目的探讨酵母多糖-角鲨烯复合佐剂对甲型肝炎(简称甲肝)和乙型肝炎(简称乙肝)抗原诱导小鼠体液免疫应答的增强作用,并对皮下和鼻腔接种的免疫效果进行比较。方法将酵母多糖与角鲨烯按一定比例混合,制成复合佐剂,将不同剂量的复合佐剂与HBsAg混合,并设生理盐水对照组、HBsAg对照组、铝佐剂对照组和酵母多糖对照组,均经皮下免疫ICR小鼠1次,分别于免疫后2、4、8、16周,采用ELISA法检测小鼠血清中抗HBsAg IgG抗体水平,筛选复合佐剂的最佳免疫剂量。以复合佐剂的最佳免疫剂量与甲肝抗原混合,并设生理盐水对照组、甲肝抗原对照组、铝佐剂对照组和酵母多糖对照组,均经皮下免疫ICR小鼠1次,分别于免疫后2、4、8周检测小鼠血清中抗甲肝抗原IgG抗体水平。以复合佐剂最佳免疫剂量分别与HBsAg和甲肝抗原混合,并设生理盐水对照组、HBsAg对照组和甲肝抗原对照组,经鼻腔免疫ICR小鼠,共免疫3次,间隔2周,分别于末次免疫后4、8周检测小鼠血清中抗HBsAg和甲肝抗原IgG抗体水平。在试验期间,对小鼠的健康状况进行观察,并对小鼠的心脏、肝脏、脾脏、肺脏、肾脏组织进行病理分析。结果酵母多糖-角鲨烯复合佐剂对HBsAg的最佳免疫剂量为2 mg酵母多糖+8μl角鲨烯,该组抗体水平明显高于铝佐剂对照组和酵母多糖对照组(P<0.05);复合佐剂对甲肝抗原也有较好的体液免疫增强作用,抗体水平显著高于铝佐剂对照组和酵母多糖对照组(P<0.05);复合佐剂通过鼻腔免疫,对HBsAg和甲肝抗原的体液免疫应答也有增强作用,但效果不如皮下免疫,甲肝抗原鼻腔免疫效果优于HBsAg(P<0.05)。在试验期内,小鼠均未出现异常反应,各器官均未发生病变。结论酵母多糖-角鲨烯复合佐剂能显著增强甲肝和HBsAg抗原对小鼠的体液免疫应答,免疫效果优于铝佐剂;皮下免疫效果优于鼻腔免疫;在免疫剂量范围内,复合佐剂安全性良好。 Objective To investigate the enhancing effect of zymosan-squalene adjuvant on the humoral immune response induced by hepatitis A (Hepatitis A) and hepatitis B (Hepatitis B) antigen in mice and to compare the immune effects of subcutaneous and nasal vaccination. Methods Zymosan and squalene were mixed in a certain proportion to make compound adjuvant. Different doses of compound adjuvants were mixed with HBsAg and normal saline control group, HBsAg control group, aluminum adjuvant control group and zymosan control group The mice were immunized subcutaneously with ICR mice once, respectively. The serum levels of anti-HBsAg IgG in mice were detected by ELISA at 2, 4, 8 and 16 weeks after immunization, and the best immunized dose of compound adjuvant was screened. The optimal adjuvant immunization dose was mixed with hepatitis A antigen, and the control group of normal saline, hepatitis A antigen, aluminum adjuvant and zymosan control group were subcutaneously immunized with ICR mice once, respectively, After 2, 4, 8 weeks, the serum level of anti-hepatitis B antigen IgG in mice was detected. The best immune dose of compound adjuvant was mixed with HBsAg and Hepatitis A antigen separately. The normal saline control group, HBsAg control group and Hepatitis A antigen control group were immunized with ICR mice via nasal cavity three times at intervals of 2 weeks. At 4 and 8 weeks after the last immunization, the serum levels of anti-HBsAg and anti-Hepatitis A antigen IgG in mice were detected. During the experiment, the mice were observed for their health status, and pathological analysis was performed on the heart, liver, spleen, lung and kidney of mice. Results The optimal immunization dose of HBsAg and yeast polysaccharide - squalene adjuvant was 2 mg zymosan + 8 μl squalene. The antibody level was significantly higher than that of aluminum adjuvant and zymosan control group (P <0.05) ; The composite adjuvant also had better enhancement of humoral immunity against hepatitis A antigen, the antibody level was significantly higher than that of aluminum adjuvant control group and zymosan control group (P <0.05) Humoral immune response also enhance the effect, but the effect is not as good as subcutaneous immunization, hepatitis A antigen nasal immunity better than HBsAg (P <0.05). During the experimental period, no abnormal reaction occurred in all mice and no lesions were observed in all organs. CONCLUSION: The zymosan-squalene adjuvant can significantly enhance the humoral immune response of hepatitis A and HBsAg antigens in mice, and the immune effect is superior to that of aluminum adjuvant. The subcutaneous immune effect is better than nasal immunity. In the range of immunological dose, Good safety.