酵母多糖-角鲨烯复合佐剂对甲型肝炎和乙型肝炎抗原诱导小鼠体液免疫应答的增强作用  

Enhancement of zymosan combined with squalene to humoral immune response induced by HBsAg and hepatitis A antigen

作　　者：王丽萍[1] 王越[1] 王海漩[1] 胡云章[1] 胡凝珠[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所疫苗研究室云南省重大传染病重点实验室云南省重大传染病疫苗工程技术研究中心,云南昆明650118

出　　处：《中国生物制品学杂志》2013年第4期473-476,共4页Chinese Journal of Biologicals

基　　金：国家高技术研究发展计划(863计划)"新型疫苗佐剂的研发及其应用研究"(2012AA02A406);云南省创新团队"中国医学科学院医学生物学研究所新型疫苗佐剂应用研究省创新团队"(2011CI140)

摘　　要：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Objective To investigate the enhancement of zymosan combined with squalene to humoral immune response induced by HBsAg and hepatitis A（HA） antigen in mice.Methods Zymosan and squalene were mixed at a certain proportion to prepare a complex adjuvant.HBsAg were mixed with the complex adjuvant at various dosages respectively,with which ICR mice were immunized once,using physiological saline,HBsAg,aluminum adjuvant and zymosan as controls.The IgG levels against HBsAg in sera of mice were determined by ELISA 2,4,8 and 16 weeks after immunization respectively,based on which the dosage of complex adjuvant was optimized.The complex adjuvant at the optimal dosage was mixed with HA antigen and immunized s.c.to ICR mice once,using physiological saline,hepatitis A antigen,aluminum adjuvant and zymosan as controls,and the serum IgG levels against HA antigen were determined 2,4 and 8 weeks after immunization.The complex adjuvant at the optimal dosage was mixed with HBsAg and HA antigen respectively,with which ICR mice were immunized i.n.for 3 times each at an interval of 2 weeks,using physiological saline,HBsAg and HA antigen as controls.The IgG levels against HBsAg and HA antigen in sera of mice were determined 4 and 8 weeks after the last immunization respectively.The mice were observed for health during the test,of which the heart,liver,spleen,lung and kidney were subjected to pathological examination.Results The optimal dosage of the complex adjuvant for HBsAg was 2 mg zymosan ＋ 8 μl squalene,with which the IgG level induced was significantly higher than those in aluminum adjuvant and zymosan control groups（P 0.05）.The complex adjuvant also enhanced the humoral immune response induced by HA antigen,with which the antibody level induced were significantly higher than those in aluminum adjuvant and zymosan control groups（P 0.05）.The complex adjuvant administered by intranasal route also enhanced the humoral immune response induced by HBsAg and HA antigen,though the effect was not as satisfatory as that by subc

关 键 词：酵母多糖 角鲨烯 佐剂 乙型肝炎表面抗原 甲型肝炎抗原 

分 类 号：R392-33[医药卫生—免疫学]