The Strategic Research About Culture And Antigen Loading Of DC Originating In PBMC

PART I The Improvement of Dendritic Cell Culture Measures in vitroObjective: Improve the measures of dendritic cell culture in vitro, to increase the rate of mature and function of DC. Method: Culture the PBMC with 1640 containing GM-CSF and IL-4, make DC mature with three conditions (1)TNF-a +CD40L(2)TNF-a +CD40L+HSP(3)TNF-a +CD40L+ligand of HSP and GM-CSF, identify the phenotype with flow cytometry and exam the bioactivity with IL-12/IFN- Y secretion. Results: DC can be cultured from monocyte and has the bioactivity, lyse corresponding target cells activity of three conditions is 28. 70%+5.84% 36.03% + 7. 28% 42. 29%+6. 74% (n=5, P=0.023) individually, IFN-r secretion of T cell is 143.92 + 42. 34,204. 24+35. 45, 241. 14 + 48. 69 (n=5, P=0.011) individually. Conclusion: HSP and ligand of HSP with GM-CSF are important factors of DC culture , can increase ability of DC motivate T cell.PART II The Research of Tumor Vaccinia With Apoptotic Tumor Cell andTumor Cell LysateObjective: To discuss apoptotic tumor cell and tumor cell lysate as antigens, presented by DC, and induce special CTL in vitro. Method: Mature DC loading with apoptotic tumor cell and tumor cell lysate to stimulate autologous T cells in order to induce special CTL and detect the special lysis of corresponding target cells by LDH, measuring IFN-r secretion of T cell by staining in cell. Results: 137Cs and MMC can make tumor cell apoptosis , apoptotic tumor cell is 93. 0+6. 20% 88. 70 +8.28% individually, apoptotic tumor cell and tumor cell lysate presented by DC, can stimulate autologous T cells to induce special CTL, and kill corresponding target cells; lysis of corresponding target cells by LDH and IFN-r secretion of T cells, measure of apoptotic tumor cell is better than tumor cell lysate. Conclusion: Apoptotic tumor cell and tumor cell lysate can induce special CTL and take as the candidate of anti-tumor peptide vaccine.