Revision of the functional analysis and structural features of immortalized dendritic cell lines derived from mice lacking both type I and type II interferon receptors.
Cell lines with dendritic morphology were obtained from several organs of mice lacking both type I and II interferon receptors after a retroviral immortalization procedure. Their surface antigen phenotype was analyzed by flow cytometry with monoclonal antibodies and their functional capabilities to induce antigen dependent specific immune response was also determined. Two representative cell lines called AG101 (skin-derived) and AG116 (brain-derived) were cloned and analyzed in more detail. Cytometric analysis showed that they constitutively expressed the cell surface markers CD45, CD1 1b, MHC class II, F4/80, N418, B7-2 and ICAM1. Despite both cell lines expressing Thy-1 only, the AG116 show CD4 but both were negative for CD8 and B220. The functional analysis showed that the cell lines were capable and very efficient at actively taking up, processing and presenting soluble antigens like Ovalbumin (OVA). The processed protein was presented by both cell lines to the OVA-peptide-specific T cell hybridoma BO97.105, which responded specifically with the production of IL-2. In addition AG101 and AG116 cells were able to induce in naive allogeneic T cells, a mixed lymphocyte reaction, determined by T cell proliferation and T cell dependent L-2 production. Moreover, the capability to prime naive syngeneic T cells was also demonstrated by loading AG101 and AG116 cells with soluble antigens, then co-culturing with naive T cells which yielded both T cell proliferation and IL-2 production. The cell lines priming capability was shown to be quite similar, as freshly isolated and cultured cutaneous dendritic cells from 129Sv/Lv mice (wtDCs) to prime naive T cells. In addition to a basal production of IL-6, the cell lines were found to increase their synthesis of IL-6 and IL-12 p40 after interaction with T cells in a similar way as mature wtDCs. Also it was determined that DC cell lines devoid of functional IFN system allow the replication of infectious agents like BDV and even are able to induce in vivo a specific humoral response against proteins of the BDV. Therefore, the cell lines AG101 and AG116 show structural and functional features of DCs. They are able to take up, process and present antigens as well as prime naive T cell in a similar manner as nontransformed DC. Therefore, these cell lines will be useful for studying the interactions between DC and the effectors cells of the immune response at the clonal level and in the absence of functional interferon receptors.
Nunez
R
R
Institute for Virology, Faculty of Veterinary Medicine, University of Zurich, Switzerland. rafaeln@vetvir.unizh.ch
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D017951:Antigen Presentation / Q000235:genetics; D001891:Borna disease virus / Q000276:immunology; D002461:Cell Line, Transformed / Q000166:cytology* / Q000276:immunology*; D002472:Cell Transformation, Viral; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D005822:Genetic Vectors; D006825:Hybridomas; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008565:Membrane Proteins; D051379:Mice; D018345:Mice, Knockout; D053633:Receptor, Interferon alpha-beta; D017471:Receptors, Interferon / Q000172:deficiency* / Q000235:genetics; D012190:Retroviridae; D013601:T-Lymphocytes / Q000378:metabolism; D014765:Viral Vaccines / Q000276:immunology; D000096743:Interferon gamma Receptor
D000914:Antibodies, Viral; D016207:Cytokines; D008565:Membrane Proteins; D053633:Receptor, Interferon alpha-beta; D017471:Receptors, Interferon; D014765:Viral Vaccines; D000096743:Interferon gamma Receptor