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 11 CD16 2 and the CD14 1 CD16 11 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 1 CD16 11 monocyte gate. For these CD14 1 CD16 11 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 1 CD16 11 monocytes (45.4 AE 13.5 cells/ll) compared with males (59.1 AE 20.3 cells/ll) (P < 0.02). Using this novel approach, we can confirm that exercise will lead to more than three-fold increase of the CD14 1 CD16 11 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. ' International Society for Advancement of CytometryKey terms monocyte heterogeneity; gender; exercise; glucocorticoids MYELOMONOCYTIC stem cells in bone marrow give rise to monocytes and these are released into blood where they circulate for a few days and then migrate into tissue where they develop into different types of macrophages (1). Monocytes were initially defined based on morphology and cytochemistry. With the advent of monoclonal antibodies and flow cytometry, human monocytes were characterized with antibodies like CD14. This technology also allowed for the definition of two monocyte subsets, i.e., the classical CD14 11 CD162 and the CD14 1 CD16 11 nonclassical monocytes (2) with the latter cells characterized by higher MHC class II expression and by higher production of the cytokine tumor necrosis factor (3). With the recognition of the potential role of the nonclassical CD14

The Contraceptive Depot Medroxyprogesterone Acetate Impairs Mycobacterial Control and Inhibits Cytokine Secretion in Mice Infected with Mycobacterium tuberculosis

“…Contraceptive doses of DMPA also influenced the recruitment and accumulation of inflammatory cells to and at the site of disease. Glucocorticoid therapies are known to deplete monocyte populations in humans (34) and limit the recruitment of eosinophils and T lymphocytes but not neutrophils in the airways of mice (27). DMPA treatment furthermore resulted in an increased bacterial burden in C57BL/6 mice infected with M. tuberculosis compared to that of untreated mice.…”

Pivotal Role for CD16+ Monocytes in Immune Surveillance of the Central Nervous System

“…1 (peripheral blood) and Supplemental Fig. 2 (CSF), was adapted from Heimbeck et al (19). Instead of an oblique line to the right of non-classical events, a straight vertical line was placed to the lefthand end of the classical monocytes.…”

Modulatory Effects of CD14+CD16++ Monocytes on CD14++CD16− Monocytes: A Possible Explanation of Monocyte Alterations in Systemic Lupus Erythematosus

“…EFFECTS OF CD14ϩCD16ϩϩ MONOCYTES ON CD14ϩϩCD16Ϫ MONOCYTESously reported ratios (6:4 ratio of CD14ϩϩCD16Ϫ cells to CD14ϩCD16ϩϩ cells) (2,18,(20)(21)(22). The final number of cells per well was 1.2 ϫ 10 5 , using 96-well plates.…”