炭疽杆菌PA83蛋白的表达及与传统疫苗免疫效果的比较研究  

Expression of PA83 protein of Bacillus anthracis and its immune effect in comparison with the conventional vaccines

作　　者：李雷斌 宋晓晖[2] 孙航 冯冰 陈玉红 何雷 陈玲玲 刘玉良[2] 王传彬[2] 李义平 孙雨[2] LI Lei-bin;SONG Xiao-hui;SUN Hang;FENG Bing;CHEN Yu-hong;HE Lei;CHEN Ling-ling;LIU Yu-liang;WANG Chuan-bin;LI Yi-ping;SUN Yu(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;China Center for Animal Disease Control and Prevention,102600 Beijing,China;Zhongshan School of Medicine,Sun Yet-sen University,510080 Guangzhou,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]中国动物疫病预防控制中心,北京102600 [3]中山大学中山医学院,广东广州510080

出　　处：《中国兽医科学》2023年第8期1010-1018,共9页Chinese Veterinary Science

基　　金：现代农业产业技术体系建设专项(CARS39)。

摘　　要：炭疽芽胞杆菌PA83蛋白是细菌识别毒素受体并寡聚化携带致病基因进入细胞质进而引起感染的重要蛋白,具有良好的免疫原性,被广泛应用于炭疽杆菌病预防和诊断技术研究。本研究对PA83基因进行密码子优化后,原核表达了PA83蛋白并通过镍柱进行纯化。将纯化的PA83蛋白和传统疫苗(Ⅱ型炭疽芽胞疫苗和无荚膜炭疽芽胞杆菌疫苗)分别免疫新西兰大白兔。对免疫兔体内血清抗体水平、细胞因子和体外淋巴细胞增殖变化等进行了检测、比较和分析。结果显示,PA83蛋白免疫后血清抗体效价最高可达1∶122880,明显高于两种传统疫苗免疫抗体水平,差异显著(P<0.01)。不过,PA83蛋白与两种传统疫苗免疫后引起的细胞因子变化以及与对照组相比,均无显著差异(P>0.05)。此外,PA83和两种传统疫苗免疫后,对体外淋巴细胞无促进生长和增值作用。本研究结果表明,PA83蛋白免疫兔后,虽然未见免疫细胞反应变化,但可激发机体产生高水平体液免疫,研究结果为炭疽芽胞杆菌候选疫苗的研制奠定了基础。PA83 of Bacillus anthracis is a key protein that recognizes toxin receptors.After oligomerization,it carries pathogenic protein into cytoplasm,leading to infection in host.PA83 has high immunogenicity,allowing it to be widely used in the prevention and diagnosis of Bacillus anthracis.In this study,the codon of PA83 gene was optimized,and the PA83 protein was expressed in prokaryotic expression system and then purified by nickel column.New Zealand white rabbits were immunized with the purified PA83 protein and conventional vaccines(Anthrax Spore Live Vaccine Strain No.2 and Anthrax Nonencapsulated Vaccine Strain),respectively.The level of serum antibody and proliferation of cytokines and lymphocyte in the immunized rabbits were measured,compared and analyzed.The results showed that,the highest serum antibody titer up to 1∶122880 was detected post-immunization of PA83,which was significantly higher than that of the two conventional vaccines(P<0.01).However,neither no significant difference in the proliferation of cytokine induced by the immunized PA83 protein when the two conventional vaccines were compared,nor the control group in comparison with the PA83 immunization group(P>0.05).In addition,there was no dramatic change in the growth and proliferation of lymphocytes in rabbits immunized with the PA83 and the two vaccines.The results in this study suggest that,the PA83 could induce robust immune response in the serum of New Zealand white rabbit,although strong cellular immune response was not detected.Our findings laid a foundation for the development of candidate vaccine against Bacillus anthracis.

关 键 词：炭疽杆菌 PA83蛋白 传统疫苗 血清抗体 淋巴细胞增值 细胞因子 

分 类 号：S852.616[农业科学—基础兽医学]