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目的探讨硫酸软骨素B(chondroitin sulfate B,CSB)佐剂对甲型肝炎病毒((hepatitis A virus,HAV)疫苗诱导小鼠体液免疫应答的影响,以评价其佐剂效果。方法将ICR小鼠随机分成9组:甲型肝炎减毒活疫苗Hep A-l18 EU+不同剂量CSB(5、25、45、65、85、105μg)组、阴性对照组(生理盐水)、抗原对照组(Hep A-l 18 EU)和铝佐剂对照组[Hep A-l 18 EU+Al(OH)3300μg],每组8只,各组均经皮下注射ICR小鼠,200μl/只。于免疫后的第4、8、12和16周,采用间接ELISA法检测小鼠血清中抗-HAV特异性Ig G抗体水平。在实验期间,观察小鼠的健康状况,并于免疫16周后,取CSB 65μg组及阴性对照组小鼠肾脏、脾脏、肝脏、肺脏、心脏各主要器官,制备病理切片,进行对比观察。结果各组小鼠免疫后第4、8、12、16周,均产生抗-HAV Ig G抗体,除阴性对照组外,抗体水平均随时间的延长逐渐升高,于第8周达峰值,此后逐渐下降。CSB 65μg组抗体可长时间维持在高水平,至免疫后第12、16周,均高于抗原对照组(P<0.05),略高于铝佐剂对照组,最佳浓度为65μg/只。实验期间,各组小鼠均未出现异常;CSB65μg组小鼠肾脏、脾脏、肝脏、肺脏、心脏组织均未观察到病变。结论 CSB可明显增强HAV抗原诱导小鼠的体液免疫应答,具有成为新型疫苗佐剂的研发价值。
Objective To investigate the effect of chondroitin sulfate B (CSB) adjuvant on humoral immune response induced by hepatitis A virus (HAV) vaccine in mice to evaluate its adjuvant effect.Methods ICR mice Randomly divided into 9 groups: live attenuated hepatitis A vaccine Hep A -18 EU + different doses of CSB (5,25,45,65,85,105μg) group, negative control group (saline), antigen control group (Hep Al 18 EU) and aluminum adjuvant control group [Hep Al 18 EU + Al (OH) 3300μg], with 8 mice in each group.All mice were injected subcutaneously with ICR mice at 200μl / And 16 weeks, the level of anti-HAV-specific Ig G antibody in serum was detected by indirect ELISA.The health status of mice was observed during the experiment and after 16 weeks of immunization, CSB 65μg group and negative control group The kidneys, spleen, liver, lung and heart organs were prepared and pathological sections were prepared for comparison.Results Anti-HAV Ig G antibodies were produced at the 4th, 8th, 12th and 16th weeks after immunization in all groups except the negative In the control group, the antibody levels increased gradually with time and peaked at the 8th week, then decreased gradually.CSB 65μg group antibody (P <0.05), slightly higher than the control group with aluminum adjuvant, the optimal concentration was 65μg / only. During the experiment, each group No abnormalities were observed in mice, and no lesions were found in kidney, spleen, liver, lung and heart of CSB65μg group.Conclusion CSB can significantly enhance the humoral immune response induced by HAV antigen in mice, and become a new vaccine adjuvant for R & D value.