Antigen-level resolution of commensal-specific B cell responses enabled by phage-display screening and B cell tetramers

Abstract

SummaryInduction of adaptive immune responses to commensal microbes is critical for tissue homeostasis, and perturbation of these responses is associated with multiple chronic inflammatory disorders. However, the mechanisms underlying the induction and regulation of mucosal B cells targeting commensal microbes remain poorly understood, in part due to a lack of tools to identify commensal-specific B cellsex vivo. To address this, we identified immunogenic protein epitopes recognized by Segmented Filamentous Bacteria (SFB)-specific serum antibodies using a whole-genome phage display screen and identified immunogenic proteins engaging IgA, IgG1and IgG2bresponses. Using these antigens, we generated B cell tetramers to identify and track SFB-specific B cell responses in the gut associated lymphoid tissue during natural andde novocolonization. We revealed a compartmentalized response in SFB-specific B cell activation between Peyer’s patches and mesenteric lymph nodes, with a gradient of IgA, IgG1and IgG2bisotypes along the small intestine, and selective production of IgG2bwith the mesenteric lymph node chain. VDJ sequencing analyses and generation of SFB-specific monoclonal antibodies identified that somatic hypermutation drives affinity maturation to SFB derived antigens under homeostatic conditions. By combining phage display screening and B cell tetramer technologies, we now enable antigen-level based studies of immunity to intestinal microbes, which will advance our understanding of the ontogeny and function of commensal-specific B cell responses in tissue immunity, inflammation and repair.