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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。这些新建的单抗有望帮助认识T细胞成熟过程中胸腺细胞与上皮细胞的密切关联
Objective To establish anti-thymic epithelial monoclonal antibody (mAb) to study the role of thymic epithelial cells in T cell differentiation. Methods Balb / c mice were immunized with thymus epithelial cell line and monoclonal antibodies against thymic epithelial cells were prepared by B lymphocyte hybridoma technique. Immunochemical staining and flow cytometry analysis were used to determine the distribution of antigens recognized by these mAbs. Immuno-electrophoresis was used to determine the molecular weight of these antigens. Results Five anti-rat thymic epithelial cell monoclonal antibodies were obtained. According to the distribution of the thymus in the immunohistochemical staining, we concluded that the RE 4D8 monoclonal antibody was CTESⅡA type 1 and the RE 12B2 monoclonal antibody was CTESV type. ; RE 5C6 monoclonal antibody CTES Ⅳ type; RE 6D6 monoclonal antibody may be CTES Ⅱ B type; RE 1D4 monoclonal antibody CTES Ⅴ type. Antigens RE 4D8, RE 6D6, and RE 12B2 correspond to antigens with molecular weights of 120 kDa, 220 kDa and 35 kDa, respectively. RE 1D4 antigen may be a newly discovered marker of cortical epithelium. The established epithelial classification of thymus epithelial cell lines and their original location within the thymus were established based on the way they were bound to these mAbs. Among these cell lines used, the TuD32 and FTE3 phenotypes were cortical while the TaD34 phenotypes were medullary. Conclusions These mAbs clearly confirm the atypia of the thymic epithelium. The antigens they recognize appear both in the cytoplasm and on the surface of the thymus epithelium. These new monoclonal antibodies are expected to help recognize the close association of thymocytes and epithelial cells during T cell maturation