Antigen-presenting epithelial cells can play a pivotal role in airway allergy Julia Arebro, MD, Lotta Tengroth, MSc, Ronia Razavi, MSc, Susanna Kumlien.

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Antigen-presenting epithelial cells can play a pivotal role in airway allergy Julia Arebro, MD, Lotta Tengroth, MSc, Ronia Razavi, MSc, Susanna Kumlien Georén, PhD, Ola Winqvist, MD, PhD, Lars-Olaf Cardell, MD, PhD Journal of Allergy and Clinical Immunology Volume 137, Issue 3, Pages 957-960.e7 (March 2016) DOI: 10.1016/j.jaci.2015.08.053 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, MHC class II expression on OVA-stimulated MNECs cocultured with T cells (4 hours). B, CD4+ T-cell counts in cocultures with OVA-stimulated MNECs (24 hours). C and D, Fraction of CD69+/CD4+ (Fig 1, C) and CD44+/CD4+ (Fig 1, D) T cells after coculture with OVA-stimulated MNECs. E, INF-γ release in cocultures (24 hours). F, Cocultures with MNECs and T cells (both from sensitized mice) with anti–MHC class II antibodies (anti-MHC II). ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001. Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 A, NECs from a patient with AR containing dextran (green) and MHC class II (red). Scale bar = 10 μm. B, Three-dimensional composite image. C, Level of CD4+ T cells after coculture with HNECs. D, Fraction of CD69+/CD4+ T cells after coculture with HNECs. E, Level IL-13 followed by 4 hours of coculture. F and G, Different cell types in nasal mucosa (Fig 2, F) and intensity of HLA-DR (Fig 2, G; n = 7). ∗P < .05. Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 A and B, Expression of HLA-DR and CD86 (open histogram) in cultured HNECs from a healthy control subject (Fig E1, A) and a patient with AR (Fig E1, B; representative data from 9 different experiments, respectively). The isotype control is a filled histogram. C, HNECs stained for MHC class II (green). Scale bar = 50 μm. D, Expression of HLA-DR and CD86 on freshly isolated HNECs from healthy control subjects in nasal biopsy specimens (n = 4) and nasal brushings (n = 6). Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 A and B, Expression of H2-IAb, CD86, and CD80 on freshly isolated MNECs from a naive mouse (Fig E2, A) and from a mouse sensitized with OVA (Fig E2, B; representative data from 25-28 animals). C, Expression of H2-IAb, CD86, and CD80 on cultured MNECs from naive and OVA-sensitized mice. *P < .05. Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Total number of CD69+/CD4+ (A) and CD44+/CD4+ (B) T cells after coculture with OVA-stimulated MNECs. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 A, Total number of CD69+/CD4+ T cells after coculture with HNECs. B, IL-13 level after 48 hours of coculture. C, Fraction of CD69+/CD4+ T cells in coculture without and with birch pollen–stimulated T cells, ruling out that the T-cell activation was mediated by MHC class II–expressing T cells. *P < .05. Journal of Allergy and Clinical Immunology 2016 137, 957-960.e7DOI: (10.1016/j.jaci.2015.08.053) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions