硫酸软骨素B佐剂对甲型肝炎病毒疫苗诱导小鼠体液免疫应答的影响

Effect of chondroitin sulfate B adjuvant on humoral immune response induced by hepatitis A virus vaccine in mice

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. Methods ICR mice were randomly divided into nine groups, eight for each. The mice in six test groups were injected s. c. with live attenuated hepatitis A vaccine HepA-l 18 EU ＋ CSB at concentrations of 5, 25, 45, 65, 85 and 105 μg respectively, while those in negative control group with physiological saline, those in antigen control group with HepA-1 18 EU alone, and those in aluminum control groups with HepA-1 18 EU ＋ 300 μg aluminum hydroxide, 200 μ1 for each. The specific IgG level against HAV in sera of mice at weeks 4, 8, 12 and 16 after immunization were determined by indirect ELISA. The healthy statuses of mice were observed during the study. The kidney, spleen, liver, lung and heart of mice of mice immunized with HepA-1 ＋ 65 -g CSB and those in negative control group were taken out at week 16 after immunization, and prepared into sections for pathological observation. Results Anti-HAV IgGs were induced in mice in various groups at weeks 4, 8, 12 and 16, of which the levels, except that in negative control group, increased as time went on, and reached peak values at week 8 then decreased gradually. Theanti-HAV IgG of mice immunized with HepA-1 ＋ 65 μg CSB maintained at a high level for a long time, which was significantly higher than that in antigen control group （P 〈 0. 05） and slightly higher than that in aluminum adjuvant control group at weeks 12 and 16 after immunization. The optimal dosage of CSB was 65 p.g for each mouse. No abnormality was observed in mice of various groups, while no pathological changes in the kidney, spleen, liver, lung and heart of mice immunized with HepA-1 ＋ 65 μg CSB. Conclusion CSB enhanced the humoral immune responses induced by HAV antigen in mice significantly, which might be developed as a novel adiuvant of vaccines.