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Characterization of human blood dendritic cell subsets.
- Source :
-
Blood [Blood] 2002 Dec 15; Vol. 100 (13), pp. 4512-20. Date of Electronic Publication: 2002 Aug 15. - Publication Year :
- 2002
-
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.
- Subjects :
- Adult
Antibodies, Monoclonal immunology
Antigen Presentation
Antigens, CD analysis
Antigens, Surface analysis
Blood Cells classification
Cell Separation
Cell Survival
Cells, Cultured cytology
Dendritic Cells chemistry
Dendritic Cells immunology
Flow Cytometry
Gene Expression Profiling
Humans
Immunophenotyping
Lectins metabolism
Lymphocyte Culture Test, Mixed
Monocytes cytology
Reference Standards
Dendritic Cells classification
Subjects
Details
- Language :
- English
- ISSN :
- 0006-4971
- Volume :
- 100
- Issue :
- 13
- Database :
- MEDLINE
- Journal :
- Blood
- Publication Type :
- Academic Journal
- Accession number :
- 12393628
- Full Text :
- https://doi.org/10.1182/blood-2001-11-0097