Study On Mechanism Of Anti-hepatoma Of Tumor-infiltrating Lymphocyte Activated By Dendritic Cells Hepatocellular Cancer Vaccine

Objective： Measured the DC secreted IL-2,TNF-a,IFN-y,IL-12changes before and after sensitized and detect the anti-hepatoma activity of tumor-infiltrating lymphocyte activated by dendritic cells hepatocellular cancer vaccine in vitro, and,to explore the potential meanings of DC secreted cytokine, especially the potential mechanism of tumor-infiltrating lymphocyte activated by dendritic cells hepatocellular cancer vaccine can improve the the anti-hepatoma activity.Methods:DCs and its precursor were isolated from mouse bone marrow and stimulated by granulocyte/macrophage colony stimulating factor (GM-CSF),interleukin-4(IL-4) and whole-cell hepatocellular carcinoma antigen of H22cells. The content of IL-12, IL-2, IFN-y, TNF-a in supernatant of pre-sensitized and sensitized DCs was measured by ELISA method.Meanwhile,TIL and mouse spleen lymphocytes were respectively isolated from tumor and mouse spleen,and cultured in vitro.Then, we set eight groups as effector cells which respectively is H22+DC+TIL group、 DC+TIL group、H22+TIL group、TIL group:H22+DC+Spleen lymphocytes group、DC+Spleen lymphocytes group、H22+Spleen lymphocytes group. Spleen lymphocytes group,and the H22cells and B16cells as target cells,testing the cytolytic activity of effector cells to target cells.Results:②The content of IL-12, IL-2, IFN-γ, TNF-a in supernatant of pre-sensitized DCs was19.39±0.98pg/ml,11.19±1.18pg/ml,1.03±0.19pg/ml,2.02±0.32pg/ml respectively; and80.39±1.33pg/ml,95.27±6.52pg/ml,29.59±3.15pg/ml,75.12±7.07pg/ml in supernatant of sensitized DCs.All have statistically significant (P<0.01),DCs can improve the secretion of IL-12, IL-2, IFN-y, TNF-a after sensitization.②The H22+Dc+TIL group have very strong killing activities on H22cells(81.69±3.89%),higher than its killing activities on B16cells(P<0.01),and also higher than other groups killing activities on H22cells.Conclusion:DCs secreted IL-12、IL-2、IFN-γ、TNF-a increased obviously after H22cells sensitized. TIL activated by DC and H22whole-cell antigen have very strong and special killing activities on H22cells in vitro. DCs secreted IL-12, IL-2、IFN-γ、TNF-a increased after H22cells sensitized can help TIL to activate and the improved IFN-γ、TNF-αcan inprove its directly killing activities on H22cells,which is may the potential mechanism of tumor-infiltrating lymphocyte activated by dendritic cells hepatocellular cancer vaccine can improve the the anti-hepatoma activity.