The Anti-tumor Effects Of Dendritic Cells Co-culturing With CIK Cells

Section I The proliferation profile in vitro and anti-tumor effects of dendritic cells co-culturing with CIK cellsObjective To Observe the changes of CIK(cytokine induced killer) cells after co-culturing with autologous dendritic cells(DC)in their morphology, phenotype, proliferation activity and cytotoxic actvity. Methods DC and CIK cells were generated ,respectively, by cytokines induction of culturing peripheral blood mononuclear cells (PBMC ) of healthy blood donor. Unstimulated DC and DC pulsed by A549 lung cancer cells lysate antigen were individually co-cultured with CIK cells. Expression of markers typical for these effector cells were identified at different intervals by flow cytometry analysis. The proliferation activity of these effector cells were determined. IFN-γand IL-12 secreted by CIK cells and co-cultured cells were detected by enzyme-linked immunosorbent assays (ELISA). MTT assays were used to messure the cytotoxic actvity on A549 cells and BEL-7404 cells in vitro. Results Co-culture of CIK cells with DC produced a new cell population, it proliferated and expanded 1029 fold in vitro incubation by day 19, the proliferation was much higher than 307 fold of CIK cells . Co-culture of CIK cells with DC pulsed by A549 lung cancer cells lysate antigen had a much higher proliferation for 1092 fold. Interactions between DC and CIK cells caused changes in the surface molecule expression of both populations,led to an increase of IFN-γand IL-12 secretion, especially IFN-γ. Co-culture of CIK cells led to a significant increase of DC-specific costimulatory molecules and antigen-presenting molecules in DC cells. Analogously, the cocultivation caused an increase in corresponding cell surface markers such as CD3+CD56+ ,CD8+ and an decrease of CD45RA+ on CIK cells. Co-culture of CIK cells with DC led to a significant increase in cytotoxic activity ,as messured by MTT assays . Cytotoxic activity could be further increased using DC pulsed with A549 tumor cells lysate antigen. To A549 lung cancer cells,the killing activity of CIK cells ,which were nor activated by DC, was significantly lower than that activated byDC pulsed with tumor cells lysate antigen(p<0.05). there was no significant difference in killing activity of CIK cells on BEL-7404 cells. Conclusion CIK cells co-cultured with DC is more powerful than CIK cells alone in anti-tumor reaction. DC pulsed with A549 tumor cells lysate antigen can further increase the cytotoxic activity of immunologic effector cells against A549 lung cancer cells. Section II The experimental management by dendritic cells and CIK cells for lung hematogenous metastasis of colon cancerObjective To determine the feasibility of treatment with dendritic cells and CIK cells for lung hematogenous metastasis of colon cancer.Methods Balb/c nude mice was venous injected with Lovo colon cancer cells to establish the hematogenous metastasis model. Then the tumor-bearing nude mices were treated by dendritic cells and CIK cells by day 3,7 and 10. RT-PCR was used to investigate the expression of CEA gene in Lovo colon cancer cells, and the lung, liver,renal tissues of normal nude mices and tumor-bearing nude mices.Results 　The venous injection of Lovo colon cancer cells resulted in lung metastasis and death in the control mice.In contrast ,treatment with dendritic cells and CIK cells led a stronger suppressive effect on the tumor growth ,elongatd life time of the tumor-bearing nude mices,inhibited tumor metastasis and rescued the tumor-bearing mice from death.Conculusion 　 CIK cells have a stronger significant suppression against growth of tumor cells. Dendritic cells could enhance the anti-tumor effects of CIK cells, which were potent in the control of tumor hematogenous metastasis. The result provides an experimental basis for CIK cells clinical application as an adoptive immunotherapy. Section III The anti-tumor effects of dendritic cells co-culturing with CIK after chemotherapyObjective To determine the changes of CI...