Yonsei Med J.  2011 Jan;52(1):51-58. 10.3349/ymj.2011.52.1.51.

Epithelial to Mesenchymal Transition of Mesothelial Cells in Tuberculous Pleurisy

• Kim C
• Kim DG
• Park SH
• Hwang YI
• Jang SH
• Kim CH
• Jung KS
• Min K
• Lee JW
• Jang YS

• KMID: 1106437
• DOI: http://doi.org/10.3349/ymj.2011.52.1.51

Abstract

PURPOSE
Tuberculous pleurisy is the most frequent extrapulmonary manifestation of tuberculosis. In spite of adequate treatment, pleural fibrosis is a common complication, but the mechanism has not been elucidated. This study is to determine whether epithelial to mesenchymal transition (EMT) of mesothelial cells occurs in tuberculous pleurisy.
MATERIALS AND METHODS
Normal pleural mesothelial cells, isolated from irrigation fluids during operations for primary spontaneous pneumothorax, were characterized by immunofluorescence and reverse transcription polymerase chain reaction (RT-PCR). These cells were treated in vitro with various cytokines, which were produced in the effluents of tuberculous pleurisy. The isolated cells from the effluents of tuberculous pleurisy were analyzed by immunofluorescence and RT-PCR analysis.
RESULTS
The isolated cells from the irrigation fluid of primary spontaneous pneumothorax had epithelial characteristics. These cells, with transforming growth factor-beta1 and/or interleukin-1beta treatment, underwent phenotypic transition from epithelial to mesenchymal cells, with the loss of epithelial morphology and reduction in cytokeratin and E-cadherin expression. Effluent analysis from tuberculous pleurisy using immunofluorescence and RT-PCR demonstrated two phenotypes that showed mesenchymal characteristics and both epithelial & mesencymal characteristics.
CONCLUSION
Our results suggest that pleural mesothelial cells in tuberculous pleurisy have been implicated in pleural fibrosis through EMT.

Keyword

Epithelial to mesenchymal transition; IL-1beta; mesothelial cells; pleural fibrosis; TGF-beta1; tuberculous pleurisy

MeSH Terms

Cells, Cultured
Epithelial Cells/*pathology
Epithelial-Mesenchymal Transition/*physiology
Fluorescent Antibody Technique
Humans
Pleura/*pathology
Reverse Transcriptase Polymerase Chain Reaction
Tuberculosis, Pleural/*pathology

Figure

• 

  Fig. 1 Morphologic changes in mesothelial cells from the effluent of a patient with primary spontaneous pneumothorax with IL-1β ± TGF-β1 treatment. Cells obtained from the irrigation effluent of primary spontaneous pneumothorax during operation were treated with media only, IL-1β (2 ng/mL), TGF-β1 (0.5 ng/mL), and IL-1β + TGF-β1, respectively. Phase images are in the first row, and immunofluorescence stains in the remaining rows. Cultured cells after cytokines treatment were stained with monoclonal antibodies against E-cadherin, cytokeratin, vimentin, and F-actin.

• 

  Fig. 2 Expression of snail transcription factor and the loss of E-cadherin expression in mesothelial cells with TGF-β1 ± IL-1β. Cells obtained from the irrigation effluent of a patient with primary spontaneous pneumothorax were treated with media only, TGF-β1 (0.5 ng/mL), and TGF-β1 + IL-1β (2 ng/mL), respectively. Cells were analyzed for snail and E-caherin messenger RNA expression by RT-PCR. RT-PCR, reverse transcription polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 3 Characteristics of cultured cells from effluents of tuberculous pleurisy. Cells characteristics from the effluents of five tuberculous pleurisy patients were observed under phase-contrast microscopy and stained with monoclonal antibodies against E-cadherin, cytokeratin, vimentin, and F-actin.

• 

  Fig. 4 Expression of messenger RNA encoding snail and E-cadherin in tuberculous pleurisy. Cells obtained from the seven effluents of tuberculous pleurisy were analyzed for snail and E-cadherin mRNA expression by RT-PCR. RT-PCR results of samples A) and B) from Fig. 3 are shown on columns 1 and 2 of Fig. 4, respectively. RT-PCR, reverse transcription polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

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