猪流行性腹泻病毒纤突蛋白受体结合区(PEDV-RBD)重组蛋白的免疫原性  

Immunogenicity analysis of PEDV-RBD subunit vaccine

作　　者：伊立超 郝嘉翼 邹万成 李乐天 姜宇航 张爽[1,2] 娄安钢 李昌[2] 金宁一 YI Lichao;HAO Jiayi;ZOU Wancheng;LI Letian;JIANG Yuhang;ZHANG Shuang;LOU Angang;LI Chang;JIN Ningyi(Agricultural College,Yanbian University,Yanji,Jilin 133000,China;Changchun Institute of Veterinary Medicine,Chinese Academy of Agricultural Sciences,Changchun 130122,China;College of Animal Medicine/College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]延边大学农学院,吉林延吉133000 [2]中国农业科学院长春兽医研究所,吉林长春130122 [3]吉林农业大学动物科技学院/动物医学院,吉林长春130118

出　　处：《中国兽医学报》2023年第4期643-647,共5页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2021YFD1801103-6)。

摘　　要：利用杆状病毒表达系统制备猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)纤突蛋白受体结合区(receptor binding domain,RBD)重组蛋白(PEDV-RBD),将其免疫3周龄断乳仔猪,分析其免疫原性。将PEDVRBD P3代重组杆状病毒接种到悬浮SF9细胞中,收获后离心取上清液,将上清液超速离心浓缩重组蛋白。将仔猪分为免疫组、空载体对照组和佐剂对照组,分别用制备的重组蛋白、空载体表达产物和单纯佐剂免疫仔猪。结果显示,免疫后8周,PEDV-RBD重组蛋白组产生的特异性抗体效价达1∶12800,中和抗体平均效价1∶284,最高达1∶512。流式细胞术分析CD3+CD4^(+)双阳性T淋巴细胞显著高于对照组,表明重组蛋白具有良好的免疫原性,能够激发仔猪的体液免疫和细胞免疫应答。该研究为猪流行性腹泻疫苗研制提供了理论依据。The recombinant protein of receptor binding domain(RBD)of porcine epidemic diarrhea virus(PEDV)was prepared by baculovirus expression system and was used to immunize threeweek-old weaned piglets to analyze its immunogenicity.PEDV-RBD P3generation recombinant baculovirus was inoculated to SF9cells in suspension culture.After harvest,the supernatant was centrifuged and the recombinant protein was concentrated by ultracentrifugation.The prepared recombinant protein was used to immunize piglets,and the empty vector control group and adjuvant control group were set up.The specific antibody and neutralizing antibody produced by immunized mice and piglets were significantly higher than those in the control group.After the 8th week of immunization,the specific antibody titer of PER recombinant protein group was 1∶12800,and the average titer of neutralizing antibody was 1∶284,up to 1∶512.Flow cytometry analysis showed that CD3+CD4+double positive T lymphocytes were significantly higher than those in the control group,indicating that the recombinant protein had good immunogenicity and could stimulate humoral and cellular immune responses in piglets.This study provides a theoretical basis for the development of swine epidemic diarrhea vaccine.

关 键 词：猪流行性腹泻病毒 重组蛋白 免疫原性分析 

分 类 号：S852.62[农业科学—基础兽医学]