微囊化小鼠白介素12基因工程细胞的长效抗肿瘤作用  

Long-term anti-tumor effects of microencapsulated murine interleukins-12 engineered cells

作　　者：孝作祥[1] 郑树[1] 潘月龙[1] 

机构地区：[1]浙江大学肿瘤研究所,浙江杭州310009

出　　处：《中国药理学与毒理学杂志》2002年第5期348-353,共6页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的　探讨通过微囊化能否获取长效抗肿瘤的小鼠白介素 12 (mIL 12 )基因工程细胞。方法　构建pcDNA3.1/mIL 12重组表达质粒 ,然后稳定转染CHO细胞 ,采用海藻酸钠微囊制作技术 ,将mIL 12基因修饰的CHO细胞包裹。观察微囊化mIL 12基因工程细胞的mIL 12释放 ,并将微囊化细胞植入荷瘤小鼠体内 ,测定小鼠的抗肿瘤免疫功能及抑瘤效应。结果　微囊化mIL 12基因工程细胞产生的mIL 12蛋白可自由透过微囊膜。植入荷瘤小鼠体内2 1d后 ,微囊化mIL 12基因工程细胞治疗组血清中mIL 12 ,mIL 2及mIFN γ水平分别为 (5 49± 5 3) ,(180± 2 9)和 (10 0 8± 15 6 )ng·L- 1,而mIL 4 ,mIL 10水平则显著降低。脾脏细胞毒T淋巴细胞 (CTL)活性及自然杀伤细胞 (NK)活性均显著增高 ,肿瘤生长受到显著性抑制 ,荷瘤小鼠的存活期明显延长。结论微囊化mIL 12基因工程细胞在体内可持续、稳定地释放mIL 12 ,能激发机体产生持久而强大的抗肿瘤免疫反应 ,对实验小鼠产生明显的抗肿瘤效应并延长其生存期。AIM To investigate if long term antitumoric murine interleukin 12(mIL 12) gene engineering cells could be obtained by microencapsulation. METHODS Reconstructed plasmid pcDNA3.1/mIL 12 was transfected into CHO cells stably by Superfect TM , then CHO/pcDNA 3.1/ mIL 12 cells were encapsulated in alginate microcapsules. mIL 12 release from the microencapsulated CHO/pcDNA3.1/mIL 12 cells was confirmed using ELISA assay. Microencapsulated CHO/pcDNA3.1/mIL 12 cells were transplanted subcutaneously into the tumor bearing mice with CT26 cells. The anti tumor immunoreaction responses and the anti tumor activities of the microencapsulated CHO/pcDNA3.1/mIL 12 cells were evaluated. RESULTS mIL 12 protein could release freely from the microencapsulated CHO/pcDNA3.1/mIL 12 cells. After the microencapsulated CHO/pcDNA3.1/mIL 12 cells were transplanted subcutaneously into the tumor bearing mice for 21 d, the serum average concentrations of mIL 12, mIL 2 and mIFN γ in the mice treated with microencapsulated CHO/pcDNA3.1/mIL 12 cells were(549±53), (180±29)and (1008±156)ng·L -1 ,respectively, but the serum concentrations of mIL 4 and mIL 10 were decreased significantly compared to the controls. The cytotoxicity of the CTL from the splenocytes and the NK activity were significantly higher in the mice treated with microencapsulated CHO/pcDNA3.1/mIL 12 cells. Moreover, mIL 12 released from the microencapsulated CHO/pcDNA3.1/mIL 12 cells continuously and stably brought a significantly inhibition of tumor proliferation and a prolonged survival time of tumor bearing mice. CONCLUSION The results showed that microencapsulated CHO/pcDNA3.1/mIL 12 cells expressed significant antitumor effects in the experimental tumor bearing mice by activating anti tumor immune responses in vivo.

关 键 词：白介素-12基因工程细胞 抗肿瘤作用 细胞微囊化 白介素 基因疗法 肿瘤治疗 

分 类 号：R965[医药卫生—药理学]