April 1997
Volume 38, Issue 5
Free
Articles  |   April 1997
Production of platelet-derived growth factor by interleukin-1 beta and transforming growth factor-beta-stimulated retinal pigment epithelial cells leads to contraction of collagen gels.
Author Affiliations
  • P Choudhury
    Department of Microbiology and Immunology, University of South Carolina School of Medicine, Columbia 29208, USA.
  • W Chen
    Department of Microbiology and Immunology, University of South Carolina School of Medicine, Columbia 29208, USA.
  • R C Hunt
    Department of Microbiology and Immunology, University of South Carolina School of Medicine, Columbia 29208, USA.
Investigative Ophthalmology & Visual Science April 1997, Vol.38, 824-833. doi:
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      P Choudhury, W Chen, R C Hunt; Production of platelet-derived growth factor by interleukin-1 beta and transforming growth factor-beta-stimulated retinal pigment epithelial cells leads to contraction of collagen gels.. Invest. Ophthalmol. Vis. Sci. 1997;38(5):824-833.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: In an in vitro model of the later contractile stages of proliferative vitreoretinopathy, interleukin-1 beta (IL-1 beta) and transforming growth factor-beta (TGF-beta) stimulate the contraction of collagen gels by retinal pigment epithelial (RPE) cells. This contraction occurs after a lag period and appears not to be a direct effect of the cytokines but is mediated by another factor produced in the presence of the two cytokines. The nature of this factor has been investigated. METHODS: Human RPE cells were seeded onto collagen gels in the presence of IL-1 beta and TGF-beta. After 24 hours, the conditioned medium was removed and added to new collagen gels seeded with RPE cells, and the diameter of the collagen gels was measured after various intervals. The ability of the conditioned medium to effect contraction was determined after various treatments, including size fractionation, heating, trypsin digestion, and binding to heparin-Sepharose. The involvement of platelet-derived growth factor (PDGF) as a stimulator of contraction was tested with neutralizing antibodies and by polymerase chain reaction analyses of specific mRNAs. RESULTS: IL-1 beta and TGF-beta cause RPE cells to contract after a delay of up to 24 hours, whereas conditioned medium from cytokine-treated cells results in immediate contraction in a manner similar to that of serum. The factor in the conditioned medium causing immediate contraction was found to be heat-stable, trypsin-sensitive, and resistant to extremes of pH. It has a size of between 30 and 50 kDa and binds heparin. The factor in conditioned medium from cytokine-treated cells does not act in the presence of C-kinase inhibitors or cycloheximide, suggesting that signaling is mediated by way of protein kinase C and new protein synthesis. Stimulation of contraction by conditioned medium is inhibited by anti-PDGF antibodies, and contraction is stimulated by human PDGF. CONCLUSIONS: Contraction in the presence of cytokines is mediated by the production of PDGF or a PDGF-like molecule. This factor could have implications in the pathogenesis of proliferative vitreoretinopathy.

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