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卡介苗对支气管哮喘患者外周血单个核细胞Th1／Th2平衡的影响

邹叶青,杨青,况九龙   

  1. 南昌大学第二附属医院
  • 收稿日期:2011-03-09 修回日期:2011-05-26 出版日期:2011-10-15 发布日期:2011-10-15
  • 通讯作者: 杨青

Effects of Bacillus Calmette-Guerin on Th1/Th2 balance of peripheral blood mononuclear cells from atopic asthamatics

  • Received:2011-03-09 Revised:2011-05-26 Published:2011-10-15 Online:2011-10-15

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摘要： 摘要 目的: 研究卡介苗(BCG)干预 对支气管哮喘患者外周血单个核细胞Th1/Th2平衡的影响。方法: 抽取哮喘患者外周血样本20例，同期健康献血员正常对照组外周血样本10例。分离外周血淋巴细胞并卡介苗干预培养(100μg/ml)，于培养前后采用三色流式细胞法测定外周血Th1/Th2细胞之比，RT-PCR测定培养前后外周血淋巴细胞IL-4和IFN-γ基因的表达。结果: RT-PCR检测卡介苗干预前哮喘患者IL-4 mRNA较正常对照组明显升高，而IFN-γ mRNA水平明显降低(P均<0.05)；干预后与干预前相比， IL-4 mRNA表达明显下降，IFN-γmRNA表达明显升高(P均<0.05)。流式细胞仪检测发现，哮喘病人组Th1百分率和Th1／Th2比值均显著低于正常对照组（P均＜0.05），Th2细胞百分率显著高于对照组（P＜0.05）；经卡介苗干预后，Th1百分率和Th1／Th2比值均显著升高（P均＜0.05），Th2细胞百分率显著降低（P＜0.05）。结论：哮喘患者中Th2型免疫应答占优势，卡介苗干预能在基因表达水平和细胞数量上纠正哮喘患者的Th1/Th2失衡。

关键词: 支气管哮喘, 卡介苗, Th1/Th2, 流式细胞仪, 外周血单个核细胞

Abstract: Abstract Objective:To investigate the effects of Bacillus Calmette-Guerin (BCG) on Th1/Th2 balance of human peripheral blood mononuclear cells (PBMC in asthmatic patients. Methods:PBMC were separated from 20 bronchial asthma patients and 10 non-atopic, age mathed healthy subjects as control. PBMC were cultured with BCG (100μg/ml) for 48h as postive group and PBMC were treated by saline as negative group. The mRNA expression of interleukin-4 (IL-4) and interferon-γ(IFN-γ) were measured by reverse transcription-polymerase chain reaction (RT-PCR) and the peripheral blood Th1/Th2 lymphocytes ratio by using three-color flow cytometry method. Results:The mRNA expression of IL-4 in asthmatics was significantly higher than that in healthy subjects while the mRNA expression of IFN-γwas lower significantly ( all P <0.05). And after BCG stimulation The IL-4mRNA expression decreased but IFN-γmRNA expression increased compared with untreated asthma group ( all P <0.05). The rate of Th1 and blood Th1/Th2 cells ratio were lower in asthma group compared with negative group ( all P <0.05) while the rate of Th2 was significantly higher than that in healthy gorup (P <0.05). After stimulation with BCG the rate of Th1 and Th1/Th2 ratio increased ( all P <0.05) , also the blood Th2 rate decreased compared with untreated asthma group (P <0.05). Conclusion:The Th2 immune response is stronger than Th1 immune response in bronchial asthma patients. BCG can retrieve the excursion of Th1/Th2 both on gene expression level and single-cell level.

Key words: Bronchial asthma, BCG, Th1/Th2, Flow cytometer, peripheral blood mononuclear cells.