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. 2026 May 21;27:298. doi: 10.1186/s12931-026-03734-y

Th2 cytokine-induced mucociliary remodeling in chronic rhinosinusitis: implications for antiviral defense and epithelial function

Juliana Theorell 1, Jenny Drnevich 2, Sofija Jovanovic Gasovic 1, Bridget Lawate 1, Victoria S Lee 3,4, Almudena Veiga-Lopez 1,3,✉
PMCID: PMC13435933  PMID: 42168989

Abstract

Chronic rhinosinusitis (CRS) is a sinonasal inflammatory disease, often complicated by aberrant Th2-driven immunologic responses and increased susceptibility to viral infections. Th2-induced epithelial remodeling has been proposed to facilitate viral entry and replication, thereby increasing susceptibility to infection and exacerbating inflammation in CRS. This exploratory study investigated if chronic Th2-mediated remodeling alters the transcriptional response to rhinoviral infection between individuals with and without CRS. We hypothesized that Th2 cytokine exposure of human primary nasal epithelial cells during their differentiation disrupts mucociliary function, impairing the antiviral response to rhinovirus. Primary nasal epithelial cells from patients with and without CRS were differentiated at air-liquid interface while being exposed to Th2 cytokines (IL-4, IL-13, or IL-4/13; 10 ng/mL) followed by a rhinovirus (RV-A16) infection. RNA sequencing and inflammatory cytokine profiling revealed significant downregulation of pathways involved in cilia structure, development, and function, as well as lower rhinovirus reads in Th2 cytokine-exposed cultures, with similar trends observed in CRS and non-CRS samples. Chronic Th2 cytokine exposure also altered cytokine release, shifting toward an anti-inflammatory profile. Notably, sex-specific differences were observed in unexposed cultures, with male-derived cultures exhibiting higher levels of inflammatory cytokines and accompanying more inflammatory transcriptomic profiles, thus highlighting intrinsic sex-specific immune variability. These findings underscore how Th2 cytokine–driven epithelial remodeling may compromise mucociliary function and antiviral defenses across CRS and non-CRS cultures. Understanding these mechanisms may inform therapeutic strategies aimed at restoring epithelial integrity and mitigating chronic inflammation.

Supplementary Information

The online version contains supplementary material available at 10.1186/s12931-026-03734-y.

Keywords: Chronic rhinosinusitis, Th2 cytokines, Rhinovirus, Nasal epithelial cells

Background

Chronic rhinosinusitis (CRS) is a complex inflammatory disease characterized by persistent sinonasal inflammation lasting for 12 weeks or longer, with a multifactorial pathophysiology involving both genetic predisposition and environmental factors [1]. Sinonasal epithelial cells, once considered merely as a physical barrier, are now recognized as active regulators of inflammation and upper airway integrity, serving as a critical interface that influences CRS pathogenesis [2, 3]. Chronic exposure of the airway epithelium to immune mediators, particularly those implicated in allergic inflammation, such as T-helper 2 (Th2) cytokines, induce structural changes that disrupt normal airway homeostasis [4]. Among these alterations, mucus metaplasia, a condition marked by an increased prevalence of goblet cells, leads to excessive mucus production and impaired mucociliary clearance, contributing to disease chronicity [5].

Th2 cytokines, such as IL-4 and IL-13, are central mediators of allergic inflammation and their aberrant production plays a critical role in the pathogenesis of CRS and co-existing conditions like allergic rhinitis [6]. These anti-inflammatory cytokines, produced primarily by activated T-helper 2 cells in response to allergens and immune triggers, drive mucus metaplasia and disrupt the ciliary structure and function of the nasal epithelium. This dual effect reduces the epithelial barrier’s capacity to clear pathogens and debris from the upper airway, promoting a cycle of inflammation and dysfunction [5]. Deciphering the mechanisms by which Th2 cytokines drive pathogenesis of allergic diseases has been the focus of extensive efforts dedicated towards defining the specific roles of anti-inflammatory cytokines IL-4 and IL-13 [7], particularly because IL-13-associated epithelial remodeling correlates with clinical severity in nasal polyposis [8]. While previous studies have explored these two anti-inflammatory cytokines individually [9–13], the combined effects of IL-4 and IL-13 on epithelial remodeling, particularly their influence on mucociliary dynamics of sinonasal epithelial cells derived from CRS, remain unexplored, and one of the goals of this study.

