The patients in the experimental group (n = 13) participated in the observational study on peripheral blood mononuclear cell (PBMC) analysis as a predictor of PD‐1 blockade. Written informed consent was obtained from all 13 patients included for the analysis of PBMC. Blood samples were collected at two time points before the initiation of PD‐1 blockade therapy and EGFR‐TKI rechallenge (Figure 2(b) ). PBMC analysis was performed according to our previous study.13 Cells were stained with monoclonal antibodies (mAbs) using the BD Accuri C6 plus flow cytometer (Becton Dickinson and Company) and the BD LSR Fortessa (Becton, Dickinson and Company). The following mAbs were used in the study: fluorescein isothiocyanate (FITC)‐conjugated anti‐CD3 (HIT3a) and anti‐CD4 (RPA‐T4), phycoerythrin (PE)‐conjugated anti‐CD8 (RPAT8) and anti‐CD25 (M‐A251), PE‐Cy7‐conjugated anti‐CD25 (MA251), PE‐Cy5‐conjugated anti‐CD62L (Dreg 56; all from BD Pharmingen), and FITC‐conjugated anti‐CD62L (Dreg 56; eBioscience).
Dreg 56
Manufactured by Thermo Fisher Scientific
The Dreg 56 is a laboratory equipment designed for the efficient drying and dehumidification of samples. It utilizes advanced technology to precisely control temperature and airflow, ensuring consistent and reliable drying results.
Automatically generated - may contain errors
Lab products found in correlation
Accuri c6 plus, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Red blood lysing solution, by Miltenyi Biotec (1 mentions)
Whole blood lysing reagent kit, by Beckman Coulter (1 mentions)
Ebiocb16, by Thermo Fisher Scientific (1 mentions)
Icrf44, by Thermo Fisher Scientific (1 mentions)
Macsquant analyzer 10, by Miltenyi Biotec (1 mentions)
2 protocols using dreg 56
1
Peripheral Blood Mononuclear Cell Analysis for PD-1 Blockade Response
2
Immunophenotyping of Neutrophils in IgAV Patients
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Venous blood was drawn from IgAV patients and HBDs and collected into heparin-containing tubes. Whole blood immunophenotyping was performed using 7-Color Immunphenotyping kit (Miltenyi Biotec). Briefly, 100 μl of whole blood was incubated with 10 μl immunophenotyping reagent for 10 min in the dark, at 4 °C. After incubation, erythrocytes were lysed using Red Blood Lysing Solution (Miltenyi Biotec). Neutrophil phenotyping was performed in 50 μl of whole blood, incubated for 30 min at 4 °C in the dark with mouse anti-human antibodies to CD16 (conjugated to PE, clone eBioCB16, eBioscience), CD62L (conjugated to PE-Cyanine 5, clone DREG56, eBioscience), and CD11b (conjugated to APC, clone ICRF44, eBioscience). After incubation, samples were lysed, using Whole Blood Lysing Reagent Kit (Beckman Coulter). All samples were analyzed using flow cytometer MACSQuant Analyzer 10 (Miltenyi Biotec). Analysis of flow cytometry data was performed using MACSQquantify (Analysis Software version 2.8) and FlowLogic (Flow Cytometry Analysis Package, version 700.0a).
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