脲酶B亚单位疫苗免疫对奶牛血清蛋白质组分的影响

Effect of Ure B Subunit Vaccine Immunization on Serum Proteins in Dairy Cows

为探讨脲酶B(Ure B)亚单位疫苗免疫后奶牛血清蛋白质组分的表达变化。选用8头健康的泌乳荷斯坦奶牛,分为免疫处理组和空白对照组,分别于胯部两侧肌肉注射Ure B亚单位疫苗(含抗原蛋白0.2 mg)和生理盐水,每隔2周进行加强免疫,并于免疫后第56天采集奶牛血样,用间接ELISA法测定血清中特异性IgG、IgM和IgA效价,用二维凝胶电泳(2-DE)结合MALDI-TOF-TOF质谱技术研究血清蛋白质组表达变化。免疫处理组奶牛血清中特异性IgG效价显著高于对照组(P<0.05),但Ure B免疫对奶牛血清中特异性IgM和IgA效价无显著影响(P>0.05)。血清蛋白质组分析发现6个差异表达蛋白点,其中3个蛋白点得到有效鉴定。结合珠蛋白前体和其他3个蛋白点在免疫Ure B亚单位疫苗奶牛血清中表达量增加,而Kakapo蛋白的表达量降低。Ure B亚单位疫苗免疫引起奶牛血清中结合珠蛋白和Kakapo等蛋白表达发生变化,为进一步研究脲酶B亚单位疫苗免疫对奶牛机体的影响提供了依据。

This study was to investigate the changes of serum proteins in dairy cows immunized with Ure B subunit vaccine.Eight healthy cows were divided into 2 groups and immunized with 0.2 mg antigen protein or saline by intramuscular injection on both sides of the hip,once every two weeks,and total 4 times for immunization.Serum samples were collected on 56 d after immunization with Ure B subunit vaccine.Specific antibody in blood samples were determined by ELISA methods.Serum proteomic analysis were performed using two-dimensional gel electrophoresis（2-DE） coupled with MALDI-TOF-TOF mass spectrometer.Specific IgG were significantly higher after the cows immunized with Ure B subunit vaccine than control groups（P0.05）,whereas IgM and IgA was not significant（P0.05）.Proteomic analysis revealed 6 protein spots were differentially expressed,and 3 spots were successfully identified.Haptoglobin precursor and 3 unknown spots were up-regulated in cows immunized with Ure B subunit vaccine,while Kakapo was down-regulated.The current study showed that cows injection with Ure B subunit vaccine can result in the change of haptoglobin,Kakapo and other proteins,which provide the basis for further research of the physiology effects of Ure B subunit vaccine on dairy cows.