Finnern R, Bye J M, Dolman K M, Zhao M H, Short A, Marks J D, Lockwood M C, Ouwehand W H
University of Cambridge, Division of Transfusion Medicine, UK.
Clin Exp Immunol. 1995 Dec;102(3):566-74. doi: 10.1111/j.1365-2249.1995.tb03854.x.
Recently it has been demonstrated that human antibody fragments with binding activities against self antigens can be isolated from repertoires of rearranged V genes from non-immunized humans. We have applied phage display technology to study the B cell repertoire for antibody activity against neutrophil cytoplasmic antigens. These antibodies may play an important role in Wegener's granulomatosis (WG) and related forms of vasculitides. Autoantibodies in patients with WG are directed against proteinase 3. The immunodominant antigen in other forms of vasculitis is myeloperoxidase, but the B cell response can also be directed against other neutrophil enzymes, e.g. lysozyme, human neutrophil elastase, lactoferrin and cathepsin G. We show here that anti-self reactivity against neutrophil cytoplasmic antigens can be detected in the rearranged V gene repertoire of healthy individuals and that the reactivity can be directed against structural related epitopes which are present on different neutrophil cytoplasmic antigens. The scFv with binding activities were sequenced and the V gene usage, the level of somatic mutations and the immunoserological characteristics of the antibody fragments are discussed. Further evidence is presented that antibody fragments consisting only of a heavy chain variable domain can recognize neutrophil cytoplasmic antigens in a specific manner. These single-domain antibody fragments were used in experiments designed to establish the relative role of the light chain variable domains in antigen binding.
最近有研究表明，具有针对自身抗原结合活性的人抗体片段可从未免疫人群重排的V基因库中分离得到。我们应用噬菌体展示技术研究了针对中性粒细胞胞浆抗原的抗体活性的B细胞库。这些抗体可能在韦格纳肉芽肿（WG）及相关血管炎形式中发挥重要作用。WG患者的自身抗体针对蛋白酶3。其他形式血管炎中的免疫显性抗原是髓过氧化物酶，但B细胞反应也可针对其他中性粒细胞酶，如溶菌酶、人中性粒细胞弹性蛋白酶、乳铁蛋白和组织蛋白酶G。我们在此表明，在健康个体重排的V基因库中可检测到针对中性粒细胞胞浆抗原的自身反应性，且该反应性可针对存在于不同中性粒细胞胞浆抗原上的结构相关表位。对具有结合活性的单链抗体片段进行了测序，并讨论了V基因的使用情况、体细胞突变水平以及抗体片段的免疫血清学特征。进一步的证据表明，仅由重链可变域组成的抗体片段能够以特异性方式识别中性粒细胞胞浆抗原。这些单域抗体片段被用于实验，以确定轻链可变域在抗原结合中的相对作用。
