Cells with dendritic cell morphology and immunophenotype, binuclear morphology, and immunosuppressive function in dendritic cell cultures.
Culturing of human peripheral blood CD14 positive monocytes is a method for generation of dendritic cells (DCs) for experimental purposes or for use in clinical grade vaccines. When culturing human DCs in this manner for clinical vaccine production, we noticed that 5-10% of cells within the bulk culture were binuclear or multiple nuclear, but had typical dendritic cell morphology and immunophenotype. We refer to the cells as binuclear cells in dendritic cell cultures (BNiDCs). By using single cell PCR analysis of mitochondrial DNA polymorphisms we demonstrated that approximately 20-25% of cells in DC culture undergo a fusion event. Flow sorted BNiDC express low HLA-DR and IL-12p70, but high levels of IL-10. In mixed lymphocyte reactions, purified BNiDC suppressed lymphocyte proliferation. Blockade of dendritic cell-specific transmembrane protein (DC-STAMP) decreased the number of binuclear cells in DC cultures. BNiDC represent a potentially tolerogenic population within DC preparations for clinical use.
Unit of Molecular Haematology and Cancer Biology, UCL Institute of Child Health, London, UK.
Rong
R
Dong
Dale
D
Moulding
Nourredine
N
Himoudi
Stuart
S
Adams
Gerben
G
Bouma
Ayad
A
Eddaoudi
B Piku
BP
Basu
Sophie
S
Derniame
Prabhjoat
P
Chana
Andrew
A
Duncan
John
J
Anderson
D048868:Adaptor Proteins, Signal Transducing; D000906:Antibodies / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000276:immunology; D002459:Cell Fusion; D002467:Cell Nucleus; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000096:biosynthesis / Q000276:immunology; D006801:Humans; D007108:Immune Tolerance; D007109:Immunity; D007150:Immunohistochemistry; D016130:Immunophenotyping; D007165:Immunosuppression Therapy / Q000379:methods*; D016753:Interleukin-10 / Q000096:biosynthesis / Q000276:immunology; D018664:Interleukin-12 / Q000096:biosynthesis / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D009000:Monocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D059010:Single-Cell Analysis
D048868:Adaptor Proteins, Signal Transducing; D000906:Antibodies; C501589:DCSTAMP protein, human; D006684:HLA-DR Antigens; D018950:Lipopolysaccharide Receptors; D008565:Membrane Proteins; D016753:Interleukin-10; D018664:Interleukin-12