白细胞介素-12基因再修饰基因瘤苗免疫小鼠的脾细胞过继治疗肝癌

The adoptive theraputic effect of interleukin-12 gene transfected splenocytes from the mice immunized by immunogene tumor vaccine on implanted liver cancer

目的 探讨白细胞介素 (IL) 2和B7 1双免疫基因转染肝癌细胞瘤苗免疫小鼠后获得的脾淋巴细胞经mIL 12基因修饰后治疗小鼠肝癌的可行性和疗效。方法 用 2 0 0PFU 细胞滴度的重组腺病毒载体 (Adv)将人IL 2和B7 1基因共同转染小鼠肝癌Hepal 6细胞株 ,经 80mg L丝裂霉素 (MMC)处理制备成肝癌细胞瘤苗 ,免疫同系小鼠后分离其脾淋巴细胞 (SLC) ,转染mIL 12基因 (Adv滴度 2 0 0PFU 细胞 )后注射入直径 1cm的小鼠皮下移植肝癌内 ,观察抗瘤效果。结果 转mIL 12基因SLC治疗组小鼠瘤体增加值最小 ( 0 .0 8± 0 .0 5 )cm3,与各对照组比差异有显著性 [(转染BGFP基因SLC、未转染SLC和生理盐水注射组分别为 ( 3 .46± 0 .15 ) ,( 3 .5 6± 0 .2 3)和( 8.12± 0 .5 4)cm3,P <0 .0 5 ]。结论 IL 2和B7 1双基因修饰肝癌瘤苗诱导小鼠产生的免疫脾淋巴细胞可能成为一种新的过继免疫治疗效应细胞及携带IL 12基因的载体细胞 ;基因治疗、特异性主动免疫和过继性细胞免疫治疗结合将有更优越的抗瘤效果。

Objective To find the feasibility and antitumor effect of mIL-12 gene transfected splenocytes derived from IL-2 and B7-1 geneco-transfected liver cancer vaccines immunized mice.Methods The murine liver cancer cell strain Hepal-6 cotransfected with both human IL-2 and B7-1 gene via racombinant adenovirus vectors (200 PFU/cell)were treated with 80 mg/L mytomycin (MMC) to prepare liver cancer cell vaccines.The splenocytes isolated from the mice immunized with these vaccines were transfected with mIL-12 gene or BGFP gene or nontransfected and injected into the mice xenograft liver cancer with tumor diameter being 1 cm) respectively.The adoptive anticancer effect was investigated.Results The increasing volume of the tumor injected with mIL-12 gene transfected splenocytes intratumorally was significantly smaller than in the control groups [The increasing volume of the tume in the mIL-12 gene transfected,BGFP gene transfected,nontransfected splenocytes and saline injected groups was (0.08±0.05),(3.46±0.15),(3.56±0.23) and (8.12±0.54) cm3,respectively, P <0.05].Conclusion Significant tumor inhibitory effect was demonstrated in the immuno-activated splenocytes derived from IL-2 and B7-1 gene co-transfected liver cancer vaccines immunized mice and was further improved while modified with mIL-12 gene.It is suggested that it would be more efficiency if gene therapy,specific active immunotherapy in combination with adopive cellular immunotherapy were adopted.