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Characterization of human memory CD4+ T-cell responses to the dog allergen Can f 4  Aino L. Rönkä, MD, Tuure T. Kinnunen, MD, PhD, Amélie Goudet, BSc,

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Presentation on theme: "Characterization of human memory CD4+ T-cell responses to the dog allergen Can f 4  Aino L. Rönkä, MD, Tuure T. Kinnunen, MD, PhD, Amélie Goudet, BSc,"— Presentation transcript:

1 Characterization of human memory CD4+ T-cell responses to the dog allergen Can f 4 
Aino L. Rönkä, MD, Tuure T. Kinnunen, MD, PhD, Amélie Goudet, BSc, Marja A. Rytkönen-Nissinen, PhD, Joni Sairanen, BM, Anssi H.T. Kailaanmäki, MSc, Jukka T. Randell, MD, PhD, Bernard Maillère, PhD, Tuomas I. Virtanen, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 136, Issue 4, Pages e10 (October 2015) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Frequencies and functional avidities of the Can f 4–specific TCLs of memory origin from allergic (solid circles) and nonallergic (open circles) subjects. A, Frequencies of specific TCLs per 106 CD4+CD45RO+ memory T cells. B, Functional avidities of specific TCLs assessed as EC50 values calculated from the titrated response curves of each TCL. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Phenotypic characterization of the Can f 4–specific memory TCLs. A, Production of the cytokines IL-4, IL-5, IFN-γ, and IL-10 by the Can f 4–specific TCLs of allergic (solid circles) and nonallergic (open circles) subjects on stimulation with Can f 4 at 100 μg/mL. B, Cytokine phenotypes of the TCLs from allergic (A) and nonallergic (NA) subjects based on IFN-γ/IL-5 ratio (TH1, >5; TH2, <0.01; TH0, ). C, Mean fluorescence intensities (MFI; ± SEMs) of the cell-surface markers CD25 and CXCR3 on Can f 4–specific CD4+ T cells. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 T-cell epitope regions (gray boxes) recognized by the Can f 4–specific TCLs of allergic (A) and nonallergic (NA) subjects. A proliferative response with an SI of greater than 2 by a specific TCL to at least 2 consecutive 16-mer peptides (exception being the terminal p ) was regarded as epitope specific. The most stimulatory epitope regions (I-IV) are highlighted in red. The most prevalently recognized region (ie, amino acids 43-67) is marked with a red box. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Alignment of the T-cell epitope core sequences and predicted HLA-binding motifs on the Can f 4 sequence. Lines above the Can f 4 sequence represent the core sequences recognized by each TCL obtained from allergic (red) and nonallergic (green) subjects. Below the sequence are presented the predicted HLA-binding motifs (the length of 9 amino acids, with P1 anchor residues colored in red). Black lines indicate the most frequently recognized regions (I-IV) of Can f 4. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Binding affinities of the Can f 4 peptides to HLA-DR molecules. The binding affinities of 15 peptides covering the 4 most prevalently recognized epitope regions (I-IV) of Can f 4 to a panel of 6 common HLA-DR molecules are indicated as relative binding ratios. Lower numbers correspond to higher binding affinities. Green, Moderate binding (IC50 ratio, ); red, strong binding (IC50 ratio, <20). aa, Amino acids. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E1 Correlation between serum IgE reactivity to rCan f 4 (OD: 450 nm, x-axis) and the number of Can f 4−specific TCLs per 106 CD4+CD45RO+ T cells obtained from 13 allergic subjects (y-axis). Spearman r = , P = .28. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E2 Full map of the epitope regions recognized by each Can f 4–specific TCL obtained. A T-cell response with SI of greater than 2 to at least 2 consecutive peptides was considered epitope specific. Black boxes, Proliferation of ΔCPM of greater than 5000; dark gray boxes, ΔCPM of 1000 to 5000; light gray boxes, ΔCPM of less than 1000. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E3 Fine epitope mapping. Five minimal T-cell core epitopes within the core of the immunodominant epitope region of amino acids 46 to 64 were discovered by stimulating 12 TCLs from 6 different subjects specific to the region with a panel of 16 peptides of 9 to 16 amino acids truncated at the N- or C-terminus. An SI of greater than 2 and a ΔCPM of greater than 1000 (dotted line) was considered a positive response. The minimal core epitopes are indicated with vertical lines. Journal of Allergy and Clinical Immunology  , e10DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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