摘要
目的探讨BPI_(700)-Fcγ1_(700)嵌合基因导入,对最小致死量E.coli感染小鼠的保护作用机制。方法采用常规菌落计数法、内毒素和细胞因子检测试剂盒、H-E染色法检测菌落数、内毒素和细胞因子含量及主要器官病理学改变。结果(1)基因导入小鼠血清中的目的基因产物对E.coli具有杀伤作用,可中和内毒素和介导吞噬细胞产生交叉调理吞噬作用;(2)基因导入组小鼠血清、脏器中细菌数明显低于对照组小鼠;(3)基因导入组小鼠血清中内毒素和促炎细胞因子含量显著低于对照组小鼠;(4)对照组濒死小鼠主要脏器血管扩张、淤血明显,与内毒素休克死亡引发的病理学改变相符。结论基因导入组小鼠可通过表达目的基因产物即BPI1-199-Fcγ1嵌合抗菌蛋白,对致死量E.coli感染及其引发的内毒素休克死亡产生抵抗作用。
Objective To identify the resistance mechanism of mice to minimal lethal dose （MIX）） of E.coli by transfection of BPI700-Fcγ1 700 chimeric gene. Methods Endotoxin neutralization, opsonization test, bacteria counting and pathological microscopy of vascular damage in test animals were carried out by routine technology as referenced. Result （1） The chimeric target protein in serum of the target gene transferred mice killed E.coli and neutralized endotoxin, and showed activity of opsonizafion. （2） The bacterium number in serum and main viscera, endotoxin level in serum of the transferred mice were markedly less than those of controls. （3） Twelve hours after MLD E.coli challenge, the endotoxin level and the production of proinflammstory cytokines （TNF-α, IL-1β）in the serum of the control mice were markedly higher than that of transferred mice. （4） Pathological profile was proved as found in dead mice. Conclusion The mice transferred by AAV2-BPI700-Fcγ1 700 chimetic gene can resist MLD E. coli infection and endotoxin induced by this infection through expressing chimeric target protein BPI1-199-Fcγ1 . Our findings support the clinical application of anfimicrobial gene therapy on bacterial infection, and offer an experimental basis for the application of gene therapy to individuals at high risk of infection.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第2期150-154,共5页
Chinese Journal of Microbiology and Immunology
基金
北京市自然科学基金项目-北京市教委委员会科技发展计划重点项目(KZ200410025)
