microRNA-155对异氰酸酯诱发的哮喘中固有淋巴细胞的作用
2021/09/24
摘要
背景：职业性哮喘是由工作场所暴露于低分子量（LMW）药物如甲苯2，4二异氰酸酯（TDI）引起的，给患者和社会带来了巨大的负担。人们对TDI诱导的哮喘中的固有淋巴细胞（ILC）知之甚少。而ILC功能的关键调节因子是microRNA-155，一种与哮喘相关的microRNA。
目的：确定TDI暴露是否会改变肺组织中ILC的数量，以及microRNA-155是否参与了TDI诱导的气道炎症和高反应性。
方法：用TDI或溶媒致敏并激发C57BL/6野生型和microRNA-155基因敲除小鼠。通过流式细胞术检测支气管肺泡灌洗液（BAL）中ILC和T细胞内细胞因子的表达水平。在肺组织中观察支气管周围嗜酸性粒细胞和杯状细胞的变化，用强迫震荡法测定气道高反应性。应用免疫组织化学方法在TDI诱发的职业性哮喘患者的支气管活检中鉴别推测的ILC2细胞。人支气管上皮细胞暴露于TDI或溶媒。
结果：TDI暴露小鼠气道杯状细胞、BAL嗜酸性粒细胞、CD4+T淋巴细胞和ILC数量更多，以2型反应为主，并有气道高反应倾向。在TDI暴露的microRNA-155基因敲除小鼠中，炎症和气道高反应性减弱。TDI暴露诱导人支气管上皮细胞和小鼠肺中IL-33的表达，且在小鼠中IL-33的表达是microRNA-155依赖性的。支气管活检中存在推测为ILC2的GATA3+CD3-细胞。
结论：TDI暴露于ILC数量增加有关。促炎性microRNA-155在TDI哮喘小鼠模型中起关键作用，提示其参与LMW药物所引起的职业性哮喘的发病机制。
Innate lymphoid cells in isocyanate-induced asthma: role of microRNA-155
Evy E Blomme, Sharen Provoost , Erica Bazzan, Hannelore P Van Eeckhoutte , Mirjam P Roffel , Lore Pollaris , Annelies Bontinck , Matteo Bonato , Louise Vandenbroucke , Fien Verhamme , Guy F Joos , Manuel G Cosio , Jeroen A J Vanoirbeek, Guy G Brusselle, Marina Saetta , Tania Maes
Abstract
Background: Occupational asthma, induced by workplace exposures to low molecular weight agents such as toluene 2,4-diisocyanate (TDI), causes a significant burden to patients and society. Little is known about innate lymphoid cells (ILCs) in TDI-induced asthma. A critical regulator of ILC function is microRNA-155, a microRNA associated with asthma.
Objective: To determine whether TDI exposure modifies the number of ILCs in the lung and whether microRNA-155 contributes to TDI-induced airway inflammation and hyperresponsiveness.
Methods: C57BL/6 wild-type and microRNA-155 knockout mice were sensitised and challenged with TDI or vehicle. Intracellular cytokine expression in ILCs and T-cells was evaluated in bronchoalveolar lavage (BAL) fluid using flow cytometry. Peribronchial eosinophilia and goblet cells were evaluated on lung tissue, and airway hyperresponsiveness was measured using the forced oscillation technique. Putative type 2 ILCs (ILC2) were identified in bronchial biopsies of subjects with TDI-induced occupational asthma using immunohistochemistry. Human bronchial epithelial cells were exposed to TDI or vehicle.
Results: TDI-exposed mice had higher numbers of airway goblet cells, BAL eosinophils, CD4+ T-cells and ILCs, with a predominant type 2 response, and tended to have airway hyperresponsiveness. In TDI-exposed microRNA-155 knockout mice, inflammation and airway hyperresponsiveness were attenuated. TDI exposure induced IL-33 expression in human bronchial epithelial cells and in murine lungs, which was microRNA-155 dependent in mice. GATA3+CD3- cells, presumably ILC2, were present in bronchial biopsies.
Conclusion: TDI exposure is associated with increased numbers of ILCs. The proinflammatory microRNA-155 is crucial in a murine model of TDI asthma, suggesting its involvement in the pathogenesis of occupational asthma due to low molecular weight agents.
