In Vitro Inhibitory Effect Of Lymphocytes Which Are Activated By Dendritic Cells Charged With MAGE-1 Nonapeptide On Liver Cancer Cells

As the most powerful antigen presenting cell, dendritic cells (DCs) play an important role on the tumor immunity therapy. A nonapeptide, (H-Glu-Ala-Asp-Pro-Thr-Gly-His-Ser-Tyr-OH, EADPTGHSY) is selected for gene destination from MAGE1, one of the most highly expressed genes of liver cancer cell. This method is more effective than the previous anti-tumor researches on the ability of tumor cell inhibition. The main strategy of this research is as following: firstly, peripheral blood is used to induce differentiated DCs; secondly, DCs are charged with nonapeptide gene of MAGE1 (H-Glu-Ala-Asp-Pro-Thr-Gly-His-Ser-Tyr-OH); thirdly, external inhibition experiments are conducts on SMMC7721 liver cancer cells; fourthly, the immune response of T cells which are activated by DCs charged with MAGE1 nonapeptide is studied for further research on the internal experiment of this vaccine and to provide both clinic application foundation and the theoretical directions.Methods: Peripheral blood of health donates is used to obtain mononuclear cell through adherent culture method. The mononuclear cell is external differentiated into DCs by rhGM-CSF and rhIL-4. After charging DCs with nonapeptide of MAGE1, external inhibition effect on the SMMC7721 liver cancer cells is conducted. High magnification microscopy is used to observe the morphological characters of ripe DCs and immunohistochemical method is applied to characterizated the surface molecules expression. DCs charged with nonapeptide of MAGE1 is observed and the T cells activated by DC are mixed with SMMC7721 liver cancer cells. MTT chromatometry is conducted to examine the inhibition ability of CTLs activated by nonapeptide of MAGE1 charged DCs.Results: Mononuclear cell obtained from peripheral blood of health donates can be directionally differentiated into create DCs with typical characters. After charged with nonapeptide of MAGE1, DCs can activate CTLs which has highly inhibition effects on liver cancer cells. MTT chromatometry of liver cancer cell inhibition rate results show that the T cells activated by DC charged with MAGE1 nonapeptide has higher inhibitory effect than the T cells activated by DC without MAGE1 charging.Conclusion:DCs with typical characters and cell surface morphology can be obtained from the mononuclear cell from peripheral blood through rhGM-CSF or rhIL-4 cell factor inducing culture. The mixing result of SMMC7721 liver cancer cells with T cells which are activated by DCs sensitized with nonapeptide of MAGE1 suggests those MAGE-DCs activated T cell has obvious higher inhibition effects on SMMC7721 cells than the DCs group without the sensitization of nonapeptide of MAGE1.