Generation of a single-cell B cell atlas of antibody repertoires and transcriptomes to identify signatures associated with antigen specificity

oleh: Andreas Agrafiotis, Daniel Neumeier, Kai-Lin Hong, Tasnia Chowdhury, Roy Ehling, Raphael Kuhn, Ioana Sandu, Victor Kreiner, Tudor-Stefan Cotet, Danielle Shlesinger, Daria Laslo, Stine Anzböck, Dale Starkie, Daniel J. Lightwood, Annette Oxenius, Sai T. Reddy, Alexander Yermanos

Format: Article
Diterbitkan: Elsevier 2023-03-01

Deskripsi

Summary: Although new genomics-based pipelines have potential to augment antibody discovery, these methods remain in their infancy due to an incomplete understanding of the selection process that governs B cell clonal selection, expansion, and antigen specificity. Furthermore, it remains unknown how factors such as aging and reduction of tolerance influence B cell selection. Here we perform single-cell sequencing of antibody repertoires and transcriptomes of murine B cells following immunizations with a model therapeutic antigen target. We determine the relationship between antibody repertoires, gene expression signatures, and antigen specificity across 100,000 B cells. Recombinant expression and characterization of 227 monoclonal antibodies revealed the existence of clonally expanded and class-switched antigen-specific B cells that were more frequent in young mice. Although integrating multiple repertoire features such as germline gene usage and transcriptional signatures failed to distinguish antigen-specific from nonspecific B cells, other features such as immunoglobulin G (IgG) subtype and sequence composition correlated with antigen specificity.