Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 in patients with eosinophilic disorders: Receptor expression and targeting using chimeric antibodies

Fanny Legrand, Yun Cao, Joshua Brian Wechsler, Xiang Zhu, Nives Zimmermann, Shakuntala Rampertaap, Joseph Monsale, Kimberly Romito, Bradford A. Youngblood, Emily C. Brock, Michelle A. Makiya, Nenad Tomasevic, Christopher Bebbington, Irina Maric, Dean D. Metcalfe, Bruce Scott Bochner, Amy D. Klion

Research output: Contribution to journalArticle

Abstract

Background: Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 is selectively expressed on eosinophils, mast cells, and basophils and, when engaged on eosinophils, can cause cell death. Objective: We sought to characterize surface and soluble Siglec-8 (sSiglec-8) levels in normal donors (NDs) and eosinophilic donors (EOs) and assess the efficacy of anti–Siglec-8 antibodies in inducing eosinophil cell death in vitro. Methods: Eosinophil expression of Siglec-8 was assessed by using flow cytometry and quantitative PCR. Serum sSiglec-8 levels were measured by means of ELISA. Induction of eosinophil death by IgG 4 (chimeric 2E2 IgG 4 ) and afucosylated IgG 1 (chimeric 2E2 IgG 1 [c2E2 IgG 1 ]) anti–Siglec-8 antibodies was evaluated in vitro by using flow cytometry and in vivo in humanized mice. Results: Siglec-8 was consistently expressed on eosinophils from NDs and EOs and did not correlate with absolute eosinophil count or disease activity. sSiglec-8 levels were measurable in sera from most donors unrelated to absolute eosinophil counts or Siglec-8 surface expression. c2E2 IgG 1 and chimeric 2E2 IgG 4 were equally effective at inducing cell death (Annexin-V positivity) of purified eosinophils from NDs and EOs after overnight IL-5 priming. In contrast, killing of purified eosinophils without IL-5 was only seen in EOs, and natural killer cell–mediated eosinophil killing was seen only with c2E2 IgG 1 . Finally, treatment of humanized mice with anti-Siglec antibody led to robust depletion of IL-5–induced eosinophilia in vivo. Conclusions: Siglec-8 is highly expressed on blood eosinophils from EOs and NDs and represents a potential therapeutic target for eosinophilic disorders. Enhanced killing of eosinophils in the presence of IL-5 might lead to increased efficacy in patients with IL-5–driven eosinophilia.

Original languageEnglish (US)
JournalJournal of Allergy and Clinical Immunology
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

Saliva
Eosinophils
Lectins
Immunoglobulins
Antibodies
Tissue Donors
Immunoglobulin G
Interleukin-5
Sialic Acid Binding Immunoglobulin-like Lectins
Cell Death
Eosinophilia
Flow Cytometry
Unrelated Donors
Basophils
Annexin A5
Serum
Mast Cells
Cause of Death
Enzyme-Linked Immunosorbent Assay

Keywords

  • Siglec-8
  • apoptosis
  • eosinophil
  • eosinophilic gastrointestinal disease
  • hypereosinophilic syndrome
  • inhibitory receptor
  • mast cell
  • mastocytosis
  • monoclonal antibody
  • soluble receptor

