【摘要】 目的通过观测rAAV2-BPI1-199-Fcγ1重组病毒(rAAV-BF)感染对不同品系小鼠的抗细菌感染保护作用。方法采用dot blot、Western blot、免疫组化、改良ELISA方法和内毒素检测试剂盒,检测重组BPI1-199-Fcγ1(rBF)蛋白在CHO细胞和小鼠骨骼肌细胞中的表达及血清中内毒素含量的变化;建立最小致死量(MLD)E.coli致死模型,检测rBF基因转染小鼠对MLDE.coli感染的抵抗作用。结果(1)rBF蛋白在CHO细胞和两品系小鼠骨骼肌细胞中成功表达,血清中rBF蛋白具有中和内毒素和杀伤E.coli的作用;(2)在MLDE.coli腹腔感染后,两品系小鼠rAAV-BF感染组存活率均显著高于空载体对照组小鼠;基因转染组小鼠血清内毒素含量明显低于空载体对照组;rAAV-BF感染与头孢呋辛钠联合应用具有协同抗菌作用,两品系小鼠存活率明显高于单纯rAAV-BF感染组。结论rAAV-BF感染组小鼠对致死性E.coli感染具有抵抗作用。更多还原

【Abstract】 Objective To observe the protective effect of rAAV-BF infected mice against lethal dose of E.coli.Methods The rBF protein in CHO cells,mice were injected and muscle and serum were detected by dot blot,Western blot,immunohistochemistry assay and a modified enzyme linked immunosorbent assay(ELISA),the level of endotoxin was measured by reagent kits,establishing MLD E.coli causing death model for the detection of the mice resistant to MLD E.coli infection after administration with rAAV-BF.Results(1)rBF protein was detected in CHO cells and the injected mice muscle,serum from the gene transfected mice could neutralize endotoxin and kills E.coli.(2)The survival rate of BALB/c and C57BL/6 mice of rAAV group(30%～40%)was significantly higher than control group.Serum endotoxin level of rAAV-BF group mice was significantly lower than that of control group.rAAV-BF plus antibiotics have synergistic effect:the survival rate of rAAV-BF group was significantly higher than that of control group;the survival rate(85%) of rAAV-BF infected C57BL/6 mice was significantly higherthan that of BALB/c mice(65%).Conclusion rAAV-BF infected mice are restiste to lethal dose of E.coli,which might provide a base to protect clinically high risk patients from infection by bacteria and accordingly to reduce mortality.更多还原