摘要
Background Survivin is a rather specific gene in tumor tissue. We transfected dendritic cells （DCs） with recombinant adenovirus （Ad） containing survivin gene and granulocyte-macrophage colony-stimulating factor （GM-CSF） gene and tested the inducing effect of the transfected DCs on cytotoxic T lymphocytes （CTL） to kill leukemic cells. Methods After derived from the peripheral, DCs was assayed by mixed leukocyte reaction （MLR） tests. Lactate dehydrogenase （LDH） release test was used to evaluate cytotoxicity of CTL. Results Expression of survivin in transfected DCs was confirmed by Western blotting analysis. GM-CSF expression was confirmed by enzyme-linked immunosorbent assay （ELISA）. In MLR assay, DCs coinfected with Ad-survivin and Ad-GM-CSF induced higher allogeneic lymphocyte reaction than control DCs at ratios of 1：5, 1：10, 1：50 and 1：100. DCs coinfected with Ad-survivin and Ad-GM-CSF had much higher activity of CTL to HL-60 cells than DCs infected with Ad-survivin only, Ad-GM-CSF only, or control DCs. Levels of interleukin-12 （IL-12） and interferon gamma （IFN-γ） in lymphocyte supernatants containing DCs coinfected with Ad-survivin and Ad-GM-CSF were significantly higher than those in the control group. Conclusion DCs coinfected with Ad-survivin and Ad-GM-CSF induce much higher anti-leukemic response in vitro than those infected with either factor. Therefore, adenovirus vectors containing survivin and GM-CSF genes may be promising vaccine candidates for leukemia therapy.
Background Survivin is a rather specific gene in tumor tissue. We transfected dendritic cells （DCs） with recombinant adenovirus （Ad） containing survivin gene and granulocyte-macrophage colony-stimulating factor （GM-CSF） gene and tested the inducing effect of the transfected DCs on cytotoxic T lymphocytes （CTL） to kill leukemic cells. Methods After derived from the peripheral, DCs was assayed by mixed leukocyte reaction （MLR） tests. Lactate dehydrogenase （LDH） release test was used to evaluate cytotoxicity of CTL. Results Expression of survivin in transfected DCs was confirmed by Western blotting analysis. GM-CSF expression was confirmed by enzyme-linked immunosorbent assay （ELISA）. In MLR assay, DCs coinfected with Ad-survivin and Ad-GM-CSF induced higher allogeneic lymphocyte reaction than control DCs at ratios of 1：5, 1：10, 1：50 and 1：100. DCs coinfected with Ad-survivin and Ad-GM-CSF had much higher activity of CTL to HL-60 cells than DCs infected with Ad-survivin only, Ad-GM-CSF only, or control DCs. Levels of interleukin-12 （IL-12） and interferon gamma （IFN-γ） in lymphocyte supernatants containing DCs coinfected with Ad-survivin and Ad-GM-CSF were significantly higher than those in the control group. Conclusion DCs coinfected with Ad-survivin and Ad-GM-CSF induce much higher anti-leukemic response in vitro than those infected with either factor. Therefore, adenovirus vectors containing survivin and GM-CSF genes may be promising vaccine candidates for leukemia therapy.
基金
This study was supported by the grants from the Natural Science Foundation of Fujian Province （No. C0810045） and the Medical Innovation Program of Fujian Province （No. 2007-CXB-16）.
