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CMV was grown and harvested from MRC-5 fibroblast cultures (Biowhittaker, Walkersville, MD) and concentrated by ultra-centrifugation (28000 g for 60 minutes at 4°C).
THP-1 (generously provided by Dr P Guyre, Dartmouth Medical School), Jurkat (ATCC), and 293 cell lines were cultured in RPMI complete medium (CM), consisting of RPMI 1640 with 10% heat-inactivated fetal bovine serum, 0.1 ml of sodium pyruvate, 2.0 mmol/l of L-glutamine, 25 mmol/l of 1 M HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), 0.1 mmol/l of nonessential amino acids, 0.25 μg/ml of amphotericin, 50 μg/ml gentamicin (all from BioWhittaker, Walkersville, MD, USA) and 5 × 10 -5M 2-mercaptoethanol (Sigma, St Louis, MO, USA).
Purified bovine CL-derived microvascular endothelial cell populations were generated commercially by Cambrex Biosciences (BioWhittaker, Walkersville, MD) as described in detail [ 35 ] . In the present study, CLENDO cells from frozen aliquots (passage 3) were seeded (5,000 cells/cm 2) in 6- or 24-well plates or 10 cm culture dishes.
CV-1 cells were maintained in 10% fetal bovine serum (Bio-Whittaker, Walkersville, MD), 100 u/ml penicillin, 100 μg/ml streptomycin in DMEM.
The FLS cultures were established from synovial tissues obtained during joint replacement surgery in RA patients [ 6 ] . The FLS were cultured in DMEM plus 10% heat-inactivated human AB serum (BioWhittaker, Walkersville, MD, USA), 10% fetal bovine serum, penicillin, streptomycin, and L-glutamine.