氢氧化锌与硫酸乙酰肝素复合佐剂对狂犬病疫苗诱导的小鼠体液免疫应答的作用

Effect of zinc hydroxide and heparan sulfate compound adjuvant on humoral immunity induced by rabies vaccine in mice

目的探讨氢氧化锌与硫酸乙酰肝素(Heparan sulfate,HS)复合佐剂对狂犬病疫苗诱导的小鼠体液免疫应答的影响。方法取64只ICR小鼠随机分为8组,每组8只,分别为复合佐剂(0.27 mg氢氧化锌,100μg HS,0.125 IU狂犬病疫苗)1、2、3次免疫组,狂犬病疫苗1、2、3次免疫组,狂犬病疫苗常规5次免疫组,生理盐水对照组。均经小鼠胫骨前肌免疫,除狂犬病疫苗常规5次免疫组于0、3、7、14、28 d进行免疫外,其他各组均为隔周免疫。分别于初免疫后1、2、3、4、8、12、16周经尾静脉采血,分离血清,ELISA法检测小鼠血清中抗-狂犬病毒(Rabies virus,RABV)IgG水平。结果初免后1周,除生理盐水对照组小鼠血清未检测到抗-RABV IgG外,各实验组均产生抗-RABV特异性IgG;所有复合佐剂组在初免后3周均可产生高水平的IgG,初免后12周反弹性升高达到峰值,第16周仍维持较高水平。初免后1、2、3、4、8、12、16周,复合佐剂1、2、3次免疫组IgG水平均高于狂犬病疫苗相同次数免疫组,差异均有统计学意义(P均<0.05),且IgG水平持续时间较长。初免后2周,复合佐剂2、3次免疫组的IgG水平均高于狂犬病疫苗常规5次免疫组(3次免疫后);初免后3周,复合佐剂3次免疫组的IgG水平高于狂犬病疫苗常规5次免疫组(4次免疫后)(P均<0.05)。结论氢氧化锌和HS复合佐剂能增强狂犬病疫苗诱导的小鼠体液免疫应答。

Objective To investigate the effect of zinc hydroxide and heparan sulfate（HS） compound adjuvant on humoral immunity induced by rabies vaccine in mice.Methods Sixty-four ICR mice were randomly divided into one control and seven test groups,8 for each.The mice in test groups 1,2 and 3 were immunized i.m.with 1,2 and 3 doses of 0.125 IU rabies vaccine containing compound adjuvant consisting of 0.27 mg zinc hydroxide and 100 μg HS,while those in test groups 4,5 and 6 with 1,2,3 doses of adjuvant-free rabies vaccine,respectively,each at an interval of one week.The mice in test group 7 were immunized with 5 doses of adjuvant-free rabies vaccine on days 0,3,7,14 and 28 respectively,while those in control group with physiological saline.Serum samples were collected 1,2,3,4,8, 12 and 16 weeks after the first immunization and determined for IgG level against rabies virus（RABV）by ELISA.Results Anti-RABV IgG levels were detected in the sera of mice in 7 test groups one week after the first immunization,while was not detected in those in control group.Three weeks after the first immunization,high antibody levels were induced in test groups 1,2 and 3,which reached the peak values 12 weeks and still maintained at a higher level at week 16.The IgG levels of mice 1,2,3,4,8,12 and 16 weeks after the first immunization were significantly higher in test groups 1, 2 and 3 than in test groups 4,5 and 6,respectively（each P〈0.05）,and lasted for a long time.Two weeks after the first immunization,the IgG levels in test groups 2 and 3 were significantly higher than that in test group 7（after the third dose was given）.However,3 weeks after the first immunization,the IgG level in test group 3 was significantly higher than that in test group 7（after the forth dose was given）（each P〈0.05）.Conclusion Zinc hydroxide and HS compound adjuvant enhanced the humoral immunity induced by rabies vaccine in mice.