Mononuclear cells were isolated from PBL by ficoll density gradient centrifugation as previously described35 (link). NP samples were minced using a scalpel, placed in a 50 ml tube with 8 ml DMEM + Liberase TL 125 ug/mL (Roche) + DNAse I 100 ug/ml (SIGMA) + antibiotic/antimycotic (GIBCO), incubated at 37 °C for 45 min and vortexed at low speed each 10 min. After the incubation the suspension was filtered through a sterile 70 um mesh.
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