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目的比较甲型乙型肝炎联合疫苗(Hepatitis A and B Combined Vaccine,HepAB)与重组乙型肝炎(乙肝)疫苗(酿酒酵母)(Hepatitis B Vaccine Made by Recombinant Deoxyribonucleic Acid Techniques in SaccharomycesCerevisiae Yeast,HepB-SCY)、甲型肝炎(甲肝)灭活疫苗(Hepatitis A Virus Inactivated Vaccine,HepA-Ⅰ)诱导小鼠细胞免疫和体液免疫应答的特点。方法 4组无特定病原体的BALB/c小鼠(H-2d,6～8周龄,16～18克,雌性),分别皮下接种HepAB[含甲肝抗原150单位(Unit,U)、乙肝抗原3微克(μg)]、HepB-SCY(含乙肝抗原3μg)、HepA-Ⅰ(含甲肝抗原150U)及铝佐剂对照,于免疫后14d分离脾单个核细胞(Mononuclear Cell,MNC),应用酶联免疫斑点试验(Enzyme-linked Immunospot Assay,ELISPOT)测定MNC体外刺激后所产生的细胞因子干扰素(Interferon,IFN)-γ、白细胞介素(Interleukin,IL)-2斑点形成细胞数(Spots-forming Cell,SFC);同时应用酶联免疫吸附试验(Enzyme-linked Immunosorbent Assay,ELISA)检测免疫后脾MNC培养上清中诱导分泌抗原特异性IFN-γ细胞因子的含量;分别于免疫后14d、28d检测小鼠血清抗甲肝病毒抗体(Antibody to Hepatitis AVirus,Anti-HAV)IgG和抗乙肝病毒表面抗原抗体(Antibody to Hepatitis B Virus Surface Antigen,Anti-HBs)水平。结果应用ELISPOT法检测,HepAB免疫小鼠后诱导乙肝抗原特异性IL-2分泌水平(50%)高于HepB-SCY(20%),HepAB组诱导甲肝抗原特异性IFN-γ阳转率(70%)高于HepA-Ⅰ组(30%),且HepAB诱导乙肝抗原特异性IFN-γ分泌水平(80%)均显著高于HepB-SCY组(30%)(P=0.03489)。应用ELISA法检测HepAB免疫后脾MNC培养诱导分泌甲肝抗原特异性IFN-γ阳转率(80%),也显著高于HepA-Ⅰ组(20%)(P=0.01151)。HepAB组于免疫14d、28d诱导的Anti-HBs抗体滴度分别为93.6毫国际单位/毫升(mIU/ml)和476.8 mIU/ml,均高于HepB-SCY组的26.5 mIU/ml和237.8 mIU/ml。结论两种细胞免疫评价方法结果均表明,HepAB诱导抗原特异性细胞免疫应答反应优于HepB-SCY、HepA-Ⅰ;HepAB与HepB-SCY相比较,在诱导Anti-HBs水平方面也同样具有优势。
Objective To compare Hepatitis A and B Combined Vaccine (HepAB) with Hepatitis B Vaccine Made by Recombinant Deoxyribonucleic Acid Techniques in Saccharomyces Cerevisiae Yeast (HepB-SCY) , Hepatitis A (Hepatitis A) inactivated Vaccine (HepA-Ⅰ) induced cellular and humoral immune response in mice. Methods 4 BALB / c mice without specific pathogen (H-2d, 6-8 weeks old, 16-18 g, female) were inoculated subcutaneously with HepAB [Unit: U containing Hepatitis A antigen 150 (M g), HepB-SCY (containing 3μg of Hepatitis B antigen), HepA-Ⅰ (containing hepatitis A antigen 150U) and aluminum adjuvant. The mononuclear cells (MNC) The levels of Interferon (IFN) -γ, Interleukin (IL) -2 and Spots-forming (MPS) in vitro induced by MNC were measured by Enzyme-linked Immunospot Assay (ELISPOT) Cell, SFC). Meanwhile, the levels of IFN-γ secreting antigen in the supernatant of MNCs were detected by Enzyme-linked Immunosorbent Assay (ELISA) The level of Antibody to Hepatitis B Virus Surface Antigen (Anti-HBs) was detected by ELISA. Results The level of IL-2 secretion induced by HepAB in mice was higher than that of HepB-SCY (50%) by ELISPOT. The positive rate of Hepatitis A antigen-specific IFN-γ %) Was higher than HepA-Ⅰ group (30%). HepAB-induced hepatitis B antigen-specific IFN-γ secretion was significantly higher than that of HepB-SCY group (30%) (P = 0.03489). The positive rate of hepatitis A antigen-specific IFN-γ induction (80%) induced by HepA-immunized HepA-immunized mice was also significantly higher than that of HepA-Ⅰ group (20%) (P = 0.01151) by ELISA. Anti-HBs antibody titer in HepAB group was 93.6 mIU / ml and 476.8 mIU / ml at 14d and 28d after immunization, which were higher than that of HepB-SCY group at 26.5 mIU / ml and 237.8 mIU / ml. Conclusions The results of the two methods of cellular immunity evaluation showed that HepAB induced antigen-specific cellular immune response better than HepB-SCY and HepA-I. HepAB also had the advantage of inducing Anti-HBs level compared with HepB-SCY.