The Application Of Sapylin To Prepare Dendritic-cell-based Anticancer Vaccines From Umbilical Cord Blood

Objective To eveluate the effect of replacing fetal cattle serum with umbilical cord plasma and application of sapylin(OK432) on dendritic cells from umbilical cord blood in vitro so as to offer a new technical way of preparation high-powered dendritic-cell-based vaccines for the cancer immunotherapy later. Methods Human cord blood mononuclear cells(CBMC) were cultured in RPMI1640 containing 10% autologous plasma, GM-CSF and IL-4, some of which were supplemented with tumor lysates and/ or sapylin, and then observed by the optical microscope and the electron microscope. Method of flow cytometry was used to determine surface antigens of cells before and after culture and MTT assay was applied to measure the antigen presenting ability of DC in allo-MLR. Killing rates of autologous T lymphocytes induced by DC on different target cells were measured by LDH method.Results Cells appeared typical morphology of DCs and expressed much more antigens CD la and CD83 on the surface after culture and it was reinforced by pulsed with tumor lysate. The maturest DCs were acquired by means of treating with both of tumor lysates and Sapylin. Moreover, the allostimulate capacity of DCs and the CTL response in vitro were also enhanced by the addition of Sapylin. Conclustion Mature DCs can be induced from human cord blood mononuclear cells by this means with fewer cytokines and less time. The tumor lysate antigens could be captured by DCs which were treated with both of tumor lysates and Sapylin and then presented to lymphocytes successfully. So it would be a kind of dendritic-cell-based vaccines fit for clinical gastric cancer immunotherapy with further researches and .lay the foundation of preparation of dendritic-cell-based vaccines for the other cancers.