Cd20 brilliant violet 570

Single cell isolations from biopsies were analyzed by flow cytometry immediately after isolation. Biopsies and blood or isolated neutrophils from the bacteria stimulations were stained using the following surface antigen mouse anti-human antibodies with clone denoted in (), from Becton Dickinson, and Co. (BD) Biosciences (Franklin, NJ) unless otherwise stated: CD45 PE-CF594 (HI30),CD11b APC-Cy7 (ICRF44), CD66b PE (Biolegend, G10F5), CD49d PE/Cy5 (Biolegend, 9F10), CD20 Brilliant Violet 570 (Biolegend, 2H7), CD3 Brilliant Violet 570 (Biolegend, UCHT1), CD16 BV605 (3G8), CD15 BV650 (HI98), and CD14 BV786 (M5E2). Following surface staining, cells were permeabilized using Cytofix/Cytoperm (BD Biosciences). Intracellular active Caspase-3 was stained using a v450-conjugated rabbit anti-human antibody (BD Biosciences, C92-605). Stained samples were fixed in 1% paraformaldehyde and collected on an LSR II (BD Biosciences, La Jolla, California). Analysis was performed in FlowJo (version 9.7.6, Treestar Inc., Ashland, Oregon). Samples with less than 100 events in the neutrophil gate were not included in analyses due to an inability to ensure adequate fluorescence separation of populations and therefore accurate gating of the neutrophil cell population.

CD3 BB700 (clone SP34-2, Beckton Dickenson) or CD3 H7APC (clone SK7, BD Biosciences); Ki67 Brilliant Violet 421 (clone B56, BD Horizon); CD20 Brilliant Violet 570 (clone 2H7, Biolegend); CD4 Brilliant Violet 650 (clone SK3, BD Biosciences); PD-1 Brilliant Violet 711 (clone EH12.2H7, Biolegend); Bcl-6 PE (clone K112-91, BD Pharmingen); CD57 Alexa Fluor 594 (clone NK-1, Novus Biotechne); CXCR5 (CD185) Cy7PE (clone MU5UBEE, Thermo Fisher); CD8 Pacific Blue (clone RPA-T8, BD Pharmingen); CD19 FITC (clone J3-119, Beckman-Coulter); CD38 Brilliant Violet 786 (clone HIT2, Biolegend); IgG1 BUV395 (clone G17-1, BD Biosciences); H2AX (pS139) Alexa Fluor 647 (clone N1-431, BD Pharmingen).