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- W2358746880 abstract "Objective:To assess the properties of exosomes secreted by fusion cells of hepatocellular carcinoma (HCC) patient-derived dendritic cells(DCs)and HCC cell line (HepG2) and to evaluate the function of these exosomes to trigger efficient T cell responses against HepG2 cells in vitro. Methods: Peripheral blood mononuclear cells (PBMC) from HCC patients were isolated by blood separator. In the presence of recombinant human granulocyte/macrophage-clone stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4), PBMC were cultured in vitro for one week to induce dendritic cells (DC). Fusion cells of DC with HepG2 cells (DC-HepG2) were achieved by polyethylene glycol (PEG) and were isolated by magnetic device with HLA Class II Dynabeads. Exosomes were then prepared from the supernatants of the fusion cells by ultra filtration centrifugation and sucrose gradient ultracentrifugation. Transmission electron microscopy was used to observe their structures. The expressions of several proteins were investigated by flow cytometry and ELISA assay. T lymphocytes were pulsed with exosomes directly in the presence of IL-2. After co-cultured with target cells, the stimulated lymphocytes were tested by IFN-γ release assay and cytotoxicity capacity assay. Results:After fusion, DC-HepG2 expressed high level of DC surface molecules, including CD83 87.4%, CD80 94.9%, CD86 90.13% and HLA-DR 98.62%. Application of the isolation procedure to fusion cells also revealed exosome vesicles by transmission electron microscopy. Protein analysis by FCM was performed and revealed that the costimulatory molecule CD86 and AFP protein was detectable. AFP protein was also detectable by ELISA assay. The fusion cell- derived exosomes could directly activate T lymphocytes which lysed HepG2 target cells much more effectively than T cells alone did (P0.05). To determine whether the lysis was HepG2 tumor cell-specific, SMMC7721 and K562 cells were used as target cells and nearly no lysis of these cells was observed. Furthermore, IFN-γ in the culture supernatants of exosomes-activated T cells was significantly higher than that in normal control T cells (P0.05). Conclusion:Exosomes could be purified from the supernatants of fusion cells. These exosomes could activate T cells directly and could induce effective HepG2 secific CTL responses. DC-HepG2 fusion cells-derived exosomes may represent as a promising approach of immunotherapy for HCC." @default.
- W2358746880 created "2016-06-24" @default.
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- W2358746880 date "2006-01-01" @default.
- W2358746880 modified "2023-10-03" @default.
- W2358746880 title "Specific immune responses against hepatocellular carcinoma induced by dendritic cell-HepG2 fusion cells derived-exosomes" @default.
- W2358746880 hasPublicationYear "2006" @default.
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