目的：运用活体红色荧光成像技术及影像学特征评估Ag85A DNA 疫苗和Ag85B DNA 疫苗对膀胱癌的免疫治疗效果。方法：构建稳定转染香菇珊瑚红色荧光蛋白（discosomasp red fluorescent protein, DsRed）基因的小鼠膀胱癌BTT细胞（BTTDsRed），建立BTTDsRed细胞移植瘤小鼠模型，将建模成功的24 只小鼠随机分为pVAX1Ag85A DNA 疫苗组、pVAX1Ag85B DNA 疫苗组和生理盐水治疗组，各组分别于肿瘤细胞接种后的第6天肌内注射pVAX1Ag85A、pVAX1Ag85B及生理盐水，然后用活体荧光成像系统检测移植瘤的生长和转移情况。结果：成功制备了稳定转染DsRed基因的BTTDsRed细胞，BTTDsRed接种小鼠建立可视化红色荧光膀胱癌移植瘤模型。重组质粒pVAX1Ag85A和pVAX1Ag85B治疗后的第2周，活体荧光成像显示pVAX1Ag85B 治疗组小鼠肿瘤荧光强度明显低于生理盐水组（P<0.05）；治疗后第3周，pVAX1Ag85A 和pVAX1Ag85B组小鼠肿瘤荧光强度都明显低于生理盐水组（P<0.01），但pVAX1Ag85B 与pVAX1Ag85A组无明显差别（P>0.05）；pVAX1Ag85B 组小鼠淋巴结转移率（25.0%）明显低于生理盐水组（87.5%，P<0.01）和pVAX1Ag85A组（625%，P<0.05）。结论：应用活体红色荧光成像技术能够动态、灵敏、可视化地评估 DNA 疫苗对小鼠膀胱癌移植瘤的疗效；Ag85A和Ag85B DNA 疫苗均具有抗肿瘤免疫疗效。

Objective:To use in vivo fluorescence image analysis system for evaluating the efficacies of pVAX1Ag85A and pVAX1Ag85B DNA vaccines in treatment of bladder cancer cellimplanted tumors in mice. :Methods: :Discosomasp red fluorescent protein (DsRed) stably transfected bladder cancer BTT cell line (BTTDsRed) was established and BTTDsRed cellimplanted mouse model was constructed. Six days later, 24 BTTDsRedbearing mice were randomly divided into pVAX1Ag85A DNA vaccine group, pVAX1Ag85B DNA vaccine group, and saline group through injecting the pVAX1Ag85A, pVAX1Ag85B, and saline into the right hind limbs of mice, respectively. The growth and metastasis of implanted BTTDsRed tumors were examined by in vivo fluorescence image analysis system. :Results: :BTT cell line stably transfected with DsRed (BTTDsRed) was successfully established. Fluorescence visible mouse model was successfully established by inoculating BTTDsRed cells into hind limbs of mice. After treatment with pVAX1Ag85A or pVAX1Ag85B for 2 weeks, the in vivo tumor fluorescence intensity in pVAX1Ag85B group was significantly lower than that in the saline group (P<0.05). After 3 weeks, tumor fluorescence intensities in both pVAX1Ag85A and pVAX1Ag85B groups were significantly lower than that in the saline group (P<0.01). But the efficacies of pVAX1Ag85A and pVAX1Ag85B groups were similar (P>0.05). The distant lymphatic metastasis rate in pVAX1Ag85B group was significantly lower than those in the saline (25.0% vs 87.5%) and pVAX1Ag85A groups (25.0% vs 62.5%) (P＜0.05). :Conclusion:: In vivo fluorescence image analysis system can dynamically, sensitively and visually evaluate the antitumor effects of DNA vaccines against bladder cancer cellimplanted tumors. Both pVAX1Ag85A and pVAX1Ag85B DNA vaccines have antitumor effects for bladder cancers.

山西省科技攻关基金资助项目（No. 20090321022）