Noy Nachmias1,2,3, Sheila Langier3, Rafael Y Brzezinski2,4, Matan Siterman3, Moshe Stark3, Sara Etkin3, Avital Avriel1, Yehuda Schwarz3, Shani Shenhar-Tsarfaty2, Amir Bar-Shai3. 1. The Division of Pulmonary Medicine, Barzilai Medical Center, Faculty of Health Sciences, Ben-Gurion University, Ashkelon, Israel. 2. Department of Internal Medicine "C, "D and "E, The Tel Aviv Sourasky Medical Center, Tel Aviv, affiliated to the Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel. 3. The Pulmonary Institute, Tel-Aviv Sourasky Medical Center and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel. 4. Neufeld Cardiac Research Institute, Sackler Faculty of Medicine, Tel Aviv University, Israel; Tamman Cardiovascular Research Institute, Leviev Heart Center, Sheba Medical Center, Tel-Hashomer, Israel.
Abstract
BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an inflammatory disease characterized by a progressive and irreversible deterioration of lung function. Exacerbations of COPD have prolonged negative effects on pulmonary function and a major impact on health status and outcomes. NLRP3 inflammasome is a cardinal component of the inflammatory response, with marked evidence in stable and exacerbations of COPD. The aim of our study was to evaluate the NLRP3 inflammasome activity during COPD exacerbation by using an in vitro model. METHODS: A549 cells were stimulated with different concentrations (10%, 4%, 2%) of cigarette smoke extract (CSE) with or without LPS (0.1μg/ml) for 24 hours. Cell viability was assessed by using XTT test. Levels of inflammatory cytokines (IL-8, MCP-1, and IL-1β) were measured by ELISA and the activity level of NLRP-3 was evaluated by flow cytometry. RESULTS: Cells exposed to CSE present an increase in inflammatory cytokines (IL-8 and MCP-1) production in a dose-dependent manner. Incubation with LPS to these cells results in higher levels of IL-8 and MCP-1 compared to stimulation of CSE alone. NLRP3 inflammasome activity and IL-1β levels were significantly increased in cells exposed to both CSE and LPS compared to CSE alone. CONCLUSIONS: NLRP3 inflammasome is upregulated in an in-vitro model of COPD and COPD exacerbation. Our findings provide novel biomarkers for COPD exacerbation and may present new targets for future research.
BACKGROUND:Chronic obstructive pulmonary disease (COPD) is an inflammatory disease characterized by a progressive and irreversible deterioration of lung function. Exacerbations of COPD have prolonged negative effects on pulmonary function and a major impact on health status and outcomes. NLRP3 inflammasome is a cardinal component of the inflammatory response, with marked evidence in stable and exacerbations of COPD. The aim of our study was to evaluate the NLRP3 inflammasome activity during COPD exacerbation by using an in vitro model. METHODS:A549 cells were stimulated with different concentrations (10%, 4%, 2%) of cigarette smoke extract (CSE) with or without LPS (0.1μg/ml) for 24 hours. Cell viability was assessed by using XTT test. Levels of inflammatory cytokines (IL-8, MCP-1, and IL-1β) were measured by ELISA and the activity level of NLRP-3 was evaluated by flow cytometry. RESULTS: Cells exposed to CSE present an increase in inflammatory cytokines (IL-8 and MCP-1) production in a dose-dependent manner. Incubation with LPS to these cells results in higher levels of IL-8 and MCP-1 compared to stimulation of CSE alone. NLRP3 inflammasome activity and IL-1β levels were significantly increased in cells exposed to both CSE and LPS compared to CSE alone. CONCLUSIONS:NLRP3 inflammasome is upregulated in an in-vitro model of COPD and COPD exacerbation. Our findings provide novel biomarkers for COPD exacerbation and may present new targets for future research.
Authors: Don D Sin; Zsuzsanna Hollander; Mari L DeMarco; Bruce M McManus; Raymond T Ng Journal: Am J Respir Crit Care Med Date: 2015-11-15 Impact factor: 21.405
Authors: N S Pauwels; K R Bracke; L L Dupont; G R Van Pottelberge; S Provoost; T Vanden Berghe; P Vandenabeele; B N Lambrecht; G F Joos; G G Brusselle Journal: Eur Respir J Date: 2011-05-26 Impact factor: 16.671
Authors: Jeremy A Hirota; Simon A Hirota; Stephanie M Warner; Dorota Stefanowicz; Furquan Shaheen; Paul L Beck; Justin A Macdonald; Tillie-Louise Hackett; Don D Sin; Stephan Van Eeden; Darryl A Knight Journal: J Allergy Clin Immunol Date: 2012-01-09 Impact factor: 10.793
Authors: Rosa Faner; Patricia Sobradillo; Aina Noguera; Cristina Gomez; Tamara Cruz; Alejandra López-Giraldo; Eugeni Ballester; Nestor Soler; Juan I Arostegui; Pablo Pelegrín; Roberto Rodriguez-Roisin; Jordi Yagüe; Borja G Cosio; Manel Juan; Alvar Agustí Journal: ERJ Open Res Date: 2016-07-11