口服携带人IL-12、GM-CSF基因的减毒沙门菌抗肿瘤作用的试验  

Study on the anti-tumor effect of an attenuated live Salmonella containing human IL-12 and GM-CSF genes administrated by oral route

作　　者：李玉华[1] 陈慧[2] 郭坤元[1] 解咏梅[2] 杜江[1] 黄建生[2] 任大明[2] 

机构地区：[1]第一军医大学附属珠江医院血液科 [2]复旦大学生命科学院遗传学研究所遗传工程国家重点实验室

出　　处：《中华微生物学和免疫学杂志》2001年第6期610-614,共5页Chinese Journal of Microbiology and Immunology

摘　　要：目的　探讨以减毒沙门菌作为口服基因治疗载体的可行性。方法　通过电转化法将真核表达载体pCMVhIL 12、pCMVhGM CSF、EGFPN1导入减毒鼠伤寒沙门菌SL32 6 1中 ,经由胃管喂给BALB c和C5 7BL 6小鼠。 6周后分别用 4T1乳腺癌细胞和Lewis肺癌细胞进行攻击。通过流式细胞仪、共聚焦显微镜检测绿色荧光蛋白在小鼠各组织中的表达 ,通过PCR和ELISA法检测IL 12、GM CSF基因的整合和表达情况。并考察肿瘤的受抑情况和小鼠的生存期。结果　在小鼠的肝、脾、小肠、肾脏和肿瘤中可检测到绿色荧光蛋白的表达和相应细胞因子基因的整合。血清中相应的细胞因子水平较对照组明显升高 (P <0 .0 5 ) ,生存期远远超过对照组小鼠 (P <0 .0 5 )。结论　减毒沙门菌可作为口服基因治疗载体 ,可能为肿瘤的治疗提供一条简便、安全。Objective To study the anti tumor potentiality of oral gene therapy using attenuated live Salmonella . Methods An attenuated live AraA - autotrophic mutant of Salmonella typhimurium (SL3261) was used as a carrier of eukaryotic expression vectors (EGFPN1, pCMVhIL 12, pCMVhGM CSF) and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T1 or Lewis tumor cells respectively. Flow cytometer and confocal microscopy were used to detect the expression of GFP in murine tissues. PCR and ELISA were used to detect the integration and expression of hIL 12, hGM CSF gene. The survival time of mice was also investigated. Results GFP expression and hIL 12, hGM CSF gene integration were identified in animals′ liver, spleen, intestinal, kidney and tumor. Serum level of hIL 12?mIFN γ increased significantly in the hIL 12 orally treated mice ( P <0.05) and serum level of hGM CSF also increased in hGM CSF orally treated mice ( P <0.05); both of them are believed to result in the prolongation of the survival time of those mice in comparison with that of the control ( P <0.05). Conclusion Oral gene therapy using attenuated live Salmonella has the potential to be developed as a simple, effective and safe therapy against tumor. [

关 键 词：基因治疗 减毒沙门菌 人白细胞介素-12 人粒细胞-巨噬细胞集落刺激因子 抗肿瘤作用 

分 类 号：R392[医药卫生—免疫学]