过敏性哮喘中人中性粒细胞一氧化氮的产生和细胞外捕获器的形成耦合作用
2024/03/26
摘要
背景：一氧化氮（NO）在过敏性哮喘患者的气道和血清中升高，表明它在哮喘中起着重要作用。NO的产生广泛归因于经典的诱导型一氧化氮合酶（iNOS）。已经付出了大量努力来抑制这种酶，结果有两种：没有哮喘改善；部分NO减少，这表明存在iNOS独立来源的参与。
目的：中性粒细胞在炎症条件下会产生 NO，但它们在哮喘中的作用一直被忽视。本研究分析了中性粒细胞作为 NO 来源的可能作用。
方法：在 99 名间歇性支气管哮喘过敏性患者和 26 名健康供体中测试了我们的假设。通过Griess试剂法、流式细胞术和共聚焦显微镜评估了血液和痰液中中性粒细胞对变应原、抗IgE和抗IgE受体抗体的NO产生。通过Western印迹和共聚焦显微镜定量测定了作为NO产生可能结果的细胞外陷阱（ETs）形成，以及通过增亮氧化物化学发光测定了反应性氧化物。
结果：在过敏性哮喘患者的血液和痰液颗粒细胞中，只有中性粒细胞通过一种IgE依赖的机制产生NO。这种产生不依赖于NOS，而依赖于L-精氨酸和NOX2来源的反应性氧化物之间的反应。NO和ETosis同时诱导，并且NO增强了ETs形成，这是哮喘的关键介质
结论：我们的发现揭示了中性粒细胞作为过敏原/IgE依赖性NO来源的新角色，增强了ETs形成。这些结果表明，中性粒细胞产生的NO在过敏性哮喘的治疗中需要进一步考虑。
Human Neutrophils Couple Nitric Oxide Production and Extracellular Traps Formation in Allergic Asthma
Chacón P, Vega-Rioja A, Doukkali B, Del Valle Rodriguez A, Fernández-Delgado L, Domínguez-Cereijo L, Segura C, Pérez-Machuca BM, Perkins JR, El Bekay R, Cornejo-García JA, Hajji N, Monteseirín J, Rivas-Pérez D
Abstract
Background:Nitric oxide (NO) is elevated in the airways and serum of allergic asthmatic patients, suggesting an important role in asthma. NO production has been widely attributed to the canonical inducible nitric oxide synthase (iNOS). Much effort has been made to inhibit this enzyme with two outcomes: no asthma improvement; and partial NO reduction, suggesting the involvement of an iNOS-independent source.
Objectives:Neutrophils produce NO under inflammatory conditions and their role in asthma has been overlooked. The present study analyzes their possible role as source of NO.
Methods:Our hypothesis was tested in 99 allergic patients with intermittent bronchial asthma and 26 healthy donors. NO production by blood and sputum neutrophils in response to allergens, anti-IgE, and anti-IgE receptors Abs was assessed by Griess, flow cytometry and confocal microscopy. Extracellular traps (ETs) formation, as a possible consequence of NO production, was quantified by western blot and confocal microscopy, and reactive oxygen species by luminol-enhanced chemiluminescence.
Results:Among blood and sputum granulocytes from allergic asthmatic patients, only neutrophils, produce NO by an IgE-dependent mechanism. This production is independent of NOS, but dependent on a reaction between L-arginine and reactive oxygen species from NOX2. NO and ETosis are induced in parallel, and NO amplifies ETs formation, which is a key mediator in asthma.
Conclusion:Our findings reveal a novel role of neutrophils as the unique allergen/IgE-dependent NO source in allergic asthma enhancing ETs formation. These results suggest that NO produced by neutrophils needs further consideration in the treatment of allergic asthma.
