Anti-Tumor Activity of Dendritic Cell/Myeloma Vaccine in Vitro / 实用儿科临床杂志

ABSTRACT

Objective To induce mice bone marrow(BM) mono-nuclear cells into dendritic cells(DC) in vitro,and then produce function vaccine of anti-myeloma by bone marrow derived dendritic cells(BM-DC).Methods Mono-nuclear cells separated from mice bone marrow were cultured in tissue culture plastics which supplemented granulocyte-macrophage clonystimulating factor(GM-CSF),IL-4.During the 8 days,there were numerous of mature DCs outgrown,and then fused with SP 2/0 myeloma cells in logarithmic growth phase cultured with 8-azaguaine by polyethylene(PEG).The resulting purified DC/myeloma hybrids were generated by selecting with hypoxanthine-aminopterin-thymidine(HAT).The specific anti-tumor activity was examined in vitro.Results Mono-nuclear cells derived from BM cultured by supplement propriate cytokines milieu in tissue culture plastics,and then exposed to lipopolysaccharide(LPS).There were plenty of mature DCs appear as early as day 8,and exhibit dramatic veils of cytoplasm and extensive dendrites in their surfaces,high express CD11c(83.19%),major histocompatibility complex(MHC)-Ⅱ(95.25%),CD86(89.24%),their capacity to stimulate primary T cell responses in mixed leukocyte reaction(MLR) would be shown strongly.Hybrids could grow well through PEG fusion and HAT selection.These hybrids could stimulate naive T cells into cytotoxic T-lymphocyte(CTLs) directed against SP 2/0 myeloma,and exhibit strong specific killed ability.Conclusions BM-DCs is fused with tumor cells ensure the tumor associated antigens could be processed and presented effectively,and stimulate specific anti-tumor immunity so that prevent or cure this kind of tumor.So it's feasible in practice to produce anti-tumor vaccine by BM-DCs.