| Objective:To investigate the efficacy of the MUC1-2VNTR vaccine transfected dendritic cells (DC) in inducing specific anti-tumor CTL immunity against multiple myeloma cells in vitro.Methods:(1) a synthetic target gene was cloned into the vector pcDNA3.1/ myc-hisA.(2)Plasmid DNA was amplified and extracted by Endofree Plasmid Mini Kit with alkaline lysis method.(3) DCs isolated from the blood mononuclear cells of healthy people,in vitro which stimulated by recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) and recombinant human interleukin 4 (rhIL-4) induced into mature DC cells;(4) DCs were transfected with the DNA vaccine MUC1-2VNTR (DC-2VNTR) by liposome mediate. The empty plasmid (DC-pcDNA3.1) and the lipidosome-treated DCs (DC-Lipo) were used as control;(5) The expression of MUC 1-2VNTR was confirmed by Western blot;(6) In vitro the homologous T cell was stimulated and the multiple myeloma cells were used as the target cell to check the specific cytotoxity of the CTLs induced by DC-2VNTR.Results:(1)The recombinant plamid vctors pcDNA3.1-2VNTR/myc-hisA were successfully established.(2)The peripheral blood DC cells were cultivated and identified.(3) After transfection, the expression of MUC1-2VNTR was confirmed by Western blot;(4) DC-2VNTR CTL could significantly kill multiple myeloma cells, and much higher than the group of DC-pcDNA3.1 and DC-Lipo (P<0.05);Conclusions:The MUC1-2VNTR transfected DC could induce the specific CTLs, that effectively kill the multiple myeloma cells. |
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