背景：一氧化氮（NO）在过敏性哮喘患者的气道和血清中升高，表明它在哮喘中起着重要作用。NO的产生广泛归因于经典的诱导型一氧化氮合酶（iNOS）。已经付出了大量努力来抑制这种酶，结果有两种：没有哮喘改善；部分NO减少，这表明存在iNOS独立来源的参与。
目的：中性粒细胞在炎症条件下会产生 NO，但它们在哮喘中的作用一直被忽视。本研究分析了中性粒细胞作为 NO 来源的可能作用。
方法：在 99 名间歇性支气管哮喘过敏性患者和 26 名健康供体中测试了我们的假设。通过Griess试剂法、流式细胞术和共聚焦显微镜评估了血液和痰液中中性粒细胞对变应原、抗IgE和抗IgE受体抗体的NO产生。通过Western印迹和共聚焦显微镜定量测定了作为NO产生可能结果的细胞外陷阱（ETs）形成，以及通过增亮氧化物化学发光测定了反应性氧化物。
结果：在过敏性哮喘患者的血液和痰液颗粒细胞中，只有中性粒细胞通过一种IgE依赖的机制产生NO。这种产生不依赖于NOS，而依赖于L-精氨酸和NOX2来源的反应性氧化物之间的反应。NO和ETosis同时诱导，并且NO增强了ETs形成，这是哮喘的关键介质
结论：我们的发现揭示了中性粒细胞作为过敏原/IgE依赖性NO来源的新角色，增强了ETs形成。这些结果表明，中性粒细胞产生的NO在过敏性哮喘的治疗中需要进一步考虑。
(中日友好医院呼吸与危重症医学科 沈焜路 摘译 林江涛 审校)
(Am J Respir Crit Care Med. 2024 Mar 6. DOI: 10.1164/rccm.202305-0889OC)
(Am J Respir Crit Care Med. 2024 Mar 6. DOI: 10.1164/rccm.202305-0889OC)
Human Neutrophils Couple Nitric Oxide Production and Extracellular Traps Formation in Allergic Asthma
Chacón P, Vega-Rioja A, Doukkali B, Del Valle Rodriguez A, Fernández-Delgado L, Domínguez-Cereijo L, Segura C, Pérez-Machuca BM, Perkins JR, El Bekay R, Cornejo-García JA, Hajji N, Monteseirín J, Rivas-Pérez D
Abstract
Background:Nitric oxide (NO) is elevated in the airways and serum of allergic asthmatic patients, suggesting an important role in asthma. NO production has been widely attributed to the canonical inducible nitric oxide synthase (iNOS). Much effort has been made to inhibit this enzyme with two outcomes: no asthma improvement; and partial NO reduction, suggesting the involvement of an iNOS-independent source.
Objectives:Neutrophils produce NO under inflammatory conditions and their role in asthma has been overlooked. The present study analyzes their possible role as source of NO.
Methods:Our hypothesis was tested in 99 allergic patients with intermittent bronchial asthma and 26 healthy donors. NO production by blood and sputum neutrophils in response to allergens, anti-IgE, and anti-IgE receptors Abs was assessed by Griess, flow cytometry and confocal microscopy. Extracellular traps (ETs) formation, as a possible consequence of NO production, was quantified by western blot and confocal microscopy, and reactive oxygen species by luminol-enhanced chemiluminescence.
Results:Among blood and sputum granulocytes from allergic asthmatic patients, only neutrophils, produce NO by an IgE-dependent mechanism. This production is independent of NOS, but dependent on a reaction between L-arginine and reactive oxygen species from NOX2. NO and ETosis are induced in parallel, and NO amplifies ETs formation, which is a key mediator in asthma.
Conclusion:Our findings reveal a novel role of neutrophils as the unique allergen/IgE-dependent NO source in allergic asthma enhancing ETs formation. These results suggest that NO produced by neutrophils needs further consideration in the treatment of allergic asthma.
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儿童鼻咽微生物群与哮喘严重恶化有关
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哮喘患者气道上皮细胞对RSV的反应主要受损于杯状细胞和多纤毛细胞
