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Allergen-enhanced thrombomodulin (blood dendritic cell antigen 3, CD141) expression on dendritic cells is associated with a TH2-skewed immune response 

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Presentation on theme: "Allergen-enhanced thrombomodulin (blood dendritic cell antigen 3, CD141) expression on dendritic cells is associated with a TH2-skewed immune response "— Presentation transcript:

1 Allergen-enhanced thrombomodulin (blood dendritic cell antigen 3, CD141) expression on dendritic cells is associated with a TH2-skewed immune response  Stephanie T. Yerkovich, PhD, Marjut Roponen, PhD, Miranda E. Smith, BSc (Hons), Kathy McKenna, PhD, Anthony Bosco, PhD, Lily S. Subrata, PhD, Emilie Mamessier, PhD, Matthew E. Wikström, PhD, Peter Le Souef, MD, FRACP, Peter D. Sly, MD, FRACP, DSc, Patrick G. Holt, DSc, John W. Upham, FRACP, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 1, Pages e4 (January 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Microarray analysis of HDM-stimulated DCs. A, Microarrays were performed on pooled RNA samples and are expressed as a microarray plot; the vertical axis displays the stimulated (HDM stimulated/unstimulated) fold change of atopics to nonatopics, and the horizontal axis displays the average signal. B, Thrombomodulin (TM) mRNA expression levels after HDM stimulation from the same donors used in the microarray are presented as a ratio to control. Note the log scale. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Thrombomodulin mRNA expression in HDM and LPS-stimulated DCs. DCs from an independent set of atopic (n = 7) and nonatopic (n = 8) individuals were stimulated with HDM or LPS for 6 hours, RNA extracted, and qRT-PCR performed. Expression levels were normalized to UBE2D2 and are presented as the ratio to control (mean ± SE). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Thrombomodulin protein expression on HDM and LPS-stimulated DCs. Thrombomodulin (CD141) expression levels were measured on DCs from atopic (n = 8) and nonatopic (n = 9) individuals. A, Representative plot showing the percentage of CD141 positive DC and MFI on unstimulated DCs. B, MFI expressed as ratio to control (equal to 1.0, mean ± SE) of atopic (closed symbols, closed line) or nonatopic (open symbols, dashed line) DCs after stimulation. ∗P < .022 versus control. PerCP, Peridinin chlorophyll protein. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Percentage of mDC2s in peripheral blood and thrombomodulin expression during an acute asthmatic attack. A,top left, Correlation between the percentage of mDC2s and HDM SPT wheal size with the line of best fit (n = 36); top right, The percentage of mDC2s from nonatopic subjects without asthma (n = 19), atopic subjects without asthma (n = 6), and subjects with atopic asthma (n = 11). Comparisons are also made between subjects with (n = 17) and without (n = 19) atopy and with (n = 11) and without (n = 25) atopic asthma. B, Thrombomodulin mRNA expression measured during an acute asthmatic attack and presented as a ratio to the convalescent sample (mean ± SE; n = 25). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 HDM-stimulated cytokine production after coculture with thrombomodulin (TM)+ DCs or thrombomodulin− DCs. A, TM+(white bars) and TM−(shaded bars) sorted DCs were cocultured with autologous PBMCs and stimulated with HDM (n = 9 atopic and n = 6 nonatopic). Significant differences between the TM+ and TM− sorted DCs are indicated by horizontal bars, whereas ∗ indicates P < .04 between atopic and nonatopic donors. B, The ratio of IFN-γ:IL-13 is shown for each atopic (A) or nonatopic (NA) donor. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Representative FACS staining and gating strategy
Representative FACS staining and gating strategy. A, Peripheral blood DC subsets were identified after exclusion of erythrocytes and debris, as well as CD19+ B cells (which also express CD1c) and CD14+ monocytes (which also express CD141). DCs were characterized as positive for CD1c (mDC1) (B), CD303 (plasmacytoid DC, data not shown), or highly positive for CD141 (mDC2) (C). Numbers indicate the percentage relative to total PBMCs. A minimum of 235,000 PBMCs (mean, 490,000 ± 80,000) were acquired to obtain sufficient numbers of DC subsets for accurate enumeration. SSC-H, Side scatter; PerCP, peridinin chlorophyll protein; PE, phycoerythrin; FITC, fluoroscein isothiocyanate. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Percentage of myeloid mDC1s in peripheral blood
Percentage of myeloid mDC1s in peripheral blood. The percentages of mDC1s from nonatopic subjects without asthma (n = 19), atopic subjects without asthma (n = 6), and subjects with atopic asthma (n = 11) are shown. Comparisons are also made between subjects with (n = 17) and without (n = 19) atopy and with (n = 11) and without (n = 25) atopic asthma. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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