腺病毒介导突变Rip2基因对膀胱上皮细胞清除克雷伯杆菌的影响

The Klebsiella Pneumoniae Clearance Enhanced by Adenovirus Vector Encoding Mutant Rip2 Gene in Human Urinary Epithelial Cell

目的探讨受体相互作用蛋白2(Rip2)在克雷伯杆菌所致的尿路感染中的作用。方法使用含有突变Rip2基因(mRip2)、绿色荧光蛋白(GFP)基因的复制缺陷腺病毒(Ad-mRip2-GFP)感染人膀胱上皮癌细胞T24细胞,表达GFP为转染成功标志,观察转染率,同时感染含有GFP基因腺病毒做对照,利用克雷伯杆菌临床分离株Kp5与细胞37℃孵育4、24、48小时,用平板培养菌落计数活菌数。结果刺激后,在感染Ad-mRip2-GFP的T24细胞清除胞内Kp5能力下降,与Ad-GFP组比较P<0.01。结论Rip2可以阻断膀胱上皮细胞的清除克雷伯杆菌作用,提示Rip2在膀胱上皮细胞的天然免疫中可能起重要作用。

Objective To investigate the function of Rip2 gene on urinary infections induced by Klebsiella pneumoniae. Methods Hunman bladder carcinoma cell line T-24 was transfected with adenovirus expression vector containing both mutant Rip2 gene and green fluorescent protein （GFP） gene, and the cells transfected with adenovirus expression vector merely containing GFP gene used as control. Their transfected efficiency was indicated by GFP expression. Klebsiella pneumoniae clinical isolated 5 is incubated with these transfected cells at 37℃. At 4, 24 and 48 hour after initial incubation, viable intracellular bactria were quantified by plating appropriate dilution on LB agar plates. Results Compared with the T-24 transfected with GFP gene,the ability of clearing Kp5 in T-24 transfected with mutant Rip2 gene decreased significantly（P〈0. 01）. Conclusion Mutant Rip2 gene could resist the effect of clearing Klebsiella pneumoniae in bladder epithelial cell, implying that Rip2 gene may serve as an innate immune receptor recognizing urinary pathogens.