Alzahrani Abdulrahman, Hakeem Jameel, Biddle Michael, Alhadian Fahad, Hussain Aamir, Khalfaoui Latifa, Roach Katy M, Tliba Omar, Bradding Peter, Amrani Yassine
Department of Respiratory Sciences, Clinical Sciences, University of Leicester, Glenfield Hospital, Leicester, United Kingdom.
Department of Applied Medical Sciences, Applied College, Albaha University, Albaha, Saudi Arabia.
Front Allergy. 2021 Dec 29;2:785100. doi: 10.3389/falgy.2021.785100. eCollection 2021.
The mechanisms underlying corticosteroid insensitivity in severe asthma have not been elucidated although some indirect clinical evidence points toward a role of mast cells. Here, we tested the hypothesis that mast cells can drive corticosteroid insensitivity in airway smooth muscle cells, a key player in asthma pathogenesis. Conditioned media from resting or FcεR1-activated human lung mast cells were incubated with serum-deprived ASM cells (1:4 dilution, 24 h) to determine their impact on the anti-inflammatory action of fluticasone on ASM cell chemokine expression induced by TNFα (10 ng/ml). Conditioned media from FcεR1-activated mast cells (but not that from non-activated mast cells or control media) significantly reduced the ability of 100 nM fluticasone to suppress ASM TNFα-dependent CCL5 and CXCL10 production at both mRNA and protein levels. In contrast, fluticasone inhibition of CXCL-8 production by TNFα was still preserved in the presence of activated mast cell conditioned media. Transcriptomic analysis validated by individual qPCR assays revealed that activated mast cell conditioned media dramatically reduced the number of anti-inflammatory genes induced by fluticasone in ASM cells. Our study demonstrates for the first time that conditioned media from FcεR1-activated mast cells blunt the anti-inflammatory action of corticosteroids in ASM cells by altering their transactivation properties. Because infiltration of mast cells within the ASM bundles is a defining feature of asthma, mast cell-derived mediators may contribute to the glucocorticoid insensitivity present in severe asthma.
尽管一些间接临床证据表明肥大细胞起了作用,但严重哮喘中皮质类固醇不敏感的潜在机制尚未阐明。在此,我们验证了一个假说,即肥大细胞可在气道平滑肌细胞(哮喘发病机制中的关键参与者)中导致皮质类固醇不敏感。将来自静息或FcεR1激活的人肺肥大细胞的条件培养基与血清饥饿的气道平滑肌细胞一起孵育(1:4稀释,24小时),以确定它们对氟替卡松对肿瘤坏死因子α(10纳克/毫升)诱导的气道平滑肌细胞趋化因子表达的抗炎作用的影响。来自FcεR1激活的肥大细胞的条件培养基(而非来自未激活的肥大细胞的条件培养基或对照培养基)显著降低了100纳摩尔氟替卡松在mRNA和蛋白质水平上抑制气道平滑肌细胞肿瘤坏死因子α依赖性CCL5和CXCL10产生的能力。相比之下,在存在激活的肥大细胞条件培养基的情况下,氟替卡松对肿瘤坏死因子α诱导的CXCL-8产生的抑制作用仍然存在。通过个体定量PCR分析验证的转录组分析表明,激活的肥大细胞条件培养基显著减少了氟替卡松在气道平滑肌细胞中诱导的抗炎基因数量。我们的研究首次表明,来自FcεR1激活的肥大细胞的条件培养基通过改变其反式激活特性削弱了皮质类固醇在气道平滑肌细胞中的抗炎作用。由于肥大细胞在气道平滑肌束内的浸润是哮喘的一个决定性特征,肥大细胞衍生的介质可能导致严重哮喘中存在的糖皮质激素不敏感。