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目的探讨氢氧化铝与黄连碱复合佐剂对甲肝疫苗诱导小鼠体液免疫的影响。方法选取6~8周龄雌性ICR小鼠49只,随机分为空白组、单纯抗原组、单纯黄连碱佐剂组、铝佐剂组及复合佐剂低、中、高剂量组,每组7只;在免疫后第4、8、12、16周,采用间接酶联免疫吸附测定法(ELISA)检测小鼠分离血清中的抗HAV IgG抗体水平。结果在免疫后第4、8、12、16周,除空白组外,各组小鼠均能检测到抗HAV IgG抗体,且抗体水平随免疫后时间的延长均先升高而后降低;在免疫后第12周,各组小鼠血清中抗HAV IgG抗体水平达到峰值,单纯抗原组、单纯黄连碱佐剂组、铝佐剂组及复合佐剂低、中、高剂量组小鼠分别为log_(10)(2.908±0.413)、log_(10)(2.960±0.315)、log_(10)(3.460±0.295)、log_(10)(3.460±0.351)、log_(10)(3.470±0.251)、log_(10)(3.470±0.251);在免疫后第4周,复合佐剂低剂量组小鼠抗HAV IgG抗体水平均高于单纯抗原组和单纯黄连碱佐剂组(均P<0.05);在免疫后第8、12、16周,复合佐剂低、中、高剂量组小鼠抗HAV IgG抗体水平均高于单纯抗原组和单纯黄连碱佐剂组(均P<0.05);在免疫后第12周,铝佐剂组小鼠抗HAV IgG抗体水平均高于单纯抗原组和单纯黄连碱佐剂组(均P<0.05);在免疫后第16周,铝佐剂组小鼠抗HAV IgG抗体水平高于单纯抗原组(P<0.05)。结论适当配比的黄连碱和氢氧化铝复合佐剂可快速、显著提高甲肝疫苗诱导小鼠的体液免疫应答,有望成为一种新的人用疫苗佐剂。
Objective To investigate the effect of aluminum hydroxide and coptisine complex adjuvant on humoral immunity induced by hepatitis B vaccine in mice. Methods 49 female ICR mice of 6-8 weeks old were randomly divided into blank group, antigen group, berberine adjuvant group, aluminum adjuvant group and compound adjuvant low, medium and high dose groups Only; at the 4th, 8th, 12th and 16th weeks after immunization, the level of anti-HAV IgG in the sera of mice was detected by indirect enzyme-linked immunosorbent assay (ELISA). Results At the 4th, 8th, 12th and 16th weeks after immunization, except for the blank group, anti-HAV IgG antibodies were detected in all groups, and the antibody levels increased first and then decreased with the prolongation of immunization. At the 12th week, the levels of anti-HAV IgG in the serum of the mice in each group reached a peak. The mice in the groups of antigen, chlortol hydrochloride, aluminum adjuvant and compound adjuvants were log_ (10) (2.908 ± 0.413), log_ (10) (2.960 ± 0.315), log_ (10) (3.460 ± 0.295), log_ (10) (3.460 ± 0.351), log_ (10) (10) (3.470 ± 0.251). At the 4th week after immunization, the levels of anti-HAV IgG in the low-dose adjuvant group were significantly higher than those in the antigen group and the Coptis-alkali group (all P <0.05) At the 8th, 12th and 16th week after immunization, the level of anti-HAV IgG in the low, medium and high dose groups of the adjuvant was higher than that of the antigen group and the berberine adjuvant alone group (all P <0.05) At week 12, the levels of anti-HAV IgG in the aluminum adjuvant group were significantly higher than those in the pure antigen group and the berberine alkaline adjuvant group (both P <0.05). At the 16th week after immunization, the mice in the aluminum adjuvant group had anti-HAV IgG antibody levels higher than simple antigen (P <0.05). Conclusion The proper ratio of coptis alkaloid and aluminum hydroxide compound adjuvant can rapidly and significantly enhance the humoral immune response induced by hepatitis A vaccine in mice and is expected to become a new vaccine adjuvant for human.