Fujimoto J, Ishii Y, Koshiba H, Matsuura A, Ogasawara M, Uede T, Kikuchi K
Am J Hematol. 1981;10(2):145-56. doi: 10.1002/ajh.2830100206.
Specific anti-human thymus xenoantiserum (ATS) was utilized for characterizing a human thymus antigen (HTA) preferentially expressed on human thymocytes. Binding of ATS with different cell types was studied by immunofluorescence and immunoperoxidase techniques, as well as by radioimmunoprecipitation (RIP) followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). ATS reacted selectively with the majority (87%) of human thymocytes among normal lymphoid cell populations tested. In the thymus, HTA was expressed largely on cortical thymocytes but not on those cells located in the thymic medulla. When 20 cases of different types of lymphatic leukemias were studied with ATS, HTA was found on acute lymphoblastic leukemia cells rosetting with sheep erythrocytes (T-ALL) but not on other lymphatic leukemia cases, including chronic lymphocytic leukemia cells rosetting with sheep red blood cells (T-CLL). SDS-PAGE analysis of immunoprecipitates formed between ATS and radiolabeled cell-surface components of human thymocytes demonstrated that ATS bound a 48,000-molecular-weight component that could be adsorbed to Sepharose 4B coupled with Les culinaris hemagglutinin and could be labeled by periodate-tritiated borohydride, indicating that HTA is a sialoglycoprotein. Similar antigen molecules were also precipitated by ATS from T-ALL cells but not from other lymphatic leukemia cells. Five T-ALL-derived cell lines were tested with ATS by immunofluorescence and by RIP and SDS-PAGE, but only two cell lines (MOLT-3 and P12/Ich) possessed HTA molecules on their cell membrane.
特异性抗人胸腺异种抗血清(ATS)被用于鉴定一种优先在人胸腺细胞上表达的人胸腺抗原(HTA)。通过免疫荧光和免疫过氧化物酶技术,以及放射性免疫沉淀(RIP)后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),研究了ATS与不同细胞类型的结合情况。在测试的正常淋巴细胞群体中,ATS与大多数(87%)人胸腺细胞发生选择性反应。在胸腺中,HTA主要在皮质胸腺细胞上表达,而不在位于胸腺髓质的细胞上表达。当用ATS研究20例不同类型的淋巴白血病时,发现与绵羊红细胞形成玫瑰花结的急性淋巴细胞白血病细胞(T-ALL)上有HTA,而在其他淋巴白血病病例中没有,包括与绵羊红细胞形成玫瑰花结的慢性淋巴细胞白血病细胞(T-CLL)。对ATS与放射性标记的人胸腺细胞表面成分形成的免疫沉淀物进行SDS-PAGE分析表明,ATS结合了一种分子量为48,000的成分,该成分可被吸附到与刀豆球蛋白A偶联的琼脂糖凝胶4B上,并可被高碘酸盐-氚化硼氢化钠标记,表明HTA是一种唾液糖蛋白。类似的抗原分子也可被ATS从T-ALL细胞中沉淀出来,但不能从其他淋巴白血病细胞中沉淀出来。用免疫荧光、RIP和SDS-PAGE对5种T-ALL衍生的细胞系进行了ATS测试,但只有两种细胞系(MOLT-3和P12/Ich)在其细胞膜上具有HTA分子。