Kissel K, Hamm S, Schulz M, Vecchi A, Garlanda C, Engelhardt B
Max-Planck-Institut für Physiologische und Klinische Forschung, Abteilung Molekulare Zellbiologie, Bad Nauheim, Germany.
Histochem Cell Biol. 1998 Jul;110(1):63-72. doi: 10.1007/s004180050266.
A novel monoclonal antibody (mAb), 8D3 (IgG2a), that specifically recognizes the murine transferrin receptor (TfR) was produced by immunizing a Lewis rat with a polyoma middle T oncogene-transformed endothelioma cell line. The 8D3 mAb was obtained by immunohistochemical screening for exclusive staining of vessels forming a blood-brain barrier (BBB), but not of other vessels. The anti-TfR mAb 8D3 recognizes the TfR also in FACS analysis and in western blots and should prove to be useful for affinity purification of the TfR. Whereas 8D3 brightly stains BBB-forming vessels in the central nervous system of mice, it does not stain the fenestrated capillaries within the choroid plexus and the circumventricular organs. In testis, where the blood-tissue barrier is located at the level of the Sertoli cells, the 8D3 mAb specifically stains Sertoli cells but not endothelial cells. Finally, in vitro, 8D3 does not interfere with iron uptake of lymphocytes as it does not influence their proliferation. Taken together, 8D3 represents a versatile new tool to study the tissue distribution of the murine TfR and TfR-mediated transcytosis across tissue barriers in the mouse.
通过用多瘤病毒中T癌基因转化的内皮瘤细胞系免疫一只Lewis大鼠，制备了一种新型单克隆抗体（mAb）8D3（IgG2a），它能特异性识别小鼠转铁蛋白受体（TfR）。8D3单克隆抗体是通过免疫组织化学筛选获得的，用于专门染色形成血脑屏障（BBB）的血管，而不染色其他血管。抗TfR单克隆抗体8D3在荧光激活细胞分选分析（FACS）和蛋白质免疫印迹中也能识别TfR，并且应该被证明可用于TfR的亲和纯化。虽然8D3能明亮地染色小鼠中枢神经系统中形成BBB的血管，但它不染色脉络丛和室周器官内的有窗孔毛细血管。在睾丸中，血组织屏障位于支持细胞水平，8D3单克隆抗体特异性染色支持细胞而不染色内皮细胞。最后，在体外，8D3不干扰淋巴细胞的铁摄取，因为它不影响淋巴细胞的增殖。综上所述，8D3是一种多功能的新工具，可用于研究小鼠TfR的组织分布以及TfR介导的跨小鼠组织屏障的转胞吞作用。
