Differentiation of human monocytes and derived subsets of macrophages and dendritic cells by the HLDA10 monoclonal antibody panel

Abstract

The mononuclear phagocyte system, consisting of monocytes, macrophages and dendritic cells (DCs), has an important role in tissue homeostasis as well as in eliciting immune responses against invading pathogens. Blood monocytes have been viewed for decades as precursors of tissue macrophages. Although the newest data show that in the steady state resident macrophages of many organs are monocyte independent, blood monocytes critically contribute to tissue macrophage and DC pools upon inflammation. To better understand the relationship between these populations and their phenotype, we isolated and differentiated human blood CD14+ monocytes in vitro into immature and mature monocyte-derived dendritic cells (MoDCs) as well as into seven different monocyte-derived macrophage subsets. We used the panel of 70 monoclonal antibodies (mAbs) submitted to the 10th Human Leukocyte Differentiation Antigen Workshop to determine the expression profiles of these 10 populations by flow cytometry. We now can compile subpanels of mAbs to differentiate the 10 monocyte/macrophage/MoDC subsets, providing the basis for novel diagnostic and therapeutic tools.

Human Intestinal Mononuclear Phagocytes in Health and Inflammatory Bowel Disease

“…One candidate could be GM-CSF. Indeed, it has been shown that blood monocytes differentiate to immature Mf-like cells through GM-CSF ex vivo (281,282), a factor that increases in Crohn's disease and UC, particularly in active lesion areas (283). Moreover, GM-CSF can act in concert with IFN-γ and TNF to reprogram blood monocytes into Mfs with an inflammatory profile (284).…”

The mannose 6-phosphate/insulin-like growth factor 2 receptor mediates plasminogen-induced efferocytosis

“…Monocytes were further purified from the PBMC fraction using CD14 MACS microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). Primary macrophages were differentiated from CD14 + monocytes with recombinant human macrophage colony‐stimulating factor (M‐CSF; from Peprotech, Rocky Hill, NJ; 50 ng/ml) as previously described . Following a 7‐day differentiation, macrophages were rested in macrophage serum free medium (Invitrogen), supplemented with 2 mmol/l l ‐glutamine and 2% FCS for 2 days prior to efferocytosis assays.…”

Impact of the body mass index on perioperative immunological disturbances in patients with hip and knee arthroplasty

“…While IL-2 is mainly expressed in Th cells, IL-15 is constitutively expressed by certain immune cell populations including monocytes, macrophages, and dendritic cells [15]. In the blood, circulating monocytes are recruited to inflamed tissues and can derive to macrophages or dendritic cells [16]. The removal of the inflammatory site due to the arthroplasty could therefore explain the post-operative reduction of peripheral NK cells.…”