Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils

Jeremy A. O'Sullivan, Yadong Wei, Daniela J. Carroll, Liliana Moreno-Vinasco, Yun Cao, Fengrui Zhang, James J. Lee, Zhou Zhu, Bruce S. Bochner

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a human cell surface protein expressed exclusively on eosinophils, mast cells, and basophils that, when engaged, induces eosinophil apoptosis and inhibits mast cell mediator release. This makes Siglec-8 a promising therapeutic target to treat diseases involving these cell types. However, preclinical studies of Siglec-8 targeting in vivo are lacking because this protein is only found in humans, apes, and some monkeys. Therefore, we have developed a mouse strain in which SIGLEC8 transcription is activated by Cre recombinase and have crossed this mouse with the eoCre mouse to achieve eosinophil-specific expression. We confirmed that Siglec-8 is expressed exclusively on the surface of mature eosinophils in multiple tissues at levels comparable to those on human blood eosinophils. Following ovalbumin sensitization and airway challenge, Siglec-8 knock-in mice generated a pattern of allergic lung inflammation indistinguishable from that of littermate controls, suggesting that Siglec-8 expression within the eosinophil compartment does not alter allergic eosinophilic inflammation. Using bone marrow from these mice, we demonstrated that, during maturation, Siglec-8 expression occurs well before the late eosinophil developmental marker C-C motif chemokine receptor 3, consistent with eoCre expression. Antibody ligation of the receptor induces Siglec-8 endocytosis and alters the phosphotyrosine profile of these cells, indicative of productive signaling. Finally, we demonstrated that mouse eosinophils expressing Siglec-8 undergo cell death when the receptor is engaged, further evidence that Siglec-8 is functional on these cells. These mice should prove useful to investigate Siglec-8 biology and targeting in vivo in a variety of eosinophilic disease models.

Original languageEnglish (US)
JournalJournal of Leukocyte Biology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Sialic Acid Binding Immunoglobulin-like Lectins
Eosinophils
Mast Cells
CXCR3 Receptors
Death Domain Receptors
Phosphotyrosine
Basophils
Hominidae
Ovalbumin
Endocytosis
Haplorhini
Ligation

Keywords

  • Apoptosis
  • Asthma
  • Knock-in
  • Pre-clinical
  • Siglec-F
  • Transgenic

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

O'Sullivan, J. A., Wei, Y., Carroll, D. J., Moreno-Vinasco, L., Cao, Y., Zhang, F., ... Bochner, B. S. (Accepted/In press). Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils. Journal of Leukocyte Biology. https://doi.org/10.1002/JLB.2HI0917-391R

Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils. / O'Sullivan, Jeremy A.; Wei, Yadong; Carroll, Daniela J.; Moreno-Vinasco, Liliana; Cao, Yun; Zhang, Fengrui; Lee, James J.; Zhu, Zhou; Bochner, Bruce S.

In: Journal of Leukocyte Biology, 01.01.2018.

Research output: Contribution to journalArticle

O'Sullivan, JA, Wei, Y, Carroll, DJ, Moreno-Vinasco, L, Cao, Y, Zhang, F, Lee, JJ, Zhu, Z & Bochner, BS 2018, 'Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils', Journal of Leukocyte Biology. https://doi.org/10.1002/JLB.2HI0917-391R
O'Sullivan, Jeremy A. ; Wei, Yadong ; Carroll, Daniela J. ; Moreno-Vinasco, Liliana ; Cao, Yun ; Zhang, Fengrui ; Lee, James J. ; Zhu, Zhou ; Bochner, Bruce S. / Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils. In: Journal of Leukocyte Biology. 2018.
@article{b1d04ebe2d9b4975a1b052f4cd44abaa,
title = "Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils",
abstract = "Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a human cell surface protein expressed exclusively on eosinophils, mast cells, and basophils that, when engaged, induces eosinophil apoptosis and inhibits mast cell mediator release. This makes Siglec-8 a promising therapeutic target to treat diseases involving these cell types. However, preclinical studies of Siglec-8 targeting in vivo are lacking because this protein is only found in humans, apes, and some monkeys. Therefore, we have developed a mouse strain in which SIGLEC8 transcription is activated by Cre recombinase and have crossed this mouse with the eoCre mouse to achieve eosinophil-specific expression. We confirmed that Siglec-8 is expressed exclusively on the surface of mature eosinophils in multiple tissues at levels comparable to those on human blood eosinophils. Following ovalbumin sensitization and airway challenge, Siglec-8 knock-in mice generated a pattern of allergic lung inflammation indistinguishable from that of littermate controls, suggesting that Siglec-8 expression within the eosinophil compartment does not alter allergic eosinophilic inflammation. Using bone marrow from these mice, we demonstrated that, during maturation, Siglec-8 expression occurs well before the late eosinophil developmental marker C-C motif chemokine receptor 3, consistent with eoCre expression. Antibody ligation of the receptor induces Siglec-8 endocytosis and alters the phosphotyrosine profile of these cells, indicative of productive signaling. Finally, we demonstrated that mouse eosinophils expressing Siglec-8 undergo cell death when the receptor is engaged, further evidence that Siglec-8 is functional on these cells. These mice should prove useful to investigate Siglec-8 biology and targeting in vivo in a variety of eosinophilic disease models.",
keywords = "Apoptosis, Asthma, Knock-in, Pre-clinical, Siglec-F, Transgenic",
author = "O'Sullivan, {Jeremy A.} and Yadong Wei and Carroll, {Daniela J.} and Liliana Moreno-Vinasco and Yun Cao and Fengrui Zhang and Lee, {James J.} and Zhou Zhu and Bochner, {Bruce S.}",
year = "2018",
month = "1",
day = "1",
doi = "10.1002/JLB.2HI0917-391R",
language = "English (US)",
journal = "Journal of Leukocyte Biology",
issn = "0741-5400",
publisher = "FASEB",

