Effect Of Transduction Of Human Wide-type P53, GM-CSF And B7-1 Genes By Recombinant Adenovirus On Glioma

Objective To explore the effect on apoptosis-inducing and immunogenecity of glioma cell line by recombinant adenovirus vector (BB-102) transducing wild type p53, granulocyte-macrophage colony-stimulating factor (GM-CSF) and B7-1 genes in vitro,and to evaluate the in vivo anti-tumor effect of the three genes modified glioma cell vacine on human glioma. The study was aimed to provide foundation for further combine gene therapy in clinic.Method In this study, We transfected glioma cell line U251 and U87 separately with recombinant adenovirus mediating green fluorescent protein(GFP) and BB-102. The transfer efficiency of adenovirus was tested. Expression of p53 gene was detected by Western Blotting, GM-CSF gene by ELISA and B7-1 gene by FITC labeled antibody in U251 and U87 cells. Also the apoptosis-inducing was observed with PI-Hoechst33342 staining and the immunogenecity was analysed by mixed lymphocyte reaction. The cytotoxic T lymphocyte reaction was also observed in glioma cells transfected with BB-102 in vitro. The in vivo anti-tumor immunological effect of genetically modifIed U251 cell vaccine on human glioma xenografts implanted into human peripheral blood lymphocyte-nonobese diabetic/severe combined immunodeficiency (HuPBL-NOD/SCID) mice was evaluated.Results The transfer efficiencies for U251 and U87 cells are 93.03% and 90.83% at 200 MOI respectively. Expression of p53, GM-CSF and B7-1 was demonstrated in both U251 and U87 cells. Growth of the infected glioma cells was inhibited, and apoptosis was induced. Immunogenecity of glioma cells transfected with BB-102 was significantly enhanced. Specific cytotoxicity against glioma cells were also induced by the infection. BB-102 infected U251 cell vaccination significantly reduced local tumor growth compared with control in the model of HuPBL-NOD/SCID. The transgenic p53, GM-CSF and B7-1 expression produces an immune response against glioma in vivo.Conclusion These results suggest that glioma cell vaccination co-transferred with p53, GM-CSF and B7-1 genes may be a feasible and effective immunotherapeutic approach in gliomas treatments.