Allergy Asthma Immunol Res.  2010 Jan;2(1):41-47. 10.4168/aair.2010.2.1.41.

Chlamydophila pneumoniae enhances secretion of VEGF, TGF-beta and TIMP-1 from human bronchial epithelial cells under Th2 dominant microenvironment

• Park CS
• Kim TB
• Moon KA
• Bae YJ
• Lee HR
• Jang MK
• Moon HB
• Cho YS

• KMID: 2260418
• DOI: http://doi.org/10.4168/aair.2010.2.1.41

Abstract

PURPOSE
Chlamydophila pneumoniae infection in the airways is thought to be associated with the pathogenesis of asthma, especially in non-atopic severe asthma with irreversible airway obstruction that may be related to airway remodeling. Here, we investigated whether C. pneumoniae infection enhances the secretion of critical chemical mediators for airway remodeling, such as VEGF, TGF-beta, and TIMP-1, in human bronchial epithelial cells (BECs) in a Th2-dominant microenvironment.
METHODS
Human bronchial epithelial cells (BEAS-2B cells) were infected with C. pneumoniae strain TW183 and cultured in both a Th1-dominant microenvironment with INF-gamma and a Th2-dominant microenvironment with IL-4 or IL-13 added to the culture medium. The VEGF, TGF-beta, and TIMP-1 levels in the culture supernatants were measured using enzyme-linked immunosorbent assays (ELISA). The activation of NF-kappaB in each experimental condition was determined using an electrophoretic mobility shift assay.
RESULTS
Chlamydophila pneumoniae-infected BECs showed enhanced secretion of VEGF, TGF-beta, and TIMP-1 compared with non-infected BECs. The levels of cytokines secreted from BECs were increased more when IL-13 was added to the culture medium. C. pneumoniae-infected BECs also showed increased NF-kappaB activation.
CONCLUSIONS
These results suggest that C. pneumoniae plays a role in the pathogenesis of airway remodeling in asthma, revealing a Th2-dominant immune response. Further studies are required to clarify the precise mechanism of C. pneumoniae infection in airway remodeling.

Keyword

Asthma; Chlamydophila pneumonia; epithelial cells; vascular endothelial growth factor A; tissue inhibitor of metalloproteinases; transforming growth factor beta

MeSH Terms

Airway Obstruction
Airway Remodeling
Asthma
Chlamydial Pneumonia
Chlamydophila
Chlamydophila pneumoniae
Cytokines
Electrophoretic Mobility Shift Assay
Enzyme-Linked Immunosorbent Assay
Epithelial Cells
Humans
Interleukin-13
Interleukin-4
NF-kappa B
Pneumonia
Sprains and Strains
Tissue Inhibitor of Metalloproteinase-1
Tissue Inhibitor of Metalloproteinases
Transforming Growth Factor beta
Vascular Endothelial Growth Factor A
Cytokines
Interleukin-13
Interleukin-4
NF-kappa B
Tissue Inhibitor of Metalloproteinase-1
Tissue Inhibitor of Metalloproteinases
Transforming Growth Factor beta
Vascular Endothelial Growth Factor A

Figure

• 

  Fig. 1 C. pneumoniae strain TW-183 in BEAS-2B cells. (A) BEAS-2B cells grown on cover slips were infected with C. pneumoniae. After 24 hr, cells were fixed with methanol, and stained using a fluorescein conjugated, pneumonia IgG-FITC (×200), BEAS-2B cells (red), C. pneumoniae (green). Proliferation effect of BEAS-2B cells infected with C. pneumoniae. (B) BEAS-2B cells were infected with C. pneumoniae at 0 (▪), 0.1 (▴), 1 (▾), and 10 (♦) ifu and cell viability measured at different time points post infection (12, 24, 48, 72, and 96 hr). Results shown are representative of three independent experiments.

• 

  Fig. 2 Effect of C. pneumoniae infection on VEGF, TGF-β, and TIMP-1 secretion from BEAS-2B cells. BEAS-2B cells were incubated for 24 hr with LHC-9/RPMI containing 20 ng/mL IL-4, IL-13 or IFN-γ in absence or presence of C. pneumoniae infection. The levels of VEGF (A), TGF-β (B), and TIMP-1 (C) in the supernatants were determined by ELISA. Results shown are representative of three independent experiments. *P<0.05 compared to C. pneumoniae uninfected control. †P<0.05 compared with control without cytokine treatment.

• 

  Fig. 3 NF-κB activity by C. pneumoniae infection in BEAS-2B cells. Changes of NF-κB binding activity using EMSA (A) gel using g32 p-labeled oligonucleotide probes for NF-κB. Densitometry of NF-κB specific binding activity (B) depicts the densitometric evaluation of NF-κB DNA-binding activity, normalized to the level of standard reaction.

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