title: Generation of inducible immortalized dendritic cells with proper immune function in vitro and in vivo.
abstract: Dendritic cells are the professional antigen presenting cells of innate immunity and key players in maintaining the balance of immune responses. Studies with dendritic cells are mainly limited by their low numbers in vivo and their difficult maintenance in vitro. We differentiated bone marrow cells from transgenic mice expressing an inducible SV40 large T-antigen into dendritic cells. When immortalized by dexamethasone and doxycycline, these cells were stable in long-term culture. In the absence of dexamethasone and doxycycline (de-induction), dendritic cells displayed properties of primary cells, characterized by expression of classical dendritic cell surface markers CD11c, CD11b, MHCII, CD40 and CD86. Furthermore, de-induced lipopolysaccharide activated dendritic cells secreted IL-1β, IL-6, TNFα and IL-12. De-induced, Ovalbumin-loaded dendritic cells polarize CD4(+) T cells into Th1, Th17 and Th2 cells, indicating their correct antigen presenting property. Consistent with intratracheal application of Ovalbumin-loaded primary dendritic cells into mice, the application of de-induced dendritic cells resulted in recruitment of lymphocytes to the lungs. In summary, we successfully expanded dendritic cells using conditional immortalization. The generated dendritic cells demonstrate the characteristic immunophenotype of primary dendritic cells and will facilitate further studies on immunomodulatory properties of dendritic cells.
authors.affiliation: Department of Pediatrics, University Clinic Carl Gustav Carus, Technische Universitaet Dresden, Dresden, Germany. cornelia.richter@uniklinikum-dresden.de
authors.forename: Cornelia
authors.initials: C
authors.lastname: Richter
authors.forename: Sebastian
authors.initials: S
authors.lastname: Thieme
authors.forename: Joanna
authors.initials: J
authors.lastname: Bandoła
authors.forename: Magdalena
authors.initials: M
authors.lastname: Laugsch
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Anastassiadis
authors.forename: Sebastian
authors.initials: S
authors.lastname: Brenner
mesh_terms: D056704:Adaptive Immunity; D000818:Animals; D017951:Antigen Presentation; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000378:metabolism; D000954:Antigens, Surface / Q000378:metabolism; D001854:Bone Marrow Cells / Q000276:immunology / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002461:Cell Line, Transformed; D045142:Cross-Priming; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D016130:Immunophenotyping; D016086:Lentivirus / Q000235:genetics; D008168:Lung / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D010641:Phenotype; D019076:Transgenes
chemical_list: D000952:Antigens, Polyomavirus Transforming; D000954:Antigens, Surface; D016207:Cytokines