Induction and maintenance of allergen-specific FOXP3+ Treg cells in human tonsils as potential first-line organs of oral tolerance Oscar Palomares, PhD,

Slides:

Advertisements

Similar presentations

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

Advertisements

Specific immunotherapy modifies allergen-specific CD4+ T-cell responses in an epitope- dependent manner Erik Wambre, PhD, Jonathan H. DeLong, BSc, Eddie.

Comparison of the allergic and nonallergic CD4+ T-cell responses to the major dog allergen Can f 1 Suvi Parviainen, MSc, Antti Taivainen, MD, PhD, Aino.

Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood

Barbara Stanic, PhD, Willem van de Veen, PhD, Oliver F

Expression of IL-9 receptor α chain on human germinal center B cells modulates IgE secretion Lama M. Fawaz, PhD, Ehssan Sharif-Askari, PhD, Oumnia Hajoui,

Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong Th17/Th1 T-cell responses Anja Hänsel,

Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu Amanda J. Reefer, MS, Kathryn.

Human circulating group 2 innate lymphoid cells can express CD154 and promote IgE production Laura Maggi, PhD, Gianni Montaini, BSc, Alessio Mazzoni,

Induction of anergic allergen-specific suppressor T cells using tolerogenic dendritic cells derived from children with allergies to house dust mites

Cell-to-cell contact between activated CD4+ T lymphocytes and unprimed monocytes interferes with a TH1 response Miriam Wittmann, MD, Mareike Alter, Tanja.

Novel vaccines targeting dendritic cells by coupling allergoids to nonoxidized mannan enhance allergen uptake and induce functional regulatory T cells.

IgE cross-linking impairs monocyte antiviral responses and inhibits influenza-driven TH1 differentiation Regina K. Rowe, MD, PhD, David M. Pyle, MD,

Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production Mark C. Glaum, MD, PhD, Shilpi Narula, MD, Decheng Song,

Volume 135, Issue 6, Pages (December 2008)

Lack of autophagy induces steroid-resistant airway inflammation

Peanut oral immunotherapy results in increased antigen-induced regulatory T-cell function and hypomethylation of forkhead box protein 3 (FOXP3) Aleena.

T-cell regulation during viral and nonviral asthma exacerbations

Expansion of FOXP3high regulatory T cells by human dendritic cells (DCs) in vitro and after injection of cytokine-matured DCs in myeloma patients by Devi.

Infliximab induces downregulation of the IL-12/IL-23 axis in 6-sulfo-LacNac (slan)+ dendritic cells and macrophages Patrick M. Brunner, MD, Frieder Koszik,

Crosstalk of regulatory T cells and tolerogenic dendritic cells prevents contact allergy in subjects with low zone tolerance Ulrike Luckey, PhD, Talkea.

Type 3 innate lymphoid cells induce proliferation of CD94+ natural killer cells Shuo Li, PhD, Hideaki Morita, MD, PhD, Beate Rückert, Sci Tec, Tadech.

Toll-like receptor 4 ligation enforces tolerogenic properties of oral mucosal Langerhans cells Jean-Pierre Allam, MD, Wen-Ming Peng, MSc, Torsten Appel,

Glucocorticoids promote intrinsic human TH17 differentiation

Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood

Α-MSH-Stimulated Tolerogenic Dendritic Cells Induce Functional Regulatory T Cells and Ameliorate Ongoing Skin Inflammation Matteo Auriemma, Thomas Brzoska,

Human dendritic cell subset 4 (DC4) correlates to a subset of CD14dim/−CD16++ monocytes Federica Calzetti, BS, Nicola Tamassia, PhD, Alessandra Micheletti,

Human IL-31 is induced by IL-4 and promotes TH2-driven inflammation

Forkhead box protein 3 demethylation is associated with tolerance induction in peanut- induced intestinal allergy Meiqin Wang, MD, PhD, Ivana V. Yang,

Volume 137, Issue 5, Pages (November 2009)

Kathleen R. Bartemes, BA, Gail M. Kephart, BS, Stephanie J

CD94/NKG2C is a killer effector molecule in patients with Stevens-Johnson syndrome and toxic epidermal necrolysis Esther Morel, PhD, Salvador Escamochero,

Katherine G. MacDonald, BSc, Nicholas A. J

Human mast cells drive memory CD4+ T cells toward an inflammatory IL-22+ phenotype Nicolas Gaudenzio, PhD, Camille Laurent, MD, Salvatore Valitutti,

Human lung natural killer cells are predominantly comprised of highly differentiated hypofunctional CD69−CD56dim cells Nicole Marquardt, PhD, Eliisa.

