Experimental Study On The Anti-Lung Cancer Of Dendritic Cells Vaccine Loaded With Apoptotic Tumor Cells

PartⅠ:Experimental Study on the Anti-Lung Cancer of Dendritic Cells Vaccine Loaded with Apoptotic Tumor CellsObjective: To prepare dendritic cells(DCs) from healthy human peripheral blood, which were induced mature by pulsed with apoptotic tumor cells and CD40mAb;and investigate the cytotoxicity of cytotoxic T lymphocytes(CTL) against lung cancer cell activated by DCs in vitro and in vivo.Methods: PBMC(peripheral blood monocytes) were isolated from healthy human peripheral blood by density gradient centrifugation. Briefly,PBMC were incubated in tissue culture flasks in 5%CO2 at 37℃in RPMI 1640 medium supplemented with 10% FCS.After incubation for 2 hours, non-adherent cells were removed gently,and adherent cells were cultured in mixed medium contianing rhGM-CSF(100μg/L),rhIL-4(50μg/L) to generate DCs.On day 6, immature DCs were incubated with apoptotic tumor cells.On day 7,the DCs were stimulated by CD40mAb (5mg/L) for another 2 days.Then the mature DCs were collected and cocultured with autologous T cells at ratio of 1:50 for 7 days to activited antigen specific CTL.The phenotypes changes of CTL were analyzed by flow cytometry;MTT assays were used to determine the cytotoxicity of CTL in vitro;the antitumor activity in vivo was evaluated in BALB/c nude mice bearing A549 cells.Results: The expression rates of CDlα, CD80, CD83, CD86, HLA-DR were upregulated ,after DCs were treated with apoptotic tumor cells and CD40mAb.The tumor specific CTL were generated in vitro by autologous T cells co-cultured with mature DCs,and the CTL got a significant increase of the CD8+ expression(P<0.05).In...