Tuberc Respir Dis.  2008 Jan;64(1):22-27. 10.4046/trd.2008.64.1.22.

Roles of Matrix Metalloproteinases on Intracellular Staphylococcus aureus Growth in Bronchial Epithelial Cell

• Min BR
• Lee YM
• Park JS
• Choi WI
• Kwon KY

• KMID: 1478137
• DOI: http://doi.org/10.4046/trd.2008.64.1.22

Abstract

BACKGROUND: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity.
METHODS
In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor.
RESULTS
The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased.
CONCLUSION
MMPs may mediate the invasion of S. aureus into epithelial cells.

Keyword

Staphylococcus aureus; Epithelial cell; Matrix metalloproteinases

MeSH Terms

Bacteria
Blotting, Western
Cadherins
Cell Adhesion
Colon
Dipeptides
Epithelial Cells
Epithelium
Extracellular Matrix
Humans
Matrix Metalloproteinases
Membrane Proteins
Proteins
Respiratory Tract Infections
Staphylococcus
Staphylococcus aureus
Cadherins
Dipeptides
Matrix Metalloproteinases
Membrane Proteins
Proteins

Figure

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  Figure 1 Intracellular bacterial survival/replication of S. aureus. (A) 4 h, (B) 12 h. *p<0.01 versus MOI 5 and MOI 10; †<0.05 versus MOI 20 and MOI 50.

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  Figure 2 Differential expression patterns of MMPs in BEAS-2B cells infected with S. aureus. Conditioned media were subjected to 7.5% SDS-PAGE with 0.2% gelatin and gelatinolytic activities of MMP-9 and MMP-2 were determined by zymography as described in materials and methods.

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  Figure 3 Differential expression patterns of E-cadherin and Dysadherin. Whole cell lysates was prepared and used for E-cadherin, Dysadnerin, and actin Westerin blot analysis.

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  Figure 4 Intracellular bacterial survival and replication of S. aureus in BEAS-2B cells treated with MMP inhibitor, GM6001 (40uM). (A) 4 h, (B) 12 h. *p<0.05 versus control.

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