Dysregulation of proinflammatory versus anti-inflammatory human TH17 cell functionalities in the autoinflammatory Schnitzler syndrome Rebecca Noster,

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

Targeting allergen to FcγRI reveals a novel TH2 regulatory pathway linked to thymic stromal lymphopoietin receptor Kathryn E. Hulse, PhD, Amanda J. Reefer,

Tolerogenic signaling by pulmonary CD1c+ dendritic cells induces regulatory T cells in patients with chronic obstructive pulmonary disease by IL-27/IL-10/inducible.

Patients with atopic dermatitis and history of eczema herpeticum elicit herpes simplex virus–specific type 2 immune responses Stephan Traidl, Petra Kienlin,

Volume 29, Issue 3, Pages (September 2008)

A thymic stromal lymphopoietin–responsive dendritic cell subset mediates allergic responses in the upper airway mucosa Guro R. Melum, MD, Lorant Farkas,

Volume 135, Issue 1, Pages (July 2008)

Specific immunotherapy modifies allergen-specific CD4+ T-cell responses in an epitope- dependent manner Erik Wambre, PhD, Jonathan H. DeLong, BSc, Eddie.

Chronic cat allergen exposure induces a TH2 cell–dependent IgG4 response related to low sensitization Amedee Renand, PhD, Luis D. Archila, MSc, John.

Volume 28, Issue 6, Pages (June 2008)

Bet v 1–specific T-cell receptor/forkhead box protein 3 transgenic T cells suppress Bet v 1–specific T-cell effector function in an activation-dependent.

Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis Wen Hao Wu, PhD,

Role of B cells in TH cell responses in a mouse model of asthma

Decreases in human dendritic cell–dependent TH2-like responses after acute in vivo IgE neutralization John T. Schroeder, PhD, Anja P. Bieneman, BS, Kristin.

Expression of IL-9 receptor α chain on human germinal center B cells modulates IgE secretion Lama M. Fawaz, PhD, Ehssan Sharif-Askari, PhD, Oumnia Hajoui,

Demonstration of circulating allergen-specific CD4+CD25highFoxp3+ T-regulatory cells in both nonatopic and atopic individuals Laura Maggi, BSc, Veronica.

Volume 17, Issue 2, Pages (February 2009)

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

Staphylococcal enterotoxin A–activated regulatory T cells promote allergen-specific TH2 response to intratracheal allergen inoculation Wei-ping Zeng,

Increased activation-induced cell death of high IFN-γ–producing TH1 cells as a mechanism of TH2 predominance in atopic diseases Tunc Akkoc, PhD, Pieter.

IL-4 and IL-13 Alter Plasmacytoid Dendritic Cell Responsiveness to CpG DNA and Herpes Simplex Virus-1 Jurjen Tel, Ruurd Torensma, Carl G. Figdor, I.

Intravenous immunoglobulin attenuates airway inflammation through induction of forkhead box protein 3–positive regulatory T cells Amir H. Massoud, MSc,

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

Enhanced production of CCL18 by tolerogenic dendritic cells is associated with inhibition of allergic airway reactivity Iris Bellinghausen, PhD, Sebastian.

Amplification of Toll-like receptor–mediated signaling through spleen tyrosine kinase in human B-cell activation Shigeru Iwata, MD, PhD, Kunihiro Yamaoka,

Sara Paveglio, PhD, MS, Erin Bennett, MS, Kelly L. Hawley, PhD, Adam P

Impairment of T-regulatory cells in cord blood of atopic mothers

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Claudin-1 expression in airway smooth muscle exacerbates airway remodeling in asthmatic subjects Hiroyuki Fujita, MD, PhD, Maciej Chalubinski, MD, PhD,

IgG4 production is confined to human IL-10–producing regulatory B cells that suppress antigen-specific immune responses Willem van de Veen, MSc, Barbara.

