CC and CXC chemokine receptors mediate migration, proliferation, and matrix metalloproteinase production by fibroblast-like synoviocytes from rheumatoid arthritis patients

Authors

  • Rosario García-Vicuña,

    1. Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain
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    • Drs. García-Vicuña and Gómez-Gaviro contributed equally to this work.

  • Maria Victoria Gómez-Gaviro,

    1. Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain
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    • Drs. García-Vicuña and Gómez-Gaviro contributed equally to this work.

  • Maria Jesús Domínguez-Luis,

    1. Hospital Universitario de Canarias, Universidad de La Laguna, Santa Cruz de Tenerife, Spain
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  • Martina K. Pec,

    1. Hospital Universitario de Canarias, Universidad de La Laguna, Laguna, Spain, and Instituto Canario de Investigación del Cáncer, Instituto de Bio-Orgánica “Antonio González,” Universidad de La Laguna, Laguna, Spain
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  • Isidoro González-Alvaro,

    1. Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain
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  • Jose María Alvaro-Gracia,

    1. Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain
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  • Federico Díaz-González

    Corresponding author
    1. Hospital Universitario de Canarias, Universidad de La Laguna, Santa Cruz de Tenerife, Spain
    • Servicio de Reumatología, Hospital Universitario de Canarias, Universidad de La Laguna, c/Ofra s/n, La Laguna 38320, Santa Cruz de Tenerife, Spain
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Abstract

Objective

To explore the potential involvement of the chemokine system in synoviocyte-mediated tissue destruction in rheumatoid arthritis (RA), we studied the expression profile of chemokine receptors and their function in the migration, proliferation, and matrix metalloproteinase (MMP) production of cultured fibroblast-like synoviocytes (FLS) from RA patients.

Methods

The presence of CC and CXC chemokine receptors on cultured FLS was studied at the messenger RNA (mRNA) level by reverse transcriptase–polymerase chain reaction and at the cell surface expression level by flow cytometry. Variations in cytosolic calcium influx induced by chemokine stimulation were assessed by flow cytometry on Fura Red–preloaded FLS. Two-compartment transwell chambers were used for FLS chemotaxis assays. Cell growth was measured by a fluorescence-based proliferation assay. Gelatinase and collagenase activities were determined by a fibril degradation assay and zymography.

Results

FLS constitutively expressed the receptors CCR2, CCR5, CXCR3, and CXCR4, both at the cell surface and mRNA levels, but failed to express CCR3 and CCR6. Significant intracytosolic calcium influx was observed on FLS challenged with monocyte chemotactic protein 1 (MCP-1), stromal cell–derived factor 1α (SDF-1α), and interferon-inducible protein 10 (IP-10). Stimulation with MCP-1, SDF-1α, IP-10, and monokine induced by interferon-γ enhanced the migration and proliferation of FLS. These chemokines, in addition to RANTES, increased in a dose- and time-dependent manner the gelatinase and collagenase activities in cell-free supernatants of cultured FLS. Interestingly, the chemokine-mediated up-regulation of MMP activities was significantly abrogated by the presence of anti–interleukin-1β, but not anti–tumor necrosis factor α, blocking antibodies.

Conclusion

These data suggest that through modulation of the migration, proliferation, and MMP production by FLS, the chemokine system may play a more direct role in the destructive phase of RA than is currently suspected, and thus emphasize the relevance of chemokines and their receptors as potential therapeutic targets in this disease.

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