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αS1-Casein elucidate major T-cell responses in cow's milk allergy

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Presentation on theme: "αS1-Casein elucidate major T-cell responses in cow's milk allergy"— Presentation transcript:

1 αS1-Casein elucidate major T-cell responses in cow's milk allergy
L. Diego Archila, PhD, Fatima S. Khan, MD, Nupur Bhatnagar, PhD, David Robinson, MD, Mary L. Farrington, MD, William W. Kwok, PhD  Journal of Allergy and Clinical Immunology  Volume 140, Issue 3, Pages e6 (September 2017) DOI: /j.jaci Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Frequency and phenotype of Bos d-epitope–specific CD4+ T cells. A, Frequencies of Bos d-epitope–specific CD45RA−CD4+ T cells in subjects with and without CMA. B, Comparison of Bos d–specific CD4+ T-cell frequency for each epitope described (data point) between milk-allergic (n = 23; filled squares) and nonallergic subjects (n = 13; opened circles) and adults, teenagers, and children older than 8 years (n = 16; triangles) and children 8 years or younger (n = 7; downside triangles) with CMA. C, PBMCs from a nonallergic and allergic subjects were stained with phycoerythrin-labeled DRB1*10:01/Bos d tetramers and a panel of antibodies. The percentages of memory Bos d–specific T cells that expressed the surface marker of interest are as indicated. D, Phenotype of Bos d–specific T cells in allergic and nonallergic subjects and in (E) children with CMA older than 8 years and children with CMA 8 years or younger. Symbols in Fig 1, D and E, are identical to those used in Fig 1, B. A t test was used in the statistical analysis. NS, Not significant. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Cytokine profiles of Bos d–reactive T cells. A, First row, Cytokine profile in a DRB1*01:01 nonallergic subject. Second row, Cytokine profile in a DRB1*01:01 allergic adult. Third row, Cytokine profile in a DRB1*01:01 allergic child. The percentages of memory CD154+ Bos d–reactive cytokine-producing T cells are as indicated. B and C, Cytokine profiles of Bos d–reactive T cells in nonallergic (n = 13) and allergic subjects (n = 21), and between adults, teenagers, and children older than 8 (n = 14) and children 8 years or younger (n = 7) with CMA. Data are presented as the mean frequency of cytokine-producing T cells from each group in pie charts. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Tetramer-guided epitope-mapping studies of DRB1*10:01-restricted Bos d–specific CD4+ T cells. A, PBMCs from a DRB1*10:01 subject with milk allergy were stimulated with 5 pools of Bos d 11 peptides for 2 weeks and subsequently stained with corresponding DRB1*10:01/Bos d 11 pooled peptide tetramers. B, Cells that were stimulated with pool no. 1 were restained with individual peptides from the corresponding pool. The staining identified p4 (Bos d ) as DRB1*10:01-restricted Bos d 11 T-cell epitopes. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Frequencies of milk allergen reactive CD4+ T cells. A and B, Frequencies of β-lactoglobulin-, αS1-casein-, αS2-casein-, β-casein-, and κ-casein-reactive T cells in 8 subjects with milk allergy (adults, teenagers, and children >8 years of age n = 6; children ≤8 years of age n = 2) with CD154 assays after 3-hour Bos d peptide stimulation using freshly isolated PBMCs. Each data point represents the frequency of T cells reactive to each allergen. An ANOVA test (with Bonferroni correction) was used to compare all columns in the statistical analysis. NS, Not significant. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Phenotypes of CCR6+Bos d–specific T cells in milk-allergic subjects after ex vivo pMHC-II tetramer enrichment using freshly isolated PBMCs. A and B, Tetramer+CD45RA− T cells were gated against CCR4 and CCR6. Each data point represents results for surface expression in tetramer-positive T cells from 23 subjects with milk allergy: 15 adults, teenagers, and children older than 8 years (triangles) and 8 children 8 years or younger (downside triangles) with CMA. C and D, Tetramer-positive CD45RA− T cells were gated against CCR6 and CRTH2. Each data point represents results for surface expression in tetramer-positive T cells from 26 subjects with milk allergy: 18 adults, teenagers, and children older than 8 years (triangles) and 8 children 8 years or younger (downside triangles) with CMA. A t test was used in the statistical analysis. NS, Not significant. *P < .05, **P < .01, and ****P < .0001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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