背景：职业性哮喘是由工作场所暴露于低分子量（LMW）药物如甲苯2，4二异氰酸酯（TDI）引起的，给患者和社会带来了巨大的负担。人们对TDI诱导的哮喘中的固有淋巴细胞（ILC）知之甚少。而ILC功能的关键调节因子是microRNA-155，一种与哮喘相关的microRNA。
目的：确定TDI暴露是否会改变肺组织中ILC的数量，以及microRNA-155是否参与了TDI诱导的气道炎症和高反应性。
方法：用TDI或溶媒致敏并激发C57BL/6野生型和microRNA-155基因敲除小鼠。通过流式细胞术检测支气管肺泡灌洗液（BAL）中ILC和T细胞内细胞因子的表达水平。在肺组织中观察支气管周围嗜酸性粒细胞和杯状细胞的变化，用强迫震荡法测定气道高反应性。应用免疫组织化学方法在TDI诱发的职业性哮喘患者的支气管活检中鉴别推测的ILC2细胞。人支气管上皮细胞暴露于TDI或溶媒。
结果：TDI暴露小鼠气道杯状细胞、BAL嗜酸性粒细胞、CD4+T淋巴细胞和ILC数量更多，以2型反应为主，并有气道高反应倾向。在TDI暴露的microRNA-155基因敲除小鼠中，炎症和气道高反应性减弱。TDI暴露诱导人支气管上皮细胞和小鼠肺中IL-33的表达，且在小鼠中IL-33的表达是microRNA-155依赖性的。支气管活检中存在推测为ILC2的GATA3+CD3-细胞。
结论：TDI暴露于ILC数量增加有关。促炎性microRNA-155在TDI哮喘小鼠模型中起关键作用，提示其参与LMW药物所引起的职业性哮喘的发病机制。
(中日友好医院呼吸与危重症医学科 张清 摘译 林江涛 审校)
(Eur Respir J.2020 Sep 24;56(3):1901289. doi: 10.1183/13993003.01289-2019.)
(Eur Respir J.2020 Sep 24;56(3):1901289. doi: 10.1183/13993003.01289-2019.)
Innate lymphoid cells in isocyanate-induced asthma: role of microRNA-155
Evy E Blomme, Sharen Provoost , Erica Bazzan, Hannelore P Van Eeckhoutte , Mirjam P Roffel , Lore Pollaris , Annelies Bontinck , Matteo Bonato , Louise Vandenbroucke , Fien Verhamme , Guy F Joos , Manuel G Cosio , Jeroen A J Vanoirbeek, Guy G Brusselle, Marina Saetta , Tania Maes
Abstract
Background: Occupational asthma, induced by workplace exposures to low molecular weight agents such as toluene 2,4-diisocyanate (TDI), causes a significant burden to patients and society. Little is known about innate lymphoid cells (ILCs) in TDI-induced asthma. A critical regulator of ILC function is microRNA-155, a microRNA associated with asthma.
Objective: To determine whether TDI exposure modifies the number of ILCs in the lung and whether microRNA-155 contributes to TDI-induced airway inflammation and hyperresponsiveness.
Methods: C57BL/6 wild-type and microRNA-155 knockout mice were sensitised and challenged with TDI or vehicle. Intracellular cytokine expression in ILCs and T-cells was evaluated in bronchoalveolar lavage (BAL) fluid using flow cytometry. Peribronchial eosinophilia and goblet cells were evaluated on lung tissue, and airway hyperresponsiveness was measured using the forced oscillation technique. Putative type 2 ILCs (ILC2) were identified in bronchial biopsies of subjects with TDI-induced occupational asthma using immunohistochemistry. Human bronchial epithelial cells were exposed to TDI or vehicle.
Results: TDI-exposed mice had higher numbers of airway goblet cells, BAL eosinophils, CD4+ T-cells and ILCs, with a predominant type 2 response, and tended to have airway hyperresponsiveness. In TDI-exposed microRNA-155 knockout mice, inflammation and airway hyperresponsiveness were attenuated. TDI exposure induced IL-33 expression in human bronchial epithelial cells and in murine lungs, which was microRNA-155 dependent in mice. GATA3+CD3- cells, presumably ILC2, were present in bronchial biopsies.
Conclusion: TDI exposure is associated with increased numbers of ILCs. The proinflammatory microRNA-155 is crucial in a murine model of TDI asthma, suggesting its involvement in the pathogenesis of occupational asthma due to low molecular weight agents.
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重温轻度哮喘：当前知识和未来需求
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哮喘的实质破坏：固定气流阻塞和肺功能轨迹
