Xue Luzheng, Fergusson Joannah, Salimi Maryam, Panse Isabel, Ussher James E, Hegazy Ahmed N, Vinall Shân L, Jackson David G, Hunter Michael G, Pettipher Roy, Ogg Graham, Klenerman Paul
Oxford NIHR Biomedical Research Centre, Translational Immunology Laboratory, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom; Respiratory Medicine Unit, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
Peter Medawar Building for Pathogen Research, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
J Allergy Clin Immunol. 2015 May;135(5):1358-66.e1-11. doi: 10.1016/j.jaci.2014.09.006. Epub 2014 Oct 19.
Prostaglandin D2 (PGD2) and cysteinyl leukotrienes (cysLTs) are lipid mediators derived from mast cells, which activate TH2 cells. The combination of PGD2 and cysLTs (notably cysteinyl leukotriene E4 [LTE4]) enhances TH2 cytokine production. However, the synergistic interaction of cysLTs with PGD2 in promoting TH2 cell activation is still poorly understood. The receptors for these mediators are drug targets in the treatment of allergic diseases, and hence understanding their interaction is likely to have clinical implications.
We aimed to comprehensively define the roles of PGD2, LTE4, and their combination in activating human TH2 cells and how such activation might allow the TH2 cells to engage downstream effectors, such as neutrophils, which contribute to the pathology of allergic responses.
The effects of PGD2, LTE4, and their combination on human TH2 cell gene expression were defined by using a microarray, and changes in specific inflammatory pathways were confirmed by means of PCR array, quantitative RT-PCR, ELISA, Luminex, flow cytometry, and functional assays, including analysis of downstream neutrophil activation. Blockade of PGD2 and LTE4 was tested by using TM30089, an antagonist of chemoattractant receptor-homologous molecule expressed on TH2 cells, and montelukast, an antagonist of cysteinyl leukotriene receptor 1.
PGD2 and LTE4 altered the transcription of a wide range of genes and induced diverse functional responses in TH2 cells, including cell adhesion, migration, and survival and cytokine production. The combination of these lipids synergistically or additively enhanced TH2 responses and, strikingly, induced marked production of diverse nonclassical TH2 inflammatory mediators, including IL-22, IL-8, and GM-CSF, at concentrations sufficient to affect neutrophil activation.
PGD2 and LTE4 activate TH2 cells through different pathways but act synergistically to promote multiple downstream effector functions, including neutrophil migration and survival. Combined inhibition of both PGD2 and LTE4 pathways might provide an effective therapeutic strategy for allergic responses, particularly those involving interaction between TH2 cells and neutrophils, such as in patients with severe asthma.
前列腺素D2（PGD2）和半胱氨酰白三烯（cysLTs）是源自肥大细胞的脂质介质，可激活TH2细胞。PGD2和cysLTs（尤其是半胱氨酰白三烯E4 [LTE4]）的组合可增强TH2细胞因子的产生。然而，cysLTs与PGD2在促进TH2细胞活化方面的协同相互作用仍了解甚少。这些介质的受体是治疗过敏性疾病的药物靶点，因此了解它们的相互作用可能具有临床意义。
我们旨在全面确定PGD2、LTE4及其组合在激活人TH2细胞中的作用，以及这种激活如何使TH2细胞与下游效应细胞（如中性粒细胞）相互作用，中性粒细胞在过敏反应的病理过程中起作用。
通过使用微阵列确定PGD2、LTE4及其组合对人TH2细胞基因表达的影响，并通过PCR阵列、定量RT-PCR、ELISA、Luminex、流式细胞术和功能测定（包括分析下游中性粒细胞活化）来确认特定炎症途径的变化。使用TH2细胞上表达的趋化因子受体同源分子拮抗剂TM30089和半胱氨酰白三烯受体1拮抗剂孟鲁司特测试对PGD2和LTE-4的阻断作用。
PGD2和LTE4改变了多种基因的转录，并在TH2细胞中诱导了多种功能反应，包括细胞黏附、迁移、存活和细胞因子产生。这些脂质的组合协同或相加增强了TH2反应，并且引人注目的是，在足以影响中性粒细胞活化的浓度下，诱导了多种非经典TH2炎症介质（包括IL-22、IL-8和GM-CSF）的显著产生。
PGD2和LTE4通过不同途径激活TH2细胞，但协同作用以促进多种下游效应功能，包括中性粒细胞迁移和存活。联合抑制PGD2和LTE4途径可能为过敏反应提供有效的治疗策略，特别是那些涉及TH2细胞与中性粒细胞相互作用的反应，例如重度哮喘患者。
