Abstract:
Objective To investigate the immunological competence of dendritic cell loaded with CD133+ hepatocellular carcinoma cell RNA (CD133+ HCC RNA-DC) vaccine.
Methods HCC cells were separated from hepatocellular carcinoma tissues through Enzyme Digestion, and then CD133+ HCC cells were sorted by flow cytometry. CD133+ HCC RNA-DC vaccine was obtained. Flow cytometry was used to detect the phenotype of DC, and ELISA was applied to determine the level of DC's secretion of IL-12. Mixed lymphocyte reaction was applied to test the ability of DC to stimulate the proliferation of autologous T lymphocytes in vitro.
Results The expression levels of HLA-ABC, HLA-DR, CD86, CD80 and CD83 in CD133+ HCC RNA-DC group were (96.52±2.02)%, (92.17±3.04)%, (94.25±3.28)%, (55.14±1.67)% and (40.53±2.31)%, respectively. Phenotypes expression levels were not significantly different among CD133+ HCC RNA-DC, HCC-DC and mature DC. The secretion of IL-12 in CD133+ HCC RNA-DC group, HCC-DC group, mature DC group and immature DC group were (421.50±3.12), (418.20±1.10), (324.20±2.19) and (102.47±4.60) pg/ml, respectively; the difference of the former two had no statistical significance (P=0.14), and the amounts of the former two were higher than that of the latter two (P < 0.05). The proliferation of autologous T lymphocytes stimulated by CD133+ HCC RNA-DC and HCC-DC were stronger than those by mature DC and no DC (P < 0.05).
Conclusion The dendritic cell vaccine loaded with CD133+ hepatocellular carcinoma cell RNA (CD133+ HCC RNA-DC) has mature phenotype and can effectively stimulate the proliferation of autologous T lymphocytes in vitro.
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