Human peripheral blood mononuclear cells (PBMCs)-derived dendritic cells fused with K562 cells induce a p210BCR-ABL protein-specific cytotoxic T cell response in vitro
10.3321/j.issn:1000-467X.1999.06.001
- VernacularTitle:人树突状细胞与K562细胞的融合以及体外诱导p210蛋白特异性CTL的研究
- Keywords:
Dendritic cells;
K562;
Cell fusion;
P210BCR-ABL protein;
Specific CTL
- From:Chinese Journal of Cancer
1999;18(6):617-623
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate whether human dendritic cells (DCs) derived from cultured human peripheral blood mononuclear cells (PBMCs) were capable of presenting antigen to induce a p210BCR-ABL protein-specific CTL response in vitro after fusion with K562 cells which express p210BCR-ABL protein.Methods:DCs were derived from human PBMCs in the presence of granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). To show the successful fusion of DCs with K562 cells,we first used K562 cells expressing green fluorescence protein(GFP) in one group, while in the other group,DCs fused with normal K562 cells,and these fusion cells were cocultured with autologous T lymphocytes for two rounds.Results:After 5～7 days of incubation,these adherent cells appeared in clusters or dispersed,and the results of flow cytometric analysis showed the cell surface contained markers typical of dendritic cells,i.e.,they were positive for HLA-DR,HLA-A,B,C,CD1α ,CD80,CD86 and negative for CD14 (monocytes). The fusion cells demonstrated morphologically distinct cell aggregates. Moreover,most of these cells that expressed GFP, exhibited a DC morphology with veiled processes and dendrites. Furthermore,these fusion cells can induce a p210BCR-ABL protein-specific cytotoxic T cell response in vitro using the lactate dehydrogenase (LDH) release assay.Conclusions:The direct fusion of DCs with K562 cells could result in induction of an effective p210BCR-ABL (b3a2) protein-specific immunogen, either by confering sufficient DC function to K562 cells for activation of T cells or by facilitating the delivery of tumor p210BCR-ABL (b3a2) protein to DCs for processing and presentation,which may broaden the spectrum of possible DC-based clinical applications.
