GM-CSF分泌型肝癌疫苗对荷瘤鼠脾血细胞毒性T淋巴细胞杀伤活性的影响

The inhibition of GM-CSF secreting liver cancer cell vaccine on tumor growth by stimulating cytotoxic T lymphocyte activity

目的观察粒细胞巨噬细胞集落刺激因子(GM-CSF)分泌型肝癌疫苗对移植性肝癌小鼠细胞毒性T淋巴细胞(CTL)杀伤活性的影响。方法取小鼠肝癌细胞株H22细胞1×106/只注入小鼠腹腔内,接种7d形成腹水瘤后再在小鼠体内传3代。取生长旺盛且无血性的腹水,在无菌条件下制成2×107/ml的细胞悬液,以2×106细胞/0.1ml/只接种于小鼠右前肢皮下。将肝癌细胞移植瘤动物分成3组。4天后,在右侧背部皮下进行免疫治疗,即制备GM-CSF分泌型H22肝癌瘤苗并免疫ICR小鼠(H22-GM-CSF组,n=5),同时设立无GM-CSF基因修饰H22肝癌瘤苗组(H22组,n=5)和PBs对照组(PBS组,n=5),测量各组小鼠肿瘤体积;采用细胞增殖计数法检测小鼠脾血CTL杀伤活性。结果随着效/靶比增加,各组CTL杀伤活性均增强。在效/靶比为50∶1时,GM-CSF-H22组CTL杀伤活性为60±6.1%,明显高于H22组(17.4±0.9%)和PBS组(12.2±0.6%,P<0.01);GM-CSF分泌型肝癌细胞瘤苗明显抑制H22荷瘤小鼠肿瘤生长。在21天时,H22-GM-CSF组、H22组和PBS组小鼠肿瘤体积分别为0.63±0.05mm3、1.47±0.75mm3和1.79±0.34mm(3P<0.01)。结论 GM-CSF分泌型肝癌细胞瘤苗可抑制肿瘤细胞生长,增强CTL杀伤活性。

objective To observe the effects of granulocyte - monocyte colony-stimulating factor （GM-CSF） secreting liver cancer cell vaccine on the tumor growth in mice with implanted hepotoma- 22. Methods The mice were implanted with 1 ×10^6/mouse H22 cells intraperitoneally,and 7 days later,the ascites containing tumor cells （2 ×10^7/ml） were injected into the right toes subcutaneously （2 ×10^6cells/0.1ml/mouse） for tumor formation. The mice with implanted tumor were divided into three groups,and one （n=5） were immunized with GM-CSF secret-ing liver cancer cell vaccine,one （n=5） with liver cancer cell vaccine without GM-CSF secreting activity,and one（n=5） with PBS. The volume of tumors were calculated and the killing activity of CTL were determined. Re-sults With the ratio of effect to target cell increasing,the killing activity of CTL was significantly increased in all groups;the killing activity of CTL in H22-GM-CSF group was 60±6.1% when the ratio of E/T equal to 50/1, significantly increased as compared with in H22 group （17.4±0.9%） and in PBS group （12.2±0.6%,P〈0.01）;the tumor volumes in H22-GM-CSF,H22 and PBS group were 0.63±0.05mm3,1.47±0.75mm3 and 1.79±0.34mm3（P〈0.01）, respectively,at day 21. Conclusion GM-CSF secreting H22 liver cancer cell vaccine could inhibit the growth of tumor by enhancing the CTL killing activities.