Allergy and Airway |

Galectins Expression in Leucocytes From Induced Sputum Samples FREE TO VIEW

Silvia Sanchez, PhD; Rosa Girón Moreno, PhD; Rosa Gomez Punter, MD; Emma Vazquez, MD; Gonzalo Segrelles, PhD; Carolina Cisneros, PhD; Julio Ancochea Bermúdez, PhD
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Hospital Universitario de La Princesa, Madrid, Spain

Chest. 2014;145(3_MeetingAbstracts):18A. doi:10.1378/chest.1798371
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SESSION TITLE: Asthma Posters

SESSION TYPE: Poster Presentations

PRESENTED ON: Saturday, March 22, 2014 at 01:15 PM - 02:15 PM

PURPOSE: Galectins are a family of b-galactoside-binding animal lectins with functions in a variety of biological processes, including inflammation and allergic pathologies. To explore the contribution of galectins in human asthma, induced sputum samples were collected from asthma patients and healthy controls. Expression of galectin (gal)-1, gal-3 and gal-9 was analysed by flow cytometry.

METHODS: Twenty-four asthmatic subjects with stable asthma (19 women/5 men) without systemic steroids and 18 healthy controls (9 women/9 men) were included. Asthma severity was scored according to the criteria of the Global Strategy for Asthma Management and Prevention (GINA) based on current therapy. Clinical parameters: age, sex, pulmonary function, asthma severity, atopic status, ACT, FeNO, ICS, number of years since diagnosis and history of smoking, rhinitis and nasal polyps were collected. Sputum cells from both group were labelled with PO-anti-CD45, PE-anti-HLADR, PB-anti-CD16 and APC-H7-anti-CD14. For galectin detection, cells were stained with goat polyclonal anti-gal-1, anti-gal-3 or anti-gal-9 followed by Alexa Fluor 647-DAG.Surface expression of galectin proteins in sputum cells was determined by flow cytometry. Using sputum samples we determined whether galectins were expressed differentially in the main subsets of sputum leucocytes observed in cytospin. Differential expression of HLA-DR was used to distinguish macrophages (CD16+DR+) and neutrophils (CD16+DR-) and the expression of galectins was studied in both subpopulations.

RESULTS: Macrophages are the main sputum cells expressing gal-1 and gal-9. Gal-1 and gal-9 were expressed only on macrophages, while gal-3 expression was detected on both macrophages and neutrophils.

CONCLUSIONS: Sputum induction is thus a valuable, non-invasive means of obtaining viable cells from the lower airway for evaluation of airway inflammation. - Using this method to obtain airway cells, we have identified macrophages as the cells from sputum expressing gal-1 and gal-9.

CLINICAL IMPLICATIONS: The balance of pro- and antiinflammatory signals determines the final outcome of the immune response, and proteins as gal-1 and gal-9 is probably may contribute to the inflammatory response present in human asthma.

DISCLOSURE: The following authors have nothing to disclose: Silvia Sanchez, Rosa Girón Moreno, Rosa Gomez Punter, Emma Vazquez, Gonzalo Segrelles, Carolina Cisneros, Julio Ancochea Bermúdez

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