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

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

PROVIDER: PRJNA1026914 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26351562_1.fastq.gz	Fastqsanger.gz
	SRR26351562_2.fastq.gz	Fastqsanger.gz
	SRR26351563_1.fastq.gz	Fastqsanger.gz
	SRR26351563_2.fastq.gz	Fastqsanger.gz
	SRR26351564_1.fastq.gz	Fastqsanger.gz

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

Project description:Induction of adaptive immune responses to commensal microbes is critical for intestinal 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 cells ex vivo. To address this, we first sought to identify immunodominant protein epitopes recognized by Segmented Filamentous Bacteria (SFB) specific serum antibodies using a whole-genome phage display screen and identified immunogenic proteins engaging IgA, IgG1 and IgG2b responses. 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 and de novo colonization. We identified a compartmentalized response in B cell activation between Peyer’s patches and mesenteric lymph nodes, with a gradient of IgA, IgG1 and IgG2b isotypes along the small intestine, and selective production of IgG2b with 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.

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