摘要
目的初步评价肠道病毒71型(Enterovirus 71,EV71)与甲型肝炎病毒(Hepatitis A virus,HAV)灭活疫苗联合免疫小鼠诱导的特异性免疫应答水平以及两种抗原间的相互作用。方法将HAV灭活疫苗和EV71灭活疫苗以不同剂量配比制成联合疫苗,免疫ICR小鼠后,通过ELISA法检测血清HAV抗体效价;固定病毒-稀释血清法检测血清EV71中和抗体效价;流式细胞术检测脾脏中淋巴细胞亚群CD4+/CD8+的百分率;ELISPOT法检测脾脏中淋巴细胞分泌IL-4和IFNγ的水平。结果初免后28 d和加强免疫后7 d,各组小鼠血清中EV71中和抗体阳转率均达100%,单价疫苗组和联合疫苗组EV71中和抗体效价均呈剂量依赖性;初免后28 d,小鼠血清中能够产生保护性HAV抗体,加强免疫后抗体水平升高;联合疫苗组与相应的单价疫苗组间抗体水平差异无统计学意义(P>0.05),未发现EV71与HAV的相互作用;初免后28 d,各组小鼠脾脏中CD8+细胞比例高于CD4+细胞(P<0.001),加强免疫后7 d则相反(P<0.001);初免后28 d,HAV全病毒和EV71多肽均不能刺激各组小鼠脾细胞产生阳性反应,加强免疫后7 d,各组小鼠脾细胞分泌IL-4的水平均高于IFNγ(P<0.05)。结论一定剂量的HAV和EV71联合疫苗可诱导小鼠产生良好的体液免疫应答,同时诱导细胞免疫应答,但相对较弱,未发现两种抗原间存在相互干扰或协同作用。
Objective To primarily evaluate the specific immune response level induced in mice by combined immunization with inactivated enterovirus 71（EV71 ） vaccine and inactivated hepatitis A virus （HAV） vaccine as well as the interaction of EV7t and HAV antigens. Methods Combined vaccine was prepared by formulation of inactivated HAV and inactivated EVT1 vaccines at vari- ous dosage ratios, with which ICR mice were immunized and determined for serum anti-HAV titer by ELISA, for serum neutralizing antibody titer against EV71 by virus fixation-serum dilution method, for percentage of lymphocyte subsets CD4＋/CD8~ in spleen by flow cytometry, and for IL-4 and IFN＋ levels secreted by lymphocytes in spleen by ELISPOT. Results All the positive rates of neu- tralizing antibody against EV71 in sera of mice in various groups reached 100% 28 d after primary immunization and 7 d after booster immunization, while the neutralizing antibody titers were dose-dependent. Protective HAV antibody was induced in sera of mice 28 d after primary immunization, which increased after booster immunization. The antibody levels induced by combined vaccine showed no significant difference with those by the corresponding monovalent vaccine （P 〉 0. 05 ）, and no interaction between EV71 and HAV was found. The percentages of CD8＋ cells in spleens of mice in various groups were significantly higher 28 d after primary immuniza- tion （P 〈 0. 001 ）, while was significantly lower 7 d after booster immunization, than those of CD4＋ cells （P 〈 0. 001 ）. Neither whole HAV nor EV71 polypeptide stimulated the secretion of cytokines by splenocytes of mice in various groups 28 d after primary immunization, however, 7 d after booster immunization, the IL-4 levels secreted by splenocytes of mice in various groups were signif- icantly higher than those of IFN7 （P 〈 0. 05 ）. Conclusion Combined immunization with HAV and EV71 vaccines at certain dosages induced high humoral immune response and relatively low cellular immune response. However, no interference or synergism was observed between the two vaccine antigens.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第4期398-402,共5页
Chinese Journal of Biologicals
关键词
肠道病毒A型
人
肝炎病毒
甲型
灭活疫苗
联合免疫
Enterovirus type A, human
Hepatitis virus, A
Inactivated vaccine
Combined immunization
