摘要
目的对汉逊酵母表达的adr亚型重组乙型肝炎(简称乙肝)疫苗(adr疫苗)进行免疫学特性与功能学研究。方法采用adr疫苗与酿酒酵母表达的重组乙肝疫苗(adw疫苗)免疫转人HLA-A2基因小鼠,免疫剂量2μg/0.2 ml,于初免3周后进行第2次免疫。于第2次免疫1周后,采血,分离血清,经ELISA法检测小鼠血清中抗-HBs抗体水平。同时取小鼠脾脏,制备脾细胞悬液,采用HLA-A2/WLSLLVPFV五聚体荧光标记流式细胞术分析细胞中特异性抗乙肝病毒表位的CTL比例。结果 adr疫苗与adw疫苗均可诱导转基因小鼠产生抗乙肝表面抗原抗体,且抗体水平差异无统计学意义(P>0.05),2种疫苗对转基因小鼠的体液免疫应答的增强作用相似;adr疫苗诱导的乙肝表面抗原特异性CTL应答作用较adw疫苗更明显(P<0.05)。结论 adr重组乙肝疫苗(汉逊酵母)诱导转基因小鼠的体液免疫应答作用与现有adw疫苗相似,但其诱导机体产生细胞免疫应答的能力更显著,可能具有更加良好的预防HBV感染作用和前景。
Objective To evaluate the immune character and function of recombinant HB vaccine of subtype adr expressed in Hansenula polymorpha.Methods Human HLA-A2 transgenic mice were immunized with recombinant HB vaccine of subtypes adr and adw(expressed in Saccharomyces cerevisiae),each at a dosage of 2 μg / 0.2 ml,and boostered 3 weeks later.Serum samples were collected 1 weeks after the booster immunization and determined for anit-HBs level by ELISA.Meanwhile,the spleens of mice were collected and prepared into splenocyte suspension,in which the percentage of anti-HBV specific CTL was measured by flow cytometry with HLA-A2 / WLSLLVPFV pentamer staining.Results Both recombinant HB vaccines of subtypes adr and adw induced anti-HBs in mice,while the anti-HBs levels showed no significant difference(P 0.05),indicating similar effects of the vaccine of two subtypes in enhancing the humoral immune response in transgenic mice.However,The HBsAg-specific CTL response level induced with HB vaccine of subtype adr was significantly higher than that with the vaccine of subtype adw(P 0.05).Conclusion Recombinant HB vaccine of subtype adr evoked a similar humoral immunity and a high cellular immunity as compared with HB vaccine of subtype adw,which might be of good prospect in prevention of HBV infection.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第11期1410-1413,共4页
Chinese Journal of Biologicals
基金
国家自然科学基金项目(81172884)
国家基础科学人才培养基金(J1030831/J0108)
北京大学创新人才培养计划北京大学医学部新教师启动基金项目(BMU0239)
关键词
肝炎疫苗
乙型
体液免疫
细胞免疫
肝炎表面抗原
乙型
表位
Hepatitis vaccine
B
Humoral immunity
Cellular immunity
Hepatitis surface antigen
B
Epitope