Rhinovirus, the most common respiratory virus associated with CRS exacerbations, worsens sinonasal epithelial dysfunction by disrupting mucociliary clearance and compromising barrier integrity, similar to Th2 cytokine effects [14]. Th2-induced epithelial remodeling has been proposed to facilitate viral entry and replication [10], increasing susceptibility to infection and prolonging inflammation in CRS. While previous studies have suggested that Th2 inflammation impairs innate immune responses [15, 16], the mechanisms linking rhinoviral infection with CRS progression remain poorly understood. Specific to CRS, few have examined how chronic Th2-driven epithelial remodeling affects the antiviral response to rhinovirus, particularly in primary cells derived from CRS patients [16]. This knowledge gap is significant because basal cells in CRS patients are known to maintain an inflammatory epithelial memory, potentially amplifying responses upon re-exposure to Th2 cytokines [17]. Evidence from clinical and in vitro studies highlights the importance of IL-4 and IL-13 in modulating viral susceptibility. For instance, dupilumab, an inhibitor of IL-4 and IL-13 signaling and an FDA-approved medication for the treatment of CRS, has been reported to reduce respiratory infections in patients with moderate-to-severe asthma or severe chronic rhinosinusitis with nasal polyps [18]. Similarly, exposure of bronchial epithelial cells to IL-4 or IL-13 increased rhinovirus replication and impaired interferon production [15]. Together, these findings underscore the need to investigate how Th2-mediated remodeling alters antiviral defenses in the context of CRS, which could inform future approaches to developing therapeutic strategies aimed at mitigating susceptibility to infections.

Given the interplay between Th2-driven inflammation, mucociliary remodeling, and increased susceptibility to viral infections observed in CRS, the goal of the study was to evaluate if Th2-mediated remodeling influenced the transcriptional response to rhinoviral infection between individuals with and without CRS. We hypothesized that chronic exposure of human primary nasal epithelial cells to Th2 cytokines during differentiation triggers mucociliary remodeling, leading to changes in the cell type architecture and impairing the antiviral response to rhinovirus. To test this hypothesis, we used air-liquid interface (ALI) primary cultures, which closely mimic the structure and function of human airway epithelium. Sinonasal epithelial cells from patients with and without CRS were chronically exposed to Th2 cytokines (IL-4, IL-13, or IL-4/13) during differentiation and subsequently challenged with rhinovirus. Using RNA sequencing and inflammatory cytokine profiling, key pathways and cellular changes driven by Th2 inflammation on primary sinonasal epithelial cell cultures were identified.

Material & methods

A full description of the methods used in this study is available in the Supplemental Materials (Sections. 1–16, Tables S1 - S3). Sinonasal epithelial cells were collected from 9 individuals (N = 3 healthy, N = 6 CRS; demographics in Table 1, Table S4 (by sex), and Table S5 (by disease state)) undergoing endoscopic sinus surgery (IRB STUDY2021-0676) after informed consent. CRS diagnosis was based on persistent sinus inflammation (≥ 12 weeks) with cardinal symptoms and objective evidence from imaging or endoscopy. Cells were isolated using sterile cytology brushes, transported in DMEM/F-12, and processed within 1 h. Primary cultures were established following a previously described protocol [19]. Primary cells were seeded on collagen-coated Transwells and differentiated in ALI conditions with PneumaCult-ALI medium for 30 days until fully mature, with basolateral media changes every 2–3 days and periodic mucus removal. To confirm non-cytotoxic rhinovirus concentrations, a lactate dehydrogenase (LDH) assay was performed at 12, 24, 36, and 48 h post-infection using RV-A16 (1 × 10⁶ TCID₅₀/mL) in primary nasal epithelial cells (n = 4 subjects, ≥ 2 technical replicates per timepoint) (Figure S1).

Table 1.

