Mature dendritic cells derived from human monocytes within 48 hours: a novel strategy for dendritic cell differentiation from blood precursors.
It is widely believed that generation of mature dendritic cells (DCs) with full T cell stimulatory capacity from human monocytes in vitro requires 5-7 days of differentiation with GM-CSF and IL-4, followed by 2-3 days of activation. Here, we report a new strategy for differentiation and maturation of monocyte-derived DCs within only 48 h of in vitro culture. Monocytes acquire immature DC characteristics by day 2 of culture with GM-CSF and IL-4; they down-regulate CD14, increase dextran uptake, and respond to the inflammatory chemokine macrophage inflammatory protein-1alpha. To accelerate DC development and maturation, monocytes were incubated for 24 h with GM-CSF and IL-4, followed by activation with proinflammatory mediators for another 24 h (FastDC). FastDC expressed mature DC surface markers as well as chemokine receptor 7 and secreted IL-12 (p70) upon CD40 ligation in the presence of IFN-gamma. The increase in intracellular calcium in response to 6Ckine showed that chemokine receptor 7 expression was functional. When FastDC were compared with mature monocyte-derived DCs generated by a standard 7-day protocol, they were equally potent in inducing Ag-specific T cell proliferation and IFN-gamma production as well as in priming autologous naive T cells using tetanus toxoid as a model Ag. These findings indicate that FastDC are as effective as monocyte-derived DCs in stimulating primary, Ag-specific, Th 1-type immune responses. Generation of FastDC not only reduces labor, cost, and time required for in vitro DC development, but may also represent a model more closely resembling DC differentiation from monocytes in vivo.
Dauer
Marc
M
Division of Clinical Pharmacology and Section of Gastroenterology, Medizinische Klinik Innenstadt, University of Munich, Munich, Germany.
Obermaier
Bianca
B
Herten
Jan
J
Haerle
Carola
C
Pohl
Katrin
K
Rothenfusser
Simon
S
Schnurr
Max
M
Endres
Stefan
S
Eigler
Andreas
A
D023201:CD40 Ligand / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000276:immunology; D002478:Cells, Cultured; D054421:Chemokine CCL21; D054377:Chemokine CXCL12; D019742:Chemokines, CC / Q000378:metabolism / Q000494:pharmacology; D019743:Chemokines, CXC / Q000378:metabolism / Q000494:pharmacology; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D003911:Dextrans / Q000276:immunology / Q000378:metabolism; D015232:Dinoprostone / Q000494:pharmacology; D004338:Drug Combinations; D004705:Endocytosis / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D016130:Immunophenotyping; D018836:Inflammation Mediators / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis; D007375:Interleukin-1 / Q000494:pharmacology; D018664:Interleukin-12 / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology; D015847:Interleukin-4 / Q000494:pharmacology; D007399:Interphase / Q000276:immunology; D008024:Ligands; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D054400:Receptors, CCR7; D019718:Receptors, CXCR4 / Q000378:metabolism; D019707:Receptors, Chemokine / Q000378:metabolism; D015290:Second Messenger Systems / Q000276:immunology; D012995:Solubility; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
C515478:CCL21 protein, human; C515370:CCR7 protein, human; C515255:CXCL12 protein, human; D054421:Chemokine CCL21; D054377:Chemokine CXCL12; D019742:Chemokines, CC; D019743:Chemokines, CXC; D003911:Dextrans; D004338:Drug Combinations; D018984:Epitopes, T-Lymphocyte; D018836:Inflammation Mediators; D007375:Interleukin-1; D008024:Ligands; D054400:Receptors, CCR7; D019718:Receptors, CXCR4; D019707:Receptors, Chemokine; D013745:Tetanus Toxoid; D014409:Tumor Necrosis Factor-alpha; D023201:CD40 Ligand; D018664:Interleukin-12; D015847:Interleukin-4; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D015232:Dinoprostone