大鼠肝癌模型脾内转染IL-2和(或)IL-12基因对NK细胞的激活作用

目的 :研究脾内直接注射携带 IL- 2和 (或 ) IL- 1 2基因的逆转录病毒包装细胞株对血 IL- 2和 IL- 1 2以及 NK细胞活性的影响。方法 :构建携带 h IL- 2和 (或 ) m IL- 1 2基因的逆转录病毒载体。将肝癌细胞 CBRH3注入大鼠腹腔 ,形成肿瘤 ,断颈处死 ,剖腹取出肿瘤组织 ,剪碎后接种于 5 0只 Wistar大鼠肝脏一叶 ,制备肝癌模型。模型大鼠随机分为生理盐水对照组、空载体对照组、IL- 1 2基因治疗组、IL- 2基因治疗组及 IL- 2 / IL- 1 2联合基因治疗组 ,每组 1 0只。含 IL- 2和 (或 ) IL- 1 2基因的包装细胞于肝癌接种后 1、3、5、7d进行脾内注射转染脾细胞。 EL ISA法检测大鼠血 IL- 2和 IL- 1 2浓度 ,放射性活度测定法检测 NK细胞活性 ,并进行病理学和免疫组化检查。结果 :IL基因治疗后 3d,IL - 2 / IL - 1 2联合基因组血清 h IL - 2与单基因治疗组相比无显著差异。病理示治疗后肝癌组织中淋巴细胞浸润明显增多。 IL治疗组 NK细胞活性较对照组显著增高 (P<0 .0 1 )。治疗后 3d血清 IL达高峰 ,以后逐步下降。 IL - 2 / IL - 1 2联合基因组较 IL单基因组增高 (P<0 .0 5 )。 结论 :脾内直接注射携带 IL -2和 (或 ) IL - 1 2基因的逆转录包装细胞株可明显增强 NK细胞活性 ,IL联合基因治疗优于 IL单基因

Activation of NK cell by transfecting interleukin-2 and/or interleukin-12 genes into spleen in rat liver cancer

Objective:To study the effects of retroviral packaging cells encoding mouse interleukin-12(mIL-12)and hu-man interleukin-2(hIL-2)genes on serum IL and NK cell activity in rat liver cancer model.Methods:Rats were randomized into5groups:normal saline control,blank vector control,hIL-2gene,mIL-12gene and hIL-2/mIL-12fused gene group.Hepatoma CBRH3cells were injected into abdominal cavity.After rats were sacrificed,tumors were cut into pieces and were implanted into one lobe of50rats liver to establish liver cancer model.The retroviral vectors encoding IL-2and/or IL-12genes were constructed.The retroviral packaging cells transfected by the vectors were injected into rat spleen1,3and7d after implantation.The serum IL-2and IL-12levels,NK cell activity,pathology changes and toxic reaction were all investigated.Results:Levels of hIL-2and mIL-12in IL-2/IL-12group and single gene group had no significant changes.There were a number of lymphocytes infiltrated into liver tissues in IL-2/IL-12gene group.Compared with control,NK cell activity was increased markedly in IL group.The activity of NK cell peaked at d3and decreased gradually afterwards.IL-2/IL-12gene ac-tivated NK cell significantly compared with IL group alone(P<0.05).Conclusion:Both IL-2and IL-12genes injected direct-ly into spleen can increase the NK cell activity,and IL-2/IL-12gene is more efficient in activating NK cells than IL gene alone.