Demographics

Healthy CRS
N 3 6
Age (years; mean ± SEM) 42.7 ± 8.7a 48.7 ± 5.1a
Race
 White 0 1
 Black or African American 2 3
 Asian 0 2
 Native Hawaiian or Pacific Islander 1 0
Sex (male/female) 1/2a 3/3a
Body Mass Index (mean ± SEM) 33.9 ± 1.1a 27.5 ± 3.2a
Asthma (%) 0a 16.7a
Aspirin sensitivity (%) 0a 0a
Current smoking status (tobacco) (%) 0a 50a
Current smoking status (marijuana) (%) 0a 33a
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CRS Chronic rhinosinusitis

Different letters represent statistical differences at p < 0.05

To evaluate the effect of chronic type-2 cytokine exposure during epithelial cell differentiation, primary nasal epithelial cells derived from healthy (n = 3) or CRS (n = 6) individuals were cultured under control conditions (control group) or supplemented with IL-4, IL-13, or IL-4/13 (10 ng/mL each) in the basolateral compartment starting on day 3 of differentiation (Fig. 1A). Cytokine doses were chosen based on prior studies demonstrating that this concentration induces intrinsic inflammatory memory in human respiratory epithelial progenitor cells [17]. The media in the basolateral compartment was replaced every 2–3 days until the end of the differentiation period. After differentiation, cultures were either mock-infected or infected with RV-A16 at 1 × 106 TCID50/mL for 2 h. Unbound viral particles were removed by washing the membranes eight times with PBS, and cells were then incubated at 37 °C with 5% CO2 until 36 h post-infection. Thereafter, media in the basolateral compartment and total RNA were collected, aliquoted, and stored at -80 °C for cytokine analysis and transcriptome analysis, respectively.

Fig. 1.

Fig. 1

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A Experimental Design. Primary sinonasal epithelial cells were air-lifted and cultured for 30 days under the following conditions: (1) control (C; cell culture media only), (2) IL-4 (10 ng/mL), (3) IL-13 (10 ng/mL), or (4) a combination of IL-4 and IL-13 (10 ng/mL each). On day 30, cells were either mock-infected or infected with rhinovirus (RV-A16). Following a 36-h infection period, basolateral compartment media and cells were collected for RNA sequencing and cytokine analyses. B Effect of Th2 cytokine exposure and rhinoviral infection on the transcriptomic profile of primary sinonasal epithelial cells. Heatmap displaying the transcriptomic profiles of sinonasal epithelial cells from 9 individuals (3 healthy, 6 CRS) after 30 days of Th2 cytokine exposure with or without RV-A16 infection. Supervised clustering was applied to centered and scaled data (Z score), and Euclidean distance clustering was used to group similar genes while samples were put in a pre-defined order to help with interpretability of the many factors. Color keys in the heatmap legend indicate disease status, infection status, and cytokine exposure. Hierarchical clustering of differentially expressed genes revealed distinct expression profiles across treatment groups. Genes with similar patterns across conditions were annotated into clusters (A–G) for interpretability. These clusters highlight shared trends such as Th2-associated repression or infection-induced upregulation. Statistical significance among treatment groups was determined using one-way ANOVA with an FDR-adjusted p < 0.001

Library preparation and RNA sequencing were performed at the Roy J. Carver Biotechnology Center University of Illinois Urbana-Champaign in two batches. RNA sequencing libraries were sequenced on one 10B lane for 151 cycles from both ends of the fragments on a NovaSeq X Plus with V1.0 sequencing kits. Reads were quasi-mapped using Salmon [20] to a combined human and rhinovirus type 16 transcriptome (Figure S2). Follow up analyses were structured as follows: (1) Primary Analyses: To accommodate the complex experimental design and limited sample size, transcriptomic analysis was initially structured around disease × infection × Th2 cytokine exposure model, resulting in 16 experimental groups. To adjust for inter-individual variability and unmodeled confounders (e.g., sex, age, race, batch), the Remove Unwanted Variation (RUV) method [21] was applied to estimate latent factors and improve statistical robustness [22]. Differential expression was assessed using a limma-voom model incorporating 7 RUV-derived factors, and multiple contrasts were tested with global FDR correction for multiple comparisons. To evaluate functional relevance, Gene Ontology and IPA enrichment analyses were conducted on differentially expressed gene sets identified from one-way ANOVA, cytokine effects, and disease-associated contrasts. (2) Exploratory analyses: To assess potential sex-specific effects, an additional model was developed incorporating sex × infection × Th2 cytokine interaction terms (with IL-4/13 treatments grouped). This secondary model used the same RUV-adjusted limma-voom approach with 6 factors and was applied to both differential expression and pathway enrichment analyses.

