Induction of tumor-specific cytotoxic T lymphocytes in vitro by dendritic cells pulsed with different glioma antigens / 中国病理生理杂志

AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-? release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies.

An experimental study of dendritic cell-based vaccine therapy for intracranial gliomas / 中国免疫学杂志

Objective:To investigate the therapeutic efficacy of immunotherapy with dendritic cells-based vaccine against intracranial gliomas in rats.Methods:C6 glioma cells were injected into the brain of Wistar rat under stereotactic monitor to establish an animal model of glioma.The precursors of dendritic cells were isolated from bone marrow of rats,stimulated in vitro with recombinant rat granulocyte-macrophage colony-stimulating factor(rrGM-CSF)and interleukin-4(rrIL-4).Cultured cell populations were confirmed to be functional DCs.These DCs were then pulsed ex vivo with C6 tumor-lysates prepared by three cycles of freezing to -80℃ and thawing to 0℃ and subsequently injected subcutaneously into rats harboring intracranial C6 tumor.Rats from different group were treated with five weekly subcutaneous injections of either control media,unpulsed DCs,or DCs pulsed with tumor-lysates.The animals were followed for survival,the percentage of CD8 +T cells in peripheral blood and cytotoxicity assay in vitro were determined by FACS.The level of cytokine IFN-? and IL-10 were detected by ELISA.Results:The results indicated that C6 glioma model rats treated with tumor-lysate-pulsed DCs led to prolonged survival time,increased the percentage of CD8 + T lymphocyte in peripheral blood in comparing with control group.Cytotoxicity assays suggest that vaccination with these tumor-lysate-pulsed DCs can induce specific cytotoxic T lymphocytes against C6 tumor cells compared with control group.Furthermore,significantly enhanced IFN-? and reduced IL-10 (even undetectable)were observed in rats treated with pulsed-DCs.Conclusion:Data supported the therapeutic efficacy of systemic vaccination with DCs pulsed with tumor-lysates against intracranial glioma.