摘要
目的:构建膜锚定IL-21和分泌性GM-CSF(sGM-CSF)双表达瘤苗,并对其抗肿瘤效应及其机制作初步探讨。方法:用分子生物学方法构建双表达IL-21 gpi和sGM-CSF重组质粒,将鉴定过的重组质粒以脂质体转染B16F10细胞制成瘤苗,用流式细胞仪检测转染瘤苗IL-21 gpi和sGM-CSF的表达。以瘤苗治疗荷瘤鼠,经观察小鼠肿瘤体积、生存率来分析瘤苗的抗瘤性,并检测了瘤苗治疗鼠的细胞免疫活性。结果:正确构建了pRSC/IL-21 gpi-sGM-CSF重组质粒,转染细胞可很好地表达膜锚定IL-21和分泌性GM-CSF,制备的瘤苗能有效地发挥抗肿瘤效应,其机制与瘤苗治疗鼠的脾细胞增殖活性、NK细胞及CD8+细胞细胞毒活性增强有关。结论:成功构建了具有抗肿瘤活性的膜锚定IL-21和分泌性GM-CSF双表达瘤苗,为进一步抗肿瘤免疫治疗研究奠定了基础。
Objective：To construct the tumor vaccine that can coexpress membrane IL-21 and secrete GM-CSF,and to evaluate its anti-tumor effect and mechanisms. Methods： After the recombinant pRSC/IL-21 gpi-sGM-CSF was constructed and transfected into B16F10 cells by the molecular biological method, the expression of membrane IL-21 and the secreted GM-CSF were respectively determined by flow cytometry （FCM） .The therapeutic effect was evaluated depending on the tumor size and the survival of tumor-bearing mice after the inactivated 1 × 10^7 B16F10/IL-21 gpi-sGM-CSF tumor vaccine was inoculated into s. c C57BL/6 mice which was challenged by 1 × 10^5 B16F10 cell three days ago. The cell-mediated immune activities, such as proliferation activity of the spleen cells, the activity of NK cells, and the activity of CD8^＋ cytotoxic lymphocytes, were respectively detected. Results：The recombinant pRSC/IL-21 gpi-sGM-CSF was correctly constructed, which could express membrane IL-21 and secreted GM-CSF. The tumor vaccine had good anti-tumor effect, and the cell-mediated immunity had been improved markedly in the mice treated with the IL-21 gpi-sGM-CSF tumor vaccine, compared with the other tumor vaccines, especially of the CD8^＋ cytotoxic activity. Conclusion： The tumor vaccine that can coexpress membrane anchored IL-21 and secreted GM-CSF is constructed successfully and has a powerful anti-tumor effect,which provides a good foundation for studying anti-tumor immunity in future.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2009年第6期508-512,共5页
Chinese Journal of Immunology
基金
江苏省六大人才高峰项目(D14)
国家自然基金项目(90406023)
