以壳聚糖为佐剂的幽门螺杆菌疫苗诱导的细胞免疫反应  

Cellular immunity induced by H.pylori vaccine with chitosan as adjuvant

作　　者：龚燕锋[1] 谢勇[1] 周南进[2] 陈江[1] 周小江[1] 吕农华[1] 王崇文[1] 

机构地区：[1]南昌大学第一附属医院消化研究所 [2]江西省医学科学研究所,江西南昌330006

出　　处：《细胞与分子免疫学杂志》2007年第7期595-599,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30460052);江西省学科带头人培养项目(K010501);江西卫生厅重点招标项目(31070);江西省教育厅资助项目

摘　　要：目的:探讨以壳聚糖为佐剂的幽门螺杆菌(Hp)疫苗诱导的细胞免疫反应及其在免疫保护中的作用。方法:BALB/c小鼠随机分为空白对照组(PBS溶液)、壳聚糖酸溶液组、壳聚糖颗粒组、Hp抗原组、Hp抗原+壳聚糖酸溶液组、Hp抗原+壳聚糖颗粒组、Hp抗原+霍乱霉素(CT)组、Hp抗原+壳聚糖酸溶液+CT组及Hp抗原+壳聚糖颗粒+CT组,各组于第0、7、14、21天灌胃各免疫1次,末次免疫后4周给予1×1012CFU/L的SS1Hp菌液0·5mL/只进行攻击,隔日1次,共2次。在攻击前后分批处死小鼠,采用Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用定量ELISA法检测胃黏膜内IL-2、IL-4和IL-10含量;HE染色进行胃黏膜病理学检测。结果:①以壳聚糖为佐剂的Hp疫苗的免疫保护率达60%,与以CT为佐剂的Hp疫苗的免疫保护率(58·33%)相似,显著高于单纯Hp抗原组及其他不含Hp抗原组(P<0·05)。②胃黏膜内IL-2的水平攻击后含佐剂组显著高于对照组(P<0·05)。IL-10水平攻击前以壳聚糖为佐剂组高于无佐剂组,攻击后以CT+壳聚糖颗粒为佐剂组高于其他组(P<0·05)。IL-4水平攻击前以壳聚糖为佐剂组高于无佐剂组(P<0·05),攻击后以壳聚糖颗粒为佐剂组高于以CT为佐剂组(P<0·05),以壳聚糖溶液为佐剂组高于对照组、无佐剂组及佐剂中含CT组(P<0·05)。③胃黏膜炎症程度在单纯壳聚糖组和以壳聚糖颗粒为佐剂组显著低于以CT为佐剂组(P<0·05)。结论:①以壳聚糖为佐剂的Hp疫苗对Hp感染具有免疫保护作用。②以壳聚糖为佐剂的Hp疫苗可促进Th1和Th2的混合免疫反应,并逆转Hp感染所致Th2反应的抑制,使Th1和Th2反应达到平衡,从而发挥其免疫保护作用。③以壳聚糖为佐剂的Hp疫苗所致的免疫后胃炎显著低于以CT为佐剂的Hp疫苗。AIM： To study the cellular immunity induced by H. pylori vaccine with chitosan as adjuvant and the mechanism of immunological protection. METHODS： BALB/c mice were randomly divided into nine groups and immunized by （1）PBS alone,（2）chitosan solution alone, （3）chitosan particles alone, （4） H. pylori antigen alone, （5） H. pylori antigen plus chitosan solution, （6）H. pylori antigen plus chitosan particles, （7）H. pylori antigen plus cholera toxin （CT）, （8）H. pylori antigen plus chitosan solution and CT, （9）H. pylori antigen plus chitosan particles and CT orally respectively once a week for four weeks. At 4 weeks after the last immunization, mice were challenged by alive H. pylori（ 1 × 10^9 CFU/mL） twice at two days intervals. Before and after the challenge, mice were killed in batches and the H. pylori-infection in gastric mucosa was detected by H. pylori culture and Giemsa stain. ELISA and HE stain were used to detect IL-2, IL-4, IL-10 levels and pathologic change in gastric mucosa. RESULTS： （1）ln the groups with chitosan as an adjuvant, 60% mice could achieve immunological protection, which was consistnt with using CT as an adjuvant （58.33%）, and was more than that when using H. pylori antigen alone or without H. pylori antigen. （2）After challenge, the IL-2 levels in gastric mucosa in the groups with adjuvants were significantly higher than those in the control group（ P 〈 0.001 - 0.05）. Moreover, IL-2 levels in the groups with adjuvants after challenge were significantly higher than these before challenge（P〈0.05）. Before challenge, the IL-10 and IL-4 levels in gastric mucosa were significantly higher in the groups with chitosan as adjuvant than those in non-adjuvant groups （P〈0.05）. After challenge, IL-10 levels were significantly higher in the groups with chitosan particles as adjuvant than those in other groups（ P 〈0.05） IL-4 levels were significantly higher in the groups with chitosan particles as an adjuvant than those in the

关 键 词：幽门螺杆菌 壳聚糖 疫苗 免疫佐剂 

分 类 号：R392.7[医药卫生—免疫学]