Anti hla abc pe

The purity of generated DCs was assessed as CD14⁻CD11C⁺ by staining 1 × 10⁶ cells with anti-CD11C-APC and anti-CD14-FITC (BD Biosciences, San Diego, CA, USA).
Immature moDCs were resuspended to 1 × 10⁶ cells/ml and loaded with peptides±HB_100-108 (40 μg/ml for each peptide) or 40 μg/ml of free HB_100-108 at 37°C, 5% CO₂. After one hour, pulsed DCs were washed and cultured overnight. The following monoclonal antibodies (mAbs) were used to characterize the maturation and activation status of DCs: anti-HLA-ABC-PE, anti-HLA-DR-PE/Cy7, anti-CD80-PE, anti-CD86-APC, anti-CD83-APC, anti-CD40-Alexa Fluor 700, and anti-CCR7-PE/Cy7 (BioLegend, San Diego, CA, USA). Isotype-matched fluorescent antibodies were used as negative controls. Cells were incubated with antibodies at 4°C for 30 minutes. After washing, samples were detected by a FACSCalibur analyzer (BD Biosciences, San Jose, CA, USA). The data were analyzed by FlowJo software (TreeStar).

Muse‐AT cells were incubated at 4°C for 30 minutes in the presence of the following fluorochrome‐conjugated antibodies: anti‐SSEA3‐Alexa Fluor 647 (BD Biosciences), anti‐CD105‐APC, anti‐CD90‐FITC, anti‐CD73‐PE, anti‐CD29‐PE, anti‐CD34‐PE, anti‐CD73‐PE, anti‐CD45‐FITC, anti‐HLA‐ABC‐PE, anti‐HLA‐DR‐PE, and anti‐CD44‐PE (all from BioLegend). As a negative control, isotype‐matched irrelevant monoclonal antibodies were used. Analysis (10⁴ events per run) was performed using the FACSCantoII flow cytometer and DIVA6 software (BD Biosciences).