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Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy  Erik Wambre, PhD, Jonathan.

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Presentation on theme: "Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy  Erik Wambre, PhD, Jonathan."— Presentation transcript:

1 Differentiation stage determines pathologic and protective allergen-specific CD4+ T-cell outcomes during specific immunotherapy  Erik Wambre, PhD, Jonathan H. DeLong, BSc, Eddie A. James, PhD, Rebecca E. LaFond, PhD, David Robinson, MD, William W. Kwok, PhD  Journal of Allergy and Clinical Immunology  Volume 129, Issue 2, Pages e7 (February 2012) DOI: /j.jaci Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Frequencies of Aln g 1–specific T cells. A, Ex vivo DRB1∗07:01 and DRB1∗15:01/Aln g 1–pMHCII tetramer staining of total live CD3+ T cells. Data are representative of at least 3 subjects per group. B, Ex vivo frequencies of Aln g 1–specific CD4+ T cells during the alder pollen season. C, Percentage of memory (CD45RO+) cells within Aln g 1–specific CD4+ T cells. Fig 1, B and C, Each data point represents a single epitope in a single donor. Differences between groups were analyzed by using the Mann-Whitney U test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Dynamics of the Aln g 1–specific T-cell response. A and B, Ex vivo longitudinal analysis of Ki-67 (Fig 2, A) and CD38 (Fig 2, B) expression within Aln g 1–specific CD4+ T cells in allergic subjects. C, Percentage of Aln g 1–specific CD4+ T cells expressing CD38. D, Ex vivo frequencies of Aln g 1–specific CD4+ T cells in 10 allergic subjects both outside and during the alder pollen season. E, Ex vivo Aln g 1–specific CD4+ T-cell frequencies plotted over time for 3 representative allergic subjects. Fig 2, A-C, Each data point in each time period represents a single epitope in a single donor. Fig 2, C and D, Differences between groups were analyzed by using the Wilcoxon matched-pairs test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Phenotype of Aln g 1–specific T cells. A and B, Representative flow cytometric plots showing ex vivo multicolor phenotyping of Aln g 1–specific CD4+ T cells in allergic (Fig 3, A) and nonallergic (Fig 3, B) subjects. C, Ex vivo phenotype of Aln g 1–specific CD4+ T cells. D, Cytokine production by Aln g 1–specific CD4+ T cells. Fig 3, A and B, Percentages of tetramer-positive cells expressing the given marker are indicated in the upper right quadrant. Fig 3, C and D, Data are means ± SEMs from at least 6 subjects per group. Differences between groups were analyzed by using the Mann-Whitney U test. ∗P < .001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 CD27 expression defines 2 distinct subsets of allergen-specific T cells. A, Percentage of CD27+, CCR7+, and CD7+ cells among Aln g 1–specific CD4+ T cells within the allergic (gray) and nonallergic (black) groups. Each square represents a single donor. B, CRTH2 expression (open histogram) by CD27+ and CD27− Aln g 1–specific CD4+ T cells in allergic subjects. C, Ex vivo phenotype of CD27− and CD27+ Aln g 1–specific CD4+ T cells in allergic subjects. D, Cytokine production by CD27− and CD27+ Aln g 1–specific CD4+ T cells in allergic subjects. Data are means ± SEMs from at least 7 subjects. Differences between groups were analyzed by using the Mann-Whitney U test. ∗P < .001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Effect of SIT on Aln g 1–specific CD4+ T cells. A, Average frequencies of CD27− and CD27+ Aln g 1–specific CD4+ T cells in 12 allergic and 6 nonallergic subjects and 7 subjects after successful SIT. B and C, Ex vivo phenotypes (Fig 5, B) and cytokine production (Fig 5, C) in Aln g 1–specific CD4+ T cells between nonallergic subjects and subjects after successful SIT. Data are means ± SEMs from at least 6 subjects per group. Differences between groups were analyzed by using the Mann-Whitney U test. ∗P < .001. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E1 Alder pollen counts in the Seattle area in The count indicated refers to the number of pollen grains per cubic meter according to the National Allergy Bureau. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E2 Representative flow cytometric plots showing coproduction of IFN-γ and IL-4 (left) or IFN-γ and IL-10 (right) by Aln g 1–specific CD4+ T cells from allergic (A) or nonallergic (B) subjects. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E3 A, Representative overlay of CD27+ Aln g 1–specific CD4+ T cells (black) on total CD4+ T cells (gray) showing ex vivo staining with a panel of antibodies in allergic subjects. B, Representative overlay of CD27+ Aln g 1–specific CD4+ T cells (black) on total CD4+ T cells (gray) showing cytokine production in allergic subjects. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E4 SIT is associated with a decrease in the CD27−/CD27+ allergen-specific CD4+ T-cell ratio. A, Means ± SEMs of the CD27−/CD27+ allergen-specific CD4+ T-cell ratio in allergic subjects (open circles) and SIT-treated patients (solid circles) at multiple time points. B, Ex vivo longitudinal frequency of CD27− (solid symbols) and CD27+ (open symbols) Aln g 1–specific CD4+ T cells from patient dSIT-1 (circles) and patient dSIT-2 (squares) during SIT. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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