TITLE

Modulation of expression in BEAS-2B airway epithelial cells of α- l-fucosidase A1 and A2 by Th1 and Th2 cytokines, and overexpression of α- l-fucosidase 2

AUTHOR(S)
Sobkowicz, Anna; Gallagher, Mary; Reid, Colm; Crean, Daniel; Carrington, Stephen; Irwin, Jane
PUB. DATE
May 2014
SOURCE
Molecular & Cellular Biochemistry;May2014, Vol. 390 Issue 1/2, p101
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Chronic Th2-driven airway inflammation with excessive mucus production occurs in asthma. The regulation of FUCA1 and FUCA2 gene expression and enzyme activity in response to asthma-associated Th2 cytokines and, for contrast, Th1 cytokine IFN-γ, were investigated in a human airway cell line. BEAS-2B cells were supplemented with Th2-derived cytokines (IL-13, IL-4, IL-5) or/and IFN-γ. RNA and cell supernatants from stimulated and unstimulated cells were collected over a period of 3 h. Alpha- l-fucosidase A1 and A2 gene expression were assessed using real time RT-PCR, while enzymatic activities were measured using a fluorescent assay. To characterise α- l-fucosidase A2, CHO-K1 and BEAS-2B cell lines were transiently transfected, the FUCA2 gene was overexpressed, and the protein was immunoprecipitated. The transcription of FUCA1 was upregulated ( p < 0.01) in response to IFN-γ, suggesting that FUCA1 transcription and fucosidase activity are regulated in a Th1-dependent manner. The gene expression was the highest for 30 min after IFN-γ stimulation (>twofold induction), whereas secreted enzyme activity in BEAS-2B cells was significantly increased 1 h after IFN-γ addition. IL-4, IL-5 and IL-13 had no effect on FUCA1 and FUCA2 expression and activity. The IFN-γ-induced increase in expression and activity was repressed by the presence of the Th2 cytokine IL-5. Enzymatically active α- l-fucosidase 2 was immunoprecipitated from BEAS-2B cells, with highest activity at pH 4.9. IL-13, IL-4 and IL-5 have no effect on the expression of FUCA1 and FUCA2, but its expression is upregulated by IFN-γ, a Th1 cytokine. Active α- l-fucosidase 2 was overexpressed in BEAS-2B cells.
ACCESSION #
95321073

 

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