Publication

Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans

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Last modified
  • 05/15/2025
Type of Material
Authors
    Nicholas Giovannone, Brigham & Women's HospitalAristotelis Antonopoulos, Imperial College LondonJennifer Liang, Brigham & Women's HospitalJenna Geddes Sweeney, Brigham & Women's HospitalMatthew R. Kudelka, Beth Israel Deaconess Medical CenterSandra L. King, Brigham & Women's HospitalGi Soo Lee, Harvard Medical SchoolRichard Cummings, Emory UniversityAnne Dell, Imperial College LondonSteven R. Barthel, Brigham & Women's HospitalHans R. Widlund, Brigham & Women's HospitalStuart M. Haslam, Imperial College LondonCharles J. Dimitroff, Brigham & Women's Hospital
Language
  • English
Date
  • 2018-12-14
Publisher
  • Frontiers Media
Publication Version
Copyright Statement
  • Copyright © 2018 Giovannone, Antonopoulos, Liang, Geddes Sweeney, Kudelka, King, Lee, Cummings, Dell, Barthel, Widlund, Haslam and Dimitroff.
License
Final Published Version (URL)
Title of Journal or Parent Work
ISSN
  • 1664-3224
Volume
  • 9
Start Page
  • 2857
End Page
  • 2857
Grant/Funding Information
  • This research was funded by an American Association of Immunologists Careers in Immunology Fellowship (to NG and CD); an Albert J. Ryan foundation fellowship (to NG); NIH grant NIH/NIAID R21AI125476 (to CD) ; and NIH/NCI R01 CA173610 (to CD); a Young Investigator Award from the Merck-Melanoma Research Alliance (to SB); a Research Scholar Grant from the V Foundation for Cancer Research (to SB); a Melanoma Research Scholar Award from the Rochester Melanoma Action Group/Outrun the Sun (to SB); a Biotechnology and Biological Sciences Research Council grant BBF0083091 (AD and SH) and BBK0161641 (AD and SH); and a Wellcome Trust grant (082098 to AD).
Supplemental Material (URL)
Abstract
  • Germinal centers (GC) are microanatomical niches where B cells proliferate, undergo antibody affinity maturation, and differentiate to long-lived memory B cells and antibody-secreting plasma cells. For decades, GC B cells have been defined by their reactivity to the plant lectin peanut agglutinin (PNA), which binds serine/threonine (O-linked) glycans containing the asialylated disaccharide Gal-β1,3-GalNAc-Ser/Thr (also called T-antigen). In T cells, acquisition of PNA binding by activated T cells and thymocytes has been linked with altered tissue homing patterns, cell signaling, and survival. Yet, in GC B cells, the glycobiological basis and significance of PNA binding remains surprisingly unresolved. Here, we investigated the basis for PNA reactivity of GC B cells. We found that GC B cell binding to PNA is associated with downregulation of the α2,3 sialyltransferase, ST3GAL1 (ST3Gal1), and overexpression of ST3Gal1 was sufficient to reverse PNA binding in B cell lines. Moreover, we found that the primary scaffold for PNA-reactive O-glycans in B cells is the B cell receptor-associated receptor-type tyrosine phosphatase CD45, suggesting a role for altered O-glycosylation in antigen receptor signaling. Consistent with similar reports in T cells, ST3Gal1 overexpression in B cells in vitro induced drastic shortening in O-glycans, which we confirmed by both antibody staining and mass spectrometric O-glycomic analysis. Unexpectedly, ST3Gal1-induced changes in O-glycan length also correlated with altered binding of two glycosylation-sensitive CD45 antibodies, RA3-6B2 (more commonly called B220) and MEM55, which (in humans) have previously been reported to favor binding to naïve/GC subsets and memory/plasmablast subsets, respectively. Analysis of primary B cell binding to B220, MEM55, and several plant lectins suggested that B cell differentiation is accompanied by significant loss of O-glycan complexity, including loss of extended Core 2 O-glycans. To our surprise, decreased O-glycan length from naïve to post-GC fates best correlated not with ST3Gal1, but rather downregulation of the Core 2 branching enzyme GCNT1. Thus, our data suggest that O-glycan remodeling is a feature of B cell differentiation, dually regulated by ST3Gal1 and GCNT1, that ultimately results in expression of distinct O-glycosylation states/CD45 glycoforms at each stage of B cell differentiation.
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Keywords
Research Categories
  • Health Sciences, Immunology

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