Expression of GM-CSF receptor and "in vitro" effects of GM-CSF on human fibroblasts

Life Sci. 1998;63(5):327-36. doi: 10.1016/s0024-3205(98)00281-1.

Abstract

In the present study the effects of Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) on fibroblast growth and activity have been studied. In this regard the AA have evaluated in primary cultures of human gengival normal fibroblasts (PG1 cells): a)-the expression of GM-CSF receptor (GM-CSFR) (alfa unit) on the cell surface; b)-the in vitro effects of different doses of GM-CSF on the GM-CSFR expression and on the proliferation and activity of fibroblasts. PG1 cells have been stimulated in vitro with different concentrations of GM-CSF (10, 50, 80, 100 and 150 ng/ml) using promonocytic cell line U937 as positive control for GM-CSFR expression. GM-CSFR was investigated by flow cytometry, with mouse monoclonal antibody (mAb) against the alfa chain of the human GM-CSFR and fluorescein-conjugated goat antimouse immunoglobulin G (IgG). At high GM-CSF concentration (80 ng/ml) the AA observed: 1)-A marked increase of GM-CSFR expression evaluated as fluorescence intensity (about three fold in respect to the controls); 2)-Maximal increase of PG1 cells proliferation. Moreover immunofluorescence on fibroblasts obtained from culture plates showed increased actin stress fibers and fibronectin production with low stimulation by GM-CSF, while higher concentration of this cytokine determined increased proliferation of cells, but a decreased formation of actine fibers and vinculin plaques. These results demonstrate: 1)-The presence of GM-CSFR on the surface of fibroblasts; 2)-The proliferation and the synthesis activity of these cells (in vitro) are modulated by different concentration of GM-CSF. We hypothesize that GM-CSF until 80 ng/ml can upregulate the expression of the receptor. Therefore, on the basis of previous findings of high serum levels of GM-CSF in course of scleroderma, a disease characterized by fibroblast hyperactivity, a possible role of this cytokine in the pathogenic process of this disease can be hypothesized.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Cell Division / drug effects
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Fibronectins / metabolism
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gingiva / cytology
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
  • Humans
  • Mice
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / metabolism*
  • Recombinant Proteins / pharmacology
  • Tumor Cells, Cultured / metabolism
  • Up-Regulation
  • Vinculin / metabolism

Substances

  • Antibodies, Monoclonal
  • Fibronectins
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor
  • Recombinant Proteins
  • Vinculin
  • Granulocyte-Macrophage Colony-Stimulating Factor