Allergy Asthma Immunol Res.  2014 Mar;6(2):149-155. 10.4168/aair.2014.6.2.149.

Effects of Fungi and Eosinophils on Mucin Gene Expression in Rhinovirus-Infected Nasal Epithelial Cells

• Shin SH
• Ye MK
• Kim JK

• KMID: 2260260
• DOI: http://doi.org/10.4168/aair.2014.6.2.149

Abstract

PURPOSE
Fungi, rhinoviruses (RVs), and eosinophils are associated with upper respiratory diseases. We evaluated the effects of fungal stimulation and eosinophil co-culture on the expression of mucin genes in RV-infected nasal polyp epithelial cells.
METHODS
Nasal polyp epithelial cells were obtained from chronic rhinosinusitis patients. Cultured epithelial cells were stimulated with Alternaria and Aspergillus with or without RV-16 infection. The epithelial cells were co-cultured with eosinophils for 16 h. MUC4, MUC5AC, MUC5B, and MUC8 mRNA expressions in the epithelial cells were quantified using real-time RT-PCR. To determine the underlying mechanism, nuclear factor-kappaB (NF-kappaB), activator protein-1 (AP-1), and mitogen-activated protein kinase (MAPK) inhibitors were used to inhibit mucin gene expression.
RESULTS
Fungi and RV-16 induced mucin gene expression in nasal polyp epithelial cells. However, there was no synergistic increase in mucin gene expression, with the exception of MUC4 mRNA expression stimulated by 25 microg/mL Aspergillus. When RV-16-infected epithelial cells were stimulated with fungi and then co-cultured with eosinophils, MUC4, MUC5B, and MUC8 mRNA expressions increased. Mucin gene expression was inhibited by NF-kappaB inhibitors.
CONCLUSIONS
RV-16, airborne fungi, and eosinophils may exacerbate the inflammatory process in nasal mucosal diseases by enhancing mucin gene expression.

Keyword

Rhinovirus; nasal epithelial cell; mucin gene; fungus; eosinophil; nuclear factor-kappaB

MeSH Terms

Alternaria
Aspergillus
Coculture Techniques
Eosinophils*
Epithelial Cells*
Fungi*
Gene Expression*
Humans
Methods
Mucins*
Nasal Polyps
NF-kappa B
Protein Kinases
Rhinovirus
RNA, Messenger
Transcription Factor AP-1
Mucins
NF-kappa B
Protein Kinases
RNA, Messenger
Transcription Factor AP-1

Figure

• 

  Fig. 1 Quantification of mucin gene expression in nasal polyp epithelial cells stimulated with Alternaria and Aspergillus with or without rhinovirus-16 (RV-16) infection for 24 h. MUC4, MUC5AC, and MUC8 mRNA expression levels were increased significantly by Alternaria; however, only MUC5AC mRNA expression was increased by Aspergillus. RV-16 also induced MUC4, MUC5AC, and MUC8 mRNA expression. When RV-infected epithelial cells were stimulated by fungi, MUC4, MUC 5AC, and MUC8 mRNA expression levels were increased. Values are expressed as means±SD of 6 independent experiments. Alt: Alternaria (µg/mL), Asp: Aspergillus (µg/mL), RV: rhinovirus-16-infected, (-): unstimulated. *significantly higher than unstimulated cells, †significantly higher than RV-infected cells.

• 

  Fig. 2 Quantification of mucin gene expression in nasal polyp epithelial cells co-cultured with eosinophils and stimulated with Alternaria and Aspergillus with or without rhinovirus-16 (RV-16) infection for 16 h. Values are expressed as the means±SD of 6 independent experiments. Alt: Alternaria (µg/mL), Asp: Aspergillus (µg/mL), RV: rhinovirus-16-infected, EO: eosinophils, (-): unstimulated. *significantly higher than unstimulated cells, †significantly higher than RV-infected cells.

• 

  Fig. 3 Effects of curcumin, SB 203580 and BAY 11-7082 on the expression of mucin genes. SB 203580 (MAPK inhibitor) inhibited MUC4 and MUC5AC mRNA expression and BAY 11-7082 (NF-κB inhibitor) inhibited MUC4, MUC5AC, MUC5B, and MUC8 mRNA expression. Values are expressed as the means±SD of 5 independent experiments. Alt: Alternaria (µg/mL), Asp: Aspergillus (µg/mL), V: rhinovirus-16 infected, (-): non-stimulated. *P<0.05.

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