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- Title
Characterization of exosome-like vesicles released from human tracheobronchial ciliated epithelium: a possible role in innate defense.
- Authors
Kesimer, Mehmet; Scull, Margaret; Brighton, Brian; DeMaria, Genevieve; Burns, Kimberlie; O'Neal, Wanda; Pickles, Raymond J.; Sheehan, John K.
- Abstract
Airway mucus forms the structural basis of the local innate immune defense mechanism. It is an integrated, active, viscoelastic gel matrix evolved to protect the exposed lung from physical, chemical, and pathological erosion. Exosomes are biologically active vesicles secreted by different cell types including epithelial, hematopoietic, and some tumor cells. They are also present in some biological fluids soda as serum, urine, breast mill and bronchoalveolar lavage fluid. In rids study, we demonstrate for the first time that exosome-like vesicles with antiviral properties are present in human tracheobronchial epithelial (HTBE) cell culture secretions. These vesicles have been isolated by differential centrifugation and are characterized further by mass spectrometry, flow cytometry, immunoblotting, electron microscopy, and light-scattering methods. HTBE vesicles exhibited characteristic exosomal size (30-100 nm) and morphology (cup-shaped) with a buoyant density in sucrose of 1.12-1.18 g/ml. Biochemical characterization further revealed typical surface, cytoskeletal, and cytoplasmic proteins characteristic of exosomes, including the multivesicular and late endosomal membrane markers Tsg101 and CD63. The presence of RNA was also observed. The epithelial mucins MUC1, MUC4, and MUC16 also contributed to the vesicles' structure. Notably, α-2,6-linked sialic acid was associated with these mucin molecules and subsequent functional analysis showed that these vesicles have a neutralizing effect on human influenza virus, which is known to bind sialic acid. Taken together, these findings suggest that airway epithelial cells release exosome-like vesicles mad that these structures may be involved in diverse physiological processes in airway biology, including innate mucosal defense.
- Subjects
EPITHELIUM; MUCINS; INFLUENZA viruses; MUCUS; SIALIC acids; CELL culture; RNA; MASS spectrometry
- Publication
FASEB Journal, 2009, Vol 23, Issue 6, p1858
- ISSN
0892-6638
- Publication type
Article
- DOI
10.1096/fj.08-119131