ABSTRACT
Objective:To investigate the strategies and methods of pancreatic cancer immunotherapy adopt cytotoxic T lymphocytes activated by dendritic cells(DCs),which modified by heat shock protein 70 peptide complex(HSP70-PC).Methods:HSP70 peptide complex from lumps of tumor-bearing mouse innoculated with pancreas cancer cell(MPC83)were purified by ConA-Sepharose and ADP-Agarose affinity chromatography.Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL-4.MTT method was applied to analyses the proliferation activities of DCs.To achieve the specific T lymphocyte cells,DCs modified with HSP70 peptide complex were coculture with murine spleen lymphocytes for 48 hours.Then these activated lymphocytes were cocultured with MPC83 at ratio of effect-target 10∶1 and 20∶1 in vitro and the killing activities were detected by MTT method.Results:High purify HSP70-PC was obtained;104 DCs need 50～100ng of HSP70-PC to modify,100?g of HSP70-PC peptide complex can be extracted from per 1g lump;T cells stimulated by HSP70-PC modify DCs showed more specific cytotoxic activity to MPC83 than lymphocytes activated by DCs without HSP70-PC in vitro.Conclusion:We can extract high purity HSP70-PC for clinical individual immunotherapy from pancreatic cancer lump by using hypobaric affinity chromatography;DCs vaccine modified with HSP70-PC has significant kill effect against pancreatic cancer cell in vitro.