探讨Ag85A和Ag85B DNA疫苗对大鼠膀胱癌免疫治疗的效果。方法：致癌剂N 甲基亚硝基脲(methyl nitrosourea，MNU)膀胱灌注雌性Wistar大鼠制备膀胱癌模型。将建模成功的48只大鼠随机分为生理盐水组、空白质粒组、卡介苗组、Ag85A DNA疫苗组、Ag85B DNA疫苗组、Ag85A+Ag85B DNA疫苗组（每组各8只），建模后第7、14、21天于大鼠右后肢肌内注射相应药物。第28天处死大鼠，流式细胞仪检测各组大鼠脾脏细胞的CD4 + 和CD8 + T细胞亚群数量并计算CD4 +/CD8 + 比值； ELISA法检测大鼠血清中IFN γ分泌水平；剥离膀胱肿瘤进行组织病理学检查。结果：成功建立大鼠膀胱癌模型，致瘤率100%。经疫苗治疗后，Ag85A组、Ag85B组和Ag85A+Ag85B组膀胱肿瘤体积均有所减小、病理分级也有所减轻，但效果不及卡介苗组。Ag85A组、Ag85B组、Ag85A+Ag85B组和卡介苗组CD4 +T细胞亚群数量分别为（17.27±295）%、（23.15±1.56）%、（30.80±1.83）%、（38.05±1.48）%；CD8 + T细胞亚群数量分别为（9.03±1.06）%、（10.28±039）、（1129±074）、（13.14±1.24）；CD4 +/CD8 + 比值分别为（1.90±0.10）、（2.25±0.08）、（2.73±0.19）、（2.97±023）；血清IFN γ含量分别为（96.94±12.38）、（131.03±26.68）、（179.20±28.88）、（240.53±32.17）pg/ml；与生理盐水、空白质粒组相比，Ag85A组、Ag85B组、Ag85A+Ag85B组能够显著提高以上4项检测指标（ P< 0.01），但均低于卡介苗组；Ag85A+Ag85B组效果强于Ag85B组、更强于Ag85A组，差别有统计学意义（ P< 0.05）。 结论： 应用Ag85A和Ag85B DNA疫苗均可提高膀胱癌大鼠的免疫功能，其效果为Ag85A+Ag85B >Ag85B>Ag85A，但总体上都达不到卡介苗的抗癌免疫效果。

To explore the immunotherapeutic effect of Ag85A DNA vaccine and Ag85B DNA vaccine in treatment of bladder tumor in rats. Methods: The bladders of female Wistar rats were irrigated with carcinogen methyl nitrosourea (MNU) to create the model of bladder tumor. Totally 48 model rats were evenly randomized into 6 groups: normal saline (NS), pcDNA3.1, bacille Calmette Guérin(BCG), Ag85A DNA vaccine, Ag85B DNA vaccine and Ag85A+ Ag85B DNA vaccine groups. The corresponding drugs were injected into the right hind limbs of rats intramuscularly on day 7, 14, and 21 after model establishment. Animals were sacrificed on day 28, and the spleens were removed aseptically. The percentages of CD4 +T cells and CD8 +T cells in splenocytes were measured by flow cytometry and the value of CD4 +/CD8 + was calculated. Level of serum IFN γ was assayed by ELISA and pathological examination was done. Results: The rat model of bladder tumor was successfully constructed, with a tumorigenesis rate of 100%. The bladder tumor sizes in Ag85A DNA vaccine group, Ag85B DNA vaccine group and Ag85A+ Ag85B DNA vaccine group were decreased after treatment, and the tumor pathological grades were also improved, but the outcomes were not better than those of the BCG group. The percentages of CD4 +T cells in the 4 groups were (17.27±2.95)%， (23.15±156)%， (3080±183)%， (38.05±1.48)%，respectively; and the percentages of CD8 +T cells were (9.03±106)%, (1028±039)%, (11.29±0.74)%, (1314±1.24)%, respectively; the relative values of CD4 +/CD8 + were 1.90±0.10, 2.25±0.08, 2.73±0.19, 297±0.23, respectively; the production of IFN γ in Ag85A DNA vaccine group, Ag85B DNA vaccine group, Ag85A+ Ag85B DNA vaccine group, BCG group were (96.94±1238), (131.03±26.68) , (179.20±28.88)，(240.53±32.17) pg/ml, respectively. The above 4 parameters in the Ag85A DNA vaccine group, Ag85B DNA vaccine group and Ag85A+ Ag85B DNA vaccine group were obviously improved compared with the pcDNA3.1 and NS groups, but were still poorer than those of the BCG group ( P<0.01). The effects of Ag85A+ Ag85B DNA vaccine group were better than those of the Ag85A and Ag85B DNA vaccine group( P< 0.05). Conclusion: Ag85A DNA vaccine and Ag85B DNA vaccine can improve the immune response of rats with bladder tumor; a combination of both has better outcome than they are used alone, but even the combination can not reach the effect of BCG.

山西省自然科学基金资助项目(NO.20051099)