摘要
Background Objective evaluation of the antitumor effect of interleukin-12 （IL-12） gene-transfected dendritic cell （DC）vaccine on laryngeal carcinoma requires in vivo and in vitro tests. The aim of this study was to investigate the function of IL-12 gene transfected DC at initiating specific immune response to laryngeal carcinoma in vitro.Methods Recombinant adenovirus with IL-12 gene was constructed. DCs were isolated from the peripheral blood of patients with laryngeal carcinoma, pulsed with tumor lysate of laryngeal carcinoma cells （DC＋Ag）, and transfected with IL-12 （DC-IL-12＋Ag）. The cells pheotypes including CD83, CD86 and HLA-DR on surface of DCs were assayed by flow cytometry （FCM）. The concentration of IL-12 in culture supernatant of DCs and interferon γ （IFN-γ） in culture supernatant of T cells cocultured with DCs were quantified by ELISA. Methyl thiazolys tetrazolium （MTT） was used to evaluate proliferation of autologous T lymphocytes and activation of cytotoxic T lymphocytes （CTL） stimulated by IL-12-transfected DCs pulsed with tumor lysate against laryngeal carcinoma cells.Results The recombinant adenovirus expressing IL-12 gene was constructed successfully. Gene-transfected DC plused with tumor lysate with IL-12 （DC-IL-12＋Ag） expressed higher level of CD83, CD86 and produced higher level of IL-12 than untransfected DCs （DC＋Ag） （CD83： （60.2±1.8）% vs. （50.7±1.2）%, P ＜0.05; CD86： （88.9±2.1）% vs.（78.2±3.9）%, P ＜0.05; IL-12： （262.5±3.0） ng/L vs. （103.8±5.1） ng/L, P ＜0.05）. The proliferation of autologous T lymphocytes and production of IFN-γ stimulated by DC transfected with IL-12 were more obviously than untransfected DCs. Cytotoxicity of CTL stimulated by IL-12-transfected DC pulsed with tumor lysate against laryngeal carcinoma cells were significantly stronger than stimulated by untransfected DC.Conclusion It is a promising approach for IL-12-transfected DC pulsed with tumor lysate to increase the antitumoral effect.
Background Objective evaluation of the antitumor effect of interleukin-12 （IL-12） gene-transfected dendritic cell （DC）vaccine on laryngeal carcinoma requires in vivo and in vitro tests. The aim of this study was to investigate the function of IL-12 gene transfected DC at initiating specific immune response to laryngeal carcinoma in vitro.Methods Recombinant adenovirus with IL-12 gene was constructed. DCs were isolated from the peripheral blood of patients with laryngeal carcinoma, pulsed with tumor lysate of laryngeal carcinoma cells （DC＋Ag）, and transfected with IL-12 （DC-IL-12＋Ag）. The cells pheotypes including CD83, CD86 and HLA-DR on surface of DCs were assayed by flow cytometry （FCM）. The concentration of IL-12 in culture supernatant of DCs and interferon γ （IFN-γ） in culture supernatant of T cells cocultured with DCs were quantified by ELISA. Methyl thiazolys tetrazolium （MTT） was used to evaluate proliferation of autologous T lymphocytes and activation of cytotoxic T lymphocytes （CTL） stimulated by IL-12-transfected DCs pulsed with tumor lysate against laryngeal carcinoma cells.Results The recombinant adenovirus expressing IL-12 gene was constructed successfully. Gene-transfected DC plused with tumor lysate with IL-12 （DC-IL-12＋Ag） expressed higher level of CD83, CD86 and produced higher level of IL-12 than untransfected DCs （DC＋Ag） （CD83： （60.2±1.8）% vs. （50.7±1.2）%, P ＜0.05; CD86： （88.9±2.1）% vs.（78.2±3.9）%, P ＜0.05; IL-12： （262.5±3.0） ng/L vs. （103.8±5.1） ng/L, P ＜0.05）. The proliferation of autologous T lymphocytes and production of IFN-γ stimulated by DC transfected with IL-12 were more obviously than untransfected DCs. Cytotoxicity of CTL stimulated by IL-12-transfected DC pulsed with tumor lysate against laryngeal carcinoma cells were significantly stronger than stimulated by untransfected DC.Conclusion It is a promising approach for IL-12-transfected DC pulsed with tumor lysate to increase the antitumoral effect.
