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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 on the humoral and cellular immune responses induced by HBsAg in mice. Methods HS and zinc hydroxide were combined with different doses and mixed with HBsAg (2μg), and ICR mice were immunized with negative control, antigen control, aluminum adjuvant control, single HS and single zinc hydroxide control group, a total of 23 groups . Blood samples were collected at 4, 8, 12, 16, 20 and 24 weeks after the last immunization, respectively. Serum anti-HBsAg antibody levels were detected by ELISA. BALB / c mice were immunized with the best humoral immunity Eight weeks after the immunization, cell killing activity was detected by lactate dehydrogenase method. Results In the negative control group, IgG antibody was not detected at all time points. The antibody titers of all the other groups reached their peak values at the 8th week after the last immunization. The antibody levels showed a decreasing trend with time. Among them, the antibody of group 23 (100μg HS + 1.0 mg zinc hydroxide, 2 doses of immunization) had the highest level of antibody for longer duration. The enhancement of humoral immunity to HBsAg at 8 weeks after the last immunization was significantly better than that of aluminum adjuvant and HS Single adjuvant (P <0.05), better than zinc single adjuvant, but the difference was not statistically significant (P> 0.05). Combination adjuvant can induce specific CTL cellular immune response. Conclusion Both HS and zinc hydroxide adjuvant can effectively enhance the specific humoral immune response induced by HBsAg in mice and induce CTL cellular immune response significantly.