Expression of programmed cell death ligand-1 in mastocytosis correlates with disease severity Anja Rabenhorst, PhD, Silke Leja, MTA, Juliana Schwaab,

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Expression of programmed cell death ligand-1 in mastocytosis correlates with disease severity Anja Rabenhorst, PhD, Silke Leja, MTA, Juliana Schwaab, MD, Manuela Gehring, MTA, Anja Förster, PhD, Michel Arock, PharmD, PhD, Andreas Reiter, MD, Ulrike Raap, MD, Karin Hartmann, MD Journal of Allergy and Clinical Immunology Volume 137, Issue 1, Pages 314-318.e5 (January 2016) DOI: 10.1016/j.jaci.2015.07.006 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, PD-L1 serum levels are significantly elevated in adult patients with mastocytosis (Mastocytosis) compared with healthy controls (Control). Data are presented as box plots (min to max) with medians (left) and scatter dot plots with means (right). B and C, Patients with advanced mastocytosis categories (SM-AHNMD, ASM, and MCL) show increased PD-L1 levels compared to those with non-advanced categories (MIS and ISM). D, PD-L1 levels (y-axis) positively correlate with tryptase levels (x-axis) in adult patients with mastocytosis. E, PD-L1 levels in supernatants of cultured MCs. PD-L2 (F) and PD-1 (G) serum levels in adult patients with non-advanced and advanced mastocytosis compared with healthy controls. Journal of Allergy and Clinical Immunology 2016 137, 314-318.e5DOI: (10.1016/j.jaci.2015.07.006) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Paraffin-embedded BM (A-C) and skin biopsies (D-F) of patients with systemic mastocytosis analyzed by immunofluorescence. Expression of tryptase is shown in red (left panel), and expression of PD-L1 (Fig 2, A and D), PD-L2 (Fig 2, B and E), and PD-1 (Fig 2, C and F) is shown in green (middle panel). Colocalization is demonstrated by yellow labeling of the cells in merged pictures (right panel). Journal of Allergy and Clinical Immunology 2016 137, 314-318.e5DOI: (10.1016/j.jaci.2015.07.006) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 As expected, tryptase serum levels are elevated in adult patients with mastocytosis (All) compared with adult healthy controls (Control). Patients with advanced mastocytosis categories show increased tryptase levels compared with non-advanced categories. Data are presented as box plots (min to max) with medians (left) and scatter dot plots with means (right). Journal of Allergy and Clinical Immunology 2016 137, 314-318.e5DOI: (10.1016/j.jaci.2015.07.006) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Heat map showing a hierarchical comparison between PD-L1 serum levels and tryptase, PD-L2, and PD-1 serum levels for individual patients. A green-red color scale depicts the amount of measured serum levels (green, minimum; red, maximum). Journal of Allergy and Clinical Immunology 2016 137, 314-318.e5DOI: (10.1016/j.jaci.2015.07.006) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Serum levels of tryptase (A and B), PD-L1 (C and D), PD-L2 (E and F), and PD-1 (G and H) measured in pediatric patients with mastocytosis (Fig E3, B, D, F, and H) and pediatric and adult controls (Fig E3, A, C, E, and G). Data are presented as box plots (min to max) with medians (left) and scatter dot plots with means (right). Journal of Allergy and Clinical Immunology 2016 137, 314-318.e5DOI: (10.1016/j.jaci.2015.07.006) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions