Lee Yu-Ching, Tsai Keng-Chang, Leu Sy-Jye, Wang Tuan-Jen, Liu Chia-Yu, Yang Yi-Yuan
The Institute for Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei 110, Taiwan ; Antibody and Hybridoma Core Facility, Taipei Medical University, Taipei 110, Taiwan.
National Research Institute of Chinese Medicine, Taipei 110, Taiwan.
ScientificWorldJournal. 2013 Dec 30;2013:516516. doi: 10.1155/2013/516516. eCollection 2013.
We have previously isolated several IgG rheumatoid factors (RFs) from patients with both rheumatoid arthritis and idiopathic thrombocytopenia purpura using phage display system. To study IgG RFs in patients with other autoimmune diseases, phage display antibody libraries from a hepatitis C virus infected patient with Sjögren's syndrome were constructed. After panning, a specific clone RFL11 was isolated for characterization in advance. The binding activity and specificity of RFL11 to IgG Fc fragment were comparable to those of RFs previously isolated. The analysis with existed RF-Fc complex structures indicated the homology model of RFL11 is similar to IgM RF61 complex with high binding affinity of about 6 × 10⁻⁸ M. This effect resulted from longer complementarity-determining region (CDR) combining key somatic mutations. In the RFL11-Fc interfaces, the CDR-H3 loop forms a finger-like structure extending into the bottom of Fc pocket and resulting in strong ion and cation-pi interactions. Moreover, a process of antigen-driven maturation was proven by somatically mutated VH residues on H2 and H3 CDR loops in the interfaces. Taken together, these results suggested that high affinity IgG RFs can be generated in patients with Sjögren's syndrome and may play an important role in the pathogenesis of this autoimmune disease.
我们之前使用噬菌体展示系统从类风湿性关节炎和特发性血小板减少性紫癜患者中分离出了几种IgG类风湿因子(RFs)。为了研究其他自身免疫性疾病患者中的IgG RFs,构建了来自一名丙型肝炎病毒感染的干燥综合征患者的噬菌体展示抗体文库。淘选后,预先分离出一个特异性克隆RFL11进行表征。RFL11与IgG Fc片段的结合活性和特异性与先前分离的RFs相当。对现有RF-Fc复合物结构的分析表明,RFL11的同源模型与IgM RF61复合物相似,具有约6×10⁻⁸ M的高结合亲和力。这种效应源于更长的互补决定区(CDR)结合关键的体细胞突变。在RFL11-Fc界面中,CDR-H3环形成一个手指状结构,延伸到Fc口袋底部,导致强烈的离子和阳离子-π相互作用。此外,界面中H2和H3 CDR环上的体细胞突变VH残基证明了抗原驱动的成熟过程。综上所述,这些结果表明干燥综合征患者可产生高亲和力的IgG RFs,且可能在这种自身免疫性疾病的发病机制中起重要作用。