Sarmiento U M, Valli V E
Can J Vet Res. 1987 Jan;51(1):110-6.
A murine monoclonal antibody (DT200) was raised against a 210,000-dalton (210 K) lymphocyte surface protein (a member of the lymphocyte antigen known as T200) which was purified from a canine lymphoid tumor by preparative slab gel electrophoresis. In immunoblotting studies of electrophoretically separated plasma membranes from five cases of canine lymphoma, the antibody detected two antigenically intact peptides at 95 and 110 K which, based on previous polyclonal anti-210 K antiserum immunoblotting and peptide mapping studies, may represent the protease-resistant fragment of the canine T200 molecule. Since DT200 retains its reactivity in formalin-fixed, paraffin-embedded tissue sections, 13 dogs with malignant lymphoma and a panel of normal lymphoid and nonlymphoid tissues were studied using an indirect immunoperoxidase technique. The antigen was localized predominantly to the surface membrane of lymphoid cells. DT200 reacted strongly with all five histological subtypes of lymphoma tested while moderate reactivity was detected in normal B and T cell areas of lymph node, spleen and tonsil. Thymocytes and selected hemopoietic precursors were weakly reactive with DT200 while plasma cells, mature granulocytes, red cells and megakaryocytes were unstained. It was concluded that DT200 is a useful reagent for the diagnosis of malignant lymphoma particularly in extranodal sites and may prove valuable in the investigation of the structure and function of T200 in the dog.
一种鼠单克隆抗体(DT200)是针对一种210,000道尔顿(210K)的淋巴细胞表面蛋白(一种被称为T200的淋巴细胞抗原成员)产生的,该蛋白通过制备性平板凝胶电泳从犬类淋巴瘤中纯化得到。在对5例犬淋巴瘤的电泳分离质膜进行免疫印迹研究时,该抗体在95K和110K处检测到两种抗原完整的肽段,根据先前的多克隆抗210K抗血清免疫印迹和肽图谱研究,这可能代表犬T200分子的蛋白酶抗性片段。由于DT200在福尔马林固定、石蜡包埋的组织切片中仍保持其反应性,因此使用间接免疫过氧化物酶技术对13只患有恶性淋巴瘤的犬以及一组正常淋巴组织和非淋巴组织进行了研究。抗原主要定位于淋巴细胞的表面膜。DT200与所检测的所有5种淋巴瘤组织学亚型均发生强烈反应,而在淋巴结、脾脏和扁桃体的正常B细胞和T细胞区域检测到中等反应性。胸腺细胞和选定的造血前体细胞与DT200反应较弱,而浆细胞、成熟粒细胞、红细胞和巨核细胞未被染色。得出的结论是,DT200是诊断恶性淋巴瘤特别是结外部位恶性淋巴瘤的有用试剂,并且可能在研究犬T200的结构和功能方面具有重要价值。