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

Jones, Rhys Christopher (2009) Effects of IL-33 on the interaction between HMC-1 cells and human airway smooth muscle cells. MSc(R) thesis, University of Glasgow.

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Abstract

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.

Item Type: Thesis (MSc(R))
Qualification Level: Masters
Keywords: 15-HETE 15-hydroxy-eicosatetraenoic acid ADA adalimumab ADAM33 a disintegrin and metalloproteinase 33 AHR airways hyper-responsiveness ANOVA analysis of variance ASM airway smooth muscle BAL broncholalveolar lavage BEC bronchial epithelial cell bFGF-2 basic fibroblast growth factor 2 C5 complement factor 5 CCL chemokine (C-C motif) ligand CD cluster differentiation COSHH Control of substances hazardous to health CTGF connective tissue growth factor CTLA cytotoxic T-lymphocyte antigen CXCL chemokine (C-X-C motif) ligand CXCR chemokine, (C-X-C) motif receptor DC dendritic cells DPP10 dipeptidyl peptidase 10 EAR early-phase asthmatic reactions ECM extracellular matrix ECP eosinophil cationic protein EDN eosinophil-derived neurotoxin EMTU epithelial-mesenchymal trophic unit ENT etanercept EPO eosinophil peroxidase FACS fluorescent activated cell sorting FCS foetal calf serum FGF fibroblast growth factor GBRC Glasgow biomedical research centre GDNF glial-derived neurotrophic factor GERD gastroesophageal reflux GM-CSF granulocyte macrophage colony stimulating factor HASM human airway smooth muscle HB-EGF heparin-binding epidermal growth factor HLMC human lung mast cell HMC human mast cell HMC-1 human mastocytosis cell line-1 HUCBMC human umbilical cord blood-derived mast cell ICAM intracellular adhesion molecule IDMEM Iscove’s modified Dulbecco modified eagle medium IFN interferon Ig immunoglobulin IL interleukin IL-1RAcP IL-1 receptor accessory protein LAR late-phase asthmatic reactions LFA lymphocyte function associated antigen LT leukotriene MAP kinase mitogen-activated protein kinase MBP major basic protein MCP monocyte chemotactic protein MFI mean fluorescence intensity MHC major histocompatibility complex MIP macrophage inflammatory protein MMP matrix metalloproteinase mRNA messenger ribonucleic acid NF-κB nuclear factor kappa-light-chain-enhancer of activated B cells NGF nerve growth factor NO nitric oxide PAF platelet activating factor PAMP pathogen associated molecular patterns PBMC peripheral blood mononuclear cells PBS phosphate buffered saline PDGF platelet-derived growth factor PE phycoerytherin PG prostaglandin PHF11 plant homeodomain finger protein-11 RANTES regulated on activation normal T-cell expressed and secreted rpm revolutions per minute SCF stem cell factor SNP single nucleotide polymorphism TGF transforming growth factor Th T helper cells TIM1 T-cell immunoglobulin mucin-domain TLR toll like receptor TNF tumour necrosis factor TSLC tumour suppressor in lung cancer VCAM vascular cell adhesion molecule VEGF vascular endothelial growth factor VLA Very late activating antigen β2-agonists β2-adrenoceptors agonists β-agonists beta adrenoceptor agonists
Subjects: R Medicine > RB Pathology
Q Science > QP Physiology
Q Science > Q Science (General)
Colleges/Schools: College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Supervisor's Name: Shepherd, Dr. Malcolm
Date of Award: 2009
Depositing User: Mr Rhys C Jones
Unique ID: glathesis:2009-1325
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 27 Nov 2009
Last Modified: 10 Dec 2012 13:37
URI: http://theses.gla.ac.uk/id/eprint/1325

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