Carreno M, Fuller L, Zucker K, Yang W C, Burke G, Nery J, Gomez C, Esquenazi V, Miller J
Department of Surgery, University of Miami School of Medicine, FL 33136.
Hum Immunol. 1992 Apr;33(4):249-58. doi: 10.1016/0198-8859(92)90332-h.
The administration of murine mAb specific for the CD3 epsilon subunit of the TCR complex (OKT3) has been demonstrated to engender in humans an anti-OKT3 idiotypic cascade. This study used murine-derived anti-OKT3 (Ab2) as a bioreagent to determine whether this Ab2 and polyclonal anti-(anti-OKT3) (Ab3) generated in some human kidney transplant patients are idiotypically connected. Two anti-OKT3 mAbs G-880 (IgG1) and M-12 (IgM) were derived by immunizing BALB/c mice with the OKT3-secreting hybridoma. The two mAbs exhibited specificity for OKT3 F(ab)'2 idiotypic determinants. Both mAbs were tested for their ability to inhibit OKT3 induced mitogenesis and to block FITC-OKT3 binding to cell surface CD3 epsilon chain. The M-12 mAb inhibited OKT3-induced mitogenesis and blocked (approximately 60%) the binding of OKT3 to peripheral blood (PBL) T-cell CD3 epsilon chain in flow cytometry. In contrast, the G-880 mAb did not inhibit mitogenesis and only weakly blocked OKT3 binding to CD3 epsilon chain (approximately 12%). Sera of kidney transplant recipients who received OKT3 antirejection therapy and who developed antiidiotypic anti-OKT3 antibodies could be divided into two subgroups exhibiting anti-OKT3 activity: (a) those who had similar specificity as M-12 and failed to enhance the M-12 inhibition of OKT3 binding to PBL T-cell CD3 epsilon chain when added as a third component (n = 3), and (b) those with anti-OKT3 antibodies with idiotype specificity dissimilar to M-12 and who were able to increase the (maximum 60%) inhibition obtained with M-12 in the OKT3 to T-cell CD3-binding assay (n = 4). From these observations, we conclude that M-12 had the characteristics of an Ab2 beta and G-880 that of an Ab2 alpha. Additionally, there was an idiotypic connectivity of mouse-derived M-12 anti-OKT3 (Ab2) and OKT3-engendered human polyclonal anti-(anti-OKT3) (Ab3), in that three of seven patients examined had human serum IgG antibodies that specifically recognized M-12 idiotypic determinants as demonstrated in ELISA.
已证明给予针对T细胞受体复合物CD3ε亚基的鼠单克隆抗体（OKT3）可在人体内引发抗OKT3独特型级联反应。本研究使用鼠源抗OKT3（Ab2）作为生物试剂，以确定该Ab2与一些人类肾移植患者体内产生的多克隆抗（抗OKT3）（Ab3）是否存在独特型联系。通过用分泌OKT3的杂交瘤免疫BALB/c小鼠，获得了两种抗OKT3单克隆抗体G - 880（IgG1）和M - 12（IgM）。这两种单克隆抗体对OKT3 F(ab)'2独特型决定簇具有特异性。检测了这两种单克隆抗体抑制OKT3诱导的有丝分裂以及阻断FITC - OKT3与细胞表面CD3ε链结合的能力。M - 12单克隆抗体抑制了OKT3诱导的有丝分裂，并在流式细胞术中阻断了（约60%）OKT3与外周血（PBL）T细胞CD3ε链的结合。相比之下，G - 880单克隆抗体未抑制有丝分裂，仅微弱地阻断了OKT3与CD3ε链的结合（约12%）。接受OKT3抗排斥治疗并产生抗独特型抗OKT3抗体的肾移植受者血清可分为两个具有抗OKT3活性的亚组：（a）那些具有与M - 12相似特异性且作为第三成分添加时未能增强M - 12对OKT3与PBL T细胞CD3ε链结合的抑制作用的患者（n = 3），以及（b）那些具有与M - 12独特型特异性不同的抗OKT3抗体且能够在OKT3与T细胞CD3结合试验中增加M - 12所获得的（最大60%）抑制作用的患者（n = 4）。基于这些观察结果，我们得出结论，M - 12具有Ab2β的特征，G - 880具有Ab2α的特征。此外，鼠源M - 12抗OKT3（Ab2）与OKT3诱导产生的人多克隆抗（抗OKT3）（Ab3）存在独特型联系，因为在酶联免疫吸附测定（ELISA）中，所检测的7名患者中有3名患者的人血清IgG抗体能够特异性识别M - 12独特型决定簇。
