Weber E, Lorenzoni P, Cavina N, Rossi A, Sacchi G
Dipartimento di Neuroscienze, Sezione di Medicina Molecolare, University of Siena, Italy.
Histochem J. 2000 Nov;32(11):653-8. doi: 10.1023/a:1004159214913.
The difficulty of identifying and differentiating lymphatic and blood microvessels in tissue sections can be overcome by a monoclonal antibody specific for lymphatic endothelium. Unfortunately, the only known antibody also reacts with the endothelium of some blood vessels. The technique of double immunization (passive, with an antiserum to blood endothelium, and active, with a suspension of lymphatic endothelial cells) was, therefore, used to increase the chances of recognizing specific lymphatic antigens by the mouse immune system. The monoclonal antibody obtained, LyMAb, a G1 immunoglobulin, reacted strongly with the endothelium of bovine thoracic duct, mesenteric collecting vessels and lymphatic vessels of gallbladder and lymph nodes and moderately with those of the intestinal wall. Blood vessels (intercostal arteries, azygos vein and blood microvessels of all organs tested) were consistently negative. The antibody was species-specific and did not react with formalin-fixed, paraffin-embedded sections. Cross-reactivity was limited to some connective tissue fibres and scattered cells in the lymph node parenchyma, intestinal villi and hepatic lobules.
通过一种针对淋巴管内皮的单克隆抗体,可以克服在组织切片中识别和区分淋巴管与血管微血管的困难。不幸的是,唯一已知的抗体也会与一些血管的内皮发生反应。因此,采用了双重免疫技术(被动免疫,使用抗血管内皮血清;主动免疫,使用淋巴管内皮细胞悬液),以增加小鼠免疫系统识别特定淋巴管抗原的机会。所获得的单克隆抗体LyMAb是一种G1免疫球蛋白,它与牛胸导管、肠系膜集合血管、胆囊和淋巴结的淋巴管内皮强烈反应,与肠壁的淋巴管内皮中度反应。血管(肋间动脉、奇静脉以及所有测试器官的微血管)始终呈阴性。该抗体具有物种特异性,不与福尔马林固定、石蜡包埋的切片发生反应。交叉反应仅限于一些结缔组织纤维以及淋巴结实质、肠绒毛和肝小叶中的散在细胞。