Cd141 allophycocyanin

To determine the size of peripheral blood leukocyte and BM nucleated cell populations, cells were labeled with the following antibodies against human cell surface markers: anti-human cluster of differentiation (CD)235a-phycoerythrin (PE), CD14-PE-cyanin 5, CD16-PE, CD20-PE, CD203c-PE, CD3-PE-cyanin 5, CD4-PE-cyanin 5, CD4-PE-cyanin 7, CD8-PE-cyanin 5, and CD56-PE (all from BioLegend, San Diego, CA, USA); CD71-PE-cyanin 5, CD3-PE, and CD11c-PE (all from BD Biosciences); and CD141-allophycocyanin (Miltenyi Biotec, Bergisch Gladbach, Germany). The total lymphocyte population was determined from forward and side scatter, and CD4+ and CD8+ T cells, natural killer (NK) cells, and B cells were separated into CD3+/CD4+, CD3+/CD8+, CD56+/CD3−, and CD20+ populations, respectively. The monocyte population was determined from forward and side scatter and by CD14 expression. The granulocyte population was gated by forward and side scatter, and neutrophils and eosinophils were separated according to CD16 expression (CD16+ and CD16−, respectively). Basophils were isolated as the CD203c+ population. Plasmacytoid DCs, CD1c+ myeloid DCs, and CD141^high myeloid DCs were isolated from peripheral blood mononuclear cells on a FACSAriaII cell sorter as previously described [39 (link)]. Erythroid cells present in the BM cell population were gated according to CD235 and CD71 (TFR1) expression.