重组可诱导共刺激分子融合蛋白治疗过敏性支气管哮喘小鼠的实验研究  

The therapeutic effect of soluble inducible costimulator fusion protein on inflammation in a murine model of allergic asthma

作　　者：王健[1] 张军[1] 龙宪连[1] 蔡刚[1] 沈茜[1] 

机构地区：[1]上海长海医院中心实验室,200433

出　　处：《中华结核和呼吸杂志》2005年第6期398-402,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家高新技术发展规划"863"课题基金资助项目(2002AA214091)

摘　　要：目的研究可诱导共刺激分子融合蛋白(ICOSIg)对过敏性支气管哮喘(简称哮喘)小鼠的治疗作用。方法32只健康雌性BALB/c小鼠用分段随机分组法将小鼠随机分成4组。哮喘组(A组)、ICOSIg治疗组(B组)、同型抗体对照组(C组)、生理盐水气雾激发对照组(D组),每组8只,采用重组真核表达技术制备ICOSIg,建立鸡卵白蛋白(OVA)免疫小鼠哮喘模型,给予ICOSIg治疗后观察其气道阻力变化、支气管肺泡灌洗液(BALF)中细胞总数、各炎性细胞百分比及白细胞介素4(IL4)、γ干扰素(IFNγ)和总免疫球蛋白E(IgE)含量的变化;采用流式细胞仪(FACS)检测T辅助细胞(Th)亚群变化;取肺组织行病理组织学分析观察肺内炎症情况。结果(1)制备的ICOSIg可与哮喘小鼠脾脏B细胞上相应配体结合。(2)B组小鼠气道压力变化为[(33±12)%],与A组[(58±10)%]比较差异有统计学意义(P<0.01)。B组BALF中细胞总数为[(5.9±3.1)×107/L],与A组[(22.6±5.3)×107/L]比较差异有统计学意义(P<0.01);B组嗜酸粒细胞数为0.020±0.020,与A组(0.070±0.030)比较差异有统计学意义(P<0.01);B组IL4水平为(77±31)ng/L,与A组[(179±44)ng/L]比较差异有统计学意义(P<0.01);B组外周血中总IgE水平为(175±33)μg/L,与A组[(282±22)μg/L]比较差异有统计学意义(P<0.01);B组Th2细胞比例为(4.5±1.0)%,与A组[(11.1±2.5)%]比较差异有统计学意义(P<0.01)。(3)与A组比较,B组小鼠肺内炎性浸润明显减轻、上皮细胞完整、管腔内少见分泌物。结论ICOSIg可体内阻断可诱导共刺激通路、减少Th2免疫反应偏移并抑制IgE的生成,对过敏性哮喘有治疗作用。Objective To examine the therapeutic effect of inducible costimulator fusion protein(ICOS-Ig) on airway inflammation in a murine model of allergic asthma. Methods Thirty-two healthy female BALB/c mice were randomly divided into four groups, namely the asthma group(group A), the ICOS-Ig treated group(group B), the control antibody group(group C), and the saline challenged group(group D), with 8 mice in each group. ICOS-Ig was produced by the eukaryotic expression technology. A murine model of allergic asthma was made by sensitizing animals with ovalbumin and exposing them to repeated ovalbumin inhalation challenges. After the mice were treated with ICOS-Ig at the time of ovalbumin challenge, the airway responsiveness, inflammatory cells and the cytokine content in bronchoalveolar lavage fluid (BALF), the level of IgE and Th1/Th2 in blood, and the lung histology were measured to observe the effect of the treatment on asthma in vivo. Results (1) FACS analysis confirmed that ICOS-Ig had the binding activity to B220 murine splenocytes. (2)Trans-pulmonary pressure change was significantly reduced in mice from group B [(33±12)%] compared with group A [(58±10)%, P<0.01]. The total cell numbers of BALF in mice from group B [(5.9±3.1)×10~7/L] were decreased compared with group A[(22.6±5.3)×10~7/L, P<0.01]. The percentage of eosinophils in BALF in mice from group B ((0.020±0.020)) was reduced significantly as compared with group A (0.070±0.030, P<0.01). IL-4 content in BALF was reduced in mice from group B[(77±31)ng/L] compared with group A [(179±44)ng/L, P<0.01]. IgE in blood was decreased in mice from group B [(175±33)μg/L] compared with[LM] group A [(282±22)μg/L, P<0.01]. Th2 cell numbers in blood were reduced in mice from group B [(4.5±1.0)% ] compared with group A [(11.1±2.5)%, P<0.01]. (3)Compared with group A, the pulmonary inflammation in group B was relieved and inflammatory changes in airway epithelium were absent. Conclusion ICOS-Ig has an inhibitory effect on inflammation in a murine model of all

关 键 词：可诱导共刺激分子 融合蛋白 实验研究 哮喘小鼠 ICOS-Ig 过敏性支气管哮喘 BALB/C小鼠 支气管肺泡灌洗液 Th2免疫反应 重组真核表达 总免疫球蛋白 气道压力变化 治疗作用 细胞总数 统计学 白细胞介素 IFN-γ T辅助细胞 

分 类 号：R562.25[医药卫生—呼吸系统]