Effect Of DC-CIK Pulsed With Tumor Lysate Antigens On Killing Activity Against SKOV3

Objective:To investigate the effect of dendritic cells(DC) pulsed with tumor lysate antigens of human ovarian cancer cell line SKOV3 co-cultured with cytokine induced killer(CIK) cells on the proliferation of DC and CIK cells and to investigate the impact on the cytotoxic activity against SKOV3 of CIK cells after its co-cultureding with DC that pulsed with tumor lysate antigens of SKOV3 or COC1.Methods:Peripheral blood mononuclear cells(PBMC) isolated from 12 patients with ovarian cancer were induced to obtain CIK cells and DC respectively with the corresponding in vitro-inducible factors.The Ag of SKOV3 and COC1 extracted using freeze-thaw method at the logarithmic growth phase were co-cultured with DC at the fifth day of its culture.Experiments included the experimental group and control group. CIK co-cultured with DC which were pulsed with tumor lysate antigens of SKOV3 were used as experimental group.(1)CIK co-cultured with DC without tumor antigens used as control group 1 (2) CIK cultured alone were used as control group 2.(3) DC cultured alone were used as control group 3. (4)DC pulsed with tumor lysate antigens of SKOV3 and COC1 were used as control group 4.Trypan blue staining was used to observe the proliferation of CIK cells and the impaction of DC and DC pulsed with tumor lysate antigens on CIK cells since the 1 st training day to the 20th day.The surface molecule expression of DC in DC group, Ag-DC group and Ag-DC-CIK group were measured by flow cytometry,and the impaction of Ag and CIK on DC was observed.The surface molecule expression of CIK in CIK group, DC-CIK group and Ag-DC-CIK group were also measured by flow cytometry,and the impaction of DC and Ag-DC on CIK was observed too; The cytotoxic activities of CIK group, DC-CIK group and Ag-DC-CIK group against SKOV3 were measured using lactate dehydrogenase(LDH) assay,and the impaction of DC, SKOV3Ag-DC and COC1Ag-DC on the killing activity of CIK were observed contrastly.Results:The proliferation rate of CIK cells co-cultured with DC was higher than the single CIK cells, while there was no statistical significance between the proliferation rate of CIK cells co-cultured with DC and the one of CIK cells co-cultured with antigen load DC(P>0.05). Compared with DC group and Ag-DC group, Ag-DC-CIK group led to a significant increase of mature phenotypes of DC(P<0.01); While the mature phenotypes of CIK in Ag-DC-CIK group also increased(P<0.01); The cytotoxic activities of SKOV3-DC-CIK group against SKOV3 cells were much higher than that of both CIK group, DC-CIK group and SKOV3-DC-CIK group (P<0.01).Conclusions:(1)This study indicates that DC pulsed with tumor lysate antigens of SKOV3 co-cultured with CIK cells can promote the maturity of both DC and CIK. Compared with DC, DC pulsed with tumor lysate antigens of SKOV3 can not raise the growth rate of CIK cells;(2)DC pulsed with tumor lysate antigens of SKOV3 co-cultured can improve the specific cytotoxic activity of CIK cells against SKOV3.