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Archive > Volume 22 Issue 5 > 2015,22(5):590-596. DOI:10.3872/j.issn.1007-385X.2015.05.007 Prev Next
Basic Research
Comparison of cotytoxic effects against liver cancer HepG2 cells by DCs loaded with AFP antigen in different ways
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Abstract:
Objective:To evaluate the cytotoxic effects of the cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) loaded with alpha-fetoprotein (AFP) in different approaches. Methods:Peripheral blood monocytes were isolated from healthy donors. The adhesive precursor DCs were cultured in the presence of rhGM-CSF and rhIL-4 for 6 d. The cytokine-treated DCs were then left untreated as a control, infected with recombinant AFP-carrying adeno-associated viral vector (rAAV/AFP), or loaded respectively with AFP peptide, HepG2 cell lysate, and HepG2-derived exosomes (T-exo). Changes in CD83, CD86, ICAM-1,CD58 And CD 40 in DCs before and after AFP loading were assessed by Western botting Autologous T cells were co-cultured with the various AFP-loaded DCs loaded at a ratio of 10∶1 for 48 h. The proliferative activity and cytotoxicity against HepG2 cells of DC-induced T cells were assessed by flow cytometry.Results: AFP antigen induced maturation of DCs. The expression of surface molecules CD83,CD86, ICAM-1,CD58 and CD40 in DCs infected with the rAAV/AFP vector was significantly increased compared with the nave DCs (P<0.05). AFP-loaded DCs increased the proliferation of T cells. The cytotoxic activity against HepG2 cells was (41.40±2.87)%, (44.9±4.12)%, (28.42±3.29)%, and (24.28±1.79)% for T cells after induction by T-exo-loaded DCs, rAAV/AFP-infected DCs, AFP peptide-loaded DCs and HepG2 lysate-treated DCs respectively (P<0.05). Conclusions：AFP and T-exsome are capable of increasing the proliferative activity of DCs and the cytotoxic activity of CTL against HepG2 cells. Our finding may have significant implications in the development of DC-based vaccines for liver cancer.
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Project supported by the National Health and Family Planning Commission Science Research Foundation-Health Education Joint Research Program of Fujian Province (No. WKJ-FJ-04)，and the Joint Research Foundation of Promoting Science and Technology Cooperation between School of Phamaceutical Sciences of Xiamen University and Fujian Provincial Cancer Hospital（No. 2014LY-2L-04）
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