Standardized single-platform assay for human monocyte subpopulations: Lower CD14+CD16++ monocytes in females.
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.
Heimbeck
Irene
I
KKG Inflammatory Lung Diseases, Helmholtz-Zentrum Muenchen and Asklepios Hospital, Muenchen Gauting and Comprehensive Pneumology Center, Ludwig-Maximilians University Munich, Helmholtz Zentrum München, Germany.
Hofer
Thomas P J
TP
Eder
Christiane
C
Wright
Adam K
AK
Frankenberger
Marion
M
Marei
Ayman
A
Boghdadi
Ghada
G
Scherberich
Jürgen
J
Ziegler-Heitbrock
Loems
L
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
D005938:Glucocorticoids; D006684:HLA-DR Antigens; D018950:Lipopolysaccharide Receptors; D017452:Receptors, IgG