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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 investigate the immunological characteristics and function of adr subtype recombinant hepatitis B vaccine (adr vaccine) expressed by Hansenula polymorpha. Methods The HLA-A2 mice were immunized with adr vaccine and recombinant hepatitis B vaccine (adw vaccine) expressed by Saccharomyces cerevisiae. The immunized mice were immunized with 2μg / 0.2 ml of immunization for 2 weeks after the initial immunization. One week after the second immunization, the blood was collected and the serum was separated. The anti-HBs antibody level in the serum of mice was detected by ELISA. At the same time, the spleen of the mice was taken out to prepare the spleen cell suspension. The proportion of CTL specific anti-HBV epitopes in the cells was analyzed by HLA-A2 / WLSLLVPFV pentamer fluorescent labeling flow cytometry. Results Both adr vaccine and adw vaccine induced anti-hepatitis B surface antigen antibody in transgenic mice with no significant difference in antibody levels (P> 0.05). The two vaccines had similar potentiating effects on humoral immune response in transgenic mice; adr Vaccine-induced hepatitis B surface antigen-specific CTL response was more pronounced than the adw vaccine (P <0.05). Conclusion The humoral immune response induced by adr recombinant Hepatitis B vaccine (Hansenula polymorpha) in transgenic mice is similar to that of the existing adw vaccine, but its ability to induce cellular immune response is more significant and may have a better effect and preventive effect on HBV infection .