Epstein-Barr病毒基因工程亚单位疫苗膜抗原gp340/220的纯化及其免疫学特性  

Isolation and purification of Epstein-Barr virus gene engineering subunit vaccine of membrane antigen gp340/220 and experimental studies on immunological characterization

作　　者：项树林[1] 肖瑶[1] 李燕[1] 朱秋屏[1] 谷淑燕[1] 

机构地区：[1]中国预防医学科学院病毒学研究所免疫研究室,北京100052

出　　处：《中华实验和临床病毒学杂志》1996年第4期344-348,共5页Chinese Journal of Experimental and Clinical Virology

摘　　要：利用病毒学研究所免疫研究室重组的表达Epstein-Barr病毒膜抗原(MA)gp340/220重组痘苗病毒感染143细胞,24小时后裂解细胞,获得的上清液经DEAE-Sepharose FF及Sephadex G200层析纯化,经免疫打点分析后收集阳性蛋白,经SDS-PAGE电泳,考马斯亮蓝R-250染色,显示为单一的蛋白带,从而获得纯化的亚单位疫苗MA gp340/220。将纯化的MA与A1(OH)_3佐剂或rIL-2+完全福氏佐剂免疫小鼠,对其免疫学特性进行测定。结果表明,MA免疫的小鼠能产生较高滴度的特异性抗体和中和抗体,诱导产生抗体依赖细胞介导的细胞毒反应(ADCC),并能刺激特异性T淋巴细胞的增殖,但未发现对细胞毒性T淋巴细胞(CTL)杀伤作用有何影响。Recombinant vaccinia virus which expressed EBV membrane antigen (MA) gp340/220 under the control of 7.5Kb promoter were used to infect 143 cells and cultured for 24 hours. The ceils expressing MA gp340/220 on the cell surface were harvested and lysised, then purified through DEAE-sepharose FF and Sephadex G200. Analysed by SDS-PAGE electrophoresis, followed by coomassic blue staining and immunodot, the final product was proved to be pure MA gp340/220. It was used to immunize Balb/C and CBA/J mice mixed with Al(OH)_3 adjuvant or rlL-2+complete Freud's adjuvant, to observe the immune response elicited by it, The results showed that the mice immunized by MA produced high titer specific antibody and neutralizing antibody, ADCC activity, and specific T lymphocytes proliferation, but no CTL response found.

关 键 词：肿瘤免疫疗法 EB病毒 亚单位疫苗 病毒抗原 

分 类 号：R730.51[医药卫生—肿瘤] R392-33[医药卫生—临床医学]