Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT 06511.
Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT 06511;
J Immunol. 2019 Oct 1;203(7):1687-1692. doi: 10.4049/jimmunol.1900666. Epub 2019 Sep 4.
B cell clonal expansion is vital for adaptive immunity. High-throughput BCR sequencing enables investigating this process but requires computational inference to identify clonal relationships. This inference usually relies on only the BCR H chain, as most current protocols do not preserve H:L chain pairing. The extent to which paired L chains aids inference is unknown. Using human single-cell paired BCR datasets, we assessed the ability of H chain-based clonal clustering to identify clones. Of the expanded clones identified, <20% grouped cells expressing inconsistent L chains. H chains from these misclustered clones contained more distant junction sequences and shared fewer V segment mutations than the accurate clones. This suggests that additional H chain information could be leveraged to refine clonal relationships. Conversely, L chains were insufficient to refine H chain-based clonal clusters. Overall, the BCR H chain alone is sufficient to identify clonal relationships with confidence.
B 细胞克隆扩增对于适应性免疫至关重要。高通量 BCR 测序能够研究这个过程，但需要计算推理来识别克隆关系。这种推理通常仅依赖于 BCR H 链，因为大多数当前的方案都没有保留 H:L 链配对。配对的 L 链在多大程度上有助于推理尚不清楚。使用人类单细胞配对 BCR 数据集，我们评估了基于 H 链的克隆聚类识别克隆的能力。在鉴定的扩增克隆中，<20%的分组细胞表达不一致的 L 链。这些聚类错误的克隆的 H 链具有更远的连接序列，并且共享的 V 片段突变比准确的克隆少。这表明可以利用更多的 H 链信息来细化克隆关系。相反，L 链不足以细化基于 H 链的克隆聚类。总体而言，单独的 BCR H 链足以有信心地识别克隆关系。
