Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis

Takumi Kiwamoto, Toshihiko Katoh, Christopher M. Evans, William J. Janssen, Mary E. Brummet, Sherry A. Hudson, Zhou Zhu, Michael Tiemeyer, Bruce Scott Bochner*

*Corresponding author for this work

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Background Sialic acid-binding, immunoglobulin-like lectin (Siglec) F is a glycan-binding protein selectively expressed on mouse eosinophils. Its engagement induces apoptosis, suggesting a pathway for ameliorating eosinophilia in the setting of asthma and other eosinophil-associated diseases. Siglec-F recognizes sialylated sulfated glycans in glycan-binding assays, but the identities of endogenous sialoside ligands and their glycoprotein carriers in vivo are unknown. Objectives To use mouse lung-derived materials to isolate, biochemically identify, and biologically characterize naturally occurring endogenous glycan ligands for Siglec-F. Methods Lungs from normal and mucin-deficient mice, as well as mouse tracheal epithelial cells, were investigated in vitro and in vivo for the expression of Siglec-F ligands. Western blotting and cytochemistry used Siglec-F-Fc as a probe for directed purification, followed by liquid chromatography-tandem mass spectrometry of recognized glycoproteins. Purified components were tested in mouse eosinophil-binding assays and flow cytometry-based cell death assays. Results We detected mouse lung glycoproteins that bound to Siglec-F; binding was sialic acid dependent. Proteomic analysis of Siglec-F binding material identified Muc5b and Muc4. Cross-affinity enrichment and histochemical analysis of lungs from mucin-deficient mice assigned and validated the identity of Muc5b as one glycoprotein ligand for Siglec-F. Purified mucin preparations carried sialylated and sulfated glycans, bound to eosinophils and induced their death in vitro. Mice conditionally deficient in Muc5b displayed exaggerated eosinophilic inflammation in response to intratracheal installation of IL-13. Conclusions These data identify a previously unrecognized endogenous anti-inflammatory property of airway mucins by which their glycans can control lung eosinophilia through engagement of Siglec-F.

Original languageEnglish (US)
Pages (from-to)1329-1340.e9
JournalJournal of Allergy and Clinical Immunology
Volume135
Issue number5
DOIs
StatePublished - May 1 2015

Fingerprint

Sialic Acid Binding Immunoglobulin-like Lectins
Mucins
Eosinophils
Polysaccharides
Apoptosis
Glycoproteins
Lung
Ligands
Eosinophilia
Histocytochemistry
Interleukin-13
N-Acetylneuraminic Acid
Tandem Mass Spectrometry
Liquid Chromatography
Proteomics
Carrier Proteins
Flow Cytometry
Cell Death
Anti-Inflammatory Agents
Asthma

Keywords

  • Eosinophil
  • Muc4
  • Muc5b
  • Siglec-F
  • airway
  • apoptosis
  • asthma
  • epithelium
  • glands
  • glycan ligands
  • lung
  • mucin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Kiwamoto, Takumi ; Katoh, Toshihiko ; Evans, Christopher M. ; Janssen, William J. ; Brummet, Mary E. ; Hudson, Sherry A. ; Zhu, Zhou ; Tiemeyer, Michael ; Bochner, Bruce Scott. / Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis. In: Journal of Allergy and Clinical Immunology. 2015 ; Vol. 135, No. 5. pp. 1329-1340.e9.
@article{e8e524513d3541c782f292cc8d1e6221,
title = "Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis",
abstract = "Background Sialic acid-binding, immunoglobulin-like lectin (Siglec) F is a glycan-binding protein selectively expressed on mouse eosinophils. Its engagement induces apoptosis, suggesting a pathway for ameliorating eosinophilia in the setting of asthma and other eosinophil-associated diseases. Siglec-F recognizes sialylated sulfated glycans in glycan-binding assays, but the identities of endogenous sialoside ligands and their glycoprotein carriers in vivo are unknown. Objectives To use mouse lung-derived materials to isolate, biochemically identify, and biologically characterize naturally occurring endogenous glycan ligands for Siglec-F. Methods Lungs from normal and mucin-deficient mice, as well as mouse tracheal epithelial cells, were investigated in vitro and in vivo for the expression of Siglec-F ligands. Western blotting and cytochemistry used Siglec-F-Fc as a probe for directed purification, followed by liquid chromatography-tandem mass spectrometry of recognized glycoproteins. Purified components were tested in mouse eosinophil-binding assays and flow cytometry-based cell death assays. Results We detected mouse lung glycoproteins that bound to Siglec-F; binding was sialic acid dependent. Proteomic analysis of Siglec-F binding material identified Muc5b and Muc4. Cross-affinity enrichment and histochemical analysis of lungs from mucin-deficient mice assigned and validated the identity of Muc5b as one glycoprotein ligand for Siglec-F. Purified mucin preparations carried sialylated and sulfated glycans, bound to eosinophils and induced their death in vitro. Mice conditionally deficient in Muc5b displayed exaggerated eosinophilic inflammation in response to intratracheal installation of IL-13. Conclusions These data identify a previously unrecognized endogenous anti-inflammatory property of airway mucins by which their glycans can control lung eosinophilia through engagement of Siglec-F.",
keywords = "Eosinophil, Muc4, Muc5b, Siglec-F, airway, apoptosis, asthma, epithelium, glands, glycan ligands, lung, mucin",
author = "Takumi Kiwamoto and Toshihiko Katoh and Evans, {Christopher M.} and Janssen, {William J.} and Brummet, {Mary E.} and Hudson, {Sherry A.} and Zhou Zhu and Michael Tiemeyer and Bochner, {Bruce Scott}",
year = "2015",
month = "5",
day = "1",
doi = "10.1016/j.jaci.2014.10.027",
language = "English (US)",
volume = "135",
pages = "1329--1340.e9",
journal = "Journal of Allergy and Clinical Immunology",
issn = "0091-6749",
publisher = "Mosby Inc.",
number = "5",

