Abstract
Dendritic cells (DCs) are professional antigen-presenting cells currently being discussed as a potent tool for antitumor vaccination strategies. The approach consisting of the in vitro generation of DC-tumor cell hybrids may be advantageous for individualized vaccines since there is no need for the determination of MHC-restricted tumor-associated antigens recognized by T cells. As recent vaccination studies gave varying results, we tested the impact of the fusion treatment on the cells used. Polyethylene glycol-induced fusion, as well as electrofusion, proved to be suitable for generating hybrid cells although at a low frequency. Of note, both methods also gave rise to DCs having phagocytosed apoptotic tumor cells. The expression of surface molecules relevant for specific T cell stimulation was not altered by the fusion procedure and the DCs were still functionally active as demonstrated by the secretion of IL-12 and the uptake of antigen. The cells were able to induce a tumor-specific T cell response in vitro and therefore deserve further investigation as potent tools for immunotherapy trials.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, CD / analysis
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B7-1 Antigen / analysis
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B7-2 Antigen
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CD83 Antigen
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Cell Fusion
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Cell Survival
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Coculture Techniques
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Dendritic Cells / cytology
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Dendritic Cells / immunology*
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Dendritic Cells / metabolism
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Enzyme-Linked Immunosorbent Assay
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Flow Cytometry
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Fluoresceins
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Fluorescent Dyes
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HLA-DR Antigens / analysis
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Humans
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Hybrid Cells / cytology
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Hybrid Cells / drug effects
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Hybrid Cells / immunology*
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Immunoglobulins / analysis
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Interleukin-12 / metabolism
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Leukocyte Common Antigens / analysis
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Membrane Glycoproteins / analysis
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Microscopy, Confocal
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Microscopy, Fluorescence
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Neoplasms / immunology
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Neoplasms / pathology
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Phagocytosis / immunology
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Polyethylene Glycols / pharmacology
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Rhodamines
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T-Lymphocytes / cytology
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T-Lymphocytes / immunology
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Tumor Cells, Cultured / immunology
Substances
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Antigens, CD
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B7-1 Antigen
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B7-2 Antigen
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Fluoresceins
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Fluorescent Dyes
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HLA-DR Antigens
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Immunoglobulins
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Interleukin-12
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Leukocyte Common Antigens
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Membrane Glycoproteins
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Polyethylene Glycols
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Rhodamines
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CD83 Antigen
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(((4-chloromethyl)benzoyl)amino)-tetramethylrhodamine
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CD86 protein, human
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5-chloromethylfluorescein