Préville X, Nicolas L, Flacher M, Revillard J
INSERM U503, Pavillon P, Hôpital E. Herriot, 5 Place d'Arsonval, 69437 Cedex 03, Lyon, France.
J Immunol Methods. 2000 Nov 1;245(1-2):45-54. doi: 10.1016/s0022-1759(00)00271-4.
Rabbit antilymphocyte or antithymocyte globulins (rATG) are currently used as immunosuppressive agents in clinical organ and bone marrow transplantation, but because of differences in antigen source and purification processes between manufacturers, determination of the dose to be administered remains empirical. Dosage may be adjusted to peripheral blood T cell counts or limited by side effects such as neutropenia or thrombocytopenia. We report here a measurement by indirect immunofluorescence of the amount of antibodies from rATG that bind to different blood cell types. A concentration-related immunofluorescence signal was obtained with reasonable replicability, allowing comparison of different rATGs by reference to the same rATG batch as internal standard. Ratios of antilymphocyte to undesirable antibodies (directed against antigens expressed on neutrophils, platelets or red cells) could be calculated for each rATG preparation. Finally, the method is applicable to the measurement of free antibodies in sera from patients treated with rATG and to the analysis of the effects of in vivo absorption of rATG during its administration. Marked differences in antibody content per mg of rIgG account for the differences in rATG dosage that were empirically established for clinical applications.
兔抗淋巴细胞或抗胸腺细胞球蛋白(rATG)目前在临床器官和骨髓移植中用作免疫抑制剂,但由于不同生产商的抗原来源和纯化工艺存在差异,给药剂量的确定仍基于经验。剂量可根据外周血T细胞计数进行调整,或受中性粒细胞减少或血小板减少等副作用的限制。我们在此报告一种通过间接免疫荧光法测量rATG中与不同血细胞类型结合的抗体量的方法。获得了具有合理可重复性的浓度相关免疫荧光信号,通过将不同的rATG与作为内标的同一rATG批次进行比较,可以对它们进行比较。可以为每种rATG制剂计算抗淋巴细胞抗体与不良抗体(针对中性粒细胞、血小板或红细胞上表达的抗原)的比率。最后,该方法适用于测量接受rATG治疗患者血清中的游离抗体,以及分析rATG在给药过程中体内吸收的影响。每毫克rIgG的抗体含量存在显著差异,这解释了临床应用中凭经验确定的rATG剂量差异。