| Objective : To investigate the differences of the anti-tumor effect through comparing study of the cell morphology, proliferation, killing activity to tumor cells and the ability of secretion of cytokines between cascade primed immune(CAPRI)cells and cytokine-induced killer(CIK) cells in vitro.Methods:Peripheral blood 50 ml were respectively obtained from three healthy volunteers, then separated three copies of the suspension of peripheral blood mononuclear cells(PBMCs) by Ficoll-Conray density gradient centrifugation.The cells were adjusted concentration of 1×106/ml, PBMCs were divided into two parts equally. CAPRI cells could be obtained from one part of the PBMCs through a series of induction, amplification and culture of cytokines, while another part obtained them through many kinds of cytokines induction. During the culture process,the proliferation capacity and the activity of this two kinds of cells were tested, and also observed the changes in cell morphology. At the 14 th day, the cells could be harvested and tested by the following process: 1.The cells were dyed and tested their activity with Trypan Blue; 2.The cells were counted, drawed the growth curve and tested its proliferation activity; 3.The killing activity of these two kinds of cells were tested with LDH release against K562 cells and MCF-7 cells; 4.Their secretion levels of IFN-γ and IL-2 were detected with the ELISPOT technology.Results:1. The amounts of CAPRI cells were lower than that of CIK cells on the 5th and14 th day of their culture(P<0.001/P<0.001), and the activities of them were both beyond 95%;2.The killing activity of CAPRI cells towards leukemia K562 cell line corresponding to the effector-target ratio 40:1, 20:1 were(55.1±3.25)% and(35.0±2.65)%, and both of the data were lower than CIK cells(60.0±3.03)%,(39.7±3.42)%(P=0.004/0.005). While CAPRI cells killing activity towards breast MCF-7 cell line were(71.5±3.06)%,(56.0±3.76)%,(40.2±2.90)%, corresponding to the effector-target ratio 40:1,20:1,10:1, and these data were higher than CIK cells(65.4±3.86)%,(49.5±3.91)%,(36.1±3.73)%(P =0.002/0.003/0.02);3.By using ELISPOT technology under different cell concentration( 1×106/ml and 5×105/ml), we drawed the conclusion that the IFN-γ secretion level of CAPRI cells were(126.2±10.31),(48.8±10.99), which were lower than that of CIK cells(409.3±7.76),(159.3±15.45)(P<0.001/P<0.001), while as to IL-2, CAPRI cells(325.1±16.24),(113.8±11.29) were higher than CIK cells(212.0±16.58),(70.7±10.57)(P<0.001/P<0.001).Conclusion:1. The proliferation of CAPRI cells was lower than CIK cells.2.The killing activity towards NK cell targeting leukemia K562 cell line, the CAPRI cells usually exhibited a lower advantage than the CIK cells, but they both had a higher killing activity towards solid tumor the breast cancer cell line MCF-7,which were extremely sensitive to NK.3. The level of secreting IFN-γ by CAPRI cells was lower than CIK cells, while the level of secreting IL-2 was higher than that of CIK cells. Experiments showed that.4. There were differences between CAPRI cells and CIK cells on proliferation, killing activity and function of cytokine secretion, presumably that was caused by the different anti-tumor mechanism of two kinds of cells. |
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