转GM-CSF基因瘤苗联合IL-12治疗T细胞淋巴瘤的实验研究

目的　探讨用转粒细胞巨噬细胞 集落刺激因子 (GM CSF)基因瘤苗 ,联合白介素 12 (IL 12 )治疗小鼠T细胞淋巴瘤的方法。方法　将小鼠GM CSF真核表达质粒 ,用电穿孔法导入小鼠T细胞淋巴瘤细胞系RMA ,有限稀释法制备单个细胞克隆 ,经RT PCR、骨髓祖细胞增殖实验和集落形成实验 ,筛选相对高表达GM CSF的RMA克隆 ,该克隆细胞用丝裂霉素灭活后免疫小鼠 ,以诱导产生抵抗RMA肿瘤细胞再攻击的能力 ,并观察其与IL 12联合应用对淋巴瘤的治疗作用。结果　获得了高表达GM CSF的小鼠T细胞淋巴瘤克隆 ,其动物体内致瘤活性降低 ,将其用丝裂霉素灭活后作为瘤苗可使小鼠产生抗肿瘤的免疫保护力 ,与IL 12联合应用增强其抗肿瘤活性。结论　转GM CSF基因瘤苗可作为有效的抗肿瘤瘤苗 ,与IL 12联合应用较其单独应用更为有效

Experimental Study of Immunotherapy Against T Cell lymphoma by Vaccination of Inactivated Autologous Tumor Cells Engineered to Secrete GM-CSF Combined with IL-12

Objective To study the method of immunotherapy against T cell lymphoma by tumor cells transfected by GM-CSF gene combined with interleukin 12(IL-12).Methods GM-CSF eukaryotic expressing plasmid was transfected into RMA cells by electroporation.After screening by G418 and cloning by limiting dilution,a relative high GM-CSF expressing cell clone was selected by RT-PCR,hematopoietic progenitor cells proliferative assay and hematopoietic progenitor cells colony formation assay.The cell clone was inactivated by mitomycin-C and vaccinated mice to induce anti-tumor immune reaction.The cell clone,combined with IL-12,were also used to treat mice T cell lymphoma.Results A GM-CSF high expressing cell line was selected,which had a low tumor formability and could induce mice to obtain anti-tumor immune protective ability.When it was combined with IL-12,its anti-tumor ability was enhanced.Conclusion Tumor cells transfected by GM-CSF gene can be used as an effective anti-tumor vaccine.Vaccination with inactivated tumor cells engineered to secrete GM-CSF induced more potent anti-tumor activity when combined with IL-12.