Takara Bio USA, Inc., San Jose, CA, USA.
Department of Pathology, Yale School of Medicine, New Haven, CT, USA.
Methods Mol Biol. 2022;2453:345-377. doi: 10.1007/978-1-0716-2115-8_19.
During the course of an immune response to a virus such as influenza, B cells undergo activation, clonal expansion, isotype switching, and somatic hypermutation (SHM). Members of an antigen-experienced B-cell clone can have different sequence features including SHM in the immunoglobulin heavy-chain V (IGHV) gene and can use the same IGVH gene in combination with different constant regions or isotypes (e.g., IgM, IgG, IgA). To study these features of expanded clones in an immune response by AIRR-seq, we provide a bulk RNA-based sequencing experimental procedure with unique molecular identifiers (UMIs) and the accompanying bioinformatics analytical workflow.
在对流感等病毒的免疫反应过程中，B 细胞经历激活、克隆扩增、同种型转换和体细胞高频突变 (SHM)。经历抗原的 B 细胞克隆的成员可以具有不同的序列特征，包括免疫球蛋白重链 V (IGHV) 基因中的 SHM，并且可以使用相同的 IGVH 基因与不同的恒定区或同种型（例如，IgM、IgG、IgA）结合。为了通过 AIRR-seq 研究免疫反应中扩增克隆的这些特征，我们提供了一种基于批量 RNA 的测序实验程序，其中包含独特的分子标识符 (UMIs) 和伴随的生物信息学分析工作流程。
