The Experimental Study Of DC/B16Malignant Melanoma Fusion Cells Induce T Cell-specific CTL

ObjectiveThis study was to explore whether DC/Malignant Melanoma Fusion Cells canenhance the induction of specific cytotoxic T lymphocytes and to strengthen thekilling effect of B16malignant melanoma cells.MethodsB16malignant melanoma cells were fused with mouse bone marrow-deriveddendritic cells (DC) with polyethylene glycol (PEG).After the fusion,we used DC cells,B16malignant melanoma cells as control groups, DC/B16malignant melanoma fusioncells as the experimental group, drew fusion cells growth curve and did the autologousT lymphocyte proliferation test stimulated by the fusion cells.Then we used DC cells,B16malignant melanoma cells, DC and B16malignant melanoma co-cultured cells ascontrol groups, DC/B16malignant melanoma fusion cells as the experimentalgroup,used ELASA method to detect the induction of CTL activity by DC/B16malignant melanoma fusion cells and its killing role to B16malignant melanoma.Experimental data was counted with x±s, We chose the one-way ANOVA andLSD’s post hoc test statistical methods to do statistical analysis by SPSS13.0software.ResultsFirst, we successfully achieved the fusion of mouse bone marrow-derived dendriticcells (DC) and B16malignant melanoma cells.Then,we found the growth of DC/B16fused cells was similar with DC cells,but notB16malignant melanoma cells. The proliferation of DC/B16fused cells wasconsiderably weaker than the B16malignant melanoma cells,show that the DC/B16fused cells was closer to DC growth and proliferation characteristics.The results of theefficiency of sensitized T cells proliferation showed the DC/B16malignant melanomafusion cells can effectively stimulate homologous T lymphocyte proliferation,and its ability to promote T-cell proliferation was significantly higher than the DC cells groupand B16cells group,there were significant differences between the groups (p <0.01).We used DC cells, B16malignant melanoma cells, DC and B16malignantmelanoma co-cultured cells as control groups, the DC/B16fused cells as theexperimental group,we determined the IFN-gamma secreted by sensitized T cells ineach group with ELISA method and found that the secretion of the DC/B16malignantmelanoma fused cells was significantly higher than the other control groups, thedifference between the groups was statistically significant (p <0.01).After vitro tests,we found the CTL induced by the DC/B16malignant melanoma fused cells hadsignificant specific killing effect on B16malignant melanoma.But,B16malignantmelanoma cells and DC cells cultured alone group, DC cells and B16malignantmelanoma cells mixed culture group were not significant. The difference between theexperiment group and control groups was statistically significant (p <0.01).ConclusionsDC/B16malignant melanoma fused cells could significantly promote theproliferation of autologous T cells, could significantly promote the INF-gammasecretion and cytotoxic T cell.The Significant specific killing effect of CTL induced byDC/B16malignant melanoma fused cells to B16malignant melanoma could play animportant role in anti-tumor immunity.