Combined use of toll-like receptor agonists and prostaglandin E(2) in the FastDC model: rapid generation of human monocyte-derived dendritic cells capable of migration and IL-12p70 production.
Phenotypical maturation, IL-12p70 production and migration upon chemokine receptor CCR7 ligation are currently proposed as requirements for the use of human monocyte-derived dendritic cells (DC) in antitumoral vaccination. We have previously described a short-term protocol for DC generation from monocytes including stimulation with TNF-alpha, IL-1beta and PGE(2) (FastDC). These "conventional" FastDC are mature, migrate in response to CCR7 ligation and effectively stimulate autologeous T cells in vitro, but are deficient in IL-12p70 production. Here, conventional FastDC were compared to FastDC activated with different TLR ligands. High levels of IL-12p70 were induced by combined activation of FastDC with TLR4 and TLR7/8 ligands. IL-12 secretion could be maximized by additional T cell-derived stimulation. However, TLR-stimulated FastDC failed to migrate upon CCR7 ligation, independent of additional activation with CD40 ligand and IFN-gamma. The presence of PGE(2) during TLR ligation fully restored migratory capacity of FastDC, but left IL-12p70 production and activation of tumor antigen-specific cytotoxic T cells unaffected, challenging previous findings obtained with standard 7-day monocyte-derived DC. The FastDC model thus not only represents an effective tool for antitumoral vaccination, but may also provide novel insights into human DC biology.
Dauer
Marc
M
Section of Gastroenterology and Division of Clinical Pharmacology, Medizinische Klinik Innenstadt, University of Munich, Ziemssenstr.1, 80336 Munich, Germany. marc.dauer@gmx.de
Lam
Veronique
V
Arnold
Hannah
H
Junkmann
Jana
J
Kiefl
Rosemarie
R
Bauer
Christian
C
Schnurr
Max
M
Endres
Stefan
S
Eigler
Andreas
A
D000951:Antigens, Neoplasm / Q000276:immunology; D002465:Cell Movement; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000502:physiology*; D015232:Dinoprostone / Q000494:pharmacology*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000096:biosynthesis*; D058965:MART-1 Antigen; D009000:Monocytes / Q000166:cytology*; D009363:Neoplasm Proteins / Q000276:immunology; D054400:Receptors, CCR7 / Q000502:physiology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D051193:Toll-Like Receptors / Q000819:agonists* / Q000502:physiology
D000951:Antigens, Neoplasm; C515370:CCR7 protein, human; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D054400:Receptors, CCR7; D051193:Toll-Like Receptors; D018664:Interleukin-12; D007371:Interferon-gamma; D015232:Dinoprostone