title: High-dimensional immune profiling of total and rotavirus VP6-specific intestinal and circulating B cells by mass cytometry.
abstract: In-depth phenotyping of human intestinal antibody secreting cells (ASCs) and their precursors is important for developing improved mucosal vaccines. We used single-cell mass cytometry to simultaneously analyze 34 differentiation and trafficking markers on intestinal and circulating B cells. In addition, we labeled rotavirus (RV) double-layered particles with a metal isotope and characterized B cells specific to the RV VP6 major structural protein. We describe the heterogeneity of the intestinal B-cell compartment, dominated by ASCs with some phenotypic and transcriptional characteristics of long-lived plasma cells. Using principal component analysis, we visualized the phenotypic relationships between major B-cell subsets in the intestine and blood, and revealed that IgM(+) memory B cells (MBCs) and naive B cells were phenotypically related as were CD27(-) MBCs and switched MBCs. ASCs in the intestine and blood were highly clonally related, but associated with distinct trajectories of phenotypic development. VP6-specific B cells were present among diverse B-cell subsets in immune donors, including naive B cells, with phenotypes representative of the overall B-cell pool. These data provide a high dimensional view of intestinal B cells and the determinants regulating humoral memory to a ubiquitous, mucosal pathogen at steady-state.
authors.affiliation: Department of Medicine, Stanford University, Stanford, California, USA.|Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.
authors.forename: N
authors.initials: N
authors.lastname: Nair
authors.affiliation: Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.|Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore, Singapore.
authors.forename: E W
authors.initials: EW
authors.lastname: Newell
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, California, USA.
authors.forename: C
authors.initials: C
authors.lastname: Vollmers
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, California, USA.
authors.forename: S R
authors.initials: SR
authors.lastname: Quake
authors.affiliation: Department of Surgery, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Morton
authors.affiliation: Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.|The Howard Hughes Medical Institute, Stanford, California, USA.|Institute for Immunity, Transplantation and Infection, Stanford University, Stanford, California, USA.
authors.forename: M M
authors.initials: MM
authors.lastname: Davis
authors.affiliation: Department of Medicine, Stanford University, Stanford, California, USA.|Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.
authors.forename: X S
authors.initials: XS
authors.lastname: He
authors.affiliation: Department of Medicine, Stanford University, Stanford, California, USA.|Department of Microbiology and Immunology, Stanford University, Stanford, California, USA.
authors.forename: H B
authors.initials: HB
authors.lastname: Greenberg
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D016175:B-Lymphocyte Subsets / Q000276:immunology* / Q000473:pathology / Q000821:virology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics / Q000276:immunology*; D002465:Cell Movement; D002522:Chlorocebus aethiops; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D004847:Epithelial Cells / Q000276:immunology / Q000821:virology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000276:immunology*; D054884:Host-Pathogen Interactions; D006801:Humans; D018928:Immunity, Mucosal; D007075:Immunoglobulin M / Q000235:genetics / Q000276:immunology; D007156:Immunologic Memory; D016130:Immunophenotyping; D007583:Jejunum / Q000276:immunology / Q000473:pathology / Q000821:virology; D025341:Principal Component Analysis; D012401:Rotavirus / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000172:deficiency / Q000235:genetics / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D016207:Cytokines; D007075:Immunoglobulin M; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; C061332:VP6 protein, Rotavirus