国产与进口重组酵母乙型肝炎疫苗免疫小鼠早期细胞免疫应答的比较  

Early cellular immune response induced by domestic and imported recombinant hepatitis B vaccines prepared with yeast cells in mice

作　　者：何鹏[1] 邱少辉[1] 方鑫[1] 钟熙[1] 梁争论[1] 沈琦[1] 胡忠玉[1] 

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国生物制品学杂志》2014年第6期737-741,746,共6页Chinese Journal of Biologicals

基　　金："艾滋病和病毒性肝炎等重大传染病防治"十二五科技重大专项(2012ZX10004701)

摘　　要：目的比较国产与进口重组酵母乙型肝炎疫苗免疫小鼠早期细胞免疫应答的差异。方法取批签发合格的国产疫苗1和2(酿酒酵母)、进口疫苗1(酿酒酵母)、国产疫苗3和4(汉逊酵母)、进口疫苗2(汉逊酵母)各3批,经背部皮下免疫BALB/c小鼠,3μg/(100μl·只),于免疫后7 d,处死小鼠,无菌取脾,制备脾细胞悬液,分离脾单个核细胞(mononuclear cell,MNC)。采用酶联免疫斑点(enzyme linked immunospot assay,ELISPOT)法测定小鼠MNC体外接受HBsAg刺激后产生的抗原特异性细胞因子IFNγ、IL-2、IL-4和IL-5水平及体外接受MHCⅠ类多肽S28-39刺激后产生的IFNγ水平。结果小鼠免疫酿酒酵母乙肝疫苗后,以HBsAg刺激MNC,国产疫苗1诱导产生的IFNγ、IL-2和IL-4水平均明显高于国产疫苗2和进口疫苗1,国产疫苗1和2诱导的IL-5水平均明显高于进口疫苗1(P均<0.05);以S28-39刺激MNC,3种疫苗诱导产生的IFNγ水平差异均无统计学意义(P>0.05)。小鼠免疫汉逊酵母乙肝疫苗后,以HBsAg或S28-39刺激MNC,进口疫苗2诱导的IFNγ和IL-4水平均明显高于国产疫苗3(P<0.05);3种汉逊酵母乙肝疫苗诱导的IL-2和IL-5水平差异均无统计学意义(P>0.05)。结论国产重组酵母乙肝疫苗诱导早期细胞免疫应答能力较好。Objective To compare the difference of domestic and imported recombinant hepatitis B （HB） vaccines pre- pared with yeast cells in induction of cellular immune response in mice. Methods Domestic HB vaccines 1 and 2 （Saccharomyces cerevisiae）, imported vaccine 1 （Saccharomyces cerevisiae）, domestic vaccines 3 and 4 （Hansenula polymorpha） and imported vaccine 2 （Hansenulapolymorpha）, three batches for each, were inoculated s. c. into BALB/c mice, 3 μg/ 100 μl for each. The mice were killed 7 d after immunization, of whom the spleens were collected aseptically and prepared into splenocyte suspension from which mononuclear cells （MNCs） were isolated. HBsAg antigen specific IFNγ IL-2, IL-4 and IL-5 levels produced by MNCs after stimulation in vitro with HBsAg as well as the IFNγ level after stimulation in vitro with MHC I polypeptide S28-39 were determined by enzyme linked immunospot assay （ELISPOT）. Results The IFNγ, IL-2 and IL-4 levels secreted by MNCs after stimulation in vitro with HBsAg in the mice inoculated with domestic vaccine 1 were significantly higher than those with domestic vaccine 2 and imported vaccine 1, while the IL-2 levels induced with domestic vaccines 1 and 2 were significantly higher than that with imported vaccine 1 （each P 〈 0：05 ）. After stimulation of MNCs with S28-39, the IFNγ levels induced with the three vaccines prepared with S. cerevisiae showed no significant difference （P 〉 0. 05）. However, after stimulation with HBsAg or S28-39, both the IFNγ and IL-4 levels induced with imported vaccine 2 were significantly higher than those with domestic vaccine 3, while the IL-2 and IL-5 levels induced by three vaccines prepared with tt. polymorpha showed no significant difference （P 〉 0. 05）. Conclusion Domestic recombinant HB vaccines prepared with S. cerevisiae induced early ce- llular immune response effectively.

关 键 词：乙型肝炎 疫苗 酶联免疫斑点法 细胞免疫 

分 类 号：R373.2[医药卫生—病原生物学] R392-33[医药卫生—基础医学]