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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
Objective: To investigate the effects of direct injection of recombinant retroviral packaging cell line carrying IL-2 and / or IL-12 gene on the activity of IL-2, IL-12 and NK cells in spleen. Methods: Construction of retroviral vector carrying h IL-2 and / or m IL-12 gene. The liver cancer cells CBRH3 were injected into the peritoneal cavity of the rats to form tumors, which were sacrificed by cervical dislocation. The tumor tissues were removed by cesarean section. The tumor tissues were excised and cut into pieces and then inoculated into one lobe of fifty Wistar rats to prepare liver cancer models. The model rats were randomly divided into saline control group, empty vector control group, IL-12 gene therapy group, IL-2 gene therapy group and IL-2 / IL-12 combined gene therapy group, with 10 rats in each group. The packaging cells containing IL-2 and / or IL-12 gene were spleen injected intraperitoneally at 1, 3, 5 and 7 days after inoculation of liver cancer. The levels of IL-2 and IL-12 in blood were detected by ELISA, the activity of NK cells was detected by radioactivity assay, and pathological and immunohistochemical examination was performed. Results: There was no significant difference in IL - 2 / IL - 12 combined with genomic hIL - 2 levels between the two groups at the third day after IL gene therapy. Pathology showed significantly increased lymphocytic infiltration in HCC tissues. The NK cell activity in IL-treated group was significantly higher than that in control group (P <0.01). Serum IL reached its peak after treatment and gradually decreased afterwards. IL - 2 / IL - 1 2 combined genome was higher than IL single genome (P <0.05). CONCLUSION: Direct injection of retroviral packaging cell line carrying IL - 2 and / or IL - 12 gene in spleen can obviously enhance the activity of NK cells. IL - combined gene therapy is better than IL -