FastDC derived from human monocytes within 48 h effectively prime tumor antigen-specific cytotoxic T cells.
Previously, we have shown that dendritic cells (DCs) with full T-cell stimulatory capacity can be derived from human monocytes after 48 h of in vitro culture (FastDC). Compared to a standard 7-day protocol, this new strategy not only reduces the time span and the amount of recombinant cytokines required, but may also resemble DC development in vivo more closely. Using a melanoma antigen model, we show here that FastDC prime CTL responses against tumor antigens as effectively as standard monocyte-derived DCs (moDCs). FastDC and moDCs derived from monocytes of HLA-A2(+) donors were loaded with the melanoma-associated, HLA-A(*)0201-restricted peptide Melan-A and cocultured with autologous CD3(+) T cells. After two weekly restimulations with freshly prepared, peptide-loaded FastDC or moDCs, binding of CD8(+) T cells to fluorescently labeled MHC-I/Melan-A-peptide complexes and intracellular cytokine staining revealed that the two DC preparations had an equal capacity to prime Melan-A-specific, IFN-gamma producing CD8(+) T cells. CTLs derived from cocultures with FastDC lysed Melan-A-loaded T2 cells even more effectively than CTLs primed by moDCs. Comparative analysis also revealed that FastDC possess an equal capacity to migrate in response to the chemokine receptor CCR-7 ligand 6Ckine. Importantly, DCs can be generated with higher yield and purity using the FastDC-protocol. The reliability and efficacy of this new strategy for DC development from monocytes may facilitate clinical investigation of DC-based tumor immunotherapy.
Section of Gastroenterology, Medizinische Klinik Innenstadt, University of Munich, 80336 Munich, Germany.
Marc
M
Dauer
Katharina
K
Schad
Jan
J
Herten
Jana
J
Junkmann
Christian
C
Bauer
Rosemarie
R
Kiefl
Stefan
S
Endres
Andreas
A
Eigler
D000951:Antigens, Neoplasm / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology*; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism
D000951:Antigens, Neoplasm; D016207:Cytokines