Nasal IL-4+CXCR5+CD4+ T follicular helper cell counts correlate with local IgE production in eosinophilic nasal polyps

doi:10.1016/j.jaci.2015.07.025

Journal of Allergy and Clinical Immunology

Volume 137, Issue 2, February 2016, Pages 462-473

https://doi.org/10.1016/j.jaci.2015.07.025 Get rights and content

Background

Locally produced IgE contributes to the initiation and development of eosinophilic inflammation in eosinophilic nasal polyps independent of systemic atopy. However, whether CXCR5⁺CD4⁺ T follicular helper (T_FH) cells are involved in local IgE production at mucosal sites remains unexplored.

Objective

We sought to explore the presence, phenotype, and function of CXCR5⁺CD4⁺ T_FH cells in eosinophilic nasal polyp tissues compared with noneosinophilic nasal polyp and control normal nasal tissues.

Methods

T_FH cell-surface phenotypes and subsets and B-cell subsets in nasal tissues and peripheral blood were studied by means of flow cytometry. Immunohistochemistry was used to detect the tissue location of T_FH cells. Sorted nasal T_FH cells and CXCR5⁻ T cells were cultured with autologous naive B cells purified from blood.

Results

Nasal T_FH cells expressed inducible costimulator, programmed cell death protein 1, and the transcription factor B-cell lymphoma 6 (Bcl-6) at an intermediate level when compared with bona fide T_FH cells in tonsils and circulating T_FH cells. Although counts of total T_FH cells and IL-21⁺, IFN-γ⁺, and IL-17⁺ T_FH cells were increased in both eosinophilic and noneosinophilic nasal polyp tissues compared with those in normal nasal tissues, IL-4⁺ T_FH cell counts were only increased in eosinophilic polyp tissues. IL-4 and IL-21 were involved in polyp T_FH cell–induced IgE production from naive B cells, and nasal IL-4⁺ T_FH cell counts correlated highly with local IgE levels in vivo. IL-4⁺Bcl-6⁺CD4⁺ T_FH cells were identified in ectopic lymphoid structures in eosinophilic nasal polyps. T_FH cells also positively correlated with germinal center B cells and plasma cells in nasal tissues.

Conclusion

Nasal IL-4⁺ T_FH cells might be involved in local IgE production in eosinophilic nasal polyps.

Section snippets

Patient population and clinical samples

This study was approved by the Ethics Committee of Tongji Hospital and conducted with written informed consent from each patient. The diagnosis of CRSwNP was made according to the current European Academy of Allergy and Clinical Immunology “European position paper on rhinosinusitis and nasal polyps 2012.”¹ CRSwNP was defined as eosinophilic when the percentage of tissue eosinophils exceeded 10% of total infiltrating cells, as reported by our previous study.² Subjects undergoing septoplasty

Enhanced local immunoglobulin levels in nasal polyp tissues

We first confirmed increased local IgE levels in eosinophilic polyps in comparison with that seen in noneosinophilic polyps and control nasal tissues (Fig 1).⁴ Moreover, we found that levels of total IgG, IgG₁, IgG₂, IgG₄, and IgA in nasal tissues were upregulated similarly in both patients with eosinophilic and those with noneosinophilic CRSwNP compared with those in control subjects (Fig 1).

Enrichment of CXCR5⁺CD4⁺ T_FH cells in nasal polyp tissues

Given the enhanced local immunoglobulin levels in nasal polyps, we next explored the presence of T_FH

Discussion

Local IgE production might play an important role in the development of mucosal eosinophilia not only in nasal polyps but also in patients with allergic rhinitis, asthma, and eosinophilic esophagitis, yet the mechanisms underlying this mucosal IgE overproduction remain poorly understood.3, 21, 22 Recently, the percentages of circulating T_FH cells have been found to be increased in patients with autoimmune diseases and correlated with serum concentrations of IgG autoantibodies9, 10; however, the

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Cited by (67)

