Sarmiento U M, Valli V E
Can J Vet Res. 1987 Jan;51(1):89-94.
High molecular weight surface proteins were examined in lymph node lymphocytes from five control dogs and 27 dogs with malignant lymphoma. Polyclonal rabbit antiserum was raised against a 210,000-dalton (210 K) membrane protein which was purified from a canine lymphoid tumor by preparative slab gel electrophoresis. Three high MW proteins at 210, 195 and 170 K and a common proteolytic fragment at 95 K were detected in the electrophoretically separated plasma membrane preparations by immunoblotting with polyclonal anti-210 K antiserum. Two antigenically distinct patterns were evident: 1) Type 1 lymphomas expressed a major 210 K peptide (with or without a minor 195 K component), and 2) Type 2 lymphomas lacked the 210 K form but had a 170 K or 195 K peptide singly or in combination. Immunoperoxidase staining of formalin-fixed, paraffin-embedded tissue sections demonstrated that the antigen was localized predominantly to the surface membrane of lymphocytes, and that canine lymphoma cells expressed a greater amount of the antigen than normal lymph node lymphocytes. It was concluded that these structurally and antigenically related high molecular weight proteins, based on their antigenic patterns, limited peptide analysis and tissue distribution, represent the canine homologue of the lymphocyte differentiation antigen known as T200.
对5只对照犬和27只患有恶性淋巴瘤的犬的淋巴结淋巴细胞中的高分子量表面蛋白进行了检测。用从犬类淋巴肿瘤中通过制备性平板凝胶电泳纯化的一种210,000道尔顿(210K)的膜蛋白制备了多克隆兔抗血清。通过用多克隆抗210K抗血清进行免疫印迹,在电泳分离的质膜制剂中检测到了210K、195K和170K的三种高分子量蛋白以及95K的一个常见蛋白水解片段。明显有两种抗原性不同的模式:1)1型淋巴瘤表达一种主要的210K肽(有或没有次要的195K成分),2)2型淋巴瘤缺乏210K形式,但单独或组合有170K或195K肽。对福尔马林固定、石蜡包埋的组织切片进行免疫过氧化物酶染色表明,该抗原主要定位于淋巴细胞的表面膜,并且犬淋巴瘤细胞表达的抗原量比正常淋巴结淋巴细胞更多。得出的结论是,基于它们的抗原模式、有限的肽分析和组织分布,这些结构和抗原相关的高分子量蛋白代表了被称为T200的淋巴细胞分化抗原的犬类同源物。