Anti cd19 clone sj25c1

Cells were first washed with PBS and stained with LIVE/DEAD fixable near-IR (dead cell stain kit, Invitrogen, Carlsbad, CA, USA) for 30 min at room temperature in the dark. Then, cells were washed with PBS supplemented with 1% bovine serum albumin. Extracellular staining was performed by incubating the cells for 30 min at room temperature in the dark with the following antibodies: Anti-CD19 (clone SJ25-C1, BD Bioscience, Franklin Lakes, NJ, USA), anti-CD20 (clone L27, BD Bioscience, Franklin Lakes, NJ, USA), anti-CD27 (clone L128, BD Bioscience, Franklin Lakes, NJ, USA; clone O323, eBioscience, San Diego, CA, USA), anti-CD38 (clone HB7, BD Bioscience, Franklin Lakes, NJ, USA) and anti-IgG (clone G18-145, BD Bioscience, Franklin Lakes, NJ, USA or clone MH16-1, Sanquin Reagents, Amsterdam, the Netherlands). Samples were measured on a FACSCanto, FACSLSRII or FACSLSR Fortessa (BD Bioscience, Franklin Lakes, NJ, USA) and analyzed using FlowJo software version 10 (Treestar, Ashland, OR, USA).

After thawing, PBMC were washed and immediately stained with the following panel: anti-CD3 (clone UCHT1; #IM2467, Beckman Coulter, Brea, CA, USA), anti-CD4 (clone SFCI12T4D11; #737660, Beckman Coulter), anti-CD8 (clone RPA-T8; #558207, BD Biosciences, Franklin Lakes, NJ, USA), anti-CD14 (cloneMφP9; #641394, BD Biosciences), anti-CD16 (clone3G8; #555406, BD Biosciences), anti-CD56 (clone N901; #A07788, Beckman Coulter), anti-CD11c (clone B-ly6; #561352, BD Biosciences), anti-CD19 (clone SJ25C1; #563036, BD Biosciences), anti-CD123 (clone 6H6; #45-1239-42, eBioscience, Affymetrix, Santa Clara, CA, USA), anti-HLA-DR (clone Immu-357; #IM3636, Beckman Coulter), and Zombie UV (#77474, Biolegend, San Diego, CA, USA). The samples were acquired on an BD Fortessa instrument equipped with the FACS Diva software. The analysis was performed with the FlowJo (FLOWJO, LLC, Ashland, OR, USA).