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Up-regulation of mucin secretion in HT29-MTX cells by the pro-inflammatory cytokines tumor necrosis factor-α and interleukin-6

Lookup NU author(s): Marina Smirnova, Professor Jeffrey PearsonORCiD

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Abstract

The pro-inflammatory cytokines IL-6 and TNF-α have been implicated in the pathogenesis of otitis media with effusion (OME). A disease where goblet cells proliferate in a modified respiratory epithelium, leading to the accumulation of a mucin-rich effusion in the middle ear cleft. The MUC5AC and MUC5B mucin gene products have been identified as components of these effusions. To determine the effect of IL-6 and TNF-α on MUC5AC and MUC5B secretion we have used HT29-MTX goblet cells, which secrete both types of mucins. MUC5AC and MUC5B mucin secretion was measured by an enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody NCL-HGM-45M1 and polyclonal antiserum TEPA, respectively. Time response (0.72 hours) and dose response (1.5-150 ng/ml) studies were carried out. IL-6 and TNF-α stimulated MUC5AC and MUC5B mucin secretion in a time dependent manner, both in pre-confluent and post-confluent cells. IL-6 (15 ng/ml and 20 ng/ml) produced a low and prolonged stimulation of mucin secretion that persisted for 72 hours, with peak response at 24 hours after induction. The IL-6-mediated mucin secretion at 24 hours was concentration-dependent, with a maximal effect at 15 ng/ml. TNF-α (20 ng/ml) induced rapid stimulation of mucin secretion within the first 24 hours, with peak response at 7 hours after induction. IL-6 and TNF-α exposure significantly increased MUC5AC secretion, but not MUC5B secretion. Maximal levels of cytokine-induced mucin secretion were detected in pre-confluent cells that showed one and a half- and two-fold increases in MUC5AC secretion after IL-6 and TNF-α stimulation, respectively, in comparison with post-confluent cells. The results presented here suggest that IL-6 and TNF-α generate a differential up-regulation of mucin secretion and thus contribute to the expression of mucin genes in inflammatory responses.

Publication metadata

Author(s): Smirnova MG; Pearson JP; Kiselev SL; Birchall JP

Publication type: Article

Publication status: Published

Journal: European Cytokine Network

Year: 2001

Volume: 12

Issue: 1

Pages: 119-125

Print publication date: 01/01/2001

ISSN (print): 1148-5493

ISSN (electronic): 1952-4005

Publisher: John Libbey Eurotext

URL: http://www.ncbi.nlm.nih.gov/pubmed/11282555

PubMed id: 11282555

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