利用小鼠模型的脲酶免疫刺激物效果评价  

Evaluateion of the effect of urease immunostimulants by using a murine model

作　　者：章玉涛[1,2] 王加启[1] 赵圣国[1] 胡小丽[1] 卜登攀[1] 周凌云[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所/动物营养学国家重点实验室,北京100193 [2]甘肃农业大学动物科技学院,兰州730070

出　　处：《中国农业大学学报》2011年第3期107-111,共5页Journal of China Agricultural University

基　　金：北京市自然科学基金资助项目(6092017)

摘　　要：建立小鼠模型对制备的脲酶免疫刺激物进行免疫效果及其安全性评价。利用IPTG诱导脲酶基因克隆菌,大量表达脲酶UreB蛋白,用SDS-PAGE及Bradford方法对表达产物进行定性和定量分析;将脲酶UreB蛋白与弗氏(不)完全佐剂混合并乳化,制备成脲酶免疫刺激物。选择健康昆明小白鼠40只,随机分为4组,分别注射脲酶免疫刺激物、弗氏(不)完全佐剂和生理盐水,同时设置非注射为空白对照。利用间接ELISA测定特异性抗体效价,利用血常规分析仪和流式细胞仪测定小鼠血常规。结果表明:构建克隆菌株可以大量表达脲酶B蛋白,质量浓度为0.39 mg/mL。小鼠血清中特异性IgG抗体效价能达到1∶10 000以上,显著高于其他3个对照组(P<0.05),而特异性抗体IgA和IgM效价与对照组比较差异不显著(P>0.05)。脲酶免疫刺激物能显著提高小鼠血中白细胞、淋巴细胞和中间细胞数及比例(P<0.05),而对血液其他指标没有显著影响(P>0.05)。本试验成功制备脲酶免疫刺激物,并能有效刺激机体免疫应答,为后期奶牛免疫试验打下基础。The object of this study is to prepare and evaluate the urease immunostimulants.The protein concentration was identified and quantified identified by SDS-PAGE and Bradford respectively after UreB was expressed.To prepare urease immunostimulants,UreB protein and Freund＇s complete（incomplete） adjuvant were mixed and emulsified.Forty healthy mice were randomly allocated to four groups,where three groups were immunized with urease immunostimulants,Freund＇s complete（incomplete） adjuvant and physiological saline.and the fourth group served as the non-immunization control.The mice were immunized once every two weeks for three times.One week after the third immunization,bloods were sampled.Specific antibody titers were determined by the method of ELISA,and the mice blood was determined by blood analyzer and flow cytometry.Urease B protein was expressed and purified successfully with the concentration of 0.39 mg/mL.Serum specificity IgG titer of mice can reach above 1：10000,which was significantly higher（P0.05） than other 3 control groups,but specific IgA and IgM titers were not a significant difference between other 3 control groups（P0.05）.The number of white blood cells,lymphocyte and intermediate cell were improved by urease immunostimulants（P0.05）,and other blood index had no significant influence（P0.05）.Urease immunostimulants was prepared,and it could lead to the immune response,this built a foundation for further experiment regarding cow＇s immunization.

关 键 词：小鼠 脲酶 免疫刺激物 抗体效价 

分 类 号：Q556.4[生物学—生物化学]