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13.
Immunogenicity of dendritic cells pulsed with cartilage derived soluble alloantigens
Maličev, Elvira ; Knežević, Miomir ; Jeras, Matjaž
Investigation of allo- and autoreactivity of cartilage components is useful not only for the potential allogeneneic transplantation but also for the treatment of tissue specific degenerative ...autoimmune diseases. We generated dendritic cells (DCs) to study articular cartilage immunogenicity, by using human peripheral blood mononuclear cells. Dendritic cells are extremely efficient antigen-presenting cells that initiate and modulate T-cell dependantimmune responses. After isolation by density centrifugation, GM-CSF and IL-4 were added to adherent mononuclear cells. After 7 days of cultivationthe cells in cell culture were: HLA-DR++, CD86+, CD:14-, CD80-, CD83, CD19- CD56-, CD3-. Dendritic cells were then pulsed with a supernatant taken from primary monolayer cell cultures of human allogeneic chondrocytes. The stimulating capacity of antigen-pulsed DCs was measured in a classical proliferation assay using tritium labelled thymidine as a marker of mitosis. Proliferative responses were significantly strong but became much stronger when DCs were concoinitaintly incubated with soluble antigens and human recombinant TNF-alpha and used subsequently as stimulators. Although TNF-alphapositively influenced the stimulating capacity of pulsed DCs, we wereunable to detect major diffrences in expression of specific DCs' phenotypemarkers assessed by flow cytometry in the absence or presence of thiscytokine.
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