布鲁氏菌104M疫苗株A抗原组分解析与候选蛋白筛选  

Analysis of the A antigen of Brucella 104M vaccine strain and candidate protein screening

作　　者：韩阔 赵鸿雁[1] 朴东日[1] 肖迪[1] 王磊[1] 范玉 徐晴晴 田国忠[1] 姜海[1] Han Kuo;Zhao Hongyan;Piao Dongri;Xiao Di;Wang Lei;Fan Yu;Xu Qingqing;Tian Guozhong;Jiang Hai(National Key Laboratory of Intelligent Tracing and Forecasting for Infectious Diseases,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]中国疾病预防控制中心传染病预防控制所,传染病溯源预警与智能决策全国重点实验室,北京102206

出　　处：《疾病监测》2024年第6期756-760,共5页Disease Surveillance

基　　金：国家重点研发计划项目(No.2020YFA0907101)。

摘　　要：目的探究布鲁氏菌104M疫苗株A抗原的主要成分,从A抗原中筛选潜在候选蛋白,为布鲁氏菌新型疫苗的研制提供基础。方法本研究利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳考马斯亮蓝染色、鲎试剂检测脂多糖、脂多糖银染对布鲁氏菌104M疫苗株A抗原的主要成分进行解析,通过与免疫绵羊血清的蛋白免疫印记实验,选取有印迹的部分切胶酶解,利用液相色谱–串联质谱对这部分蛋白进一步解析,通过抗原分析工具VaxiJen和毒力蛋白分析工具VirulentPred,筛选布鲁氏菌A抗原评分0.6以上的毒力相关蛋白作为潜在的候选疫苗。结果布鲁氏菌A抗原是一种淡黄色棉花糖状、质量较轻的物质,主要由蛋白和脂多糖构成,对A抗原的蛋白质组鉴定显示,104M A抗原共有1142种蛋白,能够与绵羊血清反应的蛋白有172种,VaxiJen抗原性评分在0.6以上的有87种蛋白,其中毒力相关蛋白有38种。结论布鲁氏菌104M疫苗株A抗原的主要成分是蛋白质和脂多糖,对筛选到的A抗原中的候选蛋白需要进一步的实验来测试这些蛋白质的免疫原性和保护水平以设计出针对布鲁氏菌病更为有效和安全的疫苗。Objective To explore the main components of the A antigen of Brucella 104 M vaccine strain and screen the potential candidate proteins from the A antigen and provide evidence for the development of novel vaccine for Brucella.Methods In this study,the main components of the A antigen of Brucella 104M vaccine strain were resolved using sodium dodecyl sulfate-polyacrylamide gel electrophoresis,Kaumas brilliant blue staining,horseshoe crab reagent detection of lipopolysaccharide,and lipopolysaccharide silver staining,and this protein was further resolved by western blotting with the serum of immunized sheep to select the part of blotted cuttings for enzymatic digestion of the proteins,and liquid chromatography-tandem mass spectrometry was used for further analysis.The virulence-associated proteins with Brucella A antigenic with score≥0.6 or higher as potential vaccine candidates were screened by using antigen analysis tool-VaxiJen and virulence protein analysis tool-VirulentPred.Results Brucella A antigen is a yellowish marshmallow-like,lightweight substance,mainly composed of proteins and lipopolysaccharides.Proteomic characterization of the A antigen showed that there were 1142 proteins in the 104 M A antigen,including 172 proteins capable of reacting with sheep serum,and 87 proteins with VaxiJen antigenicity score≥0.6,in which 38 were virulence-associated proteins.Conclusion The main components of the A antigen of Brucella 104M vaccine strain are proteins and lipopolysaccharides,and further experiments are needed to test the immunogenicity and level of protection of these proteins screened from A antigen in order to design a more effective and safer vaccine against brucellosis.

关 键 词：布鲁氏菌A抗原 脂多糖 抗原筛选 候选蛋白 疫苗 

分 类 号：R211[医药卫生—中医学] R516.7