Immunohistochemical localization of the murine transferrin receptor (TfF) on blood-tissue barriers using a novel anti-TfR monoclonal antibody
Immunohistochemical localization of the murine transferrin receptor (TfF) on blood-tissue barriers using a novel anti-TfR monoclonal antibody
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DOI：
10.1007/s004180050266
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发表时间：
1998-07-01
影响因子：
2.3
通讯作者：
Engelhardt, B
中科院分区：
文献类型：
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作者：
Kissel, K;Hamm, S;Engelhardt, B
通过用多瘤中 T 癌基因转化的内皮瘤细胞系对 Lewis 大鼠进行免疫接种，产生了一种特异性识别鼠转铁蛋白受体 (TfR) 的新型单克隆抗体 (mAb) 8D3 (IgG2a)。 8D3 mAb 通过免疫组织化学筛选获得，仅对形成血脑屏障 (BBB) 的血管进行染色，但不对其他血管进行染色。抗 TfR mAb 8D3 还可在 FAGS 分析和蛋白质印迹中识别 TfR，并且应该证明可用于 TfR 的亲和纯化。虽然 8D3 对小鼠中枢神经系统中的血脑屏障形成血管进行了明亮的染色，但它不会对脉络丛和心室周围器官内的有孔毛细血管进行染色。在睾丸中，血液组织屏障位于支持细胞水平，8D3 mAb 特异性染色支持细胞，但不染色内皮细胞。最后，在体外，8D3 不会干扰淋巴细胞的铁吸收，因为它不会影响淋巴细胞的增殖。总而言之，8D3 代表了一种多功能新工具，可用于研究小鼠 TfR 和 TfR 介导的跨胞吞作用跨组织屏障的组织分布。
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 FAGS 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.