Mast Cell-Specific Expression of Human Siglec-8 in Conditional Knock-in Mice
AbstractSialic 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. View Full-Text
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Wei, Y.; Chhiba, K.D.; Zhang, F.; Ye, X.; Wang, L.; Zhang, L.; Robida, P.A.; Moreno-Vinasco, L.; Schnaar, R.L.; Roers, A.; Hartmann, K.; Lee, C.-M.; Demers, D.; Zheng, T.; Bochner, B.S.; Zhu, Z. Mast Cell-Specific Expression of Human Siglec-8 in Conditional Knock-in Mice. Int. J. Mol. Sci. 2019, 20, 19.
Wei Y, Chhiba KD, Zhang F, Ye X, Wang L, Zhang L, Robida PA, Moreno-Vinasco L, Schnaar RL, Roers A, Hartmann K, Lee C-M, Demers D, Zheng T, Bochner BS, Zhu Z. Mast Cell-Specific Expression of Human Siglec-8 in Conditional Knock-in Mice. International Journal of Molecular Sciences. 2019; 20(1):19.Chicago/Turabian Style
Wei, Yadong; Chhiba, Krishan D.; Zhang, Fengrui; Ye, Xujun; Wang, Lihui; Zhang, Li; Robida, Piper A.; Moreno-Vinasco, Liliana; Schnaar, Ronald L.; Roers, Axel; Hartmann, Karin; Lee, Chang-Min; Demers, Delia; Zheng, Tao; Bochner, Bruce S.; Zhu, Zhou. 2019. "Mast Cell-Specific Expression of Human Siglec-8 in Conditional Knock-in Mice." Int. J. Mol. Sci. 20, no. 1: 19.
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