携抗白细胞介素-2受体α单抗靶向超声微泡黏附效能的体内外评价  

Construction of Targeted Ultrasound Microbubbles with Anti-IL-2Rα Monoclone Antibody and Evaluation of its Binding Ability in Vitro and Vivo

作　　者：吴凤林[1] 纪丽景[1] 李发友[1] 李军华[2] 李美瑜[2] 宾建平[2] 

机构地区：[1]南方医科大学南方医院超声科,广州市510515 [2]南方医科大学南方医院心内科,广州市510515

出　　处：《中国超声医学杂志》2010年第7期577-580,共4页Chinese Journal of Ultrasound in Medicine

基　　金：广东省自然科学基金(No.7005165)

摘　　要：目的构建小鼠携抗白细胞介素-2受体α(IL-2Rα)单抗靶向超声微泡(MB_(IL-2Rα))及体内外评价其靶向黏附效能。方法采用"亲和素-生物素"桥连法构建携带荧光FITC的MB_(IL-2Rα)和普通超声微泡(MB)。首先利用平行板流动腔体外评价在不同时间点、不同浓度小鼠IL-2Rα包被下MB_(IL-2Rα)结合数目。然后,分别随机经静脉将MB_(IL-2Rα)和MB弹丸式注入对照组(10只)和TNF-α处理组(10只)的小鼠提睾肌炎症模型。同时,200倍荧光显微镜直视下观察并记录5min内两组中不同微泡在提睾肌微血管中的黏附数量和黏附情况。结果体外结果显示,MB_(IL-2Rα)结合数量随着IL-2Rα包被浓度的增高而增加(P<0.05),并与结合时间呈明显相关(P<0.05);体内荧光显微镜下TNF-α处理组MB_(IL-2Rα)黏附数量[(4.6±1.1)个/200倍视野)]明显高于MB黏附数量[(3.0±0.7)个/200倍视野](P<0.05);且TNF-α处理组MB_(IL-2Rα)黏附数量分别为对照组MB_(IL-2Rα)和MB黏附数量的(2.8±0.8)和(3.7±1.4)倍(P<0.01)。结论 MB_(IL-2Rα)可与IL-2Rα特异、有效地结合,其高效黏附性能将有助于进一步对移植排斥反应的靶向超声分子显影。Objective To construct the site-targeted microbubbles carrying rat anti-mouse IL-2Rα antibody （MBIL-2Rα） and ewduate the efficacy of its targeted adhesion using parallel plate flow chamber and fluorescence microscope. Methods The site targeted microbubbles with FITC fluorescence and monoantibodies of IL- 2Rα and the con trol microbubbles with FITC fluorescence were constructed. The adhesive ability of MBIL -2Rα to mouse IL-2Rα immobi lized on a culture dish was firstly assessed in a parallel plate flow chamber. Thereafter, MBIL 2Rα, and MB were injec ted intravenously in random to 10 wild type mice with TNF α stimulus and 10 wild type mice without any treatments. And then the behaviors of MBIL-2Rα and MB in cremasterie venules were assessed by fluorescence microscope. Results In vitro, the number of MBIL-2Rα bound to mouse IL 2Rα increased with the increasing of antibody concentrations on coverslips surface （P〈0.05）, and there was a significantly positive correlation between the number of MBIL-2Rα bound to mouse IL-2Rα and time of combination （P〈0.05）. In vivo, the retentive number of MBIL-2Rα, was significantly greater than that of MB in TNFα stimulated group （P〈0.05）. The number of MBIL-2Rα amounted to 4.6±1. 1/200-fold field and the number of MB was just 3.0±0.7/200 fold field. The adhesive number of MBIL-2Rα in TNF α-stimulated mice were 2.8＋0.8 and 3.7±1.1 fold greater separately than both MBIL-2Rα and MB in wild type mice without any treatments （P〈0.01）. Conclusions MBIL-2α could adhere to mouse IL 2Rα in the physical condi tions specifically and effectively. It could be useful for the developing of targeted ultrasound molecular imaging to eval uate transplant rejection.

关 键 词：靶向超声微泡 平行板流动腔 IL-2Rα单抗 荧光显微镜 

分 类 号：R445.1[医药卫生—影像医学与核医学]