Aberrant follicular regulatory T cells associate with immunoglobulin hyperproduction in nasal polyps with ectopic lymphoid tissues
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Ectopic lymphoid tissues (eLTs) and associated follicular helper T (T_FH) cells contribute to local immunoglobulin hyperproduction in nasal polyps (NPs). Follicular regulatory T (T_FR) cells in secondary lymphoid organs counteract T_FH cells and suppress immunoglobulin production; however, the presence and function of T_FR cells in eLTs in peripheral diseased tissues remain poorly understood.
We sought to investigate the presence, phenotype, and function of T_FR cells in NPs.
The presence, abundance, and phenotype of T_FR cells in NPs were examined using single-cell RNA sequencing, immunofluorescence staining, and flow cytometry. Sorted polyp and circulating T-cell subsets were cocultured with autologous circulating naïve B cells, and cytokine and immunoglobulin production were measured by ELISA.
T_FR cells were primarily localized within eLTs in NPs. T_FR cell frequency and T_FR cell/T_FH cell ratio were decreased in NPs with eLTs compared with NPs without eLTs and control inferior turbinate tissues. T_FR cells displayed an overlapping phenotype with T_FH cells and FOXP3⁺ regulatory T cells in NPs. Polyp T_FR cells had reduced CTLA-4 expression and decreased capacity to inhibit T_FH cell–induced immunoglobulin production compared with their counterpart in blood and tonsils. Blocking CTLA-4 abolished the suppressive effect of T_FR cells. Lower vitamin D receptor expression was observed on polyp T_FR cells compared with T_FR cells in blood and tonsils. Vitamin D treatment upregulated CTLA-4 expression on polyp T_FR cells and restored their suppressive function in vitro.
Polyp T_FR cells in eLTs have decreased CLTA-4 and vitamin D receptor expression and impaired capacity to suppress T_FH cell–induced immunoglobulin production, which can be reversed by vitamin D treatment in vitro.
IL-21 induces pyroptosis of Treg cells via Akt–mTOR–NLRP3–caspase 1 axis in eosinophilic chronic rhinosinusitis
2023, Journal of Allergy and Clinical Immunology
Regulatory T (Treg) cells, which prevent inflammation-induced eosinophil infiltration, are deficient in nasal polyps (NPs) in patients with eosinophilic chronic rhinosinusitis (ECRS). It is concomitant with loss of Foxp3 after certain inflammatory stimuli.
We sought to determine the inflammatory cytokines involved in inducing the loss of Treg cells in NPs.
The abundance of cytokines in ECRS patients or mice were tested using ELISA, immunochemistry, immunofluorescence, quantitative reverse transcription PCR (qPCR), and/or flow cytometry. Expression of eosinophil cationic protein (ECP), CD4⁺ T cells, IL-4, and IL-17A and eosinophils in nasal mucosa of mouse model was investigated by immunochemistry, immunofluorescence, and hematoxylin and eosin staining. The percentage and death of induced Treg (iTreg) cells, source of IL-21 in NPs from ECRS and non-ECRS patients, and abundance of different systemic phenotypes of CD4⁺ T cells in a mouse model were studied by flow cytometry. Western blot analysis, scanning, and transmission electronic microscopy were used to detect pyroptosis of iTreg cells.
IL-21 was highly expressed in nasal mucosa of ECRS patients and mice, causing pyroptosis and preventing development of iTreg cells in vitro. The elevated IL-21 in NPs from ECRS patients was mainly produced by CD3⁺ T cells, including T follicular helper, T peripheral helper, T_H2, and T_H17 cells and CD3⁺CD4⁻ T cells. T peripheral helper cells and CD3⁺CD4⁻ T cells were the predominant source of IL-21 in NPs from non-ECRS patients. Blocking IL-21/IL-21R signaling significantly reduced the number of eosinophils and CD4⁺ T cells along with ECP, IL-4, and IL-17A expression in the nasal mucosa of ECRS mice. It also increased Treg cell percentage and systemically decreased T_H2 and T_H17 ratios. Akt-mTOR inhibition prevented IL-21–induced pyroptosis in human and mouse iTreg cells.
Elevated IL-21 drives pyroptosis and prevents Treg cell development in ECRS patients. IL-21 induced pyroptosis via activating Akt–mTOR–NLRP3–caspase 1 signaling.
Distinct subsets of innate lymphoid cells in nasal polyp
2023, Allergology International
Group 2 innate lymphoid cells (ILC2s) contribute to the pathogenesis of eosinophilic chronic rhinosinusitis with nasal polyps (CRSwNPs). However, the role of other subsets of ILCs and the differentiation of ILCs in CRSwNPs is not well understood. This study aimed to characterize the ILC subsets and evaluate the differentiation of ILCs from ILC precursors (ILCPs) in NP tissue.
ILC subsets and ILCPs were evaluated by flow cytometry in fresh sinonasal mucosa from patients with CRSwNPs and control subjects. Subsets were compared based on clinical variables and immunological features of the patients. Sorted ILCPs (Lin⁻CD127⁺CD117⁺CD45RA⁺IL1R1⁺) were cultured with cytokines.
The frequency of ILC1s and IFN-γ-producing ILC1s increased in non-eosinophilic NPs, whereas that of ILC2s and IL-5-producing ILC2s increased in eosinophilic NPs, particularly in patients with comorbid asthma. The frequency of ILC1s and IFN-γ-producing ILC1s, and frequency of ILC2s and IL-5-producing ILC2s positively correlated with that of neutrophils and eosinophils, respectively. The proportion of IFN-γ-producing ILC1s positively correlated with clinical severity and levels of IFN-γ and IL-8. The proportion of IL-5-producing ILC2s positively correlated with levels of IL-5, CCL24, and total IgE. ILCPs were identified in NP tissue and differentiated into IFN-γ-producing or IL-5-producing ILCs in response to increased IL-12 and IL-18 or IL-25 and IL-33 in non-eosinophilic NPs and eosinophilic NPs, respectively.
ILC1s and ILC2s may be associated with neutrophilic and eosinophilic inflammation in CRSwNPs, respectively. In addition, ILCPs located in the sinus mucosa could differentiate into IFN-γ- or IL-5-producing cells in response to local cytokine stimuli.
Heterogeneity, subsets, and plasticity of T follicular helper cells in allergy
2022, Journal of Allergy and Clinical Immunology
Antibody responses are critical for protection against pathogens. However, diseases such as allergic rhinitis or food allergy result from aberrant production of IgE antibodies against otherwise innocuous environmental antigens. The production of allergen-specific IgE requires interaction between B cells and CD4⁺ T cells, and a granular understanding of these interactions is required to develop novel therapies for allergic disease. CD4⁺ T cells are exceptionally heterogeneous in their transcriptional, epigenetic, and proteomic profiles, which poses significant challenges when attempting to define subsets relevant to the study of allergy among a continuum of cells. Defining subsets such as the T follicular helper (T_FH) cell cluster provides a shorthand to understand the functions of CD4⁺ T cells in antibody production and supports mechanistic experimentation for hypothesis-driven discovery. With a focus on allergic disease, this Rostrum article broadly discusses heterogeneity among CD4⁺ T cells and provides a rationale for subdividing T_FH cells into both functional and cytokine-skewed subsets. Further, it highlights the plasticity demonstrated by T_FH cells during the primary response and after recall, and it explores the possibility of harnessing this plasticity to reprogram immunity for therapeutic benefit in allergic disease.
T follicular helper cells mediate local production of allergen-specific IgE and IgG4
2022, Journal of Allergy and Clinical Immunology
IL-17A mediates pyroptosis via the ERK pathway and contributes to steroid resistance in CRSwNP
2022, Journal of Allergy and Clinical Immunology
Pyroptosis is closely related to inflammation. However, the molecular mechanisms and pathologic contributions of pyroptotic epithelial cell are not yet fully understood.
This study aimed to explore the function and molecular mechanisms of IL-17A on human nasal epithelial cell (hNEC) pyroptosis.
The expression of pyroptosis-related biomarkers and IL-17A was assessed in sinonasal mucosa from control individuals, patients with chronic rhinosinusitis without nasal polyps, and patients with chronic rhinosinusitis with nasal polyps (CRSwNP) by using quantitative RT-PCR. Their localization was analyzed via immunohistochemistry and immunofluorescence. The ultrastructural characteristics of IL-17A–induced pyroptosis in hNECs were visualized by using electron microscopy. IL-17A functional assays were performed on hNECs and airway epithelial cell lines. Cytokine levels were quantified via ELISA. The signaling pathways involved in IL-17A–induced pyroptosis were studied via unbiased RNA sequencing and Western blotting.
The expression of IL-17A and the pyroptotic biomarkers NOD-like receptor family, pyrin domain containing 3 (NLRP3), caspase-1, gasdermin D, and IL-1β was increased in nasal mucosa from patients with CRSwNP compared with in those with chronic rhinosinusitis without nasal polyps and the control subjects. IL-17A was positively correlated and colocalized with the pyroptotic biomarkers. IL-17A treatment induced pyroptosis in the hNECs and cell lines analyzed, primarily through the extracellular signal–regulated kinase (ERK)-NLRP3/caspase-1 signaling pathway, and increased IL-1β and IL-18 secretion in hNECs. Moreover, IL-17A–induced pyroptosis contributed to steroid resistance by affecting glucocorticoid receptor-α and glucocorticoid receptor-β expression, and the inhibition of pyroptotic proteins partially abolished IL-17A–induced steroid resistance in hNECs.
Elevated IL-17A level promotes pyroptosis in hNECs through the ERK-NLRP3/caspase-1 signaling pathway and contributes to glucocorticoid resistance by affecting glucocorticoid receptor homeostasis in patients with CRSwNP.

