Birk H W, Ogiermann M, Altmannsberger M, Schütterle G, Haase W, Koepsell H
Medizinische Klinik II, Justus-Liebig-Universität, Giessen, Germany.
Biochim Biophys Acta. 1993 May 14;1148(1):67-76. doi: 10.1016/0005-2736(93)90161-r.
After immunization with porcine brush-border membrane proteins, 11 monoclonal antibodies were generated which react with proximal tubules. Their antigenic polypeptides were characterized with respect to apparent molecular weight, histochemical localization in porcine and human kidney, and tissue distribution in pig. In porcine kidney, six antibodies bind selectively to the proximal tubule whereas the others also react with other nephron segments. With the exception of one antibody which reacts with the luminal and the basolateral membrane of the porcine proximal tubule, the other antibodies specific for the proximal tubule only stain the brush-border membrane. Four of them react along the entire length of the porcine proximal tubule, whereas one (R1A2) binds to the S3-segment in pig and to the entire length of the proximal tubule in man. This indicates that segment-specific expression may be species-dependent. Testing the antibodies in 21 different extrarenal tissues it was found that three of the antibodies, specific for the brush-border membrane in renal proximal tubules, only react in kidney. Two of these are specific for pig kidney whereas one also reacts with human kidney. This antibody (N4A4) is directed against a polypeptide with an apparent molecular weight of 400,000. Electron microscopic immunohistochemistry showed that N4A4 binds to the intervillus region of the brush-border membrane and to subapical vesicles.
用猪刷状缘膜蛋白免疫后，产生了11种与近端小管发生反应的单克隆抗体。对其抗原多肽的表观分子量、在猪和人肾脏中的组织化学定位以及在猪体内的组织分布进行了表征。在猪肾脏中，6种抗体选择性地结合近端小管，而其他抗体也与其他肾单位节段发生反应。除了一种与猪近端小管的管腔膜和基底外侧膜都发生反应的抗体外，其他针对近端小管的抗体仅对刷状缘膜进行染色。其中4种抗体沿猪近端小管的全长发生反应，而一种（R1A2）在猪中与S3节段结合，在人中与近端小管的全长结合。这表明节段特异性表达可能具有物种依赖性。在21种不同的肾外组织中测试这些抗体时发现，3种针对肾近端小管刷状缘膜的抗体仅在肾脏中发生反应。其中2种对猪肾脏具有特异性，而另一种也与人肾脏发生反应。这种抗体（N4A4）针对的是一种表观分子量为400,000的多肽。电子显微镜免疫组织化学显示，N4A4与刷状缘膜的绒毛间区域和顶端下小泡结合。
