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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 effects of CpG-ODN intervention on T-bet expression and IFN-γ secretion in peripheral blood mononuclear cells (PBMC) of infantile asthmatic bronchitis. Methods: A total of 28 patients with asthmatic bronchitis (asthma group) received anticoagulation venous blood in acute and convalescent phase and PBMCs were isolated and stimulated with CpG-ODN of type B at a final concentration of 10μg / ml for 48 hours respectively The expression of T-bet mRNA and protein in PBMC of PBMCs before and after CpG-ODN stimulation were detected by RT-PCR and Western blot. The changes of IFN-γ in PBMC culture supernatants were detected by ELISA, and 26 healthy infants were also established. Results: Compared with the normal control group, the expression of T-bet mRNA and protein in PBMCs were significantly decreased (P <0.05). After stimulation with CpG-ODN, the expression of T-bet in PBMC of both groups were significantly increased, but IFN- -γ, no significant increase in the control sequence ODN T-bet expression and IFN-γ secretion had no significant effect. Correlation analysis showed that there was a positive correlation between T-bet mRNA and IFN-γ secretion in children with bronchial asthma (r = 0.57, P <0.01) . CONCLUSION: T-bet decreased in PB-MC, suggesting that T-bet deficiency is involved in the immunopathology of bronchial asthma. As an immunomodulator, B-type CpG-ODN can effectively upregulate T-bet But did not induce the production of IFN-γ.