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CD4+CD25+ regulatory T cells suppress contact hypersensitivity reactions through a CD39, adenosine-dependent mechanism  Sabine Ring, PhD, Stephen J. Oliver,

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Presentation on theme: "CD4+CD25+ regulatory T cells suppress contact hypersensitivity reactions through a CD39, adenosine-dependent mechanism  Sabine Ring, PhD, Stephen J. Oliver,"— Presentation transcript:

1 CD4+CD25+ regulatory T cells suppress contact hypersensitivity reactions through a CD39, adenosine-dependent mechanism  Sabine Ring, PhD, Stephen J. Oliver, MD, Bruce N. Cronstein, MD, Alexander H. Enk, MD, Karsten Mahnke, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 6, Pages e2 (June 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Adenosine (ADO) blocks the elicitation phase of CHS and abrogates T-cell proliferation in vitro. A, Control mice were sensitized and challenged with TNCB. The other experimental groups were additionally treated with CD4+CD25− cells, Treg cells, adenosine, or Treg cells and theophylline. Shown is the mean increase of ear thickness ± SD of 3 independent experiments (n = 5; ∗P < .005). B, Proliferation of CD4+CD25− T cells (CD4) stimulated with anti-CD3/anti-CD28. C, Proliferation of CD4+CD25− T cells (CD4) stimulated with allogeneic dendritic cells (DC). Data in Fig 1, B and C, show the mean counts per minute ± SD from a typical experiment carried out in triplicates. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Adenosine (ADO) and Treg cells preactivated with anti-CD3/anti-CD28 block adherence of T cells to ECs in vitro. A and C, Merged pictures of flowing CD4+CD25− T cells (Fig 2, A) and CD8+ T cells (Fig 2, C) after 10 minutes (red) and 11 minutes (green) of flow. Nonmoving adherent cells appear yellow. B and D, Number of adherent CD4+CD25− T cells (Fig 2, B) and CD8+ T cells (Fig 2, D) on the ECs after 20 minutes of flow. Data show the fold increase of adherent cells on stimulated bEND.3 cells in comparison with that seen on unstimulated bEND.3 cells (∗∗P < .001). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 CD39+CD73+ Treg cells produce adenosine after activation with ATP. A, Expression of CD39 and CD73 on either freshly isolated or activated (anti-CD3/anti-CD μg/mL each, 24 hours) CD4+CD25+ forkhead box protein 3 (Foxp3)+ Treg cells. PE, Phycoerythrin; FITC, fluorescein isothiocyanate. B, Expression of CD39 and CD73 on CD4+CD25−Foxp3− T cells. C, Analysis of adenosine (ADO) in culture supernatants. The chromatogram shows overlay of one typical experiment. Lane 1, Treg cells; lane 2, Treg cells cultured with 1.25 mmol/L ATP; lane 3, spiking sample no. 2 with 200 μmol/L adenosine. D, Adenosine measured in the tissue culture supernatant of differently treated Treg cells. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Expression of CD39+ on Treg cells is essential for the in vivo suppression of CHS. A, Mean ear swelling ± SD of 2 independent experiments (n = 5, ∗P < .005). B, Suppressive activity of WT Treg cells and CD39 knockout (CD39KO) Treg cells in vitro. Proliferation was measured by means of tritiated thymidine incorporation. Shown is the mean proliferation in triplicates ± SD of one characteristic experiment of 3. C, Leukocyte-endothelium interaction 30 minutes after challenge. Arrows indicate examples of leukocytes attached to the vessel wall. Arrowheads indicate free-flowing leukocytes and vessel walls devoid of attached leukocytes. D, Amount of adherent cells over a period of 4 hours after challenge (mean ± SD, n = 3, ∗P < .005). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Prevention of upregulation of adhesion molecules on ECs by preactivated Treg cells and adenosine (ADO) in vitro. A, Expression of CD31 and the adhesion molecules P- and E-selectin were detected by means of flow cytometry. FITC, Fluorescein isothiocyanate; APC, allophycocyanin. B, Mean fluorescence intensity (MFI) of CD31 and percentages of P- and E-selectin in percentages of CD31+ cells ± SD (n = 5, ∗∗P < .001). Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Inflammation-induced upregulation of adhesion molecules is abrogated by Treg cell–derived adenosine (ADO). A, Expression of P-selectin and E-selectin in percentages of the CD31+ ECs in the ears of differently treated mice analyzed by means of flow cytometry. SSC-A, sidewards scatter; APC, allophycocyanin; FITC, fluorescein isothiocyanate; PE, phycoerythrin, B, Cryosections of the ears from the same experimental groups depicted in Fig 6, A, were stained with P- or E-selectin–specific antibodies and appropriate fluorescein isothiocyanate−labeled secondary reagents. Inserts show high-power magnifications. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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