Rolfe B, Cavanagh A, Forde C, Bastin F, Chen C, Morton H
J Immunol Methods. 1984 May 11;70(1):1-11. doi: 10.1016/0022-1759(84)90383-1.
The rosette inhibition test has been modified so that early pregnancy factor (EPF) in human pregnancy serum can be detected with mouse lymphocytes. Interference with the assay, which would result from incubation of lymphocytes with heterologous serum proteins, is prevented by the introduction of an ion exchange chromatography step. This provides a simple and rapid method for separating EPF from interfering serum proteins. Human pregnancy and non-pregnancy sera were assayed for EPF with human lymphocytes and results compared with those obtained with DEAE-Sephacel fractions of the same sera tested with mouse lymphocytes. The 2 systems showed good agreement but the mouse assay gave greater differentiation between positive and negative results. When monoclonal anti-T cell antibodies, Hu Ly-m1 and anti-Ly-1.1, were substituted for anti-lymphocyte sera in the human and mouse assay systems respectively, similar results were obtained. The mouse assay system has several advantages over the human assay, including stability of the anti-mouse lymphocyte serum and the ready availability of mouse lymphocytes. Moreover the modified method may be applied not only to the assay of human EPF, but also to the assay of EPF from other species. This would be of value if several species were being studied in 1 laboratory, or if sufficient quantities of lymphocytes from a particular species were not available.
玫瑰花结抑制试验已被改进,以便能用小鼠淋巴细胞检测人妊娠血清中的早孕因子(EPF)。通过引入离子交换色谱步骤,可防止淋巴细胞与异源血清蛋白孵育所导致的对该检测的干扰。这提供了一种从干扰性血清蛋白中分离EPF的简单快速方法。用人淋巴细胞检测人妊娠和非妊娠血清中的EPF,并将结果与用小鼠淋巴细胞检测的相同血清的DEAE-葡聚糖凝胶片段所得结果进行比较。这两种系统显示出良好的一致性,但小鼠检测在阳性和阴性结果之间的区分度更高。当分别在人和小鼠检测系统中用单克隆抗T细胞抗体Hu Ly-m1和抗Ly-1.1替代抗淋巴细胞血清时,得到了相似的结果。小鼠检测系统相对于人检测具有几个优点,包括抗小鼠淋巴细胞血清的稳定性以及小鼠淋巴细胞易于获得。此外,改进后的方法不仅可应用于人EPF的检测,还可应用于其他物种EPF的检测。如果在一个实验室中研究多个物种,或者如果没有足够数量的特定物种的淋巴细胞,这将很有价值。