}

Kiwamoto, T, Katoh, T, Evans, CM, Janssen, WJ, Brummet, ME, Hudson, SA, Zhu, Z, Tiemeyer, M & Bochner, BS 2015, 'Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis', Journal of Allergy and Clinical Immunology, vol. 135, no. 5, pp. 1329-1340.e9. https://doi.org/10.1016/j.jaci.2014.10.027

Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis. / Kiwamoto, Takumi; Katoh, Toshihiko; Evans, Christopher M.; Janssen, William J.; Brummet, Mary E.; Hudson, Sherry A.; Zhu, Zhou; Tiemeyer, Michael; Bochner, Bruce Scott.

In: Journal of Allergy and Clinical Immunology, Vol. 135, No. 5, 01.05.2015, p. 1329-1340.e9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis

AU - Kiwamoto, Takumi

AU - Katoh, Toshihiko

AU - Evans, Christopher M.

AU - Janssen, William J.

AU - Brummet, Mary E.

AU - Hudson, Sherry A.

AU - Zhu, Zhou

AU - Tiemeyer, Michael

AU - Bochner, Bruce Scott

PY - 2015/5/1

Y1 - 2015/5/1

N2 - Background Sialic acid-binding, immunoglobulin-like lectin (Siglec) F is a glycan-binding protein selectively expressed on mouse eosinophils. Its engagement induces apoptosis, suggesting a pathway for ameliorating eosinophilia in the setting of asthma and other eosinophil-associated diseases. Siglec-F recognizes sialylated sulfated glycans in glycan-binding assays, but the identities of endogenous sialoside ligands and their glycoprotein carriers in vivo are unknown. Objectives To use mouse lung-derived materials to isolate, biochemically identify, and biologically characterize naturally occurring endogenous glycan ligands for Siglec-F. Methods Lungs from normal and mucin-deficient mice, as well as mouse tracheal epithelial cells, were investigated in vitro and in vivo for the expression of Siglec-F ligands. Western blotting and cytochemistry used Siglec-F-Fc as a probe for directed purification, followed by liquid chromatography-tandem mass spectrometry of recognized glycoproteins. Purified components were tested in mouse eosinophil-binding assays and flow cytometry-based cell death assays. Results We detected mouse lung glycoproteins that bound to Siglec-F; binding was sialic acid dependent. Proteomic analysis of Siglec-F binding material identified Muc5b and Muc4. Cross-affinity enrichment and histochemical analysis of lungs from mucin-deficient mice assigned and validated the identity of Muc5b as one glycoprotein ligand for Siglec-F. Purified mucin preparations carried sialylated and sulfated glycans, bound to eosinophils and induced their death in vitro. Mice conditionally deficient in Muc5b displayed exaggerated eosinophilic inflammation in response to intratracheal installation of IL-13. Conclusions These data identify a previously unrecognized endogenous anti-inflammatory property of airway mucins by which their glycans can control lung eosinophilia through engagement of Siglec-F.

AB - Background Sialic acid-binding, immunoglobulin-like lectin (Siglec) F is a glycan-binding protein selectively expressed on mouse eosinophils. Its engagement induces apoptosis, suggesting a pathway for ameliorating eosinophilia in the setting of asthma and other eosinophil-associated diseases. Siglec-F recognizes sialylated sulfated glycans in glycan-binding assays, but the identities of endogenous sialoside ligands and their glycoprotein carriers in vivo are unknown. Objectives To use mouse lung-derived materials to isolate, biochemically identify, and biologically characterize naturally occurring endogenous glycan ligands for Siglec-F. Methods Lungs from normal and mucin-deficient mice, as well as mouse tracheal epithelial cells, were investigated in vitro and in vivo for the expression of Siglec-F ligands. Western blotting and cytochemistry used Siglec-F-Fc as a probe for directed purification, followed by liquid chromatography-tandem mass spectrometry of recognized glycoproteins. Purified components were tested in mouse eosinophil-binding assays and flow cytometry-based cell death assays. Results We detected mouse lung glycoproteins that bound to Siglec-F; binding was sialic acid dependent. Proteomic analysis of Siglec-F binding material identified Muc5b and Muc4. Cross-affinity enrichment and histochemical analysis of lungs from mucin-deficient mice assigned and validated the identity of Muc5b as one glycoprotein ligand for Siglec-F. Purified mucin preparations carried sialylated and sulfated glycans, bound to eosinophils and induced their death in vitro. Mice conditionally deficient in Muc5b displayed exaggerated eosinophilic inflammation in response to intratracheal installation of IL-13. Conclusions These data identify a previously unrecognized endogenous anti-inflammatory property of airway mucins by which their glycans can control lung eosinophilia through engagement of Siglec-F.

KW - Eosinophil

KW - Muc4

KW - Muc5b

KW - Siglec-F

KW - airway

KW - apoptosis

KW - asthma

KW - epithelium

KW - glands

KW - glycan ligands

KW - lung

KW - mucin

UR - http://www.scopus.com/inward/record.url?scp=84944096046&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84944096046&partnerID=8YFLogxK

U2 - 10.1016/j.jaci.2014.10.027

DO - 10.1016/j.jaci.2014.10.027

M3 - Article

VL - 135

SP - 1329-1340.e9

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

IS - 5

ER -