【摘要】 目的利用霍乱毒素B亚单位(CB)的嗅神经细胞轴浆转运载体功能,将其与抗β淀粉样蛋白(Aβ)单抗偶联,绕过血脑屏障进入小鼠大脑海马部位的效率及改善情况,为老年性痴呆的免疫治疗探索更有效途径。方法以抗Aβ单抗杂交瘤细胞株制备抗Aβ单抗(IgG),纯化后用过碘酸钠法将其与CB偶联(CB-IgG),选择31只小鼠经鼻腔给药,分为CB-IgG组、IgG组、CB组、生理盐水组,分别以3次实验完成。采用Western blot、免疫组织化学及MRI方法检测单抗进入小鼠大脑部位的量及效率,并进行比较。结果 CB-IgG组有60%的IgG进入大脑中,IgG组有12%进入大脑,CB-IgG组较IgG组效率高5倍。IgG主要进入齿状回的颗粒细胞和CA1区的椎体细胞,在齿状回CB-IgG组阳性细胞为IgG组的5倍,在CA1区CB-IgG组阳性细胞为IgG组的3倍(P<0.05,P<0.01)。弥散张量成像分析显示,CB-IgG小鼠给药前后表观弥散系数变化较IgG明显(P<0.01)。结论 CB-IgG偶联物鼻腔给药可以绕过血脑屏障将更多抗Aβ单抗进入脑内海马部位,其效率比单用IgG提高了5倍。更多还原

【Abstract】 Objective To study the effective immunotherapy for Alzheimer’s disease with anti-amyloidβ(Aβ) protein monoclonal antibodies coupled with olfactory nerve cell translocator function of cholera toxin B subunit(CB) that enter into the hippocampus of mice by bypassing their blood brain barrier.Methods Anti-Aβmonoclonal antibodies(IgG),prepared with IgG hybrid tumor cell strains,were purified and coupled with CB-IgG containing periodate sodium,and divided into CB-IgG group,IgG group,CB group,and saline group.The volume and efficiency of IgG that entered into the hippocampus of mice after intranasal administration were measured by Western blot,immunohistochemistry and MRI,respectively.Results The IgG that entered into the hippocampus was 5-fold higher in CB-IgG group than in IgG group(60%vs 12%,P<0.05).Immuno-histochemistry showed that the IgG entered into the granular cells in dentate gyrus and into the pyramidal cells in CAI region.The number of positive cells in dentate gyrus and CAI region was 5-fold and 3-fold higher in CB-IgG group than in IgG group(P<0.01).Magnetic resonance diffusion tensor imaging showed that the effect of IgG on brain was more significant in CB-IgG group than in IgG group(P<0.01).Conclusion Intranasal administration of CB-IgG coupling agents can carry more anti-Aβmonoclonal antibodies into the hippocampus by bypassing the blood brain barrier,which is 5-fold higher than that of IgG.更多还原