Blood was used for the determination of CFU counts and measurement of cytokine concentrations
Blood was used for the determination of CFU counts and measurement of cytokine concentrations. Flow cytometry Cells harvested from BAL and lungs were washed and incubated with antibodies (BD, Franklin lakes, NJ, USA) for 30 min in PBS before being washed. of cases are associated with NTHi. GSK2194069 This susceptibility to contamination involves a defective production of interleukin (IL)-22 which plays an important role in mucosal defense. Prophylactic administration of flagellin, a Toll-like receptor 5 (TLR5) agonist, protects healthy mice against respiratory pathogenic bacteria. We hypothesized that TLR5-mediated stimulation of lung immunity might prevent COPD exacerbations. Mice chronically exposed to cigarette smoke (CS), which presented COPD symptoms, were infected with NTHi and intraperitoneally treated with recombinant flagellin following a prophylactic or therapeutic protocol. Compared with control, cigarette smoke-exposed mice treated with flagellin showed a lower bacterial load in the airways, the lungs and the blood. This protection was associated with an early neutrophilia, a lower production of pro-inflammatory GSK2194069 cytokines and an increased IL-22 production. Flagellin treatment decreased the recruitment of inflammatory cells and the lung damages related to exacerbation. Morover, the protective effect of flagellin against NTHi was altered by treatment with anti-IL-22 blocking antibodies in cigarette smoke-exposed mice and in mice. The effect of flagellin treatment did not implicated the anti-bacterial peptides calgranulins and defensin-2. This study shows that stimulation of innate immunity by a TLR5 ligand is usually a potent antibacterial treatment in CS-exposed mice, suggesting innovative therapeutic strategies against acute exacerbation in COPD. Introduction Chronic obstructive pulmonary disease (COPD) is usually characterized by a progressive and irreversible decline in lung function [1]. Being the third leading cause of death worldwide, it is mainly caused by chronic exposure to cigarette smoke (CS) or pollutants [2]. Inhalation of CS essentially leads to activation of epithelial cells and macrophages responsible for the mobilization of effector and immuno-modulatory cells including neutrophils and natural killer T (NKT) cells [3,4]. The chronic inflammatory response progressively leads to airway remodeling, impaired bacterial clearance and parenchymal destruction in the lungs, further culminating in irreversible airflow limitation [5] as experienced in our murine model of chronic exposure to CS. These components are involved in the increased susceptibility of COPD patients to bacterial and viral GSK2194069 airway infections. Airway colonization with bacteria such as and contributes to the pathogenesis and clinical course of the disease [6]. This colonization is responsible for lung contamination leading to exacerbations of the disease, which have a strong impact on health status, exercise capacity, lung function, and mortality. Non-typeable (NTHi), a Gram-negative coccobacillus that lacks a polysaccharide capsule, is an important cause of COPD exacerbations and comorbidity [7,8]. Acute exacerbations in patients invariably scarred the chronic course of COPD [9]. Bacterial infections are first controlled by the innate immune system, which implicated pathogen-associated molecular pattern (PAMP) recognition by Toll-like receptors (TLR) such as those recognizing flagellin (TLR5) responsible for the mobilization of effector cells [10]. During COPD, bacterial infection is usually characterized by an increased influx of immune cells, including neutrophils, macrophages, dendritic cells (DC) and T lymphocytes [3,11,12]. However, this response is not effective enough to clear the pathogens. In this context, we recently reported a defective production of IL-22 in response to bacteria both in COPD patients and mice chronically exposed Rabbit Polyclonal to AOS1 to CS, whereas IL-17 production is only altered after contamination by [13,14]. Interestingly, the Th17 cytokines IL-17 and IL-22 promote the recruitment of neutrophils, the synthesis of antimicrobial peptides and the expression of tight junction molecules [15,16], a mechanism explaining the essential role of IL-22 in the clearance of NTHi [14]. Morover, supplementation of COPD mice with recombinant IL-22 increases the clearance of the bacteria and prevents the development of COPD exacerbations in mice. However, IL-22 expression is also promoted by exposure to CS and is involved in COPD pathogenesis [17]. Several reports showed that activation of innate receptors, including TLR, is able to elicit protective immune responses against infections [18,19]. Among them, systemic administration of flagellin, the main component of bacterial flagella and the TLR5 ligand, induces immediate production of Th17 cytokines through the activation of DC and type 3 innate lymphoid cells [20]. In this study, we hypothesized that systemic administration of recombinant form of flagellin could inhibit the development of NTHi-induced COPD exacerbation episodes through an appropriate protective.