Cd62l apc clone dreg 56

Multi-parametric flow cytometry was used for immunophenotyping. PBMC (1 × 10⁶) were stained with a T-cell panel, an innate lymphoid panel or with an isotype control panel. Before antibody labeling, cells were incubated with purified human IgG (Sigma) to reduce non-specific binding. Cells were stained with mAbs against CD3-PE Cy7 (clone SK7), CD4-BD Horizon v500 (clone RPA-T4), CD8-BD Horizon v450 (clone RPA-T8), CD45R0-PECF594 (clone UCHL1), CD62L-APC (clone DREG-56), CD25-APC Cy7 (clone M-A251), CD127-FITC (clone HIL-7R-M21), CD279-PE (clone EH12.2H7) (Biolegend), HLA-DR-PerCPCy5.5 (clone LN3) (eBioscience), CD16-APC H7 (clone 3G8), CD56-APC (clone NCAM 16.2), iNKT-PE (6B11), Vδ2-FITC (clone B6), IgG1k-APC Cy7 (clone MOPC-21), IgG1k-APC (clone MOPC-21) (Biolegend), IgG1k-FITC (clone MOPC-21) (Biolegend), IgG1k-PE (clone MOPC-21) (Biolegend), and IgG2b-PerCPCy5.5 (clone N/S) (eBioscience). Antibodies were from BD Biosciences unless stated otherwise.

Analyses of the expression of cell surface activation markers were performed using differential FACS staining. PMN were labeled with CD66b-FITC (clone 80H3, AbD Serotec, Puchheim, Germany), CD62L-APC (clone DREG-56, BioLegend, Fell, Germany), CD63-V450 (clone H5C6, BD Biosciences, Heidelberg, Germany), and CD69-PE (clone L78, BD Biosciences, Heidelberg, Germany) at 4°C for 30 min, washed with CellWash (BD Biosciences, Heidelberg, Germany), and centrifuged at 300 g for 5 min. Cells were analyzed using a FACS Canto II flow cytometer (BD Biosciences, Heidelberg, Germany). The software FlowJo 7.6.4 (BD Biosciences, Heidelberg, Germany) was used for analysis.