Exosome secretion by eosinophils: A possible role in asthma pathogenesis

• Published in: Journal of allergy and clinical immunology Vol. 135; no. 6; pp. 1603 - 1613
• Main Authors: Mazzeo, Carla, PhD, Cañas, José Antonio, MSc, Zafra, Maria Paz, MSc, Rojas Marco, Ainara, MD, Fernández-Nieto, Mar, MD, PhD, Sanz, Veronica, VET, Mittelbrunn, María, PhD, Izquierdo, Manuel, PhD, Baixaulli, Francesc, MSc, Sastre, Joaquín, MD, PhD, del Pozo, Victoria, PhD
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-06-2015; Elsevier Limited
• Subjects: Allergy and Immunology; Asthma; Asthma - diagnosis; Asthma - immunology; Asthma - metabolism; Asthma - pathology; biomarker; Biomarkers; Biomarkers - analysis; Biomarkers - metabolism; Case-Control Studies; CD63; Cell Fractionation; Cell Separation; Chemokine CCL11 - pharmacology; Cytokines; Disease; endosomes; eosinophils; Eosinophils - drug effects; Eosinophils - immunology; Eosinophils - metabolism; Eosinophils - ultrastructure; exosomes; Exosomes - immunology; Exosomes - metabolism; Exosomes - ultrastructure; Humans; IFN-γ; Interferon-gamma - pharmacology; lysobisphosphatidic acid; Microscopy, Electron, Transmission; multivesicular bodies; Multivesicular Bodies - immunology; Multivesicular Bodies - metabolism; Multivesicular Bodies - ultrastructure; Organelle Size; Pathogenesis; Patients; Primary Cell Culture; secretion; Studies; Western blot; CD63; Protein kinase C; Multivesicular body; LBPA; Major basic protein; EPO; lysobisphosphatidic acid; eosinophils; WB; Eeosinophil peroxidase; GFP; MFI; IFN-γ; ILV; exosomes; MBP; Mean fluorescence intensity; endosomes; PKC; biomarker; Asthma; multivesicular bodies; secretion; ECP; Transmission electron microscopy; Green fluorescent protein; MVB; Eosinophil cationic protein; Intraluminal vesicle; TEM
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2014.11.026

Background Eosinophils secrete several granules that are involved in the propagation of inflammatory responses in patients with pathologies such as asthma. Objective We hypothesized that some of these granules are exosomes, which, when transferred to the recipient cells, could modulate asthma progression. Methods Eosinophils were purified from peripheral blood and cultured with or without IFN-γ or eotaxin. Multivesicular bodies (MVBs) in eosinophils were studied by using fluorescence microscopy, transmission electron microscopy (TEM), and flow cytometry. Exosome secretion was measured and exosome characterization was performed with TEM, Western blotting, and NanoSight analysis. Results Generation of MVBs in eosinophils was confirmed by using fluorescence microscopy and flow cytometry and corroborated by means of TEM. Having established that eosinophils contain MVBs, our aim was to demonstrate that eosinophils secrete exosomes. To do this, we purified exosomes from culture medium of eosinophils and characterized them. Using Western blot analysis, we demonstrated that eosinophils secreted exosomes and that the discharge of exosomes to extracellular media increases after IFN-γ stimulation. We measured exosome size and quantified exosome production from healthy and asthmatic subjects using nanotracking analysis. We found that exosome production was augmented in asthmatic patients. Conclusion Our findings are the first to demonstrate that eosinophils contain functional MVBs and secrete exosomes and that their secretion is increased in asthmatic patients. Thus exosomes might play an important role in the progression of asthma and eventually be considered a biomarker.