Generation of Th1-polarizing dendritic cells using the TLR7/8 agonist CL075.
In this paper, we describe a new method for preparation of human dendritic cells (DCs) that secrete bioactive IL-12(p70) using synthetic immunostimulatory compounds as TLR7/8 agonists. Monocyte-derived DCs were generated using a procedure that provided mature cells within 3 d. Several maturation mixtures that contained various cytokines, IFN-gamma, different TLR agonists, and PGE(2) were compared for impact on cell recovery, phenotype, cytokine secretion, migration, and lymphocyte activation. Mixtures that included the TLR7/8 agonists R848 or CL075, combined with the TLR3 agonist polyinosinic:polycytidylic acid, yielded 3-d mature DCs that secreted high levels of IL-12(p70), showed strong chemotaxis to CCR7 ligands, and had a positive costimulatory potential. They also had excellent capacity to activate NK cells, effectively polarized CD4(+) and CD8(+) T cells to secrete IFN-gamma and to induce T cell-mediated cytotoxic function. Thereby, mature DCs prepared within 3 d using such maturation mixtures displayed optimal functions required for vaccine development.
Institute of Molecular Immunology, German Research Center for Environmental Health, Marchioninistrasse 25, 81377 Munich, Germany. stefani.spranger@helmholtz-muenchen.de
Stefani
S
Spranger
Miran
M
Javorovic
Maja
M
Bürdek
Susanne
S
Wilde
Barbara
B
Mosetter
Stefanie
S
Tippmer
Iris
I
Bigalke
Christiane
C
Geiger
Dolores J
DJ
Schendel
Bernhard
B
Frankenberger
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D019496:Cancer Vaccines / Q000138:chemical synthesis; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology*; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D016764:Cell Polarity / Q000187:drug effects / Q000276:immunology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology* / Q000637:transplantation*; D006801:Humans; D007093:Imidazoles / Q000819:agonists / Q000494:pharmacology; D016219:Immunotherapy, Adoptive / Q000379:methods; D007371:Interferon-gamma / Q000378:metabolism; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008024:Ligands; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D011070:Poly I-C / Q000378:metabolism; D011804:Quinolines / Q000494:pharmacology*; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000187:drug effects / Q000276:immunology*; D013844:Thiazoles / Q000494:pharmacology*; D051196:Toll-Like Receptor 3 / Q000378:metabolism; D051199:Toll-Like Receptor 7 / Q000819:agonists* / Q000502:physiology; D051200:Toll-Like Receptor 8 / Q000819:agonists* / Q000502:physiology
D000276:Adjuvants, Immunologic; C551788:CL 075; D019496:Cancer Vaccines; D016207:Cytokines; D007093:Imidazoles; D008024:Ligands; D011804:Quinolines; C495344:TLR3 protein, human; C495348:TLR7 protein, human; C495349:TLR8 protein, human; D013844:Thiazoles; D051196:Toll-Like Receptor 3; D051199:Toll-Like Receptor 7; D051200:Toll-Like Receptor 8; D007371:Interferon-gamma; D011070:Poly I-C; C402365:resiquimod