Percp anti human cd16

The peripheral blood mononuclear cells (PBMC) of CRC patients were isolated by centrifugation with erythrocytes lysate and were used to analyze PD1⁺CD8⁺T cells and CD3^-CD56⁺CD16⁺NK cells by flow cytometry. The PBMC were stained for 30min on ice using the following antibodies: FITC anti-human CD8 (344704, Biolegend), PE anti-human PD1(367404, Biolegend), APC anti-human CD3 (300312, Biolegend), PE anti-human CD56 (985902, Biolegend), and PerCP anti-human CD16 (302030, Biolegend). Stained cell suspensions were analyzed using the BD flow cytometer (BD Accuri C6 Plus), and data analysis was performed using FlowJo_v10.8.1.

Activated B cells (ABC) and antibody secreting cells (ASC) were measured in blood as previously described [25 (link)]. Briefly, 10⁶ PBMCs were stained using the following: PerCP anti-human CD14, PerCP anti-human CD16 and APC-Cy7 anti-human CD20 from Biolegend; PerCP anti-human CD3, PE anti-human IgD, APC anti-human CD38, FITC anti-human CD19, and PECy7 anti-human CD71 from eBiosciences; the fixable viability stain 510 (e-Biosciences) was used to discriminate dead cells. Samples were stained for 30 min on ice with the viability dye, washed and stain for 20 min at 4°C with the required antibodies. Flow cytometric analysis was performed using a LSR-FORTESSA (Becton Dickinson). Data were analyzed using FlowJo software 10 (Tree Star, USA).