}

TY - JOUR

T1 - Characterization of a novel mouse strain expressing human Siglec-8 only on eosinophils

AU - O'Sullivan, Jeremy A.

AU - Wei, Yadong

AU - Carroll, Daniela J.

AU - Moreno-Vinasco, Liliana

AU - Cao, Yun

AU - Zhang, Fengrui

AU - Lee, James J.

AU - Zhu, Zhou

AU - Bochner, Bruce S.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a human cell surface protein expressed exclusively on eosinophils, mast cells, and basophils that, when engaged, induces eosinophil apoptosis and inhibits mast cell mediator release. This makes Siglec-8 a promising therapeutic target to treat diseases involving these cell types. However, preclinical studies of Siglec-8 targeting in vivo are lacking because this protein is only found in humans, apes, and some monkeys. Therefore, we have developed a mouse strain in which SIGLEC8 transcription is activated by Cre recombinase and have crossed this mouse with the eoCre mouse to achieve eosinophil-specific expression. We confirmed that Siglec-8 is expressed exclusively on the surface of mature eosinophils in multiple tissues at levels comparable to those on human blood eosinophils. Following ovalbumin sensitization and airway challenge, Siglec-8 knock-in mice generated a pattern of allergic lung inflammation indistinguishable from that of littermate controls, suggesting that Siglec-8 expression within the eosinophil compartment does not alter allergic eosinophilic inflammation. Using bone marrow from these mice, we demonstrated that, during maturation, Siglec-8 expression occurs well before the late eosinophil developmental marker C-C motif chemokine receptor 3, consistent with eoCre expression. Antibody ligation of the receptor induces Siglec-8 endocytosis and alters the phosphotyrosine profile of these cells, indicative of productive signaling. Finally, we demonstrated that mouse eosinophils expressing Siglec-8 undergo cell death when the receptor is engaged, further evidence that Siglec-8 is functional on these cells. These mice should prove useful to investigate Siglec-8 biology and targeting in vivo in a variety of eosinophilic disease models.

AB - Sialic acid-binding immunoglobulin-like lectin (Siglec)-8 is a human cell surface protein expressed exclusively on eosinophils, mast cells, and basophils that, when engaged, induces eosinophil apoptosis and inhibits mast cell mediator release. This makes Siglec-8 a promising therapeutic target to treat diseases involving these cell types. However, preclinical studies of Siglec-8 targeting in vivo are lacking because this protein is only found in humans, apes, and some monkeys. Therefore, we have developed a mouse strain in which SIGLEC8 transcription is activated by Cre recombinase and have crossed this mouse with the eoCre mouse to achieve eosinophil-specific expression. We confirmed that Siglec-8 is expressed exclusively on the surface of mature eosinophils in multiple tissues at levels comparable to those on human blood eosinophils. Following ovalbumin sensitization and airway challenge, Siglec-8 knock-in mice generated a pattern of allergic lung inflammation indistinguishable from that of littermate controls, suggesting that Siglec-8 expression within the eosinophil compartment does not alter allergic eosinophilic inflammation. Using bone marrow from these mice, we demonstrated that, during maturation, Siglec-8 expression occurs well before the late eosinophil developmental marker C-C motif chemokine receptor 3, consistent with eoCre expression. Antibody ligation of the receptor induces Siglec-8 endocytosis and alters the phosphotyrosine profile of these cells, indicative of productive signaling. Finally, we demonstrated that mouse eosinophils expressing Siglec-8 undergo cell death when the receptor is engaged, further evidence that Siglec-8 is functional on these cells. These mice should prove useful to investigate Siglec-8 biology and targeting in vivo in a variety of eosinophilic disease models.

KW - Apoptosis

KW - Asthma

KW - Knock-in

KW - Pre-clinical

KW - Siglec-F

KW - Transgenic

UR - http://www.scopus.com/inward/record.url?scp=85044625687&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85044625687&partnerID=8YFLogxK

U2 - 10.1002/JLB.2HI0917-391R

DO - 10.1002/JLB.2HI0917-391R

M3 - Article

JO - Journal of Leukocyte Biology

JF - Journal of Leukocyte Biology

SN - 0741-5400

ER -