Presentation is loading. Please wait.

Presentation is loading. Please wait.

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

Published byIsabella Sims Modified over 5 years ago

Similar presentations

Presentation on theme: "Human circulating group 2 innate lymphoid cells can express CD154 and promote IgE production  Laura Maggi, PhD, Gianni Montaini, BSc, Alessio Mazzoni,"— Presentation transcript:

1 Human circulating group 2 innate lymphoid cells can express CD154 and promote IgE production 
Laura Maggi, PhD, Gianni Montaini, BSc, Alessio Mazzoni, PhD, Beatrice Rossettini, BSc, Manuela Capone, PhD, Maria Caterina Rossi, PhD, Veronica Santarlasci, MD, Francesco Liotta, MD, Oliviero Rossi, MD, Oreste Gallo, MD, Raffaele De Palma, MD, Enrico Maggi, MD, Lorenzo Cosmi, MD, Sergio Romagnani, MD, Francesco Annunziato, PhD  Journal of Allergy and Clinical Immunology  Volume 139, Issue 3, Pages e4 (March 2017) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Isolation and characterization of circulating human ILC2s. A, Experimental design and sorting strategy to obtain ILC2s and TH2 cells from PBMCs of healthy donors. B, Representative confocal analysis of ex vivo–derived ILC2s and TH2 cells (green = CD45, blue = TOPRO-3). Scale bar = 5 μm. C, Intracytoplasmic cytokine produced by ILC2s and TH2 cells in response to stimulation with PMA/I (means ± SEs of 11 healthy subjects). D, One representative experiment showing flow cytometric analysis of cytokines produced by PMA/I-stimulated ILC2s and TH2 cells. **P ≤ .01 and ***P ≤ .001, ILC2s vs TH2 cells. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Surface markers, cytokine production, and transcriptional factor genes profile in in vitro–expanded ILC2s and TH2 cells. A, CRTH2 and CD161 expressed by in vitro–expanded ILC2s and TH2 cells (means ± SEs of 10 experiments). B, Intracytoplasmic cytokines produced by in vitro–expanded ILC2s and TH2 cells in response to PMA/I (means ± SEs of 16 experiments for all cytokines, except IL-17 and IL-22 [means ± SEs] of 6 experiments). C, One representative experiment showing flow cytometric analysis of cytokines produced by PMA/I-stimulated ILC2 and TH2 cell lines. Red dots represent IL-13–producing cells. D, Cytokine levels in culture supernatants from ILC2 and TH2 cell lines (means ± SEs of 8 experiments). E, Real-time quantitative PCR analysis of GATA3, RORA, RORC, and TBX21 mRNA levels on ILC2, TH2, and TH1 cell lines (means ± SEs of 6 experiments). *P ≤ .05, **P ≤ .01, and ***P ≤ .001, ILC2s vs TH2 cells or indicated by bar. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Cytokine and transcriptional factor methylation status. DNA methylation status of cytokines (A) and transcriptional factors (B) on ILC2, TH2, and TH1 cell lines are represented as a colorimetric code (yellow-blue scale). Methylation levels at each CpG site are the average of 3 cell lines (left part of each panel). The distribution of the average methylation levels at each CpG site of the whole region is depicted also as a box plot for each population. Means and 25th and 75th percentiles are shown in boxes, and minimum and maximum values are shown as whiskers (right part of each panel). #Indicates the number of CpG sites. *P ≤ .05 and **P ≤ .01. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 CD154 expression and induction of immunoglobulin production by ILC2s and TH2 cells. A, Intracellular expression of CD154 on ILC2 and TH2 cell lines in response to PMA/I or IL25/IL-33 (means ± SEs of 6 experiments). B, One representative flow cytometric histogram of intracellular expression of CD154 by ILC2s and TH2 cells. C, Time-course surface expression of CD154 on ILC2s and TH2 cells in response to PMA/I or IL25/IL-33 (means ± SEs of 3 experiments). D, Immunoglobulin production levels induced by ILC2s and TH2 cells cocultured with autologous B lymphocytes in the presence of IL-25/IL-33 or anti-CD3, respectively, and in the presence or absence of blocking anti-CD154 mAb (means ± SEs of 5 experiments). E, IgE production levels induced by ILC2s and TH2 cells cocultured with autologous B lymphocytes in the presence of IL-25/IL-33 or anti-CD3, respectively, and in the presence or absence of blocking anti–IL-4 and anti–IL-13 mAbs (means ± SEs of 3 experiments). *P ≤ .05 and **P ≤ .01. