Schreuder G M, Marsh S G, Heyes J M, Moses J H, Krausa P, Bodmer J G
Tissue Antigen Laboratory, Imperial Cancer Research Fund, London, United Kingdom.
Tissue Antigens. 1990 Aug;36(2):75-80. doi: 10.1111/j.1399-0039.1990.tb01803.x.
Modifications of the standard microcytotoxicity assay make it possible to use this technique to screen both alloantisera and monoclonal antibodies with mouse L cells transfected with Class II genes. It is necessary to maintain a protein-rich environment in order to prevent nonspecific complement lysis. Selection of the complement itself is also an important factor, the best results being achieved using a commercially available complement that had previously been absorbed with mouse cells and used at a dilution of 1/8. Using this modified method with transfectants of DW2 origin we could show that alloantisera against DRw15 recognize the DRB11501 gene product, whereas broad DR2 sera react only with the DRB50101 product. This technique can be applied successfully to study the fine specificity of polymorphic monoclonal antibodies, as shown by the reactivity of HU-30 which binds to the LDR2b transfectant and not to the LDR2a, indicating that the antibody recognizes an epitope present on the DRB1 chain and not the DRB5 chain of DR2 cell lines.
对标准微量细胞毒性试验进行改进后，就能够利用该技术，用转染了Ⅱ类基因的小鼠L细胞来筛选同种异体抗血清和单克隆抗体。为防止非特异性补体溶解，保持富含蛋白质的环境很有必要。补体本身的选择也是一个重要因素，使用先前已用小鼠细胞吸收过、并以1/8稀释度使用的市售补体可获得最佳结果。使用这种改良方法对源自DW2的转染体进行检测，我们发现针对DRw15的同种异体抗血清可识别DRB11501基因产物，而宽泛的DR2血清仅与DRB50101产物发生反应。如HU - 30与LDR2b转染体反应而不与LDR2a反应所示，该技术可成功应用于研究多态性单克隆抗体的精细特异性，这表明该抗体识别的是DR2细胞系DRB1链而非DRB5链上存在的一个表位。
