Effects of IL-33 on the interaction between HMC-1 cells and human airway smooth muscle cells

RB Pathology;QP Physiology;Q Science (General)

Jones, Rhys Christopher ; Shepherd, Malcolm

University:University of Glasgow

Department:Institute of Infection Immunity and Inflammation

关键词: 15-HETE15-hydroxy-eicosatetraenoic acidADAadalimumabADAM33a disintegrin and metalloproteinase 33AHRairways hyper-responsivenessANOVAanalysis of varianceASMairway smooth muscleBALbroncholalveolar lavageBECbronchial epithelial cellbFGF-2basic fibroblast growth factor 2C5complement factor 5CCLchemokine (C-C motif) ligand CDcluster differentiationCOSHHControl of substances hazardous to healthCTGFconnective tissue growth factorCTLAcytotoxic T-lymphocyte antigen CXCLchemokine (C-X-C motif) ligandCXCRchemokine, (C-X-C) motif receptorDCdendritic cellsDPP10dipeptidyl peptidase 10EARearly-phase asthmatic reactionsECMextracellular matrixECPeosinophil cationic proteinEDNeosinophil-derived neurotoxinEMTUepithelial-mesenchymal trophic unitENTetanerceptEPOeosinophil peroxidaseFACSfluorescent activated cell sorting FCSfoetal calf serumFGFfibroblast growth factorGBRCGlasgow biomedical research centreGDNFglial-derived neurotrophic factorGERDgastroesophageal refluxGM-CSFgranulocyte macrophage colony stimulating factorHASMhuman airway smooth muscleHB-EGFheparin-binding epidermal growth factorHLMChuman lung mast cellHMChuman mast cellHMC-1human mastocytosis cell line-1HUCBMChuman umbilical cord blood-derived mast cellICAMintracellular adhesion molecule IDMEMIscove’s modified Dulbecco modified eagle mediumIFNinterferonIgimmunoglobulin ILinterleukin IL-1RAcPIL-1 receptor accessory proteinLARlate-phase asthmatic reactionsLFAlymphocyte function associated antigenLTleukotrieneMAP kinasemitogen-activated protein kinaseMBPmajor basic proteinMCPmonocyte chemotactic proteinMFImean fluorescence intensityMHCmajor histocompatibility complexMIPmacrophage inflammatory proteinMMPmatrix metalloproteinasemRNAmessenger ribonucleic acidNF-κBnuclear factor kappa-light-chain-enhancer of activated B cellsNGFnerve growth factorNOnitric oxidePAFplatelet activating factorPAMPpathogen associated molecular patternsPBMCperipheral blood mononuclear cellsPBSphosphate buffered salinePDGFplatelet-derived growth factorPEphycoerytherinPGprostaglandinPHF11plant homeodomain finger protein-11RANTESregulated on activation normal T-cell expressed and secretedrpmrevolutions per minuteSCFstem cell factorSNPsingle nucleotide polymorphismTGFtransforming growth factorThT helper cellsTIM1T-cell immunoglobulin mucin-domainTLRtoll like receptorTNFtumour necrosis factorTSLCtumour suppressor in lung cancerVCAMvascular cell adhesion moleculeVEGFvascular endothelial growth factorVLAVery late activating antigenβ2-agonistsβ2-adrenoceptors agonistsβ-agonistsbeta adrenoceptor agonists;   

Others  :  http://theses.gla.ac.uk/1325/1/2009jonesmscl.pdf

来源: University of Glasgow

【 摘 要 】

Asthma is one of the most common chronic diseases in the world, affecting approximately 300 million people worldwide.Asthma has many phenotypes, allergic asthma being the most common form.Most cases are initiated by IgE antibodies, generally referred to as IgE-mediated allergic asthma.Treatments include β2-agonists and inhaled corticosteroids, although there is no definitive cure for the disease.Asthma results in a plethora of pathological processes, present in many types of asthma.This can make differential diagnosis difficult, although several processes are characteristic of the disease pathology including; airway remodelling, airways hyperreactivity and airways inflammation.The interaction of mast cells and human airway smooth muscle (HASM) cells play an important role in the characteristic pathological processes involved with asthma. The recently discovered cytokine, interleukin 33 (IL-33), is thought to play an important role in a variety of autoimmune diseases; including asthma and atopic dermatitis.The aims of this study were to investigate the interaction of mast cells and HASM cells in vitro, analysing the adhesion molecules involved and the potential presence of chemokines relevant to the adhesion process.In addition, the effects of IL-33 on the above processes were also investigated.HMC-1 cells were analysed for the presence of several adhesion molecules, using a primary and secondary antibody, which were quantified using a fluorescently activated cell sorting machine (FACS).Once the presence of the adhesion molecules were ascertained, the effects of IL-33 on the adhesion molecule expression was performed.Anti-TNF-α therapies were also used to assess whether a change in adhesion molecule expression, on the HMC-1 cells, was apparent.The results confirm the presence of the adhesion molecules intracellular adhesion molecule 1 (ICAM-1), and Integrins α4 and β1.The presence of IL-33 (21 hours exposure) resulted in an up regulation of ICAM-1 on HMC-1 cells, being statistically different to the HMC-1 cells left untreated (99% confidence level).IL-33 seemed to have little effect on Integrin α4 or β1 expression on the HMC-1 cells.The anti-TNF-α therapies lead to a decrease in ICAM-1 expression on HMC-1 cells, after being exposed to both TNF-α and IL-33.This reduction in ICAM-1 expression was statistically different to the positive controls (HMC-1 cells treated with TNF-α alone) using both anti-TNF-α therapies with TNF-α and IL-33, at the 95% confidence level.The effects of IL-33 on HMC-1 cell/HASM cells could lead to differences in adhesion, via ICAM-1.These effects may also lead to HMC-1 cells releasing TNF-α, having an autocrine effect on the HMC-1 cells.Recommendations for future work include; analysing other adhesion molecules relevant to mast cell/HASM cell interactions, including tumour suppressor in lung cancer-1 (TSLC-1) and vascular cell adhesion molecule-1 (VCAM-1).Using mast cells derived from human umbilical cord blood stem cells would represent an improvement to using HMC-1 cells.In vivo research would also prove invaluable, quantifying and localising mast cells in asthma induced mice for example.