ABSTRACT

The autoimmune disease known as Jo-1 positive anti-synthetase syndrome (ASS) is characterized by circulating antibody titers to histidyl-tRNA synthetase (HARS), which may play a role in modulating the non-canonical functions of HARS. Monoclonal antibodies to HARS were isolated by single-cell screening and sequencing from three Jo-1 positive ASS patients and shown to be of high affinity, covering diverse epitope space. The immune response was further characterized by repertoire sequencing from the most productive of the donor samples. In line with previous studies of autoimmune repertoires, these antibodies tended to have long complementarity-determining region H3 sequences with more positive-charged residues than average. Clones of interest were clustered into groups with related sequences, allowing us to observe different somatic mutations in related clones. We postulated that these had found alternate structural solutions for high affinity binding, but that mutations might be transferable between clones to further enhance binding affinity. Transfer of somatic mutations between antibodies within the same clonal group was able to enhance binding affinity in a number of cases, including beneficial transfer of a mutation from a lower affinity clone into one of higher affinity. Affinity enhancement was seen with mutation transfer both between related single-cell clones, and directly from related repertoire sequences. To our knowledge, this is the first demonstration of somatic hypermutation transfer from repertoire sequences to further mature in vivo derived antibodies, and represents an additional tool to aid in affinity maturation for the development of antibodies.

摘要

被称为 Jo-1 阳性抗合成酶综合征 (ASS) 的自身免疫性疾病的特征是循环中针对组氨酰 tRNA 合成酶 (HARS) 的抗体滴度，这可能在调节 HARS 的非经典功能中发挥作用。HARS 的单克隆抗体是通过单细胞筛选和测序从三名 Jo-1 阳性 ASS 患者中分离出来的，并且显示出高亲和力，覆盖了不同的表位空间。免疫反应的进一步特征是来自最有生产力的供体样品的库序列。与之前对自身免疫组库的研究一致，这些抗体往往具有较长的互补决定区 H3 序列，其带正电荷的残基比平均值多。感兴趣的克隆被聚集成具有相关序列的组，允许我们观察相关克隆中的不同体细胞突变。我们假设这些已经找到了高亲和力结合的替代结构解决方案，但突变可能可以在克隆之间转移以进一步增强结合亲和力。在许多情况下，同一克隆组内抗体之间的体细胞突变转移能够增强结合亲和力，包括将突变从较低亲和力克隆有益地转移到较高亲和力克隆中。通过相关单细胞克隆之间和直接从相关库序列的突变转移，可以看到亲和力增强。据我们所知，这是首次证明体细胞超突变从库序列转移到进一步成熟 我们假设这些已经找到了高亲和力结合的替代结构解决方案，但突变可能可以在克隆之间转移以进一步增强结合亲和力。在许多情况下，同一克隆组内抗体之间的体细胞突变转移能够增强结合亲和力，包括将突变从较低亲和力克隆有益地转移到较高亲和力克隆中。通过相关单细胞克隆之间的突变转移以及直接来自相关库序列的突变转移，都可以看到亲和力增强。据我们所知，这是首次证明体细胞超突变从库序列转移到进一步成熟 我们假设这些已经找到了高亲和力结合的替代结构解决方案，但突变可能可以在克隆之间转移以进一步增强结合亲和力。在许多情况下，同一克隆组内抗体之间的体细胞突变转移能够增强结合亲和力，包括将突变从较低亲和力克隆有益地转移到较高亲和力克隆中。通过相关单细胞克隆之间的突变转移以及直接来自相关库序列的突变转移，都可以看到亲和力增强。据我们所知，这是首次证明体细胞超突变从库序列转移到进一步成熟 在许多情况下，同一克隆组内抗体之间的体细胞突变转移能够增强结合亲和力，包括将突变从较低亲和力克隆有益地转移到较高亲和力克隆中。通过相关单细胞克隆之间的突变转移以及直接来自相关库序列的突变转移，都可以看到亲和力增强。据我们所知，这是首次证明体细胞超突变从库序列转移到进一步成熟 在许多情况下，同一克隆组内抗体之间的体细胞突变转移能够增强结合亲和力，包括将突变从较低亲和力克隆有益地转移到较高亲和力克隆中。通过相关单细胞克隆之间和直接从相关库序列的突变转移，可以看到亲和力增强。据我们所知，这是首次证明体细胞超突变从库序列转移到进一步成熟体内衍生的抗体，代表了一种额外的工具，可以帮助抗体开发的亲和力成熟。