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Legrand, Fanny ; Cao, Yun ; Wechsler, Joshua Brian ; Zhu, Xiang ; Zimmermann, Nives ; Rampertaap, Shakuntala ; Monsale, Joseph ; Romito, Kimberly ; Youngblood, Bradford A. ; Brock, Emily C. ; Makiya, Michelle A. ; Tomasevic, Nenad ; Bebbington, Christopher ; Maric, Irina ; Metcalfe, Dean D. ; Bochner, Bruce Scott ; Klion, Amy D. / Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 in patients with eosinophilic disorders : Receptor expression and targeting using chimeric antibodies. In: Journal of Allergy and Clinical Immunology. 2019.
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abstract = "Background: Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 is selectively expressed on eosinophils, mast cells, and basophils and, when engaged on eosinophils, can cause cell death. Objective: We sought to characterize surface and soluble Siglec-8 (sSiglec-8) levels in normal donors (NDs) and eosinophilic donors (EOs) and assess the efficacy of anti–Siglec-8 antibodies in inducing eosinophil cell death in vitro. Methods: Eosinophil expression of Siglec-8 was assessed by using flow cytometry and quantitative PCR. Serum sSiglec-8 levels were measured by means of ELISA. Induction of eosinophil death by IgG 4 (chimeric 2E2 IgG 4 ) and afucosylated IgG 1 (chimeric 2E2 IgG 1 [c2E2 IgG 1 ]) anti–Siglec-8 antibodies was evaluated in vitro by using flow cytometry and in vivo in humanized mice. Results: Siglec-8 was consistently expressed on eosinophils from NDs and EOs and did not correlate with absolute eosinophil count or disease activity. sSiglec-8 levels were measurable in sera from most donors unrelated to absolute eosinophil counts or Siglec-8 surface expression. c2E2 IgG 1 and chimeric 2E2 IgG 4 were equally effective at inducing cell death (Annexin-V positivity) of purified eosinophils from NDs and EOs after overnight IL-5 priming. In contrast, killing of purified eosinophils without IL-5 was only seen in EOs, and natural killer cell–mediated eosinophil killing was seen only with c2E2 IgG 1 . Finally, treatment of humanized mice with anti-Siglec antibody led to robust depletion of IL-5–induced eosinophilia in vivo. Conclusions: Siglec-8 is highly expressed on blood eosinophils from EOs and NDs and represents a potential therapeutic target for eosinophilic disorders. Enhanced killing of eosinophils in the presence of IL-5 might lead to increased efficacy in patients with IL-5–driven eosinophilia.",
keywords = "Siglec-8, apoptosis, eosinophil, eosinophilic gastrointestinal disease, hypereosinophilic syndrome, inhibitory receptor, mast cell, mastocytosis, monoclonal antibody, soluble receptor",
author = "Fanny Legrand and Yun Cao and Wechsler, {Joshua Brian} and Xiang Zhu and Nives Zimmermann and Shakuntala Rampertaap and Joseph Monsale and Kimberly Romito and Youngblood, {Bradford A.} and Brock, {Emily C.} and Makiya, {Michelle A.} and Nenad Tomasevic and Christopher Bebbington and Irina Maric and Metcalfe, {Dean D.} and Bochner, {Bruce Scott} and Klion, {Amy D.}",
year = "2019",
month = "1",
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language = "English (US)",
journal = "Journal of Allergy and Clinical Immunology",
issn = "0091-6749",
publisher = "Mosby Inc.",

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Legrand, F, Cao, Y, Wechsler, JB, Zhu, X, Zimmermann, N, Rampertaap, S, Monsale, J, Romito, K, Youngblood, BA, Brock, EC, Makiya, MA, Tomasevic, N, Bebbington, C, Maric, I, Metcalfe, DD, Bochner, BS & Klion, AD 2019, 'Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 in patients with eosinophilic disorders: Receptor expression and targeting using chimeric antibodies' Journal of Allergy and Clinical Immunology. https://doi.org/10.1016/j.jaci.2018.10.066

Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 in patients with eosinophilic disorders : Receptor expression and targeting using chimeric antibodies. / Legrand, Fanny; Cao, Yun; Wechsler, Joshua Brian; Zhu, Xiang; Zimmermann, Nives; Rampertaap, Shakuntala; Monsale, Joseph; Romito, Kimberly; Youngblood, Bradford A.; Brock, Emily C.; Makiya, Michelle A.; Tomasevic, Nenad; Bebbington, Christopher; Maric, Irina; Metcalfe, Dean D.; Bochner, Bruce Scott; Klion, Amy D.

In: Journal of Allergy and Clinical Immunology, 01.01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 in patients with eosinophilic disorders

T2 - Receptor expression and targeting using chimeric antibodies

AU - Legrand, Fanny

AU - Cao, Yun

AU - Wechsler, Joshua Brian

AU - Zhu, Xiang

AU - Zimmermann, Nives

AU - Rampertaap, Shakuntala

AU - Monsale, Joseph

AU - Romito, Kimberly

AU - Youngblood, Bradford A.

AU - Brock, Emily C.

AU - Makiya, Michelle A.

AU - Tomasevic, Nenad

AU - Bebbington, Christopher

AU - Maric, Irina

AU - Metcalfe, Dean D.

