Effects of formoterol and salmeterol on cytokine release from monocyte-derived macrophages

Abstract

Pulmonary macrophages are a target for inhaled therapies. Combinations of longacting b 2 -agonists (LABA) and glucocorticosteroids have been developed for asthma and chronic obstructive pulmonary disease (COPD). This study examined two LABA, salmeterol and formoterol, and the glucocorticosteroid, budesonide, on cytokine release from monocytederived macrophages (MDM) to determine whether anti-inflammatory effects observed in patients are due to inhibition of macrophages.MDM were incubated in the absence or presence of LABA or budesonide prior to stimulation with lipopolysaccharide (LPS). Tumour necrosis factor (TNF)-a, granulocyte macrophage-colony stimulating factor (GM-CSF) and CXC chemokine ligand (CXCL)8 were measured by ELISA.Formoterol and salmeterol inhibited LPS-stimulated release of TNF-a (mean effective concentration (EC50) 2.4¡1.8 and 3.5¡2.7 nM, respectively; n511-16), GM-CSF (EC50 24.6¡2.1 and 52.4¡40.8 nM, respectively, n511-12) but not CXCL8 from LPS-stimulated MDM. Budesonide inhibited release of all three cytokines (EC50 TNF-a: 1.2¡0.4 nM; GM-CSF: 0.4¡0.2 nM; CXCL8: 0.4¡0.1 nM; n53-4). Formoterol but not salmeterol elevated cAMP in these cells. These effects were attenuated by b-adrenoceptor antagonists, propranolol and ICI118551. Salmeterol (10 -7 M) also inhibited formoterol-induced cAMP and formoterol-mediated attenuation of cytokine release. Combining budesonide (0.3 nM) with formoterol, inhibited TNF-a release additively. LABA may inhibit inflammatory cytokine release from macrophages in a cAMP-independent manner and act additively with budesonide.

Evaluation of the anti-inflammatory effects of β-adrenoceptor agonists on human lung macrophages

“…Salmeterol and indacaterol were reasonably effective inhibitors of cytokine generation from lung macrophages whereas formoterol was ineffective. These findings differ with those reported for MDM in which both formoterol and salmeterol were capable of reducing the extent of LPS-induced cytokine generation (Donnelly et al, 2010). Indeed, our own studies indicate that all the short-acting -agonists tested (isoprenaline, salbutamol and terbutaline) as well as the long-acting -agonist formoterol are considerably more effective inhibitors of cytokine generation in MDM than in lung macrophages (Fig.…”

“…Attenuating excessive macrophage activation would be expected to be benefical in the treatment of respiratory diseases. Studies investigating the effects of -agonists on monocytes, monocyte-derived macrophages (MDM) and macrophagelike cell lines suggest that these drugs may modulate a number of macrophage functions including the prevention of pro-inflammatory cytokine generation (Yoshimura et al, 1997;Izeboud et al, 1999;Donnelly et al, 2010;Shirato et al, 2013). By contrast, other studies show limited effects of -agonists in these systems (Linden, 1992;Zetturlund et al, 1998;Ezeamuzie and Shihab, 2010).…”

“…Previous studies have demonstrated that -adrenoceptor agonists are effective inhibitors of cytokine generation from MDM (Donnelly et al, 2010). The effects of both short-acting and long-acting -agonists on cytokine generation from LPS activated MDM were investigated (Fig.…”

Genome Expression Profiling-Based Identification and Administration Efficacy of Host-Directed Antimicrobial Drugs against Respiratory Infection by Nontypeable Haemophilus influenzae

“…The structure, affinity, efficacy, kinetics, and ability to elevate intracellular cAMP vary for salmeterol and formoterol (57,65), which could explain the differences observed. We also assessed the effect of increasing intracellular cAMP by inhibiting PDEs.…”

The In Vitro Pharmacology of GS-5759, A Novel Bifunctional Phosphodiesterase 4 Inhibitor and Long Acting β2-Adrenoceptor Agonist

“…The combination of a bronchodilator with an anti-inflammatory agent as a treatment of COPD has been extensively discussed and is reviewed in Phillips and Salmon (2012). The ability of b 2 -adrenoceptor agonists to also have direct anti-inflammatory activity has been previously described, including inhibition of histamine, arachidonic acid metabolites, and TNFa release from mast cells (Undem et al, 1988;Bissonnette and Befus, 1997;Chong et al, 1998) and on cytokine release from monocytes (Seldon et al, 2005;Donnelly et al, 2010). Previous studies from this laboratory and others have shown the effects of roflumilast combined with salmeterol, formoterol, or dexamethasone on lipopolysaccharide (LPS)-induced peripheral blood mononuclear cells (PBMC) cytokine production, showing additive effects for PDE4 inhibition with either a LABA or glucocorticosteroid (Seldon et al, 2005;Tannheimer et al, 2012a), suggesting an additional anti-inflammatory effect in combination or possible triple combination.…”