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Fused Late Endocytic Compartments and Immunostimulatory Capacity of Dendritic–Tumor Cell Hybridomas
Authors:Mateja Gabrijel  Martina Bergant  Marko Kreft  Matjaž Jeras  Robert Zorec
Affiliation:(1) Laboratory of Neuroendocrinology – Molecular Cell Physiology, Institute of Pathophysiology, Faculty of Medicine, University of Ljubljana, 1000 Ljubljana, Slovenia;(2) Celica Biomedical Center, Technology Park 24, 1000 Ljubljana, Slovenia;(3) Tissue Typing Center, Blood Transfusion Centre of Slovenia, 1000 Ljubljana, Slovenia
Abstract:Late endocytic compartments, containing MHC class II molecules in antigen presenting cells, fuse to each other in order to deliver antigens to these molecules. We have shown previously that fusion of late endocytic compartments takes place also in hybridomas. Therefore, we investigate here whether the level of fused late endocytic compartments affects the immunostimulatory capacity of hybridomas obtained by the electrofusion of dendritic and tumor cells. The level of fused late endocytic compartments in a single hybridoma cell was assessed and samples of electrofused cells were then cocultured with autologous T cells, resulting in the priming of naïve T cells. To test the immunostimulatory capacity of hybridoma cells, T-cell-induced cytotoxicity of tumor cells was assayed. The results demonstrate that in vitro cytotoxic T cell responses are enhanced if a higher percentage of fused late endocytic compartments is present in the cell population of electrofused hybridoma cells.
Keywords:Cancer  Confocal microscopy  Hybridoma cells  Fusion of late endocytic compartments  Quantitative three-dimensional colocalization
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