Siglec-7 is an inhibitory receptor on human mast cells and basophils

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Presentation transcript:

Siglec-7 is an inhibitory receptor on human mast cells and basophils Sa'ar Mizrahi, PhD, Bernhard F. Gibbs, PhD, Habil, Laila Karra, MSc, Micha Ben-Zimra, PhD, Francesca Levi-Schaffer, PharmD, PhD Journal of Allergy and Clinical Immunology Volume 134, Issue 1, Pages 230-233.e3 (July 2014) DOI: 10.1016/j.jaci.2014.03.031 Copyright © 2014 Terms and Conditions

Fig 1 A, FC analysis of Siglec-7 expression in CBMCs and MC cell lines. B, IF of Siglec-7 expression (green fluorescence on cell surface) on slide-fixed CBMCs and MC cell lines. C, CBMC proteins were IPed with an anti–Siglec-7 or isotype-matched antibody (mouse IgG1 [mIgG1]), resolved by SDS-PAGE, and analyzed by WB using a polyclonal goat anti–Siglec-7 antibody. Protein marker sizes are indicated. Representative results are shown from n = 10 (Fig 1, A), 2 (Fig 1, B), and 3 (Fig 1, C) separate experiments performed with different donors/cell batches. IF, Immunofluorescence; WB, Western blot. Journal of Allergy and Clinical Immunology 2014 134, 230-233.e3DOI: (10.1016/j.jaci.2014.03.031) Copyright © 2014 Terms and Conditions

Fig 2 Effects of Siglec-7 stimulation on IgE-dependent mediator release from CBMCs. Data are shown as the mean ± SEM from n = 3 separate experiments. Data shown are tryptase release (%) (A), β-hex (B), PGD2 (C), and GM-CSF (D) (see Mediator assays and calculations in the Methods section in this article's Online Repository). E, IP-WB analysis showing Siglec-7, CD300a, and co-IP of phosphorylated SHP-1 expressed in CBMCs incubated with or without orthovanadate (representative of n = 3 experiments). F, Effect of Siglec-7 stimulation on IgE-dependent β-hex release from CBMCs without co–cross-linking. Percentage of maximum release is shown, and data are expressed as the mean ± SEM from n = 2 separate experiments (see the Methods section in this article's Online Repository). Antibody concentrations are expressed as μg/mL. β-hex, β-Hexosaminidase; mIgG1, mouse IgG1; WB, Western blot. *P < .03. Journal of Allergy and Clinical Immunology 2014 134, 230-233.e3DOI: (10.1016/j.jaci.2014.03.031) Copyright © 2014 Terms and Conditions

Fig E1 A, FC analysis of Siglec-7 expression on human foreskin MC, CBMCs, NK cell line, peripheral blood NK cells, monocytes, and basophils (representative data of n = 3 experiments performed with different donors). B, WB analysis of Siglec-7 IPed from 2 different donors for CBMCs and basophils, and 1 NK cell line. C, Effects of Siglec-7 stimulation on IgE-dependent histamine secretion from primary human basophils. Data are shown as the mean ± SEM for n = 5 separate experiments (see “Cell treatments” section above). WB, Western blot; *P ≤ .04. Journal of Allergy and Clinical Immunology 2014 134, 230-233.e3DOI: (10.1016/j.jaci.2014.03.031) Copyright © 2014 Terms and Conditions