Cytotoxicity Of K-ras Induced Dendritic Cells Enhance The Effect Of Cytokine Induced Killer Cells Against Pancreatic Cancer Cells

Objective:Pancreatic cancer is exceptionally aggressive with high mortality. Since no effective therapeutic methods have been found to now,it is urgent to find more powerful method to diagnose the disease earlier and treat it more effectively. K-ras mutant peptide in 12 of pancreatic cancer in up to 75%-90%,is considered a potential immunotherapy of pancreatic cancer. Dendritic cells are the most powerful professional antigen-presenting cells in vivo. Its play a pivotal role in the tumor cells and T lymphocytes in the interaction between. Cytokine induced killer cells are a kind of non-MHC restricted the efficient dissolution of the tumor cytotoxic T-cells,which induced by several cytokines such as IFN-γ,IL-2,CD3mcab and so on. Its not only improved the value-add and antitumor activity of CIK,but also more specific to antitumor when dendritic cells co-cultured with cytokine induced killer cells. We observe the effect of cytokine induced killer cells (CIK) co-cultured with DC pulsed with K-ras mutant peptide against pancreatic cancer.for provide experimental basis for pancreatic cancer immunotherapy.Methods:Peripheral blood mononuclear cells(PBMC) were isolated from peripheral blood of healthy donors.DCs and CIK cells were induced in vitro and then cocultured.Increased number of cells were counted by tapan-blue staining, immunophenotype of CIK and DCCIK were analyzed by flow cytometry.The IFN-yand IL-12 levels of the cultured supernatants were detected by ELISA kits. The cytotoxic ability against pancreatic cells(PANC-1) in vitro of pure CIK(CIK group),CIK induced by DC(DCCIK group) and CIK induced by K-ras load DC(K-ras-DCCIK group) was detected and compared by MTT assays.Results:At the 15th day of culture,the amplification fold of DCCIK was 21.2±3.1,which was 1.31 times as many as CIK group(P<0.05).The expression of CD3+CD56+,CD3+CD8+ were identified highly when CIK were co-culture with-Kras induced dendritic cells, the level of IFN-yand IL-12 in the Kras-DCCIK group were noticeably higher than the other groups(P<0.05). The cytotoxic ability of K-ras-DCCIK group was (47.6±0.02)%,Better than the DCCIK group((40.1±0.03)%and CIK group(32.5±0.02).While the cytotoxity effects were futher enhanced at a higher effecter to targat radio.Conclusion:K-ras mutant peptide can promote DC to mature. K-ras induced DC can increase the amplification amount of CIK and enhanced the cytotoxic effect of CIK against pancreatic cancer cells(PANC-1).