For cytokine analysis, media from the basolateral compartment was analyzed for 15 inflammatory human cytokines in duplicate using Eve Technologies’ Human Focused 15-Plex Discovery Assay (MilliporeSigma, Burlington, MA, USA) using the Luminex 200 system (Luminex, Austin, TX, USA) by Eve Technologies Corp (Calgary, Alberta, Canada) according to the manufacturer’s protocol.

Results

Sinonasal epithelial cells RNA sequencing summary

Transcriptomic profiling using RNA sequencing revealed significant expression differences driven by Th2 cytokine exposure, followed by infection and disease status. Multidimensional scaling identified dimension 1 (39.4% variance) separating control and Th2-exposed samples, dimension 2 (9.2%) as distinguishing infection, and dimension 3 (4.2%) reflecting disease status (Figure S3). A one-way ANOVA test identified 13,230 genes (83.7%) with significant differences among the 16 experimental groups (FDR < 0.001). Hierarchical clustering of these DEGs revealed distinct expression patterns associated with Th2 cytokine treatment, infection, and disease status (Fig. 1B). Notably, cultures from any of the Th2 cytokine groups (IL-4, IL-13, and IL-4/13) had minimal viral RNA reads post-infection (Figure S2B-C) as well as decreased expression of several IFN receptor subtypes (Fig. 2). Th2 cytokine groups also exhibited increased expression of IL-4 receptor subtypes IL4R and IL2RG, while the IL-13 receptor subtype, IL13RA1, showed a downward trend in expression. In contrast, the decoy IL-13 receptor, IL13RA2, was upregulated (Fig. 2B).

Fig. 2.

Fig. 2

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Expression levels (mean RUV-corrected log2-transformed, counts per million (CPM) values ± SEM) of (A) rhinovirus (RV-A16), (B) IL-4 and IL-13 receptor subunits, and (C) selected rhinovirus and interferon subunit receptors in sinonasal epithelial cells after exposure to (1) control (C; cell culture media only), (2) IL-4 (10 ng/mL), (3) IL-13 (10 ng/mL), or (4) a combination of IL-4 and IL-13 (10 ng/mL each). N = 9 primary cultures per treatment group. Statistical significance was determined using one-way ANOVA among all exposure groups. Asterisks denote statistical differences at FDR-adjusted *p < 0.05, **p < 0.01, ***p < 0.001

Th2 cytokine-dependent effects on sinonasal epithelial cells’ transcriptome and cell morphology

GO analyses revealed terms associated with “differentiation” and “development” for downregulated genes and “immunity” and “antigen presentation” for upregulated genes, mediated upon Th2 cytokine exposure (Fig. 3). Several genes involved in epithelial tight junctions and adherens junctions were downregulated in cytokine-exposed groups (Figure S4). To assess the impact of Th2 cytokine exposure on mucus-secretory programming, key genes involved in this pathway for assessed. Cytokine exposure upregulated MUC5AC and MUC2, the primary gel-forming mucins, as well as the transcription factors/genes SPDEF and FOXA3, which drive goblet cell differentiation and coordinate a transcriptional program underlying mucus hypersecretion (Figure S5). Comparison of the DEGs among the three uninfected Th2 cytokine groups revealed a 91.7% similarity (Fig. 4A). Therefore, subsequent analyses were conducted only on the IL-4/13 exposed cultures.

Fig. 3.

Fig. 3

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Effect of Th2 cytokine exposure (IL-4, IL13, or IL-4/13) and rhinoviral infection on semantic similarity analysis of Biological Process (BP) GO terms. Groups of differentially expressed genes with similar expression patterns were categorized and labeled as follows (A–F): A genes downregulated following Th2 cytokine exposure, B genes exhibiting mixed effects: upregulated due to infection but downregulated in response to Th2 cytokine exposure, C genes with increased expression following Th2 cytokine exposure, D genes exhibiting mixed effect: a baseline decrease in expression due to viral infection and increased expression upon Th2 cytokine exposure, E genes with increased expression specifically in response to viral infection, and F genes with decreased expression specifically in response to viral infection. The package clusterProfiler was utilized for GO over-representation testing and the SimplifyEnrichment package reduced down GO terms based on semantic similarity using BP terms with p < 0.01

Fig. 4.

