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Changes in markers associated with dendritic cells driving the differentiation of either TH2 cells or regulatory T cells correlate with clinical benefit.

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Presentation on theme: "Changes in markers associated with dendritic cells driving the differentiation of either TH2 cells or regulatory T cells correlate with clinical benefit."— Presentation transcript:

1 Changes in markers associated with dendritic cells driving the differentiation of either TH2 cells or regulatory T cells correlate with clinical benefit during allergen immunotherapy  Claire Gueguen, PhD, Julien Bouley, PhD, Hélène Moussu, MSc, Sonia Luce, MSc, Magalie Duchateau, PhD, Julia Chamot-Rooke, PhD, Marc Pallardy, PhD, Vincent Lombardi, PhD, Emmanuel Nony, PhD, Véronique Baron-Bodo, PhD, Laurent Mascarell, PhD, Philippe Moingeon, PhD  Journal of Allergy and Clinical Immunology  Volume 137, Issue 2, Pages (February 2016) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Characterization of DC2s in comparison with Ctrl-DCs, DC1s, and DCreg cells. A, Cytokine production was analyzed with a multiplex quantification assay (n = 12). B, qPCR analysis of genes expressed in effector and regulatory DCs (n = 6). C, Cytokine production by CD4+ T cells cultured for 5 days with DCs analyzed by using a multiplex quantification assay (n = 12). Data are shown as means ± SEMs. *P < .05, **P < .01, and ***P < .001, Wilcoxon tests. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Comparative patterns of gene expression in Ctrl-DCs, DC1s, DC2s, and DCreg cells. Hierarchically clustered heat maps display median-centered log 2 intensities (based on the analysis of 6 groups of DC subsets originating from distinct donors). A and B, Genes upregulated (red) and downregulated (green), respectively, in DC2s in comparison with DC1s, with a false discovery rate P value of less than .01 and a fold difference of 4 or greater. C, Genes upregulated (green) in DCreg cells in comparison with Ctrl-DCs, with a false discovery rate P value of less than .01 and a fold difference of 4 or greater. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 2 Comparative patterns of gene expression in Ctrl-DCs, DC1s, DC2s, and DCreg cells. Hierarchically clustered heat maps display median-centered log 2 intensities (based on the analysis of 6 groups of DC subsets originating from distinct donors). A and B, Genes upregulated (red) and downregulated (green), respectively, in DC2s in comparison with DC1s, with a false discovery rate P value of less than .01 and a fold difference of 4 or greater. C, Genes upregulated (green) in DCreg cells in comparison with Ctrl-DCs, with a false discovery rate P value of less than .01 and a fold difference of 4 or greater. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 3 Validation of markers associated with DC2s by using qPCR and flow cytometry. A-C, qPCR analysis of mRNAs corresponding to DC2 markers identified by using cDNA microarrays (markers upregulated [Fig 3, A] or downregulated [Fig 3, B] by DC2s) or label-free quantitative proteomics (Fig 3, C). D and E, mRNA (Fig 3, D) and cell-surface (Fig 3, E) expression of potential type 2 markers previously identified by others27-30 were assessed by using qPCR and flow cytometry, respectively, in Ctrl-DCs and polarized DCs. Results in Fig 3, E, are expressed as mean fluorescence intensities (MFI) after subtracting signals obtained with control isotype antibodies. FACS, Fluorescence-activated cell sorting. Data are shown as means ± SEMs (n = 6). *P < .05, Wilcoxon tests. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 4 Validation of markers associated with DCreg cells by using qPCR. Expression of mRNAs corresponding to markers identified by using cDNA microarrays (A) or label-free quantitative proteomics (B) was analyzed in Ctrl-DCs and polarized DCs by using qPCR. Data are shown as means ± SEMs (n = 6). *P < .05, Wilcoxon tests. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 5 DC2-associated markers are downregulated after 4 months of AIT in PBMCs from patients with grass pollen allergy in relationship with clinical benefit. A, Changes after AIT in expression of CD141, GATA3, OX40L, and RIPK4 genes were assessed by using qPCR in PBMCs from patients of the active and placebo groups (ARs, n = 21; ANRs, n = 21; placebo responders [PR], n = 7; and placebo nonresponders [PNR], n = 31). *P < .05, **P < .01, and ***P < .001, Mann-Whitney tests. B, Spearman correlation at an individual patient level of changes after AIT in the expression of CD141, GATA3, OX40L, and RIPK4 genes with percentage improvements in ARTSSs (visit 7/visit 3) in patients from the active and placebo groups. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 6 DCreg cell–associated markers are upregulated after 4 months of AIT in PBMCs from patients with grass pollen allergy in relationship with clinical benefit. A, Changes after AIT in expression of C1QA, FcγRIIIA, FTL, and SLCO2B1 genes were assessed by using qPCR in PBMCs from patients of the active and placebo groups. *P < .05, **P < .01, and ***P < .001, Mann-Whitney test. B, Spearman correlations at the individual patient level of changes after AIT in expression of C1QA, FcγRIIIA, FTL, and SLCO2B1 mRNAs with percentage improvement in ARTSSs (visit 7/visit 3) in patients from the active and placebo groups. C, Spearman correlations of changes after 4 months of AIT in gene expression in patients from the active group (n = 42). PNR, Placebo nonresponders; PR, placebo responders. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig 7 Changes in expression of 5 combined DCreg/DC2–associated markers in PBMCs from patients with grass pollen allergy correlate with clinical efficacy after 2 and 4 months of AIT. ROC analyses of a combination of 3 DC2-associated markers (CD141, GATA3, and RIPK4) and 2 DCreg-associated markers (C1QA and FcγRIIIA) after 2 months of AIT (A) and 4 months of AIT (B). Spearman correlations at the individual patient level of changes in expression of the 5 combined markers with percentages of ARTSS improvement in patients from the active and placebo groups after 2 months (C) and 4 months (D) of AIT. AUC, Area under the ROC curve; PNR, placebo nonresponders; PR, placebo responders. *P < .05 and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig 8 Alterations in DCreg/DC2–associated markers are induced during AIT in both patients with and those without allergen-specific antibody responses. Correlation between changes after 4 months of AIT in expression of the 5 combined DCreg/DC2 markers and percentages of ARTSS improvement in patients from the active group, comprising both immunoreactive (ie, exhibiting strong allergen-specific antibody responses, black dots) and nonimmunoreactive (ie, exhibiting no or low allergen-specific antibody responses, open dots) patients. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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