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. 2001 Mar;60(3):187–193. doi: 10.1136/ard.60.3.187

Demonstration of mast cell chemotactic activity in synovial fluid from rheumatoid patients

N Olsson 1, A Ulfgren 1, G Nilsson 1
PMCID: PMC1753565  PMID: 11171676

Abstract

OBJECTIVES—The significance of the mast cell in the pathogenesis of rheumatic diseases has become more evident. Although mast cell hyperplasia is a feature of rheumatoid arthritis, the nature of mast cell chemoattractants involved in the recruitment of mast cells in joint diseases has not been studied in any detail. In this study the presence of mast cell chemotactic activity in synovial fluids was examined.
METHODS—Synovial fluids from seven rheumatoid patients were tested in a modified Boyden chamber, where a human mast cell line was used as responder. The presence of stem cell factor (SCF) and transforming growth factor β (TGFβ) was measured by enzyme linked immunosorbent assay (ELISA).
RESULTS—Six of the seven synovial fluids tested exhibited mast cell chemotactic activity. Two characterised human mast cell chemotaxins, SCF and TGFβ, were highly expressed in the synovium. Soluble SCF could be detected in all fluids analysed. Blocking antibodies against SCF or TGFβ almost completely blocked the activity in one fluid, partially blocked the activity in three, and did not affect the activity in two. Treatment of the responder cells with pertussis toxin reduced the migratory response against seven fluids, indicating the presence of chemoattractants mediating their effect through Gi coupled receptors.
CONCLUSION—These data demonstrate the presence of multiple factors in synovial fluid acting as mast cell chemoattractants, two of which are SCF and TGFβ that contribute to the effect. These findings may be of importance for developing new strategies to inhibit mast cell accumulation in rheumatic diseases.



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Figure 1  .

Figure 1  

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Immunolocalisation of mast cells and stem cell factor (SCF) in synovial tissue. Tissues were stained for tryptase (A and B) or SCF (C and D). The figures are from the same patient (RD1), but different sections. The stainings are presented at two different magnifications from the same section. Magnification for A and C is ×80, and for B and D ×200.

Figure 2  .

Figure 2  

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Migratory response of human cultured mast cells to synovial fluid. The fluid was diluted in medium and tested for mast cell chemotactic activity. Dilutions: undiluted, 5×, 25×, 125×, 250×, and 500×. Migration was measured after 150 minutes. The results depicted are from three experiments performed in triplicate. Results are given as means (SEM).

Figure 3  .

Figure 3  

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Inhibition of activity of synovial fluids in mast cell chemotaxis assay by specific antibodies against (A) SCF (10 µg/ml) and (B) TGFβ (10 µg/ml). Open bars = control, filled bars = with antibody. The results are from three experiments performed in triplicate. Results are given as means (SEM). A significant effect was obtained as indicated: *p<0.05, ***p<0.001.

Figure 4  .

Figure 4  

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Analysis of signal transduction pathways which play a part in mediating mast cell chemoattractant activity in synovial fluids. Mast cells were treated with (A) pertussis toxin, or (B) genistein for 90 minutes before chemotaxis assay. The fluids were used at dilutions giving optimal migration. Open bars = control; filled bars = with inhibitor. The results are from three experiments performed in triplicate. Results are given as means (SEM). A significant effect was obtained as indicated: **p<0.01, ***p<0.001.

Selected References

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