Fig. 4

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Comparison of differentially expressed genes (DEGs) and pathway analysis among IL-4, IL13, and IL-4/13-exposed sinonasal epithelial cell cultures. A Venn Diagram representing number of unique and overlapping DEGs among uninfected treatment groups from the heatmap A and C gene groups, which corresponded to genes with increased and decreased expression following Th2 cytokine exposure. B Volcano plot of DEGs (FC ≥ 2 or ≤ -2; FDR < 0.001) of IL-4/13 exposed cultures compared to control. C Bubble chart depicting ingenuity pathway analysis (IPA) predictions of dysregulated pathways associated with IL-4/13-exposure compared to controls. Z-score indicates pathway activation (positive) or inhibition (negative). -log(p-value) reflects pathway enrichment significance, and bubble size represents the ratio of overlap (proportion of molecules meeting cutoff criteria within a given pathway). D Heatmap displaying selected cilia assembly pathway DEGs (FC ≥ 2 or ≤ -2; FDR < 0.001) of IL-4/13 exposed cultures compared to the control (N = 9 per treatment group). Euclidean distance clustering was used to group similar profiles. Genes were annotated to the specific pathway component: BBsome signaling, intraflagellar transport, motor proteins, or primary ciliogenesis

A volcano plot of a subset of the 6,229 DEGs (FDR < 0.001) between the control and IL-4/13-exposed cells revealed a predominance of downregulated genes compared to upregulated ones in Th2-exposed cells (Fig. 4B). Furthermore, pathway enrichment analysis revealed four significantly downregulated pathways following cytokine exposure: (1) cilium assembly, (2) Bardet-Biedl Syndrome (BBS) -ome signaling, (3) molecular mechanism of cancer, and (4) phagosome formation (Fig. 4C). The effect of IL-4/13 exposure on cilia assembly, was explored by annotating genes associated with key pathway components, including the BBSome pathway, intraflagellar transport, motor proteins, and primary ciliogenesis (Fig. 4D). IPA demonstrated a widespread suppression of genes associated with ciliary machinery, including intraflagellar transport proteins, motor components, and cargo transport mechanisms (Fig. 5A-D). Representative gene targets from these pathway components were compared across all Th2 cytokine exposure groups (IL-4, IL-13, and IL-4/13) further supporting similarity of gene expression changes across Th2 cytokine exposure groups (Fig. 5E). Representative images of cell morphology revealed a complete absence of cilia in cells exposed to IL-4/13 (Figure S6) and a representative video of ciliary motion in differentiated cultures is provided in Video S1.

Fig. 5.

Fig. 5

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Overlay of differentially expressed genes (DEGs) after IL-4/13 exposure on sinonasal epithelial cells on pathways associated with the intraflagellar transport B complex A, primary cilia assembly and initiation B, the BBSome signaling pathway C, and cargo transport within cilia D. Green and red denote decreased and increased measurement of a particular transcript, respectively. E RNA expression (mean RUV-corrected log2-transformed, counts per million (CPM) values ± SEM) per sample of selected target genes from all four pathways A–D after exposure to: (1) control (C; cell culture media only), (2) IL-4 (10 ng/mL), (3) IL-13 (10 ng/mL), or (4) a combination of IL-4 and IL-13 (10 ng/mL each) (N = 9 per group). Statistical significance was determined using one-way ANOVA among all exposure groups. Asterisks denote statistical differences at FDR-adjusted ***p < 0.001

Th2 cytokine exposure modulates inflammatory cytokine release in sinonasal epithelial cells

Multiplex ELISA revealed significant alterations in inflammatory cytokine release following Th2 cytokine exposure (Fig. 6). IL-4 and IL-13 were detected in the basal media of the respective Th2 cytokine groups, but not in the control group, which served as a positive control. Exposure to IL-4, IL-13, or IL-4/13 significantly reduced IL-6 and MCP-1 concentrations in culture supernatants compared to controls. Interestingly, IL-13 uniquely induced IL-10 production, highlighting a divergent anti-inflammatory response. No significant differences were observed for GM-CSF, TNFα, IL-1β, or other pro-inflammatory cytokines (Figure S7).

Fig. 6.

