDEC205-EmIMP1C,并能在HEK-293T细胞中表达。pVAX1-scFvDEC205-EmIMP1C组的IgG抗体效价最高,达3200。第3次免疫后,pVAX1-scFvDEC205-EmIMP1C组血清中IFN-γ质量浓度显著高于其他组(P<0.05);用流式细胞仪检测试验鸡产生的CD4^(+)和CD8^(+)T细胞比例,用CCK8法刺激鸡脾淋巴细胞增殖,结果均显示pVAX1-scFvDEC205-EmIMP1C组显著高于其他实验组(P<0.05)。病理切片结果提示pVAX1-scFvDEC205-EmIMP1C组的病变程度最轻。结论:重组质粒pVAX1-scFvDEC205-EmIMP1C能有效诱导鸡产生免疫效应,scFvDEC205具有增强EmIMP1C抗原免疫原性的效果,为今后开发靶向鸡球虫疫苗给予了有力的支持。In order to explore the effect of scFvDEC205(DEC205 single-chain antibody)on the immunogenicity of dominant Eimeria maxima antigen fragment EmIMP1Cs,a recombinant plasmid pVAX1-scFvDEC205-EmIMP1C was constructed.The recombinant plasmid was transfected into HEK-293T cells to express in vitro,and also injected three times regularly into thigh muscle of chicken.Results showed that pVAX1-scFvDEC205-EmIMP1C was successfully constructed and expressed in HEK-293T cells.IgG antibody titer of pVAX1-scFvDEC205-EmIMP1C group was the highest,reaching 3200.After the third immunization,IFN-γmass concentration in sera of pVAX1-scFvDEC205-EmIMP1C group was significantly higher than that of other groups(P<0.05);the percentage of CD4^(+)and CD8^(+)T cells andD450 of CCK8 stimulating chicken spleen lymphocytes to proliferate were both significantly higher than those of other groups(P<0.05)in pVAX1-scFvDEC205-EmIMP1C group.Result of pathological sections indicated that pVAX1-scFvDEC205-EmIMP1C group had the least lesions.In conclusion,pVAX1-scFvDEC205-EmIMP1C can induce immune effects in chickens,and scFvDEC205 is effective in enhancing immunogenicity of EmIMP1C antigen,which provides strong support for development of targeted chicken coccidia vaccines in the future.

关 键 词：巨型艾美耳球虫 EmIMP1C抗原 DEC205 单链抗体 DNA疫苗 

分 类 号：S852.723[农业科学—基础兽医学]