单增李斯特菌ActA的表达与免疫原性研究  

Study of expression and immunogenicity of Listeria monocytogenes ActA

作　　者：张辉[1] 王兴龙[1] 李晓艳[1] 张爱玲[1] 张付贤[1] 崔丽瑾[1] 卜昭阳[1] 

机构地区：[1]军事医学科学院军事兽医研究所

出　　处：《中国卫生检验杂志》2008年第5期786-788,797,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的:研究单增李斯特菌ActA原核表达产物的免疫原性。方法:构建原核表达载体pGEX-3X/ActA,IPTG诱导表达并进行SDS-PAGE与Western blot分析,GST亲合层析纯化蛋白,免疫小鼠,检测小鼠体内特异性抗体水平,脾淋巴细胞增值活性及T淋巴细胞亚群,评价表达蛋白的免疫原性。结果:在大肠杆菌中表达了融合蛋白并获得了纯度较高的ActA;免疫小鼠在免疫后7周效价达到高峰,以后逐渐下降;免疫组的脾淋巴细胞增殖活性显著高于对照组(P<0.05);免疫组CD3+细胞、CD4+细胞、CD4+/CD8+比对照组分别增加了17.14%、18.03%、32.67%,CD8+细胞降低了12.38%。结论:ActA诱导了特异性体液免疫和细胞免疫。Objevtive：To investigate the immunogenicity of ActA expressed in E.coli.Methods：The prokaryotic expression vector pGEX-3X/ActA was constructed.The actA gene was induced by IPTG and analyzed by SDS-PAGE and Western blot.The ActA was purified by affinity chromatograph.To evaluate immunogenicity of ActA,BALB/c mice were immunized with ActA and antigen specific antibodies,lymphocyte proliferative response and the subsets of the T lymphocyte of immunized mice were detected.Results：The GST-ActA was expressed in E.coli and the high purity ActA was obtained.The antibody level of the immunized mice was highest at the seventh week after immunization.The proliferation response of the immunity group was higher than the control group（P〈0.05）.The CD3^＋,CD4^＋ and CD4^＋/CD8^＋ of the immunity group was increased by 17.14%,18.03% and 32.67% respectively,CD8^＋ was decreased by 12.38%.Conclusion：The ActA inductes specific humoral immunity and cellular immunity.

关 键 词：单增李斯特菌 ACTA 表达 免疫原性 

分 类 号：R378.994[医药卫生—病原生物学]