title: Standardized single-platform assay for human monocyte subpopulations: Lower CD14+CD16++ monocytes in females.
abstract: We present a novel single-platform assay for determination of the absolute number of human blood monocyte subpopulations, i.e., the CD14(++)CD16(-) and the CD14(+)CD16(++) monocytes. A four-color combination of antibodies to CD14, CD16, CD45, and HLA-DR reduces the spill-over of natural killer cells and of granulocytes into the CD14(+)CD16(++) monocyte gate. For these CD14(+)CD16(++) monocytes, the intra-assay coefficient of variation (CV) was 4.1% and the inter-assay CV was 8.5%. Looking at a cohort of 40 donors aged 18-60 years, we found no age dependence. There was however an effect of gender in that females had lower CD14(+)CD16(++) monocytes (45.4 +/- 13.5 cells/microl) compared with males (59.1 +/- 20.3 cells/microl) (P < 0.02). Using this novel approach, we can confirm that exercise will lead to more than three-fold increase of the CD14(+)CD16(++) monocytes. Also, we show that therapy with low doses of glucocorticoids will deplete these cells. This robust single-platform assay may be a useful tool for monitoring the absolute number of monocyte subpopulations in health and disease.
authors.affiliation: KKG Inflammatory Lung Diseases, Helmholtz-Zentrum Muenchen and Asklepios Hospital, Muenchen Gauting and Comprehensive Pneumology Center, Ludwig-Maximilians University Munich, Helmholtz Zentrum München, Germany.
authors.forename: Irene
authors.initials: I
authors.lastname: Heimbeck
authors.forename: Thomas P J
authors.initials: TP
authors.lastname: Hofer
authors.forename: Christiane
authors.initials: C
authors.lastname: Eder
authors.forename: Adam K
authors.initials: AK
authors.lastname: Wright
authors.forename: Marion
authors.initials: M
authors.lastname: Frankenberger
authors.forename: Ayman
authors.initials: A
authors.lastname: Marei
authors.forename: Ghada
authors.initials: G
authors.lastname: Boghdadi
authors.forename: Jürgen
authors.initials: J
authors.lastname: Scherberich
authors.forename: Loems
authors.initials: L
authors.lastname: Ziegler-Heitbrock
mesh_terms: D000293:Adolescent; D000328:Adult; D015444:Exercise; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005938:Glucocorticoids / Q000627:therapeutic use; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D007958:Leukocyte Count / Q000379:methods*; D018950:Lipopolysaccharide Receptors / Q000032:analysis / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D017452:Receptors, IgG / Q000032:analysis / Q000276:immunology*; D013194:Staining and Labeling; D055815:Young Adult
chemical_list: D005938:Glucocorticoids; D006684:HLA-DR Antigens; D018950:Lipopolysaccharide Receptors; D017452:Receptors, IgG