Influence of seasonal exposure to grass pollen on local and peripheral blood IgE repertoires in patients with allergic rhinitis Yu-Chang B. Wu, PhD,

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Presentation transcript:

Influence of seasonal exposure to grass pollen on local and peripheral blood IgE repertoires in patients with allergic rhinitis Yu-Chang B. Wu, PhD, Louisa K. James, PhD, Jason A. Vander Heiden, MSc, Mohamed Uduman, PhD, Stephen R. Durham, MD, FRCP, Steven H. Kleinstein, PhD, David Kipling, DPhil, Hannah J. Gould, PhD Journal of Allergy and Clinical Immunology Volume 134, Issue 3, Pages 604-612 (September 2014) DOI: 10.1016/j.jaci.2014.07.010 Copyright © 2014 The Authors Terms and Conditions

Fig 1 Abundance of IGHV genes in clonotypic repertoires by antibody class and sample type. A and B, Use of IGHV gene families in IgE clonotypic repertoires was compared across sample types and patient groups (Fig 1, A) or with other antibody classes (Fig 1, B). C, Use of individual IGHV genes was compared between antibody classes. *P < .05 and **P < .005. Fig 1, B and C, Groups of clonotypic sequences were combined from peripheral blood and biopsy specimens independent of atopic status. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig 2 Changes in IGHV mutation levels by antibody class and sample type. A and B, Mean mutation frequencies were compared between clonotypic sequences of different antibody classes in peripheral blood (Fig 2, A) and nasal biopsy specimens (Fig 2, B). IgA and IgG repertories were combined as one switched IgE− population. C-F, Mean mutation levels in IgE sequences were further compared between the NA and AR groups (Fig 2, C and D) or between groups of sequences taken in and out of season (Fig 2, E and F). Red line, Two percent background error rate. G, Pie charts show relative fractions of IgE clonotypes ranked by mutations in peripheral blood and nasal biopsy specimens from the NA group (inner circles), AR.OS group (middle circles), and AR.IS group (outer circles). GL, Germline like, ≤200 mutations per 10,000 bases; Mutated, >200 mutations per 10,000 bases. ****P < .00005. ns, Not significant. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig 3 Patterns of CDR-H3 length. A, Virtual CDR-H3 spectratypes show the relative distribution of clonotypic sequences by CDR-H3 length for unmutated IgM (gray bars) and IgEAR (red bars; AR.IS and AR.OS groups combined). a.a, Amino acids. B, Mean CDR-H3 length was compared between groups, as indicated. C, Nucleotide numbers for different CDR-H3 components in IgE sequences were compared between the IgENA and IgEAR groups (AR.IS and AR.OS groups combined). D, Mean CDR-H3 length for IgE clonotypic sequences was compared between groups in peripheral blood (PB) and nasal biopsy specimens (NB). *P < .05, **P < .005, and ****P < .00005. ns, Not significant. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig 4 Selection strength of IgE sequences. Mean selection strength (Σ) for CDRs and framework regions (FWRs) for IgE clones in peripheral blood (A) or nasal biopsy specimens (B) were compared between atopic statuses. Values indicate means ± 95% CIs. *P < .05, **P < .005, and ****P < .00005. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig 5 Analysis of clonal diversity using Hill's model.32 The diversity of total clones (0D) was compared across antibody classes in peripheral blood (A) and nasal biopsy specimens (B) or between IgE clones from different patient groups in peripheral blood (C) and nasal biopsy specimens (D). Values indicate means ± 95% CIs. *P < .05 and **P < .005. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig 6 Examples of lineage trees containing IgE sequences. Only representative trees are presented for illustration purposes. Sequences from peripheral blood or nasal biopsy specimens are indicated by colored circles, with the number of point mutations between 2 adjacent sequences shown next to each branch (point mutation = 1 when not indicated). Antibody classes (A, IgA; G, IgG; E, IgE; M, IgM) and subclasses (G1-G4, IgG; A1 and A2, IgA) are indicated within the circles. Only unique sequences with identifiable antibody classes are shown. Detailed information for the 12 trees containing IgE sequences and their related variants of other classes are shown in Table E6. Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E1 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E2 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E3 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E4 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E5 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions

Fig E6 Journal of Allergy and Clinical Immunology 2014 134, 604-612DOI: (10.1016/j.jaci.2014.07.010) Copyright © 2014 The Authors Terms and Conditions