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Int J Mol Sci. 2018 Dec 21;20(1). pii: E19. doi: 10.3390/ijms20010019.

Mast Cell-Specific Expression of Human Siglec-8 in Conditional Knock-in Mice.

Author information

1
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. ywei16@jhu.edu.
2
Department of Medicine, Division of Allergy and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. krishan.chhiba@northwestern.edu.
3
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. fengrui.zhang@yale.edu.
4
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. wdxjy@whu.edu.cn.
5
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. wanglh10318@163.com.
6
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. syszhang@hotmail.com.
7
Department of Medicine, Division of Allergy and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. piper.wedman@northwestern.edu.
8
Department of Medicine, Division of Allergy and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. lmoreno@deptofmed.arizona.edu.
9
Department of Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21224, USA. schnaar@jhu.edu.
10
Institute of Immunology, University of Technology Dresden, 01069 Dresden, Germany. Axel.Roers@tu-dresden.de.
11
Department of Dermatology, University of Lübeck, 23538 Lübeck, Germany. Karin.Hartmann@uksh.de.
12
Department of Molecular Microbiology and Immunology, Department of Pediatrics, Brown University Alpert Medical School, Providence, RI 02912, USA. chang-min_lee@brown.edu.
13
Department of Molecular Microbiology and Immunology, Department of Pediatrics, Brown University Alpert Medical School, Providence, RI 02912, USA. delia_demers@brown.edu.
14
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. tao_zheng@brown.edu.
15
Department of Molecular Microbiology and Immunology, Department of Pediatrics, Brown University Alpert Medical School, Providence, RI 02912, USA. tao_zheng@brown.edu.
16
Department of Medicine, Division of Allergy and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA. bruce.bochner@northwestern.edu.
17
Section of Allergy and Clinical Immunology, Yale University School of Medicine, New Haven, CT 06511, USA. zhou_zhu@brown.edu.
18
Department of Molecular Microbiology and Immunology, Department of Pediatrics, Brown University Alpert Medical School, Providence, RI 02912, USA. zhou_zhu@brown.edu.

Abstract

Sialic acid-binding Ig-like lectin 8 (Siglec-8) is expressed on the surface of human eosinophils, mast cells, and basophils-cells that participate in allergic and other diseases. Ligation of Siglec-8 by specific glycan ligands or antibodies triggers eosinophil death and inhibits mast cell degranulation; consequences that could be leveraged as treatment. However, Siglec-8 is not expressed in murine and most other species, thus limiting preclinical studies in vivo. Based on a ROSA26 knock-in vector, a construct was generated that contains the CAG promoter, a LoxP-floxed-Neo-STOP fragment, and full-length Siglec-8 cDNA. Through homologous recombination, this Siglec-8 construct was targeted into the mouse genome of C57BL/6 embryonic stem (ES) cells, and chimeric mice carrying the ROSA26-Siglec-8 gene were generated. After cross-breeding to mast cell-selective Cre-recombinase transgenic lines (CPA3-Cre, and Mcpt5-Cre), the expression of Siglec-8 in different cell types was determined by RT-PCR and flow cytometry. Peritoneal mast cells (dual FcεRI⁺ and c-Kit⁺) showed the strongest levels of surface Siglec-8 expression by multicolor flow cytometry compared to expression levels on tissue-derived mast cells. Siglec-8 was seen on a small percentage of peritoneal basophils, but not other leukocytes from CPA3-Siglec-8 mice. Siglec-8 mRNA and surface protein were also detected on bone marrow-derived mast cells. Transgenic expression of Siglec-8 in mice did not affect endogenous numbers of mast cells when quantified from multiple tissues. Thus, we generated two novel mouse strains, in which human Siglec-8 is selectively expressed on mast cells. These mice may enable the study of Siglec-8 biology in mast cells and its therapeutic targeting in vivo.

KEYWORDS:

ROSA26; Siglec-8; allergic disease; mouse mast cells

PMID:
30577572
PMCID:
PMC6337227
DOI:
10.3390/ijms20010019
[Indexed for MEDLINE]
Free PMC Article

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