Kubo R T, Roehm N
Mol Immunol. 1986 Aug;23(8):869-78. doi: 10.1016/0161-5890(86)90072-6.
A pan-reactive xenoantiserum to the mouse T-cell receptor was prepared by immunization of a rabbit with affinity purified mouse T-cell receptor material. The T-cell receptor of the chicken ovalbumin/IAd specific T-cell hybridoma, DO-11.10, was isolated by affinity chromatography using the clone-specific monoclonal antibody, KJ1-26. Immunoprecipitation with the rabbit antiserum and subsequent SDS-PAGE analysis of the material precipitated from lysates of surface radioiodinated T cells revealed the heterodimeric structure characteristic of the T-cell receptor from virtually every T-cell source examined. Flow cytofluorometric analysis of normal peripheral T cells and mature thymocytes of BALB/c and SJL mice indicated that most all T cells bear antigenic determinants recognized by the rabbit anti-mouse T-cell receptor antibodies. The AKR thymoma, BW5147, a common fusion parent used to generate functional T-cell hybridomas, notably lacks surface expression of a T-cell receptor molecule.
通过用亲和纯化的小鼠T细胞受体材料免疫兔子,制备了一种对小鼠T细胞受体具有泛反应性的异种抗血清。使用克隆特异性单克隆抗体KJ1-26通过亲和色谱法分离鸡卵清蛋白/IAd特异性T细胞杂交瘤DO-11.10的T细胞受体。用兔抗血清进行免疫沉淀,并对从表面放射性碘化T细胞裂解物中沉淀的物质进行后续SDS-PAGE分析,揭示了几乎所有所检测的T细胞来源的T细胞受体的异二聚体结构特征。对BALB/c和SJL小鼠的正常外周T细胞和成熟胸腺细胞进行的流式细胞荧光分析表明,几乎所有T细胞都带有兔抗小鼠T细胞受体抗体识别的抗原决定簇。AKR胸腺瘤BW5147是用于产生功能性T细胞杂交瘤的常见融合亲本,明显缺乏T细胞受体分子的表面表达。