Cd19 pecy5 hib19

The antibodies used were CD19-PECy5 (HIB19, Biolegend), CD56-APC-Cy7 (clone HCD56, Biolegend), CD21-FITC (Clone B-ly4, BD Pharmingen), CD23-PE (Clone EBV CS-5, Biolegend), CD20-BV510 (Clone 2H7, BD Horizon), CD16-PECy7 (Clone 3G8, BD) and CD14-APC (Clone 61D3, Invitrogen). Cell samples were washed once with PBS. Antibodies for cell surface markers and DAPI (final concentration of 2 μg/ml, Sigma-Aldrich) were employed diluted in PBS with 10μg/ml human aggregated IgG, 2% FBS and 2mM EDTA. Cells were incubated with Abs for 15min at 4°C, washed twice with PBS, resuspended in PBS with 2% FBS and 2mM EDTA and analyzed in an LSR II flow cytometer (BD Biosciences). Data were processed with FlowJo X software (TreeStar).

The following monoclonal antibodies were used for FACS: CCR7 APC-Cy7 (G043H7, Biolegend, 353211), CD69 PE (FN50, BD, 555531), CD3 APC R700 (UCHT1, BD, 565119), PD-1 BV421 (EH12.2H7, Biolegend, 329920), HLA-DR BV605 (G46-6, BD, 562845), CXCR5 BB515 (RF8B2, BD, 564624), CD14 PE-Cy5 (61D3, abcam, ab25395), CD16 PE-Cy5 (3G8, Biolegend, 3012010), CD19 PE-Cy5 (HIB19, Biolegend, 302210), CD45RA PE CF594 (HI100, BD, 562298), CD8 PE-Cy5.5 (RPA-T8, eBioscience, 35-0088-42), and CD4 PE Cy7 (RPA-T4, Biolegend, 300512).