新型重组免疫毒素DT390-mRantes治疗小鼠EAE的初步研究

目的:构建一种含有重组免疫毒素DT390-mRantes(白喉杆菌外毒素390片段-活化T细胞表达与分泌调节因子)的真核表达质粒,并用于治疗小鼠实验性自身免疫性脑脊髓炎(EAE)。方法:利用自提的MBP诱导C57BL/6小鼠产生EAE,将mRantes导入含有DT390的真核表达质粒SRα中,经阳离子纳米脂质体包被后,治疗小鼠EAE,同时对其临床症状、脑部病理改变、外周血相关细胞因子及T/B细胞比例进行检测,了解该重组免疫毒素的治疗效果。结果:成功构建真核表达质粒DT390-mRantes-SRα,治疗组小鼠临床症状减轻,脑部特异性淋巴细胞浸润减少,外周T细胞相关细胞因子表达降低,T/B细胞比例降低。结论:新型重组免疫毒素DT390-mRantes治疗小鼠EAE有较为明显的效果,为治疗人的多发性硬化(MS)提供有益的借鉴。

Primary study of the recombinant immunotoxin DT390-mRantes in EAE therapy

AIM: To construct a novel eukaryotic expression plasmid including the recombinant immunotoxin DT390-mRantes and treat experimental autoimmune encephalomyelitis (EAE) in mice. METHODS: EAE in C57BL/6 mice were induced by the extracted MBP. The mRantes fragment was inserted into the eukaryotic expression plasmid SRalpha containing DT390. Then cationic liposome-embedded plasmid DNA was injected into the muscles of the hind-limbs in mice. The effect of DT390-mRantes was evaluated by observing clinical symptoms, pathological changes of brain, relative cytokine of peripheral blood, and the proportion of T cells and B cells. RESULTS: The recombinant immunotoxin DT390-mRantes was successfully constructed. Compared the mice in treated group with those in untreated group the clinical symptoms of EAE were alleviated, the infiltration of inflammatory cells were decreased, the IFN-gamma level was fallen, and the ratio of T/B cells was decreased. CONCLUSION: The recombinant immunotoxin DT390-mRantes has distinct effects on EAE in mice, which may be used for beneficial reference to the therapy of MS.