AAV2-BPI_(700)-Fcγ1_(700)重组病毒的制备及其转染小鼠后的抗感染作用

Preparation of AAV2-BPI_(700)-Fcγ1_(700)and it′s protection of mice after transfection

目的通过观测BPI700-Fcγ1700嵌合基因导入小鼠对最小致死量E.coli感染攻击的抵抗作用,探讨基因治疗在细菌感染性疾病中应用的可能性。方法采用RT2PCR检测嵌合基因在转录水平的表达;采用免疫组化和酶联免疫吸附实验检测嵌合基因在蛋白水平的表达。结果(1)基因导入小鼠中目的嵌合基因得到表达,在注射部位肌肉组织和血清中可检测到目的嵌合蛋白;(2)在最小致死量E.coli感染攻击后,基因导入小鼠的存活率为31.43%,明显高于对照组小鼠的存活率4.62%;与小剂量头孢呋辛钠联合应用其存活率高达65%。结论BPI700-Fcγ1700嵌合基因导入小鼠对致死量E.coli感染具有较好抵抗作用,提示抗感染基因治疗在细菌感染性疾病,特别是在感染高危人群中具有广阔的临床应用前景。

Objective Toobserve the resistance of mice that were transfeeted with BPI700-Fcγ1 700 chimera gene to minimal lethal dose（MLD） of E. coli, and the potential protection against bacterial infection by gene therapy. Methods The transcription of target gene in the injected museules of mice was tested by RT-PCR. The objective protein was identified by immunohistoehemistry assay and a modified enzyme linked immunosorbent assay （ELISA）. Results （1） The chimera protein was identified from the injected muscles and the serum of BPI700-Fcγ1 700 transfeeted mice, and the presence of the objective protein was proved. （2） After E. coli challenge, the survival rate of the target gene transfeeted mice（31.43%） was significantly higher than that of the control （4.62%）. And when the AAV2-BPI700-Fcγ1 700 transfeeted mice were treated with eefuroxime sedium（25 mouse）, the survival rate as a result of synergistic bactericidal activity rose up to 65%. Conclusion AAV2- BPI700-Fcγ1 700 transfection shows definite function of protecting mice against E. coli infection, which shows a potential of gene therapy for bacterial infection.