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 TLR expression by ILC2s and effects of their stimulation with Mix TLR-ligand on CD154 expression and induction of immunoglobulin production. A, Real-time quantitative PCR analysis of TLR1 to TLR10 mRNA levels in ILC2 and TH2 cell lines (means ± SEs of 8 experiments). B, Intracellular expression of CD154 by ILC2 and TH2 cell lines in response to Mix TLR-ligand (means ± SEs of 3 experiments). C, Cytokine levels in culture supernatants from ILC2 lines in response to Mix TLR-ligand (means ± SEs of 3 experiments). D, Immunoglobulin production levels induced by ILC2s cocultured with autologous B lymphocytes in the presence of Mix TLR-ligand and in the presence or absence of blocking anti-CD154 mAb (means ± SEs of 3 experiments). E, IgE production levels induced by ILC2s cocultured with autologous B lymphocytes in the presence of Mix TLR-ligand and in the presence or absence of blocking anti–IL-4 and anti–IL-13 mAbs (means ± SEs of 3 experiments). *P ≤ .05 and **P ≤ .01, ILC2s vs TH2 cells or indicated by bar. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Circulating ILC2s in atopic subjects. A, Frequency and numbers of circulating ILC2s and TH2 cells in nonatopic and atopic subjects (means ± SEs of 11 subjects of each group). B, Correlation between serum IgE levels and the frequency or the number of circulating ILC2s and TH2 cells in 11 atopic subjects. C, Flow cytometric intracellular cytokine production by circulating ILC2s and TH2 cells in nonatopic and atopic subjects in response to PMA/I (means ± SEs of 10 nonatopic and 9 atopic subjects). D, Representative flow cytometric plots of ILC2s and TH2 cells in PB and nasal tissue of 2 allergic patients. #Indicates the patient number. *P ≤ .05. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 Activated ILC2s are able to induce IgE production by B cells. Human circulating ILC2s can express CD154 and type 2 cytokines after IL-25/IL-33 stimulation or triggering of TLR1, TLR4, and TLR6 and are able to induce IgE production by autologous B cells. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E1 Transcription factor expression on ILC2s. One representative experiment showing flow cytometric analysis of GATA-3 and RORγt on the indicated population (A) and in association with IL-13 expression on PMA/I-stimulated ILC2s (B). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E2 ILC2s express DLL1. Real-time quantitative PCR analysis of BAFF, APRIL, and DLL1 mRNA levels in ILC2 and Th2 cell lines (A; means ± SEs of 8 experiments) and in ILC2 lines cultured in the presence of IL-25/IL-33 for 6 or 12 hours (B; means ± SEs of 4 experiments). *P ≤ .05, ILC2s vs TH2 cells. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E3 Effects of ILC2s on B cells. A, Frequency of viable B cells evaluated as Annexin negative and propidium iodide (PI) negative after 24, 48, and 72 hours and cocultured with ILC2s or TH2 cells in the presence of IL-25/IL-33 or anti-CD3 mAb, respectively (means ± SEs of 3 experiments). B, Frequency of proliferating B cells evaluated after 10 days of coculture with IL-25/IL-33–stimulated ILC2s or anti-CD3 mAb–stimulated TH2 cells (means ± SEs of 4 experiments). *P ≤ .05 for each culture condition versus B cells alone in medium. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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

Similar presentations

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

Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood 
Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity  Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste,

Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity  Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste,
Barbara Stanic, PhD, Willem van de Veen, PhD, Oliver F

Barbara Stanic, PhD, Willem van de Veen, PhD, Oliver F
Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion 

Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion 
Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu  Amanda J. Reefer, MS, Kathryn.

Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu  Amanda J. Reefer, MS, Kathryn.
Induction of anergic allergen-specific suppressor T cells using tolerogenic dendritic cells derived from children with allergies to house dust mites 

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.

Cell-to-cell contact between activated CD4+ T lymphocytes and unprimed monocytes interferes with a TH1 response  Miriam Wittmann, MD, Mareike Alter, Tanja.
Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,

Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,
Increased expression of nuclear factor of activated T cells 1 drives IL-9–mediated allergic asthma  Sonja Koch, PhD, Anna Graser, PhD, Hooman Mirzakhani,

Increased expression of nuclear factor of activated T cells 1 drives IL-9–mediated allergic asthma  Sonja Koch, PhD, Anna Graser, PhD, Hooman Mirzakhani,
Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,

Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,
Pentraxin 3 deletion aggravates allergic inflammation through a TH17-dominant phenotype and enhanced CD4 T-cell survival  Jyoti Balhara, MSc, Lianyu Shan,

Pentraxin 3 deletion aggravates allergic inflammation through a TH17-dominant phenotype and enhanced CD4 T-cell survival  Jyoti Balhara, MSc, Lianyu Shan,
Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion 

Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion 
Reduction of CRKL expression in patients with partial DiGeorge syndrome is associated with impairment of T-cell functions  Mauro Giacomelli, PhD, Rajesh.

Reduction of CRKL expression in patients with partial DiGeorge syndrome is associated with impairment of T-cell functions  Mauro Giacomelli, PhD, Rajesh.
Group 2 innate lymphoid cells facilitate sensitization to local, but not systemic, TH2- inducing allergen exposures  Matthew J. Gold, BSc, Frann Antignano,

Group 2 innate lymphoid cells facilitate sensitization to local, but not systemic, TH2- inducing allergen exposures  Matthew J. Gold, BSc, Frann Antignano,
Type 3 innate lymphoid cells induce proliferation of CD94+ natural killer cells  Shuo Li, PhD, Hideaki Morita, MD, PhD, Beate Rückert, Sci Tec, Tadech.

Type 3 innate lymphoid cells induce proliferation of CD94+ natural killer cells  Shuo Li, PhD, Hideaki Morita, MD, PhD, Beate Rückert, Sci Tec, Tadech.
Induction and maintenance of allergen-specific FOXP3+ Treg cells in human tonsils as potential first-line organs of oral tolerance  Oscar Palomares, PhD,

Induction and maintenance of allergen-specific FOXP3+ Treg cells in human tonsils as potential first-line organs of oral tolerance  Oscar Palomares, PhD,
Toll-like receptor 4 ligation enforces tolerogenic properties of oral mucosal Langerhans cells  Jean-Pierre Allam, MD, Wen-Ming Peng, MSc, Torsten Appel,

Toll-like receptor 4 ligation enforces tolerogenic properties of oral mucosal Langerhans cells  Jean-Pierre Allam, MD, Wen-Ming Peng, MSc, Torsten Appel,
Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood 

Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood 
Dawn C. Newcomb, PhD, Madison G

Dawn C. Newcomb, PhD, Madison G
Cysteinyl leukotriene E4 activates human group 2 innate lymphoid cells and enhances the effect of prostaglandin D2 and epithelial cytokines  Maryam Salimi,

Cysteinyl leukotriene E4 activates human group 2 innate lymphoid cells and enhances the effect of prostaglandin D2 and epithelial cytokines  Maryam Salimi,

Similar presentations

Ads by Google