Kootte A M, Langelaar R A, Tanke H J, Van der Voort E A, Paul L C
Department of Medicine, University Hospital, Leiden, The Netherlands.
J Immunol Methods. 1988 Jun 28;111(1):75-82. doi: 10.1016/0022-1759(88)90061-0.
T cell subset analysis with monoclonal antibodies is performed in variety of clinical situations, including the monitoring of transplant recipients. We compared the percentages CD8+ cells in the peripheral blood of ten normal volunteers and 26 renal transplant patients using fluorescein isothiocyanate (FITC)- or phycoerytrin (PE)-conjugated Leu2a and Leu2b monoclonal antibodies (mAb). Furthermore, the percentage of HLA-DR+ cells within this population was determined using two anti-HLA-DR antibodies of differing isotype (designated 203 and 243) and either an FITC or a PE label. For both controls and transplant recipients, the PE-conjugated Leu2a and Leu2b mAbs gave significantly higher percentages of positive cells than the FITC-conjugated antibodies (P less than 0.01). Furthermore, the percentage of Leu2b+ cells was higher than the percentage of Leu2a+ cells, irrespective of the fluorochrome used (P less than 0.01); this was particularly true for samples with less than or equal to 10% Leu2a+ cells. Cell sorting experiments indicated that up to 30% of the Leu2a- population were able to bind OKT8 or Leu2b mAb in blood samples with a low percentage of Leu2a+ cells. The percentage of Leu2+ cells that were stained with anti-HLA-DR mAb 203 or 243 varied considerably, depending on the fluorochrome and the isotype of the antibodies. We conclude that the analysis of peripheral blood T cells with mAbs that apparently have the same specificity may give significantly different results, depending on the patient population, the fluorochrome and the isotype of the antibodies.
使用单克隆抗体进行T细胞亚群分析已应用于多种临床情况，包括对移植受者的监测。我们使用异硫氰酸荧光素（FITC）或藻红蛋白（PE）偶联的Leu2a和Leu2b单克隆抗体（mAb），比较了10名正常志愿者和26名肾移植患者外周血中CD8 +细胞的百分比。此外，使用两种不同同种型的抗HLA - DR抗体（命名为203和243）以及FITC或PE标记，测定了该群体中HLA - DR +细胞的百分比。对于对照组和移植受者，PE偶联的Leu2a和Leu2b mAb所给出的阳性细胞百分比均显著高于FITC偶联的抗体（P小于0.01）。此外，无论使用何种荧光染料，Leu2b +细胞的百分比均高于Leu2a +细胞的百分比（P小于0.01）；对于Leu2a +细胞小于或等于10%的样本尤其如此。细胞分选实验表明，在Leu2a +细胞百分比低的血样中，高达30%的Leu2a阴性群体能够结合OKT8或Leu2b mAb。用抗HLA - DR mAb 203或243染色的Leu2 +细胞百分比差异很大，这取决于荧光染料和抗体的同种型。我们得出结论，使用表面上具有相同特异性的mAb分析外周血T细胞，可能会根据患者群体、荧光染料和抗体的同种型而产生显著不同的结果。