Fig. 6

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Effect of Th2 cytokine exposure on sinonasal epithelial cells on cytokine secretion. Concentrations (mean ± SEM, pg/mL) of differentially secreted cytokines in cell culture media collected from the basolateral compartment after 30 days of exposure to: (1) control (C; cell media), (2) IL-4 (10 ng/mL), (3) IL-13 (10 ng/mL), or (4) a combination of IL-4 and IL-13 (10 ng/mL each) (N = 8–9 per group). Statistical significance was determined using one-way ANOVA between Th2 cytokine exposure and control followed by Dunnett’s multiple comparison test. Asterisks denote statistical differences at *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Sex-specific differences in cytokine release profiles sinonasal epithelial cells

Notably, baseline cytokine release patterns in control cultures (not exposed to any Th2 cytokine) revealed marked sex-specific differences. To contextualize these findings, a sex-stratified demographic summary is provided in Table S4. Principal component analysis (PCA) of cytokine profiles demonstrated that male-derived cultures produced higher levels of cytokines, including IL-1β, MCP-1, and IL-1RA, compared to female-derived cultures (Fig. 7). Trends toward elevated IL-6 (p = 0.08), IL-12p40 (p = 0.06), IL-12p70 (p = 0.09), and IFNγ (p = 0.08) were also noted in male cultures (Figure S8). Interestingly, IL-13 production did not differ by sex or disease status (Fig. 7C).

Fig. 7.

Fig. 7

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Effect of sex on cytokine secretion from the basolateral compartment in control (unexposed) sinonasal epithelial cells after 30 days of exposure to standard differentiation media. (A) Two-dimensional principal component analysis (PCA) scatter plot of the first two principal components (PC1 and PC2), capturing 47.1% and 21.7% of the total variance, respectively in control female-derived (F, pink) and male-derived (M, blue) cultures. (B) Biplot displaying the loadings for secreted cytokines, where the arrows indicate the direction and strength of each cytokine’s contribution to the PCs. (C) Concentrations (mean ± SEM, pg/mL) of differentially secreted cytokines in female-derived and male-derived cultures (N = 4–5 per group). Statistical significance was determined using unpaired t-test between female- and male-derived cultures. Asterisks denote statistical differences at *p < 0.05 and **p < 0.01

Sex-specific differences in transcriptome profiles sinonasal epithelial cells

Given these findings, sex was incorporated as a variable in the statistical models for DEG analysis to capture sex-specific changes. Pathway enrichment of sex-associated genes in control cultures revealed downregulation of retinoic acid receptor (RAR) activation pathway in male-derived cultures (Figure S9A). To further examine this, genes associated with the RAR pathway as well as inflammatory cytokines and suppressor of cytokine signaling markers were explored (Figure S9B). Correlation analysis of expression and cytokine release identified a significant correlation in IL-1β and MCP-1 (Figure S9C–D). No sex-driven differences were observed in any steroid hormone receptors in control (Figure S10) or unexposed infected cultures (Figure S11).

Disease-specific differences in transcriptome profiles of sinonasal epithelial cells

Transcriptomic profiling using RNA sequencing amongst disease states and exposure groups identified 3,504 genes with a global FDR p < 0.05 (Figure S12). GO analyses revealed terms associated with “lipid” and “immunity” (Figure S13). No significant differences were observed in the expression of Th2 cytokine receptor subunits between samples from healthy and CRS cultures under either control or Th2-exposed conditions (Figure S14). Pathway enrichment analysis of disease status in control cultures (uninfected and unexposed) revealed downregulation of eicosanoid signaling in CRS samples compared to healthy controls (Figure S15). In infected, but unexposed cultures, CRS samples showed reduced expression of genes associated with the antimicrobial peptide pathway (Figure S16A–B). Additionally, infected CRS samples exhibited lower expression of rhinoviral transcripts relative to healthy samples; however, this difference was not observed in cultures exposed to Th2 cytokines (Figure S16C-D).

