活体红色荧光成像评估Ag85A／BDNA疫苗联合抑瘤效应的实验研究

目的：探讨可视、直观评估Ag85A／BDNA疫苗联合抑瘤效应的科学方法。方法：采用红色荧光蛋白基因DsRed—Express标记小鼠膀胱癌细胞BTT739，经G418抗性筛选获得稳定表达RFP的细胞克隆BTT739-RFP，将BTT739～RFP细胞接种到615小鼠皮下，建立可视化膀胱癌移植瘤模型。将建模成功的32只小鼠随机分为pVAX1-Ag85A／B联合组：BCG组；空白质粒（pVAX1）；生理盐水（NS）。建模后第7、14、21天注射相应的药物。分别于给药前即第7和第28天进行活体荧光成像，进行统计学处理。并于第28天检测血清特异性抗体水平和脾淋巴细胞的增殖活性。结果：构建了稳定表达RFP的BTT739细胞；成功建立可视化膀胱癌模型，成瘤率100％；经治疗后，pVAX1-Ag85A／B组的荧光强度明显低于pVAX1组和NS组，与BCG组无统计学差异。pVAX1-Ag85A／B组的抗体滴度为1：1000，BCG免疫组的抗体滴度为1：800。pVAX1-Ag85A／B组和BCG组均可提高淋巴细胞增殖活性，二者差异无统计学意义。结论：应用Ag85A／BDNA疫苗可提高膀胱癌小鼠的免疫功能，其效果与BCG差异无统计学意义；活体红色荧光体外成像的评估体系与细胞水平的分析一致。

Research of the Whole-body Imaging Evaluating the Combined Effect of AR85A/B DNA Vaccine on Bladder Tumor in Mice

Objective To explore a Visible and intuitionistic philosophy to evaluate the combined effect of Ag85A/B on bladder tumor. Methods.. DsRed-Express labeled BTT739 ceils were cloned with G418 selection （BTT739-RFP）. BTT739-RFP cells were implanted into the mice subcutaneously to established Visible tumor model. Totally 32 model mice were evenly randomized into 4 groups： pVAX1-Ag85A/B,BCG, pVAX1, normal saline （NS） groups. The corresponding drugs were injected on day 7, 14, and 21 after model establishment. The whole body imaging was used to monitor the growth of the implanted tumor on day 7 and 28. Level of serum spe- cific antibody and Proliferative response of spleen lymphocyte were measured on 28th day. Results.. We have success fully got RFP-expressing cell clones and The Visible tumor model was successfully constructed, with a tumorigenesis rate of 100%. After treatment, the red fluorescence intensity in pVAX1 Ag85A/B group was obviously decreased compared with pVAX1 and NS groups, But no different with BCG group. The serum specific antibody titer of mice immunized with pVAX1-Ag85A/B is 1.. 1000, and that of mice immunized with BCG is 1： 800. pVAX1-Ag85A/B and BCG can equally improve the Proliferative response of spleen lymphocyte. Conclusions; pVAX1 Ag85A/B DNA vaccine can improve the immune response of mouse with bladder tumorl no different with BCG. The whole-body red imaging evaluation was the same as cells.