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Direct ex vivo analysis of allergen-specific CD4+ T cells

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Presentation on theme: "Direct ex vivo analysis of allergen-specific CD4+ T cells"— Presentation transcript:

1 Direct ex vivo analysis of allergen-specific CD4+ T cells
William W. Kwok, PhD, Michelle Roti, BS, Jonathan H. DeLong, BS, Venus Tan, BS, Erik Wambre, PhD, Eddie A. James, PhD, David Robinson, MD  Journal of Allergy and Clinical Immunology  Volume 125, Issue 6, Pages e1 (June 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Ex vivo staining of Fel d 1 CD4+ T cells with tetramers. A, PBMCs from subjects with cat allergy were stained with phycoerythrin-labeled Fel d 1 tetramers (Tet) followed by the anti-phycoerythrin bead enrichment protocol. Each panel shows a representative staining result from a subject with cat allergy with the indicated HLA. The frequencies of Fel d 1 CD4+ T cells (per million) were as indicated. B, Staining of PBMCs from subjects without allergy with Fel d 1 tetramers using the magnetic bead enrichment protocol. Each panel shows a representative staining result from a subject without allergy with the indicated HLA. C, Phenotypes of Fel d 1–specific CD4+ T cells. PBMCs of a DR0401 subject with cat allergy were stained with DR0401/Fel d 1 tetramer ex vivo and a panel of antibodies. D, Comparison of phenotypes of Fel d 1 T cells (●) and total CD4+ T cells (Δ; total of 5 subjects). A Student t test was used in the statistical analysis. *P < .01. E, Cytokine profiles of Fel d 1–reactive CD4+ T cells. PBMCs from subjects with cat allergy with the indicated HLA were stained with tetramers using the anti-phycoerythrin bead enrichment protocol and assayed for IL-5 and IFN-γ. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 TGEM studies of DR1101-restricted Fel d 1–reactive CD4+ T cells
TGEM studies of DR1101-restricted Fel d 1–reactive CD4+ T cells. A, PBMCs, from a DR1101 subject with cat allergy were stimulated with 4 pools of Fel d 1 peptides for 2 weeks and subsequently stained with the corresponding DR1101/Fel d 1 pooled peptide tetramers. B, Cells that were stimulated with pool 4 peptides, composed of peptides p16 (Fel d 1 chain ), p17 (Fel d 1 chain ), and p18 (Fel d 1 chain ) were restained with DR1101/Fel d 1 p16, Fel d 1 p17, and Fel d 1 p18. C, PBMCs from 2 additional DR1101 subjects with allergy and from 2 DR1101 subjects without allergy were stimulated with Fel d 1 p17. Cells were assayed with DR1101/Fel d 1 p17 tetramers on day 14. D, Staining of cells with DR1101/Fel d 1 chain and Fel d 1 chain peptides. The percentages of tetramer-positive cells were as indicated. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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