硫酸乙酰肝素与氢氧化锌联合佐剂对HBsAg诱导小鼠免疫应答的影响

Effect of Heparan Sulfate Combined with Zinc Hydroxide on Immune Response Induced by HBsAg in Mice

目的探讨硫酸乙酰肝素(Heparan sulfate,HS)与氢氧化锌联合佐剂对HBsAg诱导的小鼠体液免疫和细胞免疫应答的影响。方法将HS与氢氧化锌按不同剂量组合,与HBsAg(2μg)混合,免疫ICR小鼠,并设阴性对照、抗原对照、铝佐剂对照、单一HS及单一氢氧化锌对照组,共23组。分别于末次免疫后4、8、12、16、20、24周采血,分离血清,ELISA法检测小鼠血清中抗HBsAg抗体水平;选择体液免疫效果最佳的组免疫BALB/c小鼠,于免疫后8周,采用乳酸脱氢酶法检测细胞杀伤活性。结果阴性对照组在各时间点均未检测到IgG抗体,其余各组的抗体滴度均在末次免疫后第8周达峰值,随着时间的延长,抗体水平呈下降趋势。其中第23组(100μg HS+1.0 mg氢氧化锌,免疫2针)抗体水平最高,且持续时间较长,末次免疫后8周,其对HBsAg的体液免疫增强效应显著优于铝佐剂和HS单一佐剂(P<0.05),优于氢氧化锌单一佐剂,但差异无统计学意义(P>0.05)。联合佐剂能诱导产生特异性的CTL细胞免疫效应。结论 HS和氢氧化锌联合佐剂既能有效增强HBsAg诱导的小鼠特异性体液免疫应答,又能显著诱导CTL细胞免疫应答。

Objective To investigate the effect of heparan sulfate（HS） combined with zinc hydroxide,as an immunological adjuvant,on immune response induced by HBsAg in mice.Methods ICR mice were divided into twenty-three groups,including test groups as well as negative control,antigen control,aluminum adjuvant control,HS control and zinc hydroxide groups.The HS and zinc hydroxide,at various dosages,were combined and further mixed with 2 μg of HBsAg,then immunized to the mice in various test groups by different schedules.Blood samples of the mice were collected 4,8,12,16,20 and 24 weeks after the last immunization,from which sera were separated and determined for anti-HBs level by ELISA.BALB/c mice were immunized at the same dosage by the same schedule as those in the group which showed the most satisfactory humoral immune effect and determined for cytotoxic activity by lactate dehydrogenase method 8 weeks later.Results No IgG was detected in blank control group at any time points.However,the IgG titers in all the rest groups reached peak values 8 weeks after the last immunization and showed a decreasing tendency as time went on.The IgG level in group 23（100 μg HS ＋ 1.0 mg zinc hydroxide,immunized twice） was the highest and lasted for a long time.Eight weeks after the last immunization,the humoral immune response levels of mice immunized with HS combined with zinc hydroxide were significantly higher than those with aluminum adjuvant and with HS adjuvant alone（P 0.05）,and insignificantly higher than that with zinc hydroxide adjuvant alone（P 0.05）.HS combined with zinc hydroxide induced specific CTL cellular immune response.Conclusion HS combined with zinc hydroxide enhanced the specific humoral immune response induced by HBsAg and induced CTL cellular immune response effectively.