粒细胞-巨噬细胞集落刺激因子修饰MB49制备膀胱肿瘤疫苗及应用  

Preparation and application of GM-CSF-surface-modified MB49 cell vaccine

作　　者：梁中锟[1] 张琳[2] 谭万龙[1] 高基民[2] 陈忠[1] 黄鑫[3] 

机构地区：[1]南方医科大学附属南方医院泌尿外科,广州510515 [2]温州医学院浙江省医学遗传学重点实验室 [3]南方医科大学生命技术学院生物治疗研究所

出　　处：《中华实验外科杂志》2011年第6期841-843,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家863高技术研究发展计划资助项目（2006AA0224C4）;广东省自然科学基金资助项目（9151051501000030）

摘　　要：目的链亲和素标记的细胞因子锚定MIM9细胞和成瘤小鼠膀胱，探讨锚定率、锚定量与时间的关系。方法MB49细胞与链亲和素标记的绿色荧光蛋白（SA—GFP）体外锚定后，在荧光显微镜下观察；建立小鼠原位表浅膀胱癌模型后，实验组膀胱灌注锚定链亲和素标记的粒细胞一巨噬细胞集落刺激因子（SA—GM—CSF）；对照组小鼠不锚定直接膀胱灌注SA—GM—CSF。取膀胱作冰冻切片免疫组织化学分析。结果MB49细胞基本被SA—GFP锚定，锚定率为98．78％；冰冻切片免疫组织化学显示实验组膀胱黏膜有阳性显色，以第1天最浓，以后逐日变淡，至第7天仍可见较明显染色；对照组膀胱黏膜无阳性显色。结论链亲和素标记的细胞因子可以锚定在细胞表面，且锚定效率高、保留时间长。Objective Streptavidin （SA） -tagged cytokine was anchored to MB49 cells in vitro and biotinylated mucosal surface of bladder wall in vivo. The anchoring rate was calculated and the effects of anchoring and the relation between anchoring effect and time were observed. Methods Streptavidin （SA）- tagged green fluorescent protein （SA-GFP） was anchored to MB49 cells in vitro and observed under an fluorescence microscope. Female C57BL/6 mice with superficial bladder cancer were intravesically instilled with NHS-PEO4-Biotin, PBS, SA-GM-CSF as SA-GM-CSF group, and PBS, SA-GM-CSF as PBS group. All the mice were killed to obtain their snap-frozen bladders for cryosectioning, and then immunohistochem- istry analysis with biotin-conjugated HRP and DAB display liquid kit. Results SA-GFP was anchored to the surface of MB49 ceils under the fluorescence microscopy, and the anchoring rate was 98.78%. SA-GM-CSF was detected on the biotinylated mucosal surface of bladder wall up to 7 days after its intravesical immobilization in SA-GM-CSF group, but no SA-GM-CSF was found in PBS group. Conclusion SA-GFP can be anchored to the surface of MIM9 cells, and the anchoring rate was 98.78%.

关 键 词：膀胱肿瘤 疫苗 粒细胞-巨噬细胞集落刺激因子 MB49 膜表面修饰 

分 类 号：R737.14[医药卫生—肿瘤]