摘要
旨在评估山羊支原体山羊肺炎亚种(Mycoplasma capricolum subsp.capripneumoniae,Mccp)膜表面蛋白P4的免疫原性和作为疫苗候选抗原的潜力。前期实验室筛选出Mccp的膜表面蛋白P4,本研究利用原核表达系统制备重组蛋白,纯化后与佐剂混合乳化制备疫苗,在山羊上进行免疫攻毒实验,通过临床症状、病理变化、血清抗体、肺脏载菌量、细胞因子等评价免疫效果。结果表明:成功表达纯化rP4蛋白,能够与His标签抗体、rP4多克隆抗体和Mccp感染抗体发生特异性反应;免疫动物能够刺激山羊产生高水平的抗体,但没有减轻临床症状,山羊存活率只有40%,与PBS对照组相同;病理学观察显示无论是急性还是慢性病例,P4组都要比PBS对照组更严重;肺脏载菌量高于全菌灭活组和PBS对照组;血清细胞因子结果显示P4组免疫后上调IL-2,下调IL-4水平。本研究成功表达纯化了rP4,制备疫苗免疫山羊能够诱导高水平抗体,但并不能保护山羊免受Mccp的攻毒,甚至会加重病变。研究结果为山羊传染性胸膜肺炎新型疫苗的设计研制提供了参考。
This study aims to evaluate the immunogenicity of membrane surface protein P4 from Mycoplasma capricolum subsp.capripneumoniae(Mccp)and its potential as a candidate antigen for vaccine.Preliminary laboratory work identified the membrane surface protein P4.In this study,recombinant protein was prepared using a prokaryotic expression system,after purification,the recombinant protein was emulsified with an adjuvant to prepare vaccine.Immunization and challenge experiments were conducted on goats,and the immune efficacy was evaluated based on clinical symptoms,pathological changes,serum antibodies,bacterial load in lung,and cytokines.The results showed that the rP4 protein was successfully expressed and purified,and could specifically react with the His-tag antibody,rP4 polyclonal antibody and Mccp infection antibody.Immunization with the rP4 stimulated high levels of antibody production in goat but did not alleviate clinical symptoms,with a survival rate of only 40%,the same as the PBS control group.Pathological observations indicated that both acute and chronic cases in the P4 group were more severe than those in the control group.The average bacterial load in the lungs was higher in the P4 group than that in the inactivated vaccine group and the PBS control group.Serum cytokines analysis revealed that the P4 group had upregulated IL-2 level and downregulated IL-4 level post-immunization.In conclusion,this study successfully expressed and purified rP4 and the prepared P4 protein induced antibody production in goats but failed to protect them from Mccp challenge,even exacerbating the pathogenic effects of Mccp on the goats.The findings provide a reference for the design and development of future new type vaccines for contagious caprine pleuropneumonia.
作者
梁晋嘉
郝华芳
陈胜利
陈睿
雒明阳
张一帆
兰仕梅
储岳峰
LIANG Jinjia;HAO Huafang;CHEN Shengli;CHEN Rui;LUO Mingyang;ZHANG Yifan;LAN Shimei;CHU Yuefeng(State Key Laboratory for Animal Disease Control and Prevention,College of Veterinary Medicine,Lanzhou University,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China;Gansu Province Research Center for Basic Disciplines of Pathogen Biology,Lanzhou 730046,China)
出处
《畜牧兽医学报》
北大核心
2026年第2期1063-1073,共11页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
甘肃省科技重大专项(23ZDNA007)
甘肃省2024年度重点人才项目(2024RCXM36)
中国农业科学院创新工程项目(CAASASTIP-2021-LVRI)。
关键词
山羊传染性胸膜肺炎
山羊
疫苗
免疫保护评价
细胞因子
Contagious caprine pleuropneumonia
goat
vaccine
immune protection evaluation
cytokine
