The following extracellular, anti-human monoclonal antibodies were used for lymph node immunophenotyping: anti-CD3 PE-Cy 7, anti-CD8+PE-Cy 5, anti-CD4 FITC, anti-CD62L PE-Cy 5, anti-CD69 FITC, anti-CD152 (CTLA-4) PE, anti-CD11c PE-Cy 5, anti-CD86 PE-Cy 7, anti-HLA-DR PE, anti-CD3 FITC, anti-CD14 FITC, anti-CD16 FITC, anti-CD19 FITC, anti-CD123 PE-Cy 7, anti-CD141 PE, anti-CD68 PE-Cy 5, anti-CD20 PE, anti-CD56 PE, anti-CD279 (PD-1) APC, anti-CD11b PE, anti-CD64 FITC (BD Pharmingen, San Jose, CA).
The human monoclonal antibodies, anti-CD4 PE-Cy 5 and anti-CD25 PE, were purchased from Biolegend (San Diego, CA) and used in conjunction with intracellular staining with anti-FoxP3 FITC for the quantification of regulatory T cells. Cell death was determined by propidium iodine staining (eBioscience, San Diego, CA USA). Four-color flow cytometry was performed on a Guava 8HT flow cytometer (EMD Millipore, Billerica, MA USA) capturing 25,000 events for all samples. Results were analyzed using Guava Soft Incyte (EMD Millipore, Billerica, MA USA).
The human monoclonal antibodies, anti-CD4 PE-Cy 5 and anti-CD25 PE, were purchased from Biolegend (San Diego, CA) and used in conjunction with intracellular staining with anti-FoxP3 FITC for the quantification of regulatory T cells. Cell death was determined by propidium iodine staining (eBioscience, San Diego, CA USA). Four-color flow cytometry was performed on a Guava 8HT flow cytometer (EMD Millipore, Billerica, MA USA) capturing 25,000 events for all samples. Results were analyzed using Guava Soft Incyte (EMD Millipore, Billerica, MA USA).