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Archive > Volume 20 Issue 6 > 2013,20(6):637-644. DOI:10.3872/j.issn.1007-385X.2013.06.002 Prev Next
Prompt Report
Construction of recombinant adenovirus Ad5/F35-MAGE-A3 and its effect on maturation and apoptosis of dendritic cells in patients with melanoma
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Abstract:
Objective:To construct a recombinant adenovirus Ad5/F35 containing human melanoma-associated antigen 3 (MAGE-A3), and to observe the effect of adenovirus-mediated MAGE-A3 overexpression on the maturation and apoptosis of dendritic cells (DCs) in patients with melanoma. Methods: To choose the adenoviral vectors with the highest transfection efficiency, and then to construct recombinant adenovirus vectors and packaging adenovirus particles Ad5/F35-MAGE-A3. Immunohistochemistry and Western blotting were performed to detect the influence of Ad5/F35-MAGE-A3 infection on the expression of MAGE-A3 of DCs in healthy people and patients with kidney cancer or melanoma. Flow cytometry was used to detect the effect of Ad5/F35-MAGE-A3 infection on the maturation and apoptosis of DCs in patients with melanoma. Results: The recombinant adenovirus vector containing human MAGE-A3 was successfully constructed and adenovirus particles Ad5/F35-MAGE-A3 were packaged with the infective titer of 7.94×108 IU/ml. Ad5/F35-MAGE-A3 infection improved the MAGE-A3 expression of DCs in healthy people and kidney cancer patients (P<0.05), and it did not affect MAGE-A3 expression of DCs in melanoma patients (\[0.3352±0.1272\] vs \[0.4672±0.0704\], P＞0.05). After Ad5/F35-MAGE-A3 infection, the co-stimulatory molecule CD80 (\[20.42±0.58\]% vs \[10.22±104\]%, \[895±0.2\]%), CD86 (\[85.3±3.98\]% vs \[39.85±2.86\]%, \[34.1±4.32\]%) and HLA-DR (\[8687±436\]% vs \[63.68±3.15\]%, \[60.69±4.81\]%) which expressed on the surface of DCs was significantly higher than that of the negative control group and the blank control group (all P<0.05), but no significant difference existed in apoptosis rate (\[1.18±0.09\]% vs \[1.09±0.11\]%, P>0.05).Conclusion: Recombinant adenovirus vector can efficiently affect DCs. Ad5/F35-MAGE-A3 infection may not affect the expression of MAGE-A3 of DCs in melanoma patients, and promote the maturation of DCs without obvious cytotoxicity.
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Project Supported:
Project supported by the Key Drug Creation and Development Program of the “Eleventh Five-year Plan”（No.2009zx09503）
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