Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans
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 (P...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-12-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2018.02857/full |
| _version_ | 1819130784761511936 |
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| author | Nicholas Giovannone Nicholas Giovannone Aristotelis Antonopoulos Jennifer Liang Jenna Geddes Sweeney Jenna Geddes Sweeney Matthew R. Kudelka Matthew R. Kudelka Sandra L. King Gi Soo Lee Richard D. Cummings Richard D. Cummings Anne Dell Steven R. Barthel Hans R. Widlund Hans R. Widlund Stuart M. Haslam Charles J. Dimitroff Charles J. Dimitroff |
| author_facet | Nicholas Giovannone Nicholas Giovannone Aristotelis Antonopoulos Jennifer Liang Jenna Geddes Sweeney Jenna Geddes Sweeney Matthew R. Kudelka Matthew R. Kudelka Sandra L. King Gi Soo Lee Richard D. Cummings Richard D. Cummings Anne Dell Steven R. Barthel Hans R. Widlund Hans R. Widlund Stuart M. Haslam Charles J. Dimitroff Charles J. Dimitroff |
| author_sort | Nicholas Giovannone |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-22T09:05:07Z |
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| id | doaj.art-63a75e84f9e2408c97b0768fa5fe2f1f |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-22T09:05:07Z |
| publishDate | 2018-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-63a75e84f9e2408c97b0768fa5fe2f1f2022-12-21T18:31:36ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-12-01910.3389/fimmu.2018.02857428527Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked GlycansNicholas Giovannone0Nicholas Giovannone1Aristotelis Antonopoulos2Jennifer Liang3Jenna Geddes Sweeney4Jenna Geddes Sweeney5Matthew R. Kudelka6Matthew R. Kudelka7Sandra L. King8Gi Soo Lee9Richard D. Cummings10Richard D. Cummings11Anne Dell12Steven R. Barthel13Hans R. Widlund14Hans R. Widlund15Stuart M. Haslam16Charles J. Dimitroff17Charles J. Dimitroff18Department of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesHarvard Medical School, Boston MA, United StatesDepartment of Life Sciences, Imperial College London, London, United KingdomDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesHarvard Medical School, Boston MA, United StatesDepartment of Surgery, Beth Israel Deaconess Medical Center, Boston, MA, United StatesDepartment of Biochemistry, Emory University School of Medicine, Atlanta, GA, United StatesDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesDepartment of Otology and Laryngology, Harvard Medical School, Bostonm, MA, United StatesHarvard Medical School, Boston MA, United StatesDepartment of Surgery, Beth Israel Deaconess Medical Center, Boston, MA, United StatesDepartment of Life Sciences, Imperial College London, London, United KingdomDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesHarvard Medical School, Boston MA, United StatesDepartment of Life Sciences, Imperial College London, London, United KingdomDepartment of Dermatology, Brigham and Women's Hospital, Boston MA, United StatesHarvard Medical School, Boston MA, United StatesGerminal 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.https://www.frontiersin.org/article/10.3389/fimmu.2018.02857/fullglycosylationglycanB cellCD45peanut lectinPNA |
| spellingShingle | Nicholas Giovannone Nicholas Giovannone Aristotelis Antonopoulos Jennifer Liang Jenna Geddes Sweeney Jenna Geddes Sweeney Matthew R. Kudelka Matthew R. Kudelka Sandra L. King Gi Soo Lee Richard D. Cummings Richard D. Cummings Anne Dell Steven R. Barthel Hans R. Widlund Hans R. Widlund Stuart M. Haslam Charles J. Dimitroff Charles J. Dimitroff Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans Frontiers in Immunology glycosylation glycan B cell CD45 peanut lectin PNA |
| title | Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans |
| title_full | Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans |
| title_fullStr | Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans |
| title_full_unstemmed | Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans |
| title_short | Human B Cell Differentiation Is Characterized by Progressive Remodeling of O-Linked Glycans |
| title_sort | human b cell differentiation is characterized by progressive remodeling of o linked glycans |
| topic | glycosylation glycan B cell CD45 peanut lectin PNA |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2018.02857/full |
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