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: Supported by National Natural Science Foundation of China (NSFC) grants 81020108018 and 81325006 (to Z.L.), 81200733 (to H.W.), and 81400449 (to P.-P.C.); a grant from the Ministry of Health of China (201202005); the Program for Changjiang Scholars and Innovative Research Team in University (IRT_14R20); and the 12th 5-year science and technology support program (2014BAI07B04).

: Disclosure of potential conflict of interest: H. Wang has received a grant from the National Natural Science Foundation of China (81200733). P.-P. Cao has received a grant from the National Natural Science Foundation of China (81400449). Q. Ning has received a grant from the Ministry of Education of China (Program for Changjiang Scholars and Innovative Research Team in University [IRT_14R20]). Z. Liu has received grants from the National Natural Science Foundation of China (81020108018 and 81325006), the Ministry of Science and Technology of China (2014BAI07B04), and the Ministry of Health of China (201202005). The rest of the authors declare that they have no relevant conflicts of interest.

View full text

Rhinitis, sinusitis, and upper airway diseaseNasal IL-4+CXCR5+CD4+ T follicular helper cell counts correlate with local IgE production in eosinophilic nasal polyps

Background

Objective

Methods

Results

Conclusion

Section snippets

Patient population and clinical samples

Enhanced local immunoglobulin levels in nasal polyp tissues

Enrichment of CXCR5+CD4+ TFH cells in nasal polyp tissues

Discussion

J Allergy Clin Immunol

Immunity

Immunity

Immunity

Immunity

Trends Immunol

Acta Histochem

J Allergy Clin Immunol

Immunity

Immunity

European position paper on rhinosinusitis and nasal polyps 2012

Rhinol Suppl

Distinct immunopathologic characteristics of various types of chronic rhinosinusitis in adult Chinese

J Allergy Clin Immunol

Increased local IgE production induced by common aeroallergens and phenotypic alteration of mast cells in Chinese eosinophilic, but not non-eosinophilic, chronic rhinosinusitis with nasal polyps

Clin Exp Allergy

Local receptor revision and class switching to IgE in chronic rhinosinusitis with nasal polyps

Allergy

Activation-induced cytidine deaminase (AID)-associated multigene signature to assess impact of AID in etiology of diseases with inflammatory component

PLoS One

Rhinitis, sinusitis, and upper airway disease
Nasal IL-4⁺CXCR5⁺CD4⁺ T follicular helper cell counts correlate with local IgE production in eosinophilic nasal polyps

Enrichment of CXCR5⁺CD4⁺ T_FH cells in nasal polyp tissues