TNF-α treatment results in increased number of GWB in THP-1 and human PBMCs. (a) THP-1 cells were treated with 10 ng/ml TNF-α, IFN-α, IFN-β, IFN-γ, IL-12p70, M-CSF, IL-4, IL-10, or 25 ng/ml MCP-1 for 4 hours. IIF was performed using a human anti-GWB serum to detect GWB, and the number of GWB were counted using CellProfiler image analysis software. Average number of GWB per cell and SEM is shown. *P < 0.0001, as determined by one-way analysis of variance. (b) Human PBMCs were obtained from a healthy donor and isolated using Ficoll density-gradient centrifugation. The cells were then cultured for 4 hours in the presence of 1 ng/ml TNF-α. GWB were detected by IIF using rabbit anti-Rck/p54 antibodies. Average number of GWB and SEM is shown. *P < 0.0001, as determined by Mann-Whitney test. (c) IIF image of THP-1 and PBMCs treated with 10 ng/ml or 1 ng/ml TNF-α for 4 hours, respectively. GWB are shown in green, and nuclei are counterstained with 4',6-diamidino-2-phenylindole (DAPI; blue). Bar = 10 μm. GWP, GW or P bodies; IL, interleukin; IFN, interferon; IIF, indirect immunofluorescence; MCP, macrophage chemoattractant protein; M-CSF, macrophage colony-stimulating factor; PBMC, peripheral blood mononuclear cell; SEM, standard error of the mean; TNF, tumor necrosis factor.
Pauley et al. Arthritis Research & Therapy 2008 10:R101 doi:10.1186/ar2493