Discussion

In this exploratory study, we have evaluated the individual (IL-4 or IL-13) and combined (IL-4/13) effects of chronic Th2 cytokine exposure, particularly their influence on mucociliary dynamics and rhinoviral susceptibility of sinonasal epithelial cells derived from healthy and CRS subjects. Independent of disease status, our transcriptomic data demonstrated a significant downregulation of pathways involved in cilia structure, development, and function, as well as lower rhinovirus reads in Th2 cytokines-exposed cultures. Cytokine release was also altered by chronic Th2 cytokine exposure, reflecting an anti-inflammatory profile. Notably, the cytokine release profile derived from unexposed cultures displayed marked sex-specific differences, with male-derived cultures exhibiting an increased production of several inflammatory cytokines. Consistent with this, transcriptomic analyses revealed sex-associated differences in gene expression, particularly in pathways related to immune regulation and inflammatory signaling. Overall, these findings indicate that chronic Th2 cytokine exposure drives key molecular and structural changes that impact rhinoviral susceptibility in both CRS-derived and non-CRS cultures.

Independent of disease status, chronic Th2 cytokine exposure in sinonasal epithelial cells significantly altered major components of the cilia assembly pathway, a highly coordinated process essential for the formation, maintenance, and function of cilia. Our findings revealed a broad downregulation of key systems involved in this pathway, including the BBSome pathway, which involves a cargo adaptor complex that facilitates the transport of membrane proteins and signaling receptors throughout the cilia and the intraflagellar transport system, responsible for bidirectional movement of cargo along the axoneme [23]. Motor proteins like kinesin-2 and dynein-2, which power anterograde and retrograde transport [24], respectively, were similarly affected. Processes critical to ciliogenesis, necessary for the formation and maintenance of cilia [25], were also downregulated. Together, these disruptions indicate a systemic inability of primary sinonasal epithelial cells to form and sustain functional cilia, likely leading to compromised ciliary signaling, defective cargo transport, and structural abnormalities. Ultimately, this could contribute to the loss of ciliated cells and reduction in mucociliary clearance, a hallmark feature of sinonasal epithelial dysfunction [26]. These observations align with previous studies demonstrating chronic IL-13 exposure drives extensive mucociliary remodeling, further underscoring the critical role of Th2-driven inflammation in impairing epithelial integrity and function [27, 28].

Chronic Th2 cytokine exposure in sinonasal epithelial cells triggered transcriptional activation of a mucus-secretory program, a process that drives goblet cell differentiation and mucus production. Expression of MUC5AC and MUC2, the main gel-forming mucins [29], was upregulated, along with transcription factors SPDEF and FOXA3, which coordinate goblet cell lineage commitment [30, 31]. This transcriptional signature reflects a shift a hypersecretory epithelial phenotype, consistent with goblet cell metaplasia and excessive mucus production. These changes likely contribute to disrupted mucociliary balance and impaired epithelial function in the sinonasal tract. These findings align with previous reports showing that IL-13 exposure induces pronounced goblet cell hyperplasia and mucus remodeling [32], further emphasizing the central role of type 2 inflammation in shaping epithelial secretory responses [33]. IL-4 and IL-13 have long been thought to possess largely redundant effector profiles due to their shared signaling mechanisms and genomic proximity. Both cytokines are encoded by adjacent genes in humans, share the GATA-3 transcriptional regulatory element, and signal through a shared type II receptor complex (IL-4Rα/IL-13Rα1) [34]. Additionally, IL-4 can also signal through the type I receptor (IL-4Rα / IL-2Rγc), which is predominantly expressed in lymphocytes and myeloid cells, while the type II receptor is expressed more broadly, including on myeloid cells and non-hematopoietic cells [35]. In this study, we observed that chronic exposure to IL-4, IL-13, or a combination of both cytokines to sinonasal epithelial cells produced remarkably similar transcriptomic profiles. This similarity may reflect receptor redundancy or saturation, as both cytokines signal through the type II receptor complex (IL-4Rα/IL-13Rα1). While receptor expression levels were not directly measured, airway epithelial cells are known to modulate receptor expression as a function of differentiation [36]. For instance, airway epithelial cells have reportedly increased expression of type I receptor subunits as a function of the degree of differentiation, which could influence differential responses between IL-4 and IL-13 under certain conditions [36], such as during different stages of differentiation, individual variability, disease states, or changes in receptor availability. However, the similar transcriptomic profiles observed following IL-4, IL-13, or combined exposure in this study suggest that receptor redundancy, rather than modulation, is likely the primary driver of these shared responses.

