G-quadruplex DNA targeting alters class-switch recombination in B cells and attenuates allergic inflammation Zeinab Dalloul, MSc, Pauline Chenuet, MSc,

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G-quadruplex DNA targeting alters class-switch recombination in B cells and attenuates allergic inflammation Zeinab Dalloul, MSc, Pauline Chenuet, MSc, Iman Dalloul, MSc, François Boyer, PhD, Jean-Claude Aldigier, MD, PhD, Brice Laffleur, PhD, Yolla El Makhour, PhD, Bernhard Ryffel, MD, PhD, Valerie F.J. Quesniaux, PhD, Dieudonnée Togbé, PhD, Jean-Louis Mergny, PhD, Jeanne Cook-Moreau, PhD, Michel Cogné, MD, PhD Journal of Allergy and Clinical Immunology Volume 142, Issue 4, Pages 1352-1355 (October 2018) DOI: 10.1016/j.jaci.2018.06.011 Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 RHPS4 decreases CSR in stimulated primary mouse B cells. A, Primary B cells stimulated with LPS/IL-4 were treated with 0, 0.5, and 1 μM RHPS4, collected after 4-day culture and stained for flow cytometry. Histograms represent mean % ± SEM for 6 mice. ***P < .001 compared with untreated cells. B, Supernatants from adequately stimulated splenocytes were quantified by ELISA for the production of IgM, IgG2b, and IgG3 (top), and IgG2a, IgG1, and IgE (bottom). C, Germline Cγ1 and Cμ transcripts (left) and postswitch (right) Iμ-Cγ1 and Iμ-Cε transcripts from LPS + IL-4–stimulated B cells. Histograms represent mean fold change ± SEM for 6 mice. D, AICDA expression from LPS-stimulated cells. E, ChIP experiments showing AID recruitment to Sμ, Sε, and Sγ1 regions in LPS + IL-4–stimulated cells (right). ChIP, Chromatin immunoprecipitation; NS, not significant. *P < .05, **P < .01, ***P < .001 compared with the control untreated cells. Journal of Allergy and Clinical Immunology 2018 142, 1352-1355DOI: (10.1016/j.jaci.2018.06.011) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 RHPS4 inhibited class switching in vivo and decreased production of specific antibodies. A, Ag-specific IgM- or IgG-producing cells by ELISPOT in mice after primary immunization (left panels), and levels of Ag-specific antibodies (right panels). B, Ag-specific IgG1 and IgE antibodies after 25 days of intraperitoneal and intranasal OVA immunization. C, In an allergic asthma model after repeated Ag administration, mice receiving RHPS4 displayed less abundant cell infiltration of lung, (D) lower inflammation and mucus scores, and (E) lower counts of total cells, lymphocytes, macrophages, eosinophils, and neutrophils, together with lower protein levels, in bronchoalveolar fluid. Journal of Allergy and Clinical Immunology 2018 142, 1352-1355DOI: (10.1016/j.jaci.2018.06.011) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E7 Journal of Allergy and Clinical Immunology 2018 142, 1352-1355DOI: (10.1016/j.jaci.2018.06.011) Copyright © 2018 American Academy of Allergy, Asthma & Immunology Terms and Conditions