以壳聚糖为佐剂的Hp疫苗诱导的体液免疫应答及其免疫保护效应  

Role of humoral immune response in the protection induced by H.pylori vaccine with chitosa as adjuvant

作　　者：谢勇[1] 龚燕锋[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,江西南昌330006 [2]江西省医学科学研究所,江西南昌330006

出　　处：《中国病理生理杂志》2007年第3期438-443,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No30460052);江西省学科带头培养项目;江西省卫生厅重点招标项目;江西省教育厅资助项目

摘　　要：目的:探讨以壳聚糖为佐剂的Hp疫苗的免疫保护作用及其机制。方法:BALB/c小鼠随机分为9组:①空白对照组:PBS溶液;②壳聚糖酸溶液组;③壳聚糖颗粒组;④Hp抗原组;⑤Hp抗原+壳聚糖酸溶液组;⑥Hp抗原+壳聚糖颗粒组;⑦Hp抗原+CT组;⑧Hp抗原+壳聚糖酸溶液+CT组;⑨Hp抗原+壳聚糖颗粒+CT组,各组于第0、7、14、21d灌胃各免疫1次,免疫后4周给予1×1012CFU/L的SS1Hp菌液每只0.5mL进行攻击,隔日1次,共2次。4周后,采用定量Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用ELISA法检测血清抗HpIgG、IgG1、IgG2a及唾液和胃黏膜内抗HpIgA,用SP免疫组织化学法检测胃黏膜内分泌型IgA(sIgA)。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抗原组及其它不含Hp抗原组(P<0.01或P<0.05),同时以CT+壳聚糖为佐剂的Hp疫苗的保护率为84.62%、85.71%,其Hp的定植评分显著低于无佐剂组及以CT为佐剂组(P<0.01,P<0.05)。②含佐剂的Hp疫苗所诱导产生的HpIgG水平显著高于对照组及无佐剂组(P<0.01,P<0.05),而以CT+壳聚糖为佐剂组所产生的抗HpIgG水平显著高于仅以CT或壳聚糖为佐剂组(P<0.05)。③胃黏膜内sIgA及特异性抗HpIgA水平在壳聚糖为佐剂组与以CT为佐剂组无差别(P>0.05),显著高于无佐剂组,而壳聚糖与CT联合应用组显著高于单以CT为佐剂组(P<0.01,P<0.05)。结论:以壳聚糖为佐剂的Hp疫苗对Hp感染具有免疫保护作用,并可成功诱导黏膜局部的特异性体液免疫应答,从而发挥免疫防御作用。AIM： To study the immunological protection of H. pylori vaccine with chitosa as adjuvant. METHODS： One- grade female BALB/c mice were randomly divided into nine groups and immunized by ①PBS alone; ②chitosan solution alone; ③chitosan particles alone; ④H. pylori antigen alone; ⑤H. pylori antigen plus chitosan solution; ⑥H. pylori antigen plus chitosan particles; ⑦H. pylori antigen plus CT; ⑧H. pylori antigen plus chitosan solution and CT; ⑨H. pylori antigen plus chitosan particles and CT. At 4 weeks after the last immunization, these mice were challenged by alive H. pylori（ 1 × 10^12CFU/L） twice at two - day intervals. At 4 weeks after the last challenge, these mice were all killed and gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa staining. The other gastric mucosa were used to quantitatively culture with H. pylori. ELISA was used to detect H. pylori IgA in saliva and gastric mucosa and anti -H. pylori IgG, IgG1, IgG2a in serum, and immunohistochemical method was used to examine sIgA in gastric mucosa. RESULTS： ①In the groups with chitosan as adjuvant, 60% mice achieved immunological protection, which was according to that with CT as adjuvant （58.33%）, and was significantly higher than H. pylori antigen alone and other groups without H. pylori antigen（ P 〈 0. 01 or P 〈 0. 05 ）. While the rates of protection in the groups with chitosan plus CT as adjuvant were 84. 62% ,85.71% and the H. pylori colonization score in it was significantly lower than that in the groups with CT as an adjuvant and without adjuvants（P 〈0. 01 or P 〈0. 05）. ②The each isotype IgG levels induced by H. pylori vaccine with adjuvants were significantly higher than those in control group and non- adjuvant groups （P 〈 0. 01 or P 〈0. 05） ,while the anti -H. pyloriIgG levels in the groups with CT plus chitosan as adjuvant were significantly higher than those in the groups with CT or chitosan as an adjuvant alone（ P 〈 0. 05 ）. ③ The labeling index for sl

关 键 词：螺杆菌 幽门 壳聚糖 疫苗 佐剂 免疫 

分 类 号：R377[医药卫生—病原生物学]