This study builds upon prior research by using a non-targeted transcriptomic approach to evaluate how chronic Th2 cytokine exposure modulates sinonasal epithelial cell function in the context of CRS. Notably, our findings demonstrate that the transcriptional downregulation of the cilia assembly pathway is driven by chronic exposure to Th2 cytokines rather than by individual variations. While genetic variation could theoretically influence individual responses to Th2 cytokines [37], our results suggest that these cytokines exert a consistent effect on ciliary function in both CRS-derived and non-CRS cultures. This points to a shared molecular mechanism by which IL-4 and IL-13 drive CRS pathogenesis through ciliary dysfunction and impaired mucociliary clearance. Such Th2-mediated cilia impairment underscores the broader consequences for sinonasal epithelial barrier function, particularly its reduced capacity to clear pathogens and debris from the upper airway [38]. By linking chronic Th2-driven inflammation to a specific molecular pathway, these findings provide insight into the mechanisms underlying epithelial dysfunction in Th2-driven CRS and highlight potential therapeutic targets for restoring mucociliary integrity.

Cytokine release by epithelial cells plays a pivotal role in modulating immune responses at mucosal surfaces, influencing inflammation, barrier integrity, and pathogen clearance [39]. Chronic exposure to Th2 cytokines significantly decreased production of IL-6 and MCP-1 cytokines in the basal media, reflecting an anti-inflammatory profile. The observed suppression of IL-6 production aligns with the well-established role of Th2 cytokines in activating the JAK-STAT6 signaling pathway [7]. This pathway is known to counteract pro-inflammatory signaling cascades such as NF-κB through the phosphorylation of STAT6, which suppresses the production of inflammatory cytokines like IL-6 [40]. Interestingly, IL-13 uniquely induced the production of IL-10 compared to IL-4 or combined IL-4/13 exposure, suggesting that IL-13 may exert distinct immunomodulatory effects not shared by IL-4. IL-10, a key regulatory cytokine associated with dampening immune responses and maintaining epithelial homeostasis [41], may reflect differential receptor activity or feedback mechanisms. While receptor expression was not directly measured in this study, prior reports indicate that simultaneous stimulation with IL-4 and IL-13 may activate negative feedback loops, such as those mediated by suppressor of cytokine signaling (SOCS) proteins, limiting maximal IL-10 production compared to IL-13 alone [42]. Comparisons of cytokine release profiles between sinonasal epithelial cells derived from healthy and CRS subjects showed no significant differences, suggesting that the anti-inflammatory effects of Th2 cytokine exposure are consistent, independent of CRS pathology.

While disease status did not affect cytokine production in epithelial cultures, male-derived control (Th2 unexposed) cultures exhibited significantly, higher concentrations of IL-1β, MCP-1, and IL-1RA compared to female-derived cultures suggesting intrinsic sex-based differences in the inflammatory response of sinonasal epithelial cells. Consistent with these cytokine differences, transcriptomic analyses revealed upregulation of several canonical inflammatory cytokine genes (TNF, IL6, IL1B) and downregulation of suppressor of cytokine signaling (SOCS) transcripts in males, suggesting a more pro-inflammatory baseline in male-derived cultures compared to females. Male-derived cultures also demonstrated significant downregulation of retinoic acid receptor (RAR) activation pathway. Activated by retinoic acid (RA), RAR plays a role in immune regulation by promoting regulatory T cell differentiation, inhibiting NF-κB-mediated pro-inflammatory cytokine production (e.g., IL-6, TNF-α, IL-1β), and maintaining epithelial barrier integrity, thereby suppressing excessive immune activation and inflammation [43]. Importantly, estrogen stimulates several components of the RAR pathway [44–49], suggesting a potential mechanism linking sex-specific differences to immune regulation. While these findings are particularly relevant given that females have 6-fold greater odds of developing nasal polyposis in low circulating estradiol conditions [50], transcriptomic analyses did not reveal differential expression of steroid receptor subtypes. However, differences in receptor protein expression or sensitivity cannot be ruled out as potential contributors of the observed sex-specific cytokine release differences. Moreover, sex-specific observations in control cultures emphasize the importance of considering sex as a biological variable in immunological studies.

The impact of chronic Th2-driven epithelial remodeling on the cellular response to rhinovirus is inconsistent. Previously, chronic IL-13-induced mucous metaplasia was reported to increase the susceptibility of the airway epithelium to rhinovirus [10], while another study demonstrated lower rhinovirus replication after chronic IL