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• 作者：Anna D. Sobkowicz (1) (2)
  Mary E. Gallagher (1)
  Colm J. Reid (1)
  Daniel Crean (1)
  Stephen D. Carrington (1)
  Jane A. Irwin (1)
  
• 关键词：α ; l ; fucosidase ; Th1 ; Th2 ; Asthma ; Interferon ; gamma ; IL ; 5 ; BEAS ; 2B cells
• 刊名：Molecular and Cellular Biochemistry
• 出版年：2014
• 出版时间：May 2014
• 年：2014
• 卷：390
• 期：1-2
• 页码：101-113
• 全文大小：1,353 KB
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• 作者单位：Anna D. Sobkowicz (1) (2)
  Mary E. Gallagher (1)
  Colm J. Reid (1)
  Daniel Crean (1)
  Stephen D. Carrington (1)
  Jane A. Irwin (1)
  
  1. Veterinary Sciences Centre, School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland
  2. Cardiff University-Peking University Cancer Institute, Cardiff University School of Medicine, Henry Wellcome Building, Heath Park, Cardiff, CF14 4XN, UK
  
• ISSN：1573-4919

文摘

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.