Pe labeled anti pd 1 clone eh12.2h7

Blood samples were obtained from 56 patients with septic shock and 30 healthy volunteers. After lysing red cells with FACS-lysing solution (BD Biosciences; San Jose, CA, USA), the immune cells were stained with FITC-labeled anti-CD4 (Clone A161A1) or FITC-labeled anti-CD14 (Clone HCD14) and PE-labeled anti-PD-1 (Clone EH12.2H7) or PE-labeled anti-PD-L1 antibodies (Clone 29E.2A3) (Biolegend; San Diego, CA, USA). The stained cells were analyzed by using a FACS Calibur flow cytometer (Becton-Dickinson, BD Biosciences) and CELLQUEST software.

Blood samples were obtained from 63 patients with AP at days 1 and 3 after diagnosis with AP and from 32 healthy volunteers. After lysing red blood cells with fluorescence-activated cell sorting lysing solution (BD Biosciences, San Jose, CA, USA), cells were incubated in the dark at 4 °C for 30 minutes with the following fluoresceinated monoclonal antibodies and their isotype controls: fluorescein isothiocyanate (FITC)-labeled anti-CD4 (clone A161A1), FITC-labeled anti-CD14 (clone HCD14), phycoerythrin (PE)-labeled anti-PD-1 (clone EH12.2H7), PE-labeled anti-PD-L1 (clone 29E.2A3), and PE-labeled anti-human leukocyte antigen-DR (anti-HLA-DR) (clone L243; BioLegend, San Diego, CA, USA). Stained cells were analyzed using a FACSCalibur flow cytometer and CellQuest software (BD Biosciences). The percentage of PD-1-expressing CD4⁺ lymphocytes was calculated as the percentage of PD-1⁺ cells in the total CD4⁺ lymphocyte population, and the percentage of PD-L1/HLA-DR-expressing CD14⁺ monocytes was calculated as the percentage of PD-L1⁺/HLA-DR⁺ cells in the total CD14⁺ monocyte population. Interleukin (IL)-10 concentration was measured by enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN, USA) in accordance with the supplied instructions.