Immuno-modulatory mediators Role in asthma Table 1 : Immuno-modulatory factors produced in vivo by ASM native tissues in asthmatic patients Author Immuno-modulatory mediators Role in asthma Berkman 1996 CCL5 Chemotaxis Ghaffar 1999 CCL11 Kaur 2006 CCL19** El-Shazly 2006 CX3CL1 Kaur 2012 Thymic stromal lymphopoietin (TSLP) ** TH2 allergic response Corticosteroid Insensitivity Robertson 2002 TNF-related apoptosis-inducing ligand (TRAIL) Eosinophil survival Prefontaine 2009 IL-33** Th2 allergic response Airway remodelling Joubert 2012 CCL15 Olgart Hoglund 2002 Nerve growth factor (NGF) ASM migration Mast cell activation and survival Zhang 2012 Pentraxin (PTX3)** Chemokine secretion (ASM) Foley 2007; Lee 2006 ADAM metallopeptidase domain 33 (ADAM33)** Impaired lung function ASM contractile function Hassan 2010 Heparin-binding EGF** ASM remodelling **Denotes mediators still expressed in patients with severe asthma

Immuno-modulatory mediators Role in asthma Table 2 : Glucocorticoid-insensitive proteins induced by TNFa/IFNg in ASM cells Author Immuno-modulatory mediators Role in asthma Clarke 2010; Chachi 2013 CXCL10 Chemotaxis Chachi 2013 CCL5 Sukkar 2004; Chachi 2013 CX3CL1 Prefontaine 2009 IL-33** Th2 allergic response Airway remodelling Joubert 2012 CCL15 Tliba 2008 IRF-1 Transcription of inflammatory genes Inhibition of GRa transactivation via GRIP-1 depletion CCL11 Tliba 2006 CD38 Airway inflammation Airway hyperresponsiveness GRb Inhibition of GRa transactivation via dominant negative action Bouazza 2012; Chachi 2013 PP5 Inhibition of GRa transactivation via ser211 dephosphorylation **Denotes mediators still expressed in native ASM tissues in severe asthmatics

Alteration of ASM function Chemoattraction of inflammatory cells Figure 1 : Role of the immuno-modulatory factors produced in vivo by ASM in asthma pathogenesis Asthmatic ASM bundles Airway remodelling/ Hyper-responsiveness NGF CCL19 CCL11 Pentraxin 3 ADAM33 HB-EGF TRAIL IL-33 TSLP CCL5 CCL11 CCL19 CX3CL1 CCL15 Activation Survival GC insensitivity Alteration of ASM function Migration Proliferation Contractility Chemoattraction of inflammatory cells T cells Mast cells Eosinophils Airway remodelling Airway Inflammation

Impaired GRa phosphorylation Impaired GRa transactivation activity Figure 2 : Potential molecular mechanisms mediating cytokine-induced GC insensitivity in ASM cells TNFa/IFNg GC Airway smooth muscle KCa3.1 PP5 P GRb IRF-1 GRIP-1 Impaired GRa phosphorylation 4-6 hr Rapid mechanisms GC P GRa Impaired GRa transactivation activity 12-24 hr Delayed mechanisms Corticosteroid Insensitivity