Induction of anergic allergen-specific suppressor T cells using tolerogenic dendritic cells derived from children with allergies to house dust mites

CCL17/thymus and activation-regulated chemokine induces calcitonin gene–related peptide in human airway epithelial cells through CCR4 Kandace Bonner,

Volume 29, Issue 3, Pages (September 2008)

Comparison of the allergic and nonallergic CD4+ T-cell responses to the major dog allergen Can f 1 Suvi Parviainen, MSc, Antti Taivainen, MD, PhD, Aino.

Presentation transcript:

Induction and maintenance of allergen-specific FOXP3+ Treg cells in human tonsils as potential first-line organs of oral tolerance Oscar Palomares, PhD, Beate Rückert, Tuomas Jartti, MD, Umut Can Kücüksezer, MD, PhD, Tuomo Puhakka, MD, Enrique Gomez, PhD, Heinz B. Fahrner, MD, Andreas Speiser, MD, Andreas Jung, MD, William W. Kwok, PhD, Livije Kalogjera, MD, Mübeccel Akdis, MD, PhD, Cezmi A. Akdis, MD Journal of Allergy and Clinical Immunology Volume 129, Issue 2, Pages 510-520.e9 (February 2012) DOI: 10.1016/j.jaci.2011.09.031 Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 High numbers of functional CD4+FOXP3+ Treg cells in human palatine tonsils. A, Percentage of cells expressing the indicated surface markers in freshly isolated TMCs and PBMCs. B, FOXP3 T-cell expression in freshly isolated TMCs (n = 20) compared with PBMCs (n = 10). C, Characterization of CD3+CD4+FOXP3+ Treg cells in freshly isolated TMCs (n = 5). D, Dot plots of the indicated cytokines and FOXP3 in freshly isolated TMCs (n = 7) gated on CD3+CD4+ T cells. E, Equal suppressive capacity and no self-proliferation of tonsil and peripheral blood Treg cells. The histograms show the percentage of proliferating responder TMCs or PBMCs stimulated with plate-bound anti-CD3 for each assayed condition. Data are from 1 of at least 3 independent experiments with the same result. ∗∗∗P < .0001. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Human tonsils predominantly contain pDCs. A, CD123/CD303- and CD1c/CD11c-expressing DCs in freshly isolated TMCs. The graph displays the percentages of DC subsets in freshly isolated TMCs and PBMCs. ∗∗P < .001, ∗∗∗P < .0001. B, The pDC and mDC fractions enriched in TDCs were stained for CD123/CD303 and CD1c/CD11c. The graph displays the percentages of DC subsets in TDCs. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 FOXP3+ Treg cells colocalize with pDCs in the T-cell areas of human lingual tonsils. A, The black squares inside H&E sections (left panels) indicate the B- and T-cell areas shown on the right panels. The sections were stained for FOXP3 (green) and CD123 (red) and analyzed by confocal microscopy. B, Close contact and direct interactions between FOXP3+ (green) Treg and CD123+ (red) pDCs. C, Sections were stained for FOXP3 (green), ki-67 (red), and DAPI (blue) or for the corresponding matching isotype controls. Data are from 1 of at least 3 tissue samples with similar results. White bars, 10 μM. DAPI, 4’-6-Diamidino-2-phenylindole, dihydrochloride. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Detection of Bet v 1–specific FOXP3+ T cells in human palatine tonsils. A, Flow cytometric analysis of Bet v 1 tetramer+ CD4+ T cells and Bet v 1 tetramer+ FOXP3+ CD4+ T cells in freshly isolated TMCs and PBMCs. Class II-associated invariant chain peptide (CLIP) tetramers were used as negative control. The graphs display the percentage of Bet v 1 tetramer-positive T cells detected in CD3+CD4+ T cells and the percentage of FOXP3+ cells within the Bet v 1 tetramer+ CD4+ T cells. ∗P < .05, ∗∗P < .005. B, Bet v 1 tetramer+ T cells and Bet v 1 tetramer+ FOXP3+ T cells within the CD3+CD4+ T cells in freshly isolated TMCs and after expansion of tonsil CD4+ T cells with Bet v 1. The graphs display the percentage of Bet v 1 tetramer+ T cells and the percentage of FOXP3+ tetramer+ cells in CD3+CD4+ T cells. ∗P < .05. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 FOXP3+ Treg cells in immune tolerance to allergens in human tonsils. A, TMCs and TMCs depleted of FOXP3+ Treg cells (TMC [−Treg]) stimulated with the indicated antigens and allergens. B, TMC (−Treg) and TMC (−Treg) plus purified autologous tonsil FOXP3+ Treg cells stimulated with the indicated antigens and allergens. Proliferation was measured after 5 days of culture by [3H]-thymidine incorporation during the last 16 hours. The demonstrated counts per minute (cpm) values are those resulting after subtracting the cpm values of the corresponding unstimulated condition. ∗P < .05, ∗∗∗P < .0005. PPD, Purified protein derivative; TT, tetanus toxoid. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Tonsil pDCs drive naive CD4+ T cells to functional CD4+FOXP3+ T cells. A, Flow cytometric analysis of FOXP3/CD25 and FOXP3/CD127 expression in CD4+ T cells after 6 days of coculturing allogeneic naive CD4+ T cells with TpDCs or TmDCs activated as indicated. B, FOXP3/CD25 and FOXP3/CD127 expression of purified Treg cells induced by TpDCs and purified CD4+CD25+ T cells induced by TmDCs. One representative experiment out of 3 with similar results is displayed. C, Proliferation of CFSE-labeled PBMCs gated on CD4+ cells after 5 days of coculture with autologous purified CD4+FOXP3+ Treg cells generated by allogeneic TpDCs or TmDCs. The histograms show the percentage of proliferating responder cells stimulated with plate-bound anti-CD3. Naive CD4+ cells cultured without DC were used for control purposes. One representative experiment out of 3 with similar results is displayed. TmDC, Tonsil myeloid dendritic cells; TpDC, tonsil plasmacytoid dendritic cells. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 The frequencies of FOXP3+ Treg cell and DC subsets in tonsils from atopic and nonatopic individuals. A, Percentages of FOXP3+ Treg cells and DC subsets in TMCs from atopic and nonatopic individuals. B, Correlation analysis of the percentages of FOXP3+ Treg cells and pDCs or mDCs in TMCs from atopic and nonatopic individuals (∗P < .05). Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Purity of FOXP3+ Treg cells purified from tonsils and peripheral blood. Representative dot plots of CD3 and FOXP3 gating on CD3+CD4+ T cells in freshly isolated TMCs and the corresponding purified tonsil FOXP3+ Treg cells or freshly isolated PBMCs and the corresponding purified blood FOXP3+ Treg cells are shown. The percentage of FOXP3+ cells is displayed inside the quadrant. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 FOXP3+ Treg cells colocalize with pDCs in the T-cell areas of human palatine tonsils. A, The black squares inside H&E sections (left panels) indicate the B- and T-cell areas shown on the right panels. The sections were stained for FOXP3 (green) and CD123 (red) and analyzed by confocal microscopy. B, Close contact and direct interactions between FOXP3+ (green) Treg and CD123+ (red) pDCs. C, Sections were stained for FOXP3 (green), ki-67 (red), and DAPI (blue) or for the corresponding matching isotype controls. Data are from 1 of at least 3 tissue samples with similar results. White bars, 10 μM. DAPI, 4’-6-Diamidino-2-phenylindole, dihydrochloride. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 FOXP3+ Treg cells colocalize with pDCs and proliferate in T-cell areas of human tonsils. A, Human palatine and lingual tonsil sections stained for FOXP3 (green) and CD123 (red) and analyzed by confocal microscopy. The graph displays the percentage of FOXP3+ Treg/pDC (CD123+) pairs relative to the total FOXP3+ Treg cell population. B, Human palatine and lingual tonsil sections stained for FOXP3 (green) and ki-67 (red). Coexpression of both markers (yellow cells) is indicated by white arrows. The graph displays the percentage of FOXP3+/Ki67+ Treg cells relative to the total Treg-cell population. Data are from 1 of at least 3 to 4 tissue samples with similar results. White bars, 10 μM. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Control tetramer and FOXP3 staining in freshly isolated TMCs from a nonmatching donor for allele specificity. Representative flow cytometry dot plots of the staining with HLA-DRB1∗0701, ∗1101, and ∗1501 CLIP peptide or Bet v 1 peptide tetramers in combination with FOXP3 in freshly isolated TMCs from a HLA-DRB1∗13/∗14 typed donor. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Depletion of FOXP3+ Treg cells from TMCs. A, Representative dot plots of TMCs and TMCs depleted of FOXP3+ Treg cells. On the left side, representative dot plots of CD3 and CD4 expression in TMCs and TMCs depleted of Treg cells are shown. The black squares indicate the CD3+CD4+ T-cell fraction gated for the subsequent analysis of FOXP3 and CD25 expression (representative dot plots are on the right side). The graph displays the percentage of FOXP3+ and CD25+ within the CD3+CD4+ T-cell population in TMCs and TMCs depleted of Treg cells (n = 4). B, Representative dot plots of TMCs and TMCs depleted of FOXP3+ Treg cells for B cells (CD19 vs CD1c), mDC subsets (CD1c vs CD11c), and pDCs (CD123 vs CD303). Percentages of B cells, mDC subsets, and pDCs are displayed inside the plots (n = 4). Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E6 Purification and characterization of tonsil pDCs and mDCs. A, Different DC subsets can be visualized in the TDC fraction. On the right side, representative dot plots of CD123/CD303 of the purified TpDC and CD1c/CD11c of the obtained TmDC fraction are shown. The mean frequency (± SEM) of cells expressing the indicated markers is also displayed. B, Purified TpDCs and TmDCs were stimulated for 24 hours with the indicated stimulus. Cytokines levels in cell-free supernatants were quantified by cytometric bead array (IL-6, TNF-α, IL-10, and IL-12) or by ELISA (IFN-α). Data represent the mean (± SEM) of 3 to 6 independent experiments. C, The surface expression of HLA-DR, CD80, and CD83 on freshly purified TpDCs or TmDCs and TpDCs or TmDCs activated with the indicated stimulus for 24 hours was determined by flow cytometry. Histograms for a representative experiment are shown. Open histograms represent staining of the indicated marker, and closed histograms represent the matching isotype control. The graphs below display the mean fluorescence intensity using the entire cell population for each surface marker at different conditions. The data represent the mean ± SEM of 3 independent experiments. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E7 FOXP3+ Treg cells induced by activated TpDCs do not produce cytokines. A, FOXP3 mRNA expression determined by quantitative real-time RT-PCR after 6 days of coculturing allogeneic naive CD4+ T cells with TpDCs or TmDCs activated as indicated without any second further stimulation, n = 3. B, Representative flow cytometry dot plots of FOXP3/CD25 expression in the freshly isolated allogeneic naive CD4+ T cells used for coculture experiments with TpDCs and TmDCs. C, After 6 days of coculturing allogenic naive CD4+ T cells with TpDCs (activated as indicated), primed T cells were washed and polyclonally stimulated with a mixture of soluble anti-CD2/CD3/CD28 mAbs for 48 hours. IFN-γ, IL-4, IL-5, and IL-10 mRNA expression was determined by using quantitative real-time RT-PCR. D, After 6 days of coculturing allogenic naive CD4+ T cells with TpDCs activated as indicated, primed T cells were washed and stimulated with phorbol 12-myristate 13-acetate/ionomycin for 6 hours. FOXP3 and IL-10 expression determined by flow cytometry in CD3+CD4+ T cells. One representative example out of 3 with similar results is displayed. Journal of Allergy and Clinical Immunology 2012 129, 510-520.e9DOI: (10.1016/j.jaci.2011.09.031) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions