Zhang Ming-Qiang, Wan Yong, Jin Yang, Xin Jian-Bao, Zhang Jian-Chu, Xiong Xian-Zhi, Chen Long, Chen Gang
Department of Respiratory and Critical Care Medicine, Key Laboratory of Pulmonary Diseases of Health Ministry, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Respiratory and Critical Care Medicine WUHAN NO. 1 HOSPITAL, Wuhan, China.
PLoS One. 2014 Nov 6;9(11):e112350. doi: 10.1371/journal.pone.0112350. eCollection 2014.
CD4+ T cells in the lung are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD), although CD4+ T cell subsets and the direct effect of smoking on these cells, especially the expression of MRs, have not been comprehensively examined.
First, circulating CD4+ T cell subsets in healthy nonsmokers, patients with SCOPD and patients with AECOPD were evaluated by flow cytometry. Then, differentiation experiments were carried out using RT-PCR, and Ki-67/Annexin V antibodies were used to measure proliferation and apoptosis. We also explored the impact of CSE on the differentiation and survival of CD4+Th/Tregs and examined the expression of MRs in healthy nonsmokers and patients with SCOPD.
We found the percentages of circulating Th1 and Th17 cells were increased in patients with AECOPD, while the percentage of Th2 cells was decreased in patients with SCOPD. The percentages of Th10 cells were decreased in both patients with SCOPD and patients with AECOPD, while the percentages of Tregs were increased. In addition, the percentages of CD4+α-7+ T cells were decreased in patients with SCOPD and patients with AECOPD. However, only the decrease observed in patients with AECOPD was significant. In vitro studies also revealed MR expression affected the polarization of T cells, with different CD4+ T cell subtypes acquiring different MR expression profiles. The addition of CSE facilitated CD4+ T cell polarization towards pro-inflammatory subsets (Th1 and Th17) and affected the survival of CD4+ T cells and Treg cells by up-regulating the expression of MR3 and 5, resulting in an imbalance of CD4+ T cell subsets.
Our findings suggest an imbalance of circulating CD4+ T cell subsets is involved in COPD pathogenesis in smokers. Cigarette smoking may contribute to this imbalance by affecting the polarization and survival of Th/Tregs through the up-regulation of MR3 and MR5.
肺内的CD4+ T细胞参与慢性阻塞性肺疾病（COPD）的发病机制，尽管CD4+ T细胞亚群以及吸烟对这些细胞的直接影响，尤其是MRs的表达，尚未得到全面研究。
首先，通过流式细胞术评估健康非吸烟者、稳定期慢性阻塞性肺疾病（SCOPD）患者和慢性阻塞性肺疾病急性加重期（AECOPD）患者外周血循环中的CD4+ T细胞亚群。然后，采用逆转录聚合酶链反应（RT-PCR）进行分化实验，并用Ki-67/膜联蛋白V抗体检测增殖和凋亡情况。我们还探讨了香烟烟雾提取物（CSE）对CD4+Th/Tregs分化和存活的影响，并检测了健康非吸烟者和SCOPD患者中MRs的表达。
我们发现，AECOPD患者外周血循环中Th1和Th17细胞的百分比增加，而SCOPD患者中Th2细胞的百分比降低。SCOPD患者和AECOPD患者中Th10细胞的百分比均降低，而调节性T细胞（Tregs）的百分比增加。此外，SCOPD患者和AECOPD患者中CD4+α-7+ T细胞的百分比降低。然而，仅AECOPD患者中观察到的降低具有统计学意义。体外研究还表明，MRs的表达影响T细胞极化，不同的CD4+ T细胞亚群具有不同的MRs表达谱。添加CSE促进CD4+ T细胞向促炎亚群（Th1和Th17）极化，并通过上调MR3和MR5的表达影响CD4+ T细胞和Treg细胞的存活，导致CD4+ T细胞亚群失衡。
我们的研究结果表明，外周血循环中CD4+ T细胞亚群失衡参与吸烟者COPD的发病机制。吸烟可能通过上调MR3和MR5影响Th/Tregs的极化和存活，从而导致这种失衡。
