Murine dendritic cells generated under serum-free conditions have a mature phenotype and efficiently induce primary immune responses

Vaccination with in vitro-generated dendritic cells (DC) that present tumor-associated antigens is a promising approach for immunotherapy of malignant tumors. For optimization of DC-based vaccination protocols, preclinical tumor models that mimic the clinical situation closely are highly desirable....

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Bibliographic Details
Published in:Journal of immunological methods 2006-03, Vol.310 (1), p.1-11
Main Authors: Warncke, Max, Dodero, Anna, Dierbach, Heide, Follo, Marie, Veelken, Hendrik
Format: Article
Language:English
Subjects:
Animals
Antigens, CD - immunology
Antigens, Neoplasm - immunology
Biological and medical sciences
Cell Culture Techniques - methods
Cell Differentiation - immunology
Culture Media, Serum-Free
Dendritic cells
Dendritic Cells - cytology
Dendritic Cells - immunology
Fetal calf serum
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Immunophenotyping
Immunotherapy
Immunotherapy, Adoptive - methods
Interferon-gamma - immunology
Interleukin-12 - immunology
Mice
Mice, Inbred BALB C
Microscopy, Confocal
Molecular immunology
Murine study
Peptide Fragments - immunology
Serum-free media
Specific Pathogen-Free Organisms
Techniques
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Summary:Vaccination with in vitro-generated dendritic cells (DC) that present tumor-associated antigens is a promising approach for immunotherapy of malignant tumors. For optimization of DC-based vaccination protocols, preclinical tumor models that mimic the clinical situation closely are highly desirable. Strong non-specific T cell activation was observed in experimental immunization of mice with syngeneic DC generated in standard FCS-supplemented culture medium. To avoid deviation of the immune response to FCS-derived antigens, a serum-free culture protocol for in vitro generation of murine DC from bone marrow progenitor cells was developed. In comparison to DC differentiated with FCS supplementation, DC generated under serum-free conditions (sfDC) have a more homogeneous phenotype with higher expression of IL-12 and the differentiation and activation markers CD11c, CD40, CD80, CD83, CD86, DEC-205, and MHC class II. Demonstration of strong uptake of protein and carbohydrate antigens and analysis of the in vivo migration behaviour of sfDC also indicated excellent APC function. Vaccination of mice with peptide-pulsed sfDC efficiently induced an antigen-specific T cell response as assessed by MHC tetramer staining, IFN-γ ELISPOT and in vivo cytotoxicity assay. sfDC may therefore represent a valuable tool to improve active tumor immunotherapy in animal models.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2005.09.012