Open Access Highly Accessed Research article

c-Fms-mediated differentiation and priming of monocyte lineage cells play a central role in autoimmune arthritis

Ricardo T Paniagua12, Anna Chang12, Melissa M Mariano12, Emily A Stein12, Qian Wang12, Tamsin M Lindstrom12, Orr Sharpe12, Claire Roscow12, Peggy P Ho3, David M Lee4 and William H Robinson12*

Author Affiliations

1 Department of Medicine, Division of Immunology and Rheumatology, Stanford University School of Medicine, CCSR 4135, 269 Campus Drive, Stanford, CA 94305, USA

2 GRECC, Palo Alto VA Health Care System, 3801 Miranda Avenue, Palo Alto, CA 94304, USA

3 Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Beckman B-002, 279 Campus Drive, Stanford, CA 94305, USA

4 Department of Medicine, Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, 1 Jimmy Fund Way, Smith 552B, Boston, MA 02115, USA

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Arthritis Research & Therapy 2010, 12:R32  doi:10.1186/ar2940

Published: 24 February 2010

Additional files

Additional file 1:

GW2580 potently inhibits c-Fms kinase and does not cross-react with other imatinib-targeted kinases at clinically relevant concentrations. (a, b) Cell-free kinase activity assay with time-resolved fluorescent readout for determination of the IC50 of imatinib and GW2580 for the kinases (a) Abl and (b) c-Kit. (c) Cell-based assay for determination of the IC50 of imatinib and GW2580 for c-Fms. Human peripheral blood mononuclear cells were treated with M-CSF in the presence of 0-10 μM GW2580 or imatinib for 48 hours. Macrophages were counted and values expressed relative to M-CSF treatment alone. (d) Cell-based assay for determination of the IC50 of imatinib and GW2580 for PDGFR. Fibroblast-like synoviocytes from a human RA patient were incubated with PDGF-bb in the presence of 0-30 μM GW2580 or imatinib. After 48 hours, FLS cultures were pulsed with [3H] thymidine for 18 hours. Values are expressed relative to PDGF-bb treatment. Data shown in a-d are representative of at least 2 independent experiments.

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Additional file 2:

GW2580 does not modulate T-cell function in vivo. Splenocytes were harvested from DBA/1 mice with CIA and treated with GW2580, imatinib, or vehicle, and stimulated with 20 μg/ml heat-denatured, whole CII. [3H]thymidine incorporation was used to measure proliferation of CII-specific T cells. IFNγ, TNFα and IL-10 were measured in culture supernatants by ELISA. Values are the mean ± SEM. *P < 0.05 compared with stimulated cells from vehicle-treated CIA mice. Results are representative of 2 independent experiments.

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