Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adjuvants, Immunologic / pharmacology
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Cancer Vaccines / chemical synthesis
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Cell Differentiation / drug effects
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Cell Differentiation / immunology*
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Cell Movement / drug effects
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Cell Movement / immunology
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Cell Polarity / drug effects
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Cell Polarity / immunology*
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Cells, Cultured
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Coculture Techniques
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Cytokines / metabolism
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Dendritic Cells / cytology
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Dendritic Cells / immunology*
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Dendritic Cells / transplantation*
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Humans
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Imidazoles / agonists
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Imidazoles / pharmacology
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Immunotherapy, Adoptive / methods
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Interferon-gamma / metabolism
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Killer Cells, Natural / drug effects
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Killer Cells, Natural / immunology
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Killer Cells, Natural / metabolism
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Ligands
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Lymphocyte Activation / drug effects
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Lymphocyte Activation / immunology
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Poly I-C / metabolism
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Quinolines / pharmacology*
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T-Lymphocytes, Cytotoxic / drug effects
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T-Lymphocytes, Cytotoxic / immunology
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T-Lymphocytes, Cytotoxic / metabolism
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Th1 Cells / drug effects
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Th1 Cells / immunology*
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Thiazoles / pharmacology*
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Toll-Like Receptor 3 / metabolism
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Toll-Like Receptor 7 / agonists*
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Toll-Like Receptor 7 / physiology
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Toll-Like Receptor 8 / agonists*
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Toll-Like Receptor 8 / physiology
Substances
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Adjuvants, Immunologic
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CL 075
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Cancer Vaccines
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Cytokines
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Imidazoles
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Ligands
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Quinolines
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TLR3 protein, human
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TLR7 protein, human
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TLR8 protein, human
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Thiazoles
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Toll-Like Receptor 3
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Toll-Like Receptor 7
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Toll-Like Receptor 8
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Interferon-gamma
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Poly I-C
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resiquimod