重组hIFN-α-2b—BCG对小鼠原位膀胱肿瘤免疫效应的研究  

Immune response of recombinant IFN-α-2b-BCG treatment in mice bladder cancer

作　　者：孙二琳[1] 范晓东[1] 王玉叶[1] 韩瑞发[1] 

机构地区：[1]天津医科大学第二医院泌尿外科天津市泌尿外科研究所,300211

出　　处：《中华泌尿外科杂志》2011年第1期38-41,共4页Chinese Journal of Urology

基　　金：基金项目：国家自然科学基金（30872587）;国家科技部“十一五”重大专项新药创建课题（2009zx09103-699）;天津市科技支撑计划重大项目（07ZCKFSH03200）

摘　　要：目的探讨重组hIFN-α-2bBCG对原位膀胱肿瘤小鼠体内抗肿瘤免疫反应及其作用机制。方法构建C57BL／6原位膀胱肿瘤小鼠模型，采用流式细胞仪对小鼠膀胱灌注治疗后的淋巴细胞亚群进行分析，并测定小鼠血清中mTNF-α和mlI，12的水平，了解小鼠全身免疫状况。肿瘤组织冰冻切片进行T细胞亚群的免疫组织化学分析，免疫组化检测肿瘤Fas表达，了解膀胱局部免疫反应。结果重组BCG灌注治疗后小鼠外周血CD4＋细胞比例明显增高，CD8＋细胞比例无明显变化，CD4＋／CD8＋比例为2．63，与野生BCG组（2．10）比较差异有统计学意义（P〈0．05）。荷瘤小鼠外周血中Th1型细胞因子mTNF-α和mIL-12灌注治疗后比PBS对照组大幅提高，重组BCG组mTNF-α为806pg／ml，mIL-12为860pg／ml，与野生BCG组及野牛BCG联合IFN组比较差异无统计学意义（P〉0．05）。重组BCG组瘤组织内CD2、CD3和CD8检测强阳性，显著高于PBS对照组（P〈0．05），重组BCG组和野生BCG加IFN组瘤组织CD4＋、重组BCG组CD8＋检测值显著高于野生BCG组（P〈O．05）。BCG灌注后荷瘤小鼠膀胱肿瘤表达Fas明显高于PBS对照组（P〈0．05）。结论重组hIFN-α-2b—BCG具有在小鼠体内调节系统及局部免疫能力的作用，纠正荷瘤小鼠淋巴细胞亚群的比例失调，增强局部淋巴细胞浸润，具有增强Th1型细胞因子产生作用，上调荷瘤小鼠Fas的表达，诱导对膀胱肿瘤细胞的免疫攻击。Objective To study local and systemic immune response in an animal model treated with recombinant hIFN-α2b BCG instillation. Methods The MB49 orthotopic bladder cancer model in C57BL/6 mice was established and treated separately with rBCG, wild BCG, wild BCG combined with IFN-α-2b and PBS as the control. The changes of lymphocyte subgroups in peripheral blood were analyzed with FCM, and mTNF-α and raiL-12 in peripheral blood of mice were detected with ELISA. Immunohistochemistry was carried out to detect the local immune reaclion, T cell subsets and FAS, in bladder cancer after being treated with rBCG or wBCG. Results The content of CD8＋ T lymphocyte was up-regulated in the rBCG group. The CD4＋/CD＋ ratio of 2. 63 was up-regulated than pretreat ment, significantly different than that of wBCG group（P〈0.05）. ELISA assay showed that BCG sig nificantly up-regulated the level of mTNF-a and mIL-12 in serum of orthotopic murine bladder cancer mice. The mTNF-α 806 pg/ml, mIL-12 860 pg/ml in rBCG group, was not significantly higher than those in wBCG group and combination group. The immunocompetent cell numbers with CD4, CD3 , CD4 phenotype increased significantly in the tumor tissue of BCG treated group than the control（P〈 0.05）. The results of CD8＋ in rBCG group and the combination group, and CD＋ in rBCG group were significantly higher than that of the wBCG（P〈0.05）. The expression of Fas in tumor tissues treatedwith intravesical BCG was increased（P〈0.05）. Conclusions The recombinant IFN-α-2b-BCG can retrieve the disproportion of systemic lymphocyte subgroups, and increases Thl-type factors and local Fas expression in orthotopic murine bladder cancer. The recombinant IFN-α-2b-BCG is effective in regulating local and systemic immune reaction in orthotopic routine bladder cancer model.

关 键 词：重组BCG IFN-Α-2B 膀胱肿瘤 淋巴细胞亚群 原位膀胱肿瘤模型 

分 类 号：R737.14[医药卫生—肿瘤]