一种针对犬类CD8 +淋巴细胞的小鼠单克隆抗体（M10）的鉴定与特性分析。

Identification and characterization of a mouse monoclonal antibody (M10) directed against canine (dog) CD8+ lymphocytes.

作者信息

Voss C, Kremmer E, Hoffmann-Fezer G, Schumm M, Günther W, Kolb H J, Thierfelder S

机构信息

GSF, Institute of Immunology, Munich, Germany.

出版信息

Vet Immunol Immunopathol. 1993 Oct;38(3-4):311-25. doi: 10.1016/0165-2427(93)90090-q.

DOI:10.1016/0165-2427(93)90090-q

PMID:8291208

Abstract

Monoclonal antibodies (mAb) were produced by immunizing BALB/c mice with non-adherent dog lymphocytes. M10 was specific for a subset of dog lymphocytes. M10 belonged to the IgG1 subclass and reacted with 26% of dog peripheral blood lymphocytes, 24% of spleen lymphocytes, 81% of thymus cells, 1.2% of bone marrow cells (5.8% of bone marrow lymphocytes) and 23% of PHA-stimulated lymphocytes. Immunohistology of snap-frozen thymus and spleen showed that the spleen B-cell area stained negative, whereas the spleen T-cell area and the thymus medulla exhibited positive reaction in 20-30%. The thymus cortex was strongly positive. M10 diminished cell lysis by 58% in cell mediated lysis assays (CML). Immunoblot assays revealed that M10 recognized an antigen with a molecular weight of 76 kD under non-reducing and 33 kD under reducing conditions. Finally, M10 bound to a canine CD8 alpha transfected rat T-cell line (NB2). These findings characterize M10 as an antibody directed against the dog CD8 antigen.

摘要

通过用非贴壁犬淋巴细胞免疫BALB/c小鼠来制备单克隆抗体（mAb）。M10对犬淋巴细胞的一个亚群具有特异性。M10属于IgG1亚类，与26%的犬外周血淋巴细胞、24%的脾淋巴细胞、81%的胸腺细胞、1.2%的骨髓细胞（5.8%的骨髓淋巴细胞）以及23%的PHA刺激淋巴细胞发生反应。速冻胸腺和脾的免疫组织学显示，脾B细胞区呈阴性染色，而脾T细胞区和胸腺髓质有20 - 30%呈阳性反应。胸腺皮质呈强阳性。在细胞介导的细胞溶解试验（CML）中，M10使细胞溶解减少了58%。免疫印迹分析显示，M10在非还原条件下识别一种分子量为76 kD的抗原，在还原条件下识别一种分子量为33 kD的抗原。最后，M10与转染了犬CD8α的大鼠T细胞系（NB2）结合。这些发现表明M10是一种针对犬CD8抗原的抗体。