AU - Bochner, Bruce Scott

AU - Klion, Amy D.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Background: Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 is selectively expressed on eosinophils, mast cells, and basophils and, when engaged on eosinophils, can cause cell death. Objective: We sought to characterize surface and soluble Siglec-8 (sSiglec-8) levels in normal donors (NDs) and eosinophilic donors (EOs) and assess the efficacy of anti–Siglec-8 antibodies in inducing eosinophil cell death in vitro. Methods: Eosinophil expression of Siglec-8 was assessed by using flow cytometry and quantitative PCR. Serum sSiglec-8 levels were measured by means of ELISA. Induction of eosinophil death by IgG 4 (chimeric 2E2 IgG 4 ) and afucosylated IgG 1 (chimeric 2E2 IgG 1 [c2E2 IgG 1 ]) anti–Siglec-8 antibodies was evaluated in vitro by using flow cytometry and in vivo in humanized mice. Results: Siglec-8 was consistently expressed on eosinophils from NDs and EOs and did not correlate with absolute eosinophil count or disease activity. sSiglec-8 levels were measurable in sera from most donors unrelated to absolute eosinophil counts or Siglec-8 surface expression. c2E2 IgG 1 and chimeric 2E2 IgG 4 were equally effective at inducing cell death (Annexin-V positivity) of purified eosinophils from NDs and EOs after overnight IL-5 priming. In contrast, killing of purified eosinophils without IL-5 was only seen in EOs, and natural killer cell–mediated eosinophil killing was seen only with c2E2 IgG 1 . Finally, treatment of humanized mice with anti-Siglec antibody led to robust depletion of IL-5–induced eosinophilia in vivo. Conclusions: Siglec-8 is highly expressed on blood eosinophils from EOs and NDs and represents a potential therapeutic target for eosinophilic disorders. Enhanced killing of eosinophils in the presence of IL-5 might lead to increased efficacy in patients with IL-5–driven eosinophilia.

AB - Background: Sialic acid–binding immunoglobulin-like lectin (Siglec) 8 is selectively expressed on eosinophils, mast cells, and basophils and, when engaged on eosinophils, can cause cell death. Objective: We sought to characterize surface and soluble Siglec-8 (sSiglec-8) levels in normal donors (NDs) and eosinophilic donors (EOs) and assess the efficacy of anti–Siglec-8 antibodies in inducing eosinophil cell death in vitro. Methods: Eosinophil expression of Siglec-8 was assessed by using flow cytometry and quantitative PCR. Serum sSiglec-8 levels were measured by means of ELISA. Induction of eosinophil death by IgG 4 (chimeric 2E2 IgG 4 ) and afucosylated IgG 1 (chimeric 2E2 IgG 1 [c2E2 IgG 1 ]) anti–Siglec-8 antibodies was evaluated in vitro by using flow cytometry and in vivo in humanized mice. Results: Siglec-8 was consistently expressed on eosinophils from NDs and EOs and did not correlate with absolute eosinophil count or disease activity. sSiglec-8 levels were measurable in sera from most donors unrelated to absolute eosinophil counts or Siglec-8 surface expression. c2E2 IgG 1 and chimeric 2E2 IgG 4 were equally effective at inducing cell death (Annexin-V positivity) of purified eosinophils from NDs and EOs after overnight IL-5 priming. In contrast, killing of purified eosinophils without IL-5 was only seen in EOs, and natural killer cell–mediated eosinophil killing was seen only with c2E2 IgG 1 . Finally, treatment of humanized mice with anti-Siglec antibody led to robust depletion of IL-5–induced eosinophilia in vivo. Conclusions: Siglec-8 is highly expressed on blood eosinophils from EOs and NDs and represents a potential therapeutic target for eosinophilic disorders. Enhanced killing of eosinophils in the presence of IL-5 might lead to increased efficacy in patients with IL-5–driven eosinophilia.

KW - Siglec-8

KW - apoptosis

KW - eosinophil

KW - eosinophilic gastrointestinal disease

KW - hypereosinophilic syndrome

KW - inhibitory receptor

KW - mast cell

KW - mastocytosis

KW - monoclonal antibody

KW - soluble receptor

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DO - 10.1016/j.jaci.2018.10.066

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JO - Journal of Allergy and Clinical Immunology

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