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A flow cytometric immune function assay for human peripheral blood dendritic cells
Author(s) -
Willmann Kerstin,
Dunne John F.
Publication year - 2000
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.67.4.536
Subject(s) - biology , cd80 , cd86 , cd40 , cd14 , dendritic cell , immunology , il 2 receptor , cd11c , tumor necrosis factor alpha , flow cytometry , immune system , microbiology and biotechnology , t cell , cytotoxic t cell , biochemistry , gene , in vitro , phenotype
CD11c + and CD11c − (CD123 + ) dendritic cells (DCs) have been described in blood. Both cell types express high levels of HLA‐DR and lack the lineage markers CD3, CD14, CD19, CD20, CD16, and CD56. These immunophenotypic properties were used along with analysis of activation‐related surface antigens and intracellular staining of cytokines to characterize functional responses of these DC subsets to stimuli in whole human blood (WB). Samples from healthy donors were activated with lipopolysaccharide (LPS) or phorbol 12‐myristate 13‐acetate plus ionomycin (PMA+I). The only distinct response in CD11c − DCs was the expression of CD25 upon PMA+I activation. CD11c + cells responded to LPS stimulation by producing high levels of interleukin‐1β (IL‐1β) and tumor necrosis factor α (TNF‐α), and lower levels of IL‐6, IL‐1Ra, and IL‐8 and an increased expression of accessory molecules (CD25, CD40, CD80, CD86, HLA‐DR, and HLA‐DQ). PMA+I activation of CD11c + cells resulted in high levels of IL‐1p and lower levels of IL‐8, IL‐1Ra, and TNF‐ct and up‐regulation of CD80, CD86, HLA‐DR, and HLA‐DQ. Our data support prior observations of functional differences between peripheral blood DC subsets and demonstrate the power of multiparameter flow cytometry to characterize the pleiotropic responses of these cells to various stimuli. J. Leukoc. Biol. 67: 536–544; 2000.