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Recombinant adenovirusmediated survivin gene transfection of dendritic cells induces immune responses against renal cell carcinoma
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Abstract:
Abstract Objective:To investigate the antitumor immune responses against renal cell carcinoma induced by dendritic cells (DCs) transfected with survivin gene mediated by recombinant adenovirus.Methods: The DCs derived from human peripheral blood were infected with recombined adenovirus vector carrying the survivin gene. The expression of survivin protein in the infected DCs was examined by Western blotting assay; the surface expression of CD83, MHCⅡ, CD80 and CD86 by flow cytometry (FCM), Interleukin12 (IL12) in the supernatants of DCs and IFNγ released by the cytotoxic T lymphocytes (CTLs) by ELISA, the ability of DCs in proliferating allolymphocytes by mixed lymphocyte reaction (MLR), and specific killing activity of CTLs by MTT assay. Results: The DCs presented mature DCs phenotype after transfection with Adsvv. The expression of survivin protein in transfected DCs was confirmed by Western blotting analysis. The IL12 level in the supernatant of DCs transfected with Adsvv was significantly higher than that transfected with empty vector (AdCMV, P<0.01); and both of them were significantly higher than that in the control group(P<001). During MLR assay the DCs infected and not infected with adenovirus both stimulated allogeneic lymphocyte proliferation, with DCs infected with Adsvv having the strongest stimulating activity (P<0.01). After MLR the DCs infected and not infected with adenovirus both stimulated IFNγ secretion by T lymphocytes, with DCs infected with Adsvv having the strongest secreting activity (P<0.01). The CTL activity of DCs was significantly higher against renal cancer cell line 7860 positive for survivin than against hepatic cancer cell line L02 negative for survivin(P<0.01). DCs in the AdCMV group and empty control group had not cytotoxic effect on 7860 cells. Conclusion: Infection with the recombinant adenovirus encoding survivin can greatly enhance the antigen presenting ability of DCs and subsequently induce survivinspecific CTL activity and antitumor effect.
Keywords:
survivin gene ; dendritic cell ; renal cell carcinoma ; adenovirus ; immuno therapy ; gene therapy
Project Supported:
Surpported by the Medicine Science Research Foundation of Guangdong Province (No.A2006778)
