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Alexander S. Kim, MD, Taylor A. Doherty, MD, Maya R

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1 Regulatory B cells and T follicular helper cells are reduced in allergic rhinitis 
Alexander S. Kim, MD, Taylor A. Doherty, MD, Maya R. Karta, PhD, Sudipta Das, PhD, Rachel Baum, BS, Peter Rosenthal, BS, Andrew Beppu, BS, Marina Miller, MD, PhD, Richard Kurten, PhD, David H. Broide, MB, ChB  Journal of Allergy and Clinical Immunology  Volume 138, Issue 4, Pages e5 (October 2016) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 FACS gating strategy to detect Breg cells, CD25hi Breg cells, and TFH cells in peripheral blood. A, Circulating Breg cells and TFH-like cells were identified from the circulating lymphocyte population as CD19+CD73−CD25+CD71+ and CD4+PD-1+CXCR5+, respectively, using the gating strategy shown. B, Example of FACS of circulating Breg cells from a nonallergic individual (left) and an allergic individual (right) including isotype controls was used to quantitate their total CD25+ and CD25hi subsets. FSC-A, Forward-scatter area; SSC-A, side-scatter area. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 FACS quantitation of Breg cells, CD25hi Breg cells, and TFH-like cells in subjects with allergic rhinitis and controls. A, Peripheral blood total CD25+ (left) and CD25hi (right) Breg cells from individuals with allergic rhinitis (n = 10) and nonallergic individuals (n = 7) were quantitated as percentages of CD19+ cells. B, Circulating CD4+PD-1+CXCR5+ TFH-like cells from individuals with allergic rhinitis (n = 7) and nonallergic individuals (n = 5) were quantitated as percentages of CD4+ T cells. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 FACS gating strategy to detect Breg cells and CD25hi Breg cells in human lung lymph nodes. A, Human lung lymph node Breg cells were identified from the lung lymph node lymphocyte population as CD19+CD73−CD25+CD71+ using the gating strategy shown. B, Human lung lymph node Breg cells from an individual with asthma were analyzed using isotype controls to analyze their total CD25+ and CD25hi subsets. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 FACS gating strategy to detect TFH-like cells in human lung lymph nodes. TFH-like cells from a lung lymph node from an individual with asthma were identified from the lung lymph node lymphocyte population as CD4+CXCR5+PD-1+ using the gating strategy shown. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Cytokine production by Breg cells and TFH cells. A, IL-10 protein levels measured by ELISA in supernatants of sorted Breg cells from a nonallergic subject. B, IL-21 mRNA expression quantitated by quantitative PCR in sorted TFH cells from a nonallergic subject. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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