title: Characterization of human blood dendritic cell subsets.
abstract: Dendritic cells (DCs) are key antigen-presenting cells for stimulating immune responses and they are now being investigated in clinical settings. Although defined as lineage-negative (Lin(-)) HLA-DR(+) cells, significant heterogeneity in these preparations is apparent, particularly in regard to the inclusion or exclusion of CD14(+), CD16(+), and CD2(+) cells. This study used flow cytometry and a panel of monoclonal antibodies (mAbs), including reagents from the 7th Leukocyte Differentiation Antigen Workshop, to define the cellular composition of 2 standardized peripheral blood mononuclear cell (PBMCs)-derived Lin(-) HLA-DR(+) preparations. Lin(-) cells were prepared from PBMCs by depletion with CD3, CD14, CD19, CD11b, and either CD16 or CD56 mAbs. Analysis of the CD16-replete preparations divided the Lin(-) HLA-DR(+) population into 5 nonoverlapping subsets (mean +/- 1 SD): CD123 (mean = 18.3% +/- 9.7%), CD1b/c (18.6% +/- 7.6%), CD16 (49.6% +/- 8.5%), BDCA-3 (2.7% +/- 1.4%), and CD34 (5.0% +/- 2.4%). The 5 subsets had distinct phenotypes when compared with each other, monocytes, and monocyte-derived DCs (MoDCs). The CD85 family, C-type lectins, costimulatory molecules, and differentiation/activation molecules were also expressed differentially on the 5 Lin(-) HLA-DR(+) subsets, monocytes, and MoDCs. The poor viability of CD123(+) DCs in vitro was confirmed, but the CD16(+) CD11c(+) DC subset also survived poorly. Finally, the individual subsets used as stimulators in allogeneic mixed leukocyte reactions were ranked by their allostimulatory capacity as CD1b/c > CD16 > BDCA-3 > CD123 > CD34. These data provide an opportunity to standardize the DC populations used for future molecular, functional and possibly even therapeutic studies.
authors.affiliation: Dendritic Cell Laboratory, Mater Medical Research Institute, Mater Misericordiae Hospitals, South Brisbane, Australia.
authors.forename: Kelli P A
authors.initials: KP
authors.lastname: MacDonald
authors.forename: David J
authors.initials: DJ
authors.lastname: Munster
authors.forename: Georgina J
authors.initials: GJ
authors.lastname: Clark
authors.forename: Andrzej
authors.initials: A
authors.lastname: Dzionek
authors.forename: Juergen
authors.initials: J
authors.lastname: Schmitz
authors.forename: Derek N J
authors.initials: DN
authors.lastname: Hart
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology; D017951:Antigen Presentation; D015703:Antigens, CD / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D001773:Blood Cells / Q000145:classification; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured / Q000166:cytology; D003713:Dendritic Cells / Q000737:chemistry / Q000145:classification* / Q000276:immunology; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D016130:Immunophenotyping; D037102:Lectins / Q000378:metabolism; D007959:Lymphocyte Culture Test, Mixed; D009000:Monocytes / Q000166:cytology; D012015:Reference Standards
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000954:Antigens, Surface; D037102:Lectins