B cells differentiate in human thymus and express AIRE

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News Beyond Our Pages Journal of Allergy and Clinical Immunology

News Beyond Our Pages Journal of Allergy and Clinical Immunology

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B cells differentiate in human thymus and express AIRE Vincent Gies, Aurélien Guffroy, MD, François Danion, MD, Philippe Billaud, MD, Céline Keime, PhD, Jean-Daniel Fauny, PhD, Sandrine Susini, PhD, Anne Soley, LT, Thierry Martin, MD, PhD, Jean-Louis Pasquali, MD, PhD, Frédéric Gros, PhD, Isabelle André-Schmutz, PhD, Pauline Soulas-Sprauel, PharmD, PhD, Anne-Sophie Korganow, MD, PhD Journal of Allergy and Clinical Immunology Volume 139, Issue 3, Pages 1049-1052.e12 (March 2017) DOI: 10.1016/j.jaci.2016.09.044 Copyright © 2016 Terms and Conditions

Fig 1 A-C, Thymus immunostaining with anti-CD19 (Fig 1, A) or anti-IgM (Fig 1, B and C) antibody (red). medulla (m); cortex (c); DAPI (gray). D, B-cell subsets (percentages of CD19+ cells) in thymus (n ≥ 4) and BM (n = 3). Antibodies mentioned on axes are applied to the population defined on the top of each plot. E, Isotype frequency in sIg+ switched thymic B cells (n = 4). DAPI, 4′-6-Diamidino-2-phenylindole, dihydrochloride. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig 2 A-E, MFI (black) and isotypic controls (gray) on thymic B cells (n ≥ 4). F, AIRE protein (green) in TECs and in CD19+ or IgM+ (red) thymic B cells. G, AIRE expression in thymocytes (n = 6) and thymic B cells (n = 5). H, AIRE-related genes and genes involved in autophagy in thymic B cells (n = 5) compared with their peripheral counterpart (n = 4). CDH17 remained undetectable in periphery, except for 1 sample. I, Graphical representation of RNA sequencing data. AIRE (blue); TRAs associated with autoimmune diseases (red). MFI, Mean fluorescence intensity. *P < .05 and **P < .005. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig E1 A, Flow cytometry analysis, in the lymphoid gate, of thymic (n = 5) and BM (n = 3) CD19+ B cells B, Flow cytometry analysis of the different B-cells subsets (indicated as a percentage of total CD19+ cells) in thymus (n ≥ 4) compared with BM (n = 3). Antibodies mentioned on the axes are applied to the population defined on the top of each plot. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig E2 B-cell ontogeny and markers used for B-cell subsets gating strategy. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig E3 A, Flow cytometry analysis of CD19+ B cells in thymus, BM, and PB of P1. B and C, Frequency of immature B cells (Fig E3, B) and mature B cells (Fig E3, C) in P1. D and E, The panels represent, respectively, B cells' MFI of CD86 (Fig E3, D) and CD5 (Fig E3, E) in PB (red), BM (blue), and thymus (black) for P1. MFI, Mean fluorescence intensity; P1, patient 1. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig E4 A-C, Representative immunostaining of AIRE protein (Fig E4, A and B; green) compared with isotype control (Fig E4, C) in TECs. D-F, Representative immunostaining of AIRE protein (Fig E4, D and E; green) compared with isotype control (Fig E4, F) in IgM+ (red) thymic B cells. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions

Fig E5 Gating strategy and purity of sorted samples. Values are shown as mean ± SEM [minimum and maximum values] of the percentages of total viable sorted cells in the gates. A, Thymuses (n = 5). B, PBMC samples (n = 4). Fig E5, A and B, Antibodies mentioned on the axes are applied to the population defined on the top of each plot. Journal of Allergy and Clinical Immunology 2017 139, 1049-1052.e12DOI: (10.1016/j.jaci.2016.09.044) Copyright © 2016 Terms and Conditions