The Research Of Enhancement Of CIK Anti-tumor Effect By DCs Derived From Umbilical Cord Blood

Objective: Dendritic cell is the strongest system of antigen presenting cells that function to initiate several immune responses and CIK cells show the highest cytotoxic effect on the malignant tumor at present. To define the strong anti-tumor responses of the both and to establish the best effecprotocol for the culture of CIK cells in vitro, We collected the umbilical cord blood mononuclear cells(CBMNCs), and used antigen to stimulate DCs , then they were co-cultured with the CIK cells. We do all above to explore whether coculture of dendritic cells with CIK lead to an increase of cytotoxicity against tumor cells . Methods: 1. In order to obtain the CBMNCs, fresh aseptic cord blood was isolated by ficoll density gradient centrifugation. After washing, the CBMNCs were resuspended in complete RPMI1640 on 24 cells plate, interferon- γ (IFN- γ ), interlukin-2 (IL-2), interlukin-1 (IL-1), OKT₃, interferon-4(IFN-4), GM-CSF and TNF-α were added. We could generate a great deal of cells by combining use of the above seven types of cytokines. Then we Evaluated the proliferation rations, phenotypes by FCM and observed the configuration of the two kind of cells. 2.On the 10 days, we did the co-culture of the DCs and CIK cells devided into three groups:(1)the group of co-culture with antigen stimulation(CIK-A-DC);(2) the group of co-culture without antigen stimulation(CIK-DC);(3)the group of CIK alone(CIK). An aliquot of cells were removed from the culture at various time points. The phenotypes of CIK cells AND DCs were identified by flow cytometric analysis.