五种识别大鼠胸腺上皮细胞表面抗原的新型单克隆抗体的构建(英文)

Five novel monoclonal antibodies to thymic epithelial cell surface antigens in rats

目的 建立抗胸腺上皮细胞单克隆抗体 (单抗 )以研究胸腺上皮细胞在T细胞分化过程中的作用。方法 以胸腺上皮细胞株免疫Balb/c小鼠 ,用B淋巴细胞杂交瘤技术制备抗胸腺上皮细胞抗原的单抗。用免疫化学染色和流式细胞图谱分析来确定这些单抗所识别抗原的分布。用免疫电泳法确定这些抗原的分子量。结果 得到 5种抗大鼠胸腺上皮细胞单抗 ,根据其免疫组化染色确定的胸腺内分布状况推断 :RE 4D8单抗为CTESⅡA型 1;RE 12B2单抗为CTESV型 ,它只分布在髓质中 ;RE 5C6单抗为CTESⅣ型 ;RE 6D6单抗可能为CTESⅡB型 ;RE 1D4单抗为CTESⅤ型。抗原RE 4D8,RE 6D6 ,和RE 12B2所对应抗原的分子量分别为 12 0kDa ,2 2 0kDa和 35kDa。RE 1D4抗原可能为一新发现的皮质上皮的标识物。根据与这些单抗结合的方式 ,确立了一些已建立的胸腺上皮细胞株的上皮分类及它们在胸腺内的原始位置。所用这些细胞株中 ,TuD32和FTE3的表型为皮质型 ,而TaD34的表型为髓质型。结论 这些单抗明确证实胸腺上皮的异型性。它们所识别的抗原既可出现在细胞质中又可在胸腺上皮细胞表面出现。

OBJECTIVE: To establish monoclonal antibodies (mAbs) against thymic epithelial cells and study the function of epithelial cells during T-cell differentiation in the thymus. METHODS: Hybridomas secreting mAbs against thymic epithelial cells were derived by immunization of Balb/c mice with two thymic epithelial cell lines, TaD3 and FTE. The distribution of antigens recognized by these mAbs was detected by immunochemical staining and cytofluorographic analysis, and the molecular weight of the antigens by immunoblotting. RESULTS: Five specific monoclonal antibodies (mAb) were obtained. On the basis of their distribution in the thymus determined by immunochemical staining, mAb RE-4D8 was regarded as clusters of thymic epithelium staining (CTES) type IIA: mAb RE-12B2, which showed a unique distribution pattern only in the medulla, was CTES type V: mAb RE-5C6 was CTES type IV: mAb RE-6D6 might be CTES type IIB: and mAb RE-1D4 was classified as type V. The molecular weight (MW) of antigen RE-4D8, RE-6D6 and RE-12B2 were 120 kDa, 220 kDa and 35 kDa, respectively. Antigen RE-1D4 is a novel marker of cortical epithelium, several established thymic epithelial cell lines were classified and their original intrathymic locations were determined by these mAbs. Thymic cell lines, TuD3 and FTE were cortical phenotypes whereas TaD3 had a medullar phenotype. CONCLUSIONS: These mAbs clearly demonstrate the heterogeneity of the thymic epithelium; they could detect antigens not only in the cytoplasm but also on the surface of thymic epithelial cells. Our data suggest that these newly established mAbs may help elucidate the interaction between thymocytes and epithelial cells during T cell maturation.