ABSTRACT
Due to their high immunostimulatory ability as well as the critical role they play in the maintenance of self-tolerance, dendritic cells have been implicated in the pathogenesis of autoimmune diseases. The non-obese diabetic (NOD) mouse is an animal model of autoimmune type 1 diabetes, in which pancreatic beta cells are selectively destroyed mainly by T cell-mediated immune responses. To elucidate initiation mechanisms of beta cell-specific autoimmunity, we attempted to generate bone marrow-derived dendritic cells from NOD mice. However, our results showed low proliferative response of NOD bone marrow cells and some defects in the differentiation into the myeloid dendritic cells. NOD dendritic cells showed lower expressions of MHC class II, B7-1, B7-2 and CD40, compared with C57BL/6 dendritic cells. In mixed lymphocyte reactions, stimulatory activities of NOD dendritic cells were also weak. Treatment with LPS, INF-gamma and anti-CD40 stimulated NOD dendritic cells to produce IL-12p70. The amount of IL-12, however, appeared to be lower than that of C57BL/6. Results of the present study indicated that there may be some defects in the development of NOD dendritic cells in the bone marrow, which might have an impact on the breakdown of self tolerance.
Subject(s)
Mice , Animals , Autoimmune Diseases/pathology , Autoimmune Diseases/immunology , Bone Marrow Cells/pathology , Bone Marrow Cells/immunology , Bone Marrow Cells/chemistry , Cell Differentiation/immunology , Cell Differentiation/drug effects , Dendritic Cells/pathology , Dendritic Cells/immunology , Dendritic Cells/chemistry , Diabetes Mellitus, Type 1/pathology , Diabetes Mellitus, Type 1/immunology , Enzyme-Linked Immunosorbent Assay , Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology , Interleukin-12/analysis , Interleukin-4/pharmacology , Lipopolysaccharides/pharmacology , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred NOD , ObesityABSTRACT
Dendritic cells (DCs) are the most potent antigen presenting cells that can activate naive T cells. Mature DCs exress high levels of MHC and costimulatory molecules on their surface and have capacity to produce IL-12, a 75 kDa heterodimeric cytokine composed of p35 and p40 subunit. IL-12 is currently thought to be one of most critical determinants for skewing the immune response towards Th1. Expression of IL-12 in dendritic cells seems to be regulated by various stimuli including CD40L. In the present study we investigated expression of IL-12 in mature DCs, which were cultured from bone marrow cells in the presence of GM-CSF. Maturity of the DCs was confirmed by morphologic characteristics, immunophenotypes, and allostimulatory activities. Exprssion levels of IL-12 p40 in the DCs were measured by semi-quantitative RT-PCR. Increases in IL-12 p40 expression were observed after treatment with lipopolysaccharide (LPS), an anti-MHC class II monoclonal antibody, or an anti-CD40 monoclonal antibody. The most remarkable increases, however, were observed in the DCs treated with an anti-CD40 monoclonal antibody. These results support a previous notion that signals through CD40/CD40L interaction may be important for the production of IL-12 by DCs. Moreover, results of this study show a possibility of using monoclonal antibodies against CD40 molecules for preparing DCs producing high amount of IL-12, which can be used for anti-tumor or anti-viral immunotherapy.