Sanz I, Dang H, Takei M, Talal N, Capra J D
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
J Immunol. 1989 Feb 1;142(3):883-7.
The utilization of germline genes for the synthesis of autoantibodies has been suspected for many years based on the presence of cross-reactive idiotypes among patients as well as in some healthy first-degree relatives of patients with several autoimmune diseases including SLE. One such system of idiotypes involves anti-Sm antibodies, which are highly specific for SLE. To definitively establish the utilization of germline genes in the Sm system, we produced human-human B cell hybridomas from a patient with SLE who had circulating anti-Sm antibodies. One stable hybridoma designated 4B4 secretes an IgM-kappa mAb that binds Sm and shares idiotypic determinants with other anti-Sm antibodies. A second anti-Sm antibody (3C3), isolated from the same patient was also studied. Oligo(dT) priming was used to produce cDNA corresponding to full length IgM. Sequence analysis revealed that the VH gene segment (1-96) of 4B4 is identical to a VH sequence previously detected in a fetal liver cDNA library by Schroeder and his co-workers as well as a germline VH recently described by Berman and his associates. The identity of a lupus mAb and sequences derived from unrelated individuals provides strong evidence that this autoantibody is a direct copy of a germline gene.
多年来,基于患者之间以及一些患有包括系统性红斑狼疮(SLE)在内的多种自身免疫性疾病患者的健康一级亲属中存在交叉反应性独特型,人们一直怀疑种系基因参与了自身抗体的合成。其中一个独特型系统涉及抗Sm抗体,它对SLE具有高度特异性。为了明确确定种系基因在Sm系统中的利用情况,我们从一名患有循环抗Sm抗体的SLE患者身上制备了人-人B细胞杂交瘤。一个名为4B4的稳定杂交瘤分泌一种IgM-κ单克隆抗体,它能结合Sm并与其他抗Sm抗体共享独特型决定簇。还研究了从同一名患者分离出的第二种抗Sm抗体(3C3)。使用寡聚(dT)引物产生对应于全长IgM的cDNA。序列分析表明,4B4的VH基因片段(1-96)与Schroeder及其同事先前在胎儿肝脏cDNA文库中检测到的VH序列以及Berman及其同事最近描述的种系VH相同。狼疮单克隆抗体与来自无关个体的序列相同,这提供了有力证据,表明这种自身抗体是种系基因的直接拷贝。