硒化卡拉胶对甲型肝炎减毒活疫苗诱导的小鼠体液免疫应答的影响

摘要

目的 研究硒化卡拉胶(KSC)对甲型肝炎减毒活疫苗(HepA-1)诱导的小鼠体液免疫应答的影响.方法 随机分成7个组,每组7只ICR小鼠.分组:阴性对照组(生理盐水)、单纯抗原组(18EU HepA-1)、铝佐剂组[Al(OH)3 0.3 mg+18EU HepA-1]、KSC实验组(KSC 1 mg+18EU HepA-1、KSC 0.5 mg+18EU HepA-1、KSC 0.25 mg+18EU HepA-1、KSC 0.1 mg+18EU HepA-1),皮下多点免疫注射,注射总体积300μL,免疫1次.免疫注射后及其后4周、8周、12周、16周尾静脉采血,取血清用间接酶联免疫吸附法测定抗-甲型肝炎病毒(HAV) IgG抗体滴度.在实验期间,观察小鼠的健康状况,并于免疫后16周,取KSC最佳剂量组及阴性对照组小鼠肾脏、脾脏、肝脏、肺脏、心脏组织,制备病理切片,进行对比观察.结果 除阴性对照组外,其余各组在各个检测时间点均检测到抗HAV IgG抗体,且存在先升高后降低的趋势,铝佐剂组在12周达到峰值,其余各组在8周达到峰值.KSC 0.5 mg组在8周、12周抗体水平高于单纯抗原组(t=3.103,t=2.573,P＜0.05),KSC 0.25 mg组在12周抗体水平高于单纯抗原组(t=2.602,P＜0.05).本次实验的最佳剂量为KSC 0.5 mg.试验期内,观察小鼠各项生理指标未见异常,KSC 0.5 mg组小鼠心脏、肝脏、脾脏、肺脏、肾脏组织均未观察到病变.结论 KSC能够作为佐剂增强HepA-1诱导的小鼠体液免疫反应.%Objective To investigate the effect of kappa-selenocarrageenan (KSC) on humoral immune response induced by hepatitis A virus attenuated live vaccine (HepA-1) in mice.Methods Seven experiment groups were set up,with 7 ICR mouse per group randomly,negative control group (saline),pure antigen group (18EU HepA-1),Aluminium adjuvant group [Al(OH)3 0.3 mg+18 EU HepA-1],KSC experimental groups (KSC 1 mg+18 EU HepA-1,KSC 0.5 mg+18 EU HepA-1,and KSC 0.25 mg +18 EU HepA-1,KSC 0.1 mg+18 EU HepA-1),in which mice were injected subcutaneously with 300 μL,immunizing one time.The anti-HAV specific IgG antibody levels were tested by indirect ELISA method in the serum of mice after the first 4th,8th,12th,16th week of injection.The healthy status of mice were observed during the study.The kidney,spleen,liver,lung and heart of mice immunized with KSC+HepA-1 at the optimal dosage and those in negative control group were taken out at 16th week after immunization,and prepared into sections for pathological observation.Results HAV-specific IgG antibody of all mice were detected at four setting time point except negative control group.The antibody level of all groups increased first and then decreased.The peak of Aluminium adjuvant group at the 12th week and others at the 8th week.The level of antibody in KSC 0.5 mg+18EU HepA-1 group at 8th,12th week was higher than that in simple antigen group (t=3.103,t=2.573,P＜0.05).The level of antibody in KSC 0.25 mg +18EU HepA-1 group at week 12 was higher than that in pure antigen group(t=-2.602,P＜0.05).The optimal dosage of mice in this test was 0.5 mg.No abnormality was observed in the physiological indexes of mice in various groups,while there was no pathological changes in the kidney,spleen,liver,lung and heart of mice immunized with HepA-1 +0.5 mg KSC.Conclusion KSC can be used as an adjuvant to enhance humoral immune response induced by HepA-1 in mice.