Kerbel R S
J Immunol Methods. 1980;34(1):1-10. doi: 10.1016/0022-1759(80)90218-5.
The sensitivity of the conventional EA (Fc) rosetting assay for detecting Fc receptors could be significantly increased by coating sheep erythrocytes (SRBC) with high concentrations of a preparation of homogeneous monoclonal IgG2b anti-SRBC antibodies. This was made possible by virtue of the fact that the monoclonal antibody preparation used was completely incapable of directly hemagglutinating SRBC, although it possessed a high titer as assessed by direct hemolysis or direct hemagglutination. The monoclonal IgG2b- coated SRBC indicator cells were compared to SRBC coated with subhemagglutinating amounts of the IgG fraction of a hemagglutinating rabbit anti-SRBC antiserum: at high coupling concentrations, the monoclonal antibody-coupled SRBC gave consistently higher percentages of Fc rosettes, a large proportion of which were large and multi-layered, when various lymphoid cell populations and a variety of Fc receptor-positive cultured tumor cell line populations were tested. The increased sensitivity of the Fc rosette attained by this method may depend on the nature of the monoclonal anti-SRBC reagent used. Thus, indicator cells prepared by incubation with high concentrations of IgG1 subclass-specific non-hemagglutinating monoclonal anti-SRBC antibodies always gave significantly lower percentages of Fc rosettes than conventionally prepared indicator cells.
通过用高浓度的同源单克隆IgG2b抗绵羊红细胞（SRBC）抗体制剂包被绵羊红细胞（SRBC），检测Fc受体的传统EA（Fc）玫瑰花结试验的灵敏度可显著提高。这是因为所使用的单克隆抗体制剂完全不能直接使SRBC发生血凝，尽管通过直接溶血或直接血凝评估其具有高滴度。将单克隆IgG2b包被的SRBC指示细胞与用亚血凝量的血凝兔抗SRBC抗血清的IgG组分包被的SRBC进行比较：在高偶联浓度下，当检测各种淋巴细胞群体和多种Fc受体阳性培养肿瘤细胞系群体时，单克隆抗体偶联的SRBC产生的Fc玫瑰花结百分比始终较高，其中很大一部分是大的和多层的。通过这种方法获得的Fc玫瑰花结灵敏度的提高可能取决于所用单克隆抗SRBC试剂的性质。因此，用高浓度IgG1亚类特异性非血凝单克隆抗SRBC抗体孵育制备的指示细胞产生的Fc玫瑰花结百分比总是明显低于传统制备的指示细胞。
