布鲁氏菌病温敏凝胶活疫苗(S2株)对羊不同黏膜途径免疫效果的研究  

Immune efficacy of Brucellosis thermosensitive gel vaccine(S2 strain)in sheep with different mucosal immune routes

作　　者：刘燕[1,2] 颜成旭[3] 米吉提·莫合他尔汗 贺笋 何传雨 赵海龙 沈国顺 毛开荣[2] LIU Yan;YAN Cheng-xu;MI Ji-ti;HE Sun;HE Chuan-yu;ZHAO Hai-long;SHEN Guo-shun;MAO Kai-rong(Shenyang Agricultural University,Shenyang 110866,China;China Institute of Veterinary Drug Control,Beijing 100081,China;Xinjiang Center of Animal Disease Control,Urumqi 830000,China;Tecon Bio-technology Co.,Ltd.,Urumqi 830011,China)

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110866 [2]中国兽医药品监察所,北京100081 [3]新疆动物卫生监督所,新疆乌鲁木齐830000 [4]天康生物股份有限公司,新疆乌鲁木齐830011

出　　处：《中国预防兽医学报》2021年第6期646-650,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重点研发计划“牛羊重要人畜共患病新型疫苗研究”(2017YFD0500901)。

摘　　要：为了研究布鲁氏菌病温敏凝胶疫苗对羊不同黏膜途径免疫的效果,本研究采用冷溶法制备了用于黏膜免疫的布鲁氏菌病温敏凝胶活疫苗(S2株),将该疫苗以不同黏膜免疫途径(阴道灌注、点眼)、注射免疫途径免疫绵羊,同时以常规S2疫苗作为对照,无菌采集绵羊的免疫部位拭子后培养,通过菌落计数方法计算黏膜免疫部位排菌数,结果显示:凝胶疫苗免疫部位从免疫后第30 min开始停止排菌,常规疫苗的排菌持续到免疫后第2 d。免疫后采集各时间点绵羊血血清,采用微量试管凝集试验(MSAT)和攻菌保护试验分别测定不同免疫途径(阴道灌注、点眼、注射)免疫后产生的抗体水平和保护效果的差异,抗体检测结果显示,黏膜免疫后45 d左右抗体即降至1.50或以下,黏膜免疫后产生的抗体水平明显低于注射免疫产生的抗体,且两种黏膜免疫下降速度均比注射免疫的快;攻菌后的免疫保护结果显示,在对照组出菌指数为4.76情况下,免疫组的单位保护(对照出菌指数-免疫出菌指数)分别达到2.19、2.42、2.78,表明3种免疫方式均具有良好的免疫保护效果,其中阴道灌注的单位保护高于点眼,与注射免疫组更接近。以上结果表明:与常规疫苗相比,温敏凝胶疫苗可有效提升疫苗的安全性。相比点眼免疫,阴道灌注的免疫保护效果更好、安全性更好,虽然点眼抗体反应更弱、持续时间更短,但与阴道灌注相比,对诊断的干扰差异不大。综合以上结果表明,两种黏膜免疫方式中,阴道灌注是较理想的黏膜免疫方式,但黏膜免疫抗体持续时间短于注射免疫。因此,采用布鲁氏菌病温敏凝胶疫苗进行黏膜免疫更有利于鉴别诊断,同时也为新型布鲁氏菌病疫苗的研究提供新的思路。In order to evaluate the efficacy of brucellosis thermos-sensitive gel vaccine in different mucosal immune routes,a live Brucellosis thermosensitive gel vaccine(S2 strain)for mucosal immunization was prepared by cold dissolving method,the sheep were immunized with the vaccine through different mucosal immune routes(vaginal perfusion,dot eyes),injection,culture of sheep swabs from immunized spots of the sheep were collected using aseptic method and cultured,and the number of bacteria discharged from the mucosal immune sites was calculated by colony counting method.The gel vaccine stopped shedding the bacteria after 30 min post immunization,the conventional vaccine stopped shedding the bacteria after 2 days post immunization.Sheep sera were collected at each time point after immunization,the antibody levels and protective effects of different immunization routes(vaginal infusion,eye drops and injection)were determined by microtest tube agglutination test(MSAT)and protection test.The results showed that the level of mucosal immune antibody dropped to less or close to 1:50 about 45 days after immunization.The level of antibody produced by mucosal immunization was significantly lower than that produced by injection immunization,and the decline rate of two kinds of mucosal immunization was faster than that of injection immunization.The challenge test results showed that the bacterial index of the control group was 4.76,and the unit protection of the immunized group(control bacterial index-immunization index)was 2.19,2.42 and 2.78,respectively,indicating that all the three immunization methods had good immune protective effect,and the unit protection of vaginal perfusion was higher than that of eye-drops,which was closer to that of injection group.The results showed that compared with the conventional vaccine,the thermosensitive gel vaccine could effectively improve the safety of vaccine use.Compared with the two kinds of mucosal immune effect,vaginal perfusion had higher immune protection and better safety.Although eye

关 键 词：布鲁氏菌温敏凝胶疫苗 黏膜免疫 排菌 免疫抗体 单位保护 

分 类 号：S852.61[农业科学—基础兽医学]