CpG-ODN对婴幼儿喘息性支气管炎外周血单个核细胞T-bet表达的影响  

Effects of CpG-ODN on the expression of T-bet from peripheral blood mononuclear cells from infants with wheezing bronchitis

作　　者：王锁英[1] 许化溪[2] 杨敏[2] 许小朋[2] 邱谷风[2] 李国民[1] 张贇健[1] 马洁[2] 眭建[2] 姜旭淦[2] 胡嘉波[2] 

机构地区：[1]江苏大学附属医院儿科,镇江212001 [2]江苏大学检验医学研究所免疫研究室,镇江212001

出　　处：《中国免疫学杂志》2007年第11期1035-1039,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(30471627;30300169);江苏省自然科学基金项目(BK2004405)

摘　　要：目的:探讨CpG-ODN干预对婴幼儿喘息性支气管炎外周血单个核细胞(PBMC)T-bet表达和IFN-γ分泌的影响。方法:对28例喘息性支气管炎(喘支组)患儿于急性期和恢复期分别抽取抗凝静脉血并分离PBMC,以终浓度为10μg/ml的B型CpG-ODN刺激48小时,分别应用RT-PCR和Western blot检测CpG-ODN刺激前后患儿PBMC的T-bet mRNA和蛋白表达情况,以ELISA检测PBMC培养上清IFN-γ的变化,同时设立26例健康婴儿对照组。结果:与正常对照组比较,喘支患儿PBMC表达T-bet mRNA及蛋白明显减少(P<0.05),应用CpG-ODN刺激后,两组小儿PBMC表达T-bet均显著上调,但产生IFN-γ无明显增多,对照序列ODN对T-bet表达及IFN-γ的分泌无明显影响。相关分析表明,喘支患儿T-bet mRNA与IFN-γ分泌呈正相关(r=0.57,P<0.01),CpG-ODN刺激后,二者相关性明显下降(r=0.19,P>0.05)。结论:T-bet在喘支患儿PB-MC中表达减少,提示T-bet表达缺陷参与了喘支的免疫病理,B型CpG-ODN作为一种免疫调节剂,可有效上调T-bet的表达,但不能诱导IFN-γ的产生。Objective：To investigate the regulatory effects of synthetic oligodeoxynucleo tides containing CpG motif （CpG-ODN） on the expression of T-bet and the secretion of IFN-γ from peripheral blood mononuclear cells （PBMC） from infants with wheezing bron- chitis. Methods：28 eases of wheezing bronchitis （W. B group） were enrolled, and 26 normal infants were served as control. PBMCs were isolated from peripheral blood over a Ficoll-Hypaque gradient reagent according to standard procedures and incubated with CpG-ODN （10μg/ml） at 37℃, 5%CO2 for 48 hours. The mRNA and protein expression levels of T-bet in PBMC were detected by beth reverse transcription PCR （RT-PCR） and Western blot. IFN-γ levels in culture supematant of PBMC were also tested. Results：The mB.NA and protein levels of T-bet from PBMC in infants with wheezing bronchitis were lower than those of control, and were up-regulated after the stimulation of CpG-ODN at 10μ/ml. The IFN-γ levels secreted by PBMC were not affected by the treatment of control-ODN stimulating. Conclusion：The mRNA and protein levels of T-bet in PBMC are lower in infants suffered from wheezing bronchitis. It is suggested that the decreased expression of T-bet may play an important role in immunopathology of the disease. As an immunoregulator, CpG-ODN can up-regulate T-bet mRNA and protein in PBMC remarkably, despite of not driving PBMC to express more IFN-γ.

关 键 词：CPG-ODN 喘息 支气管炎 T-BET 

分 类 号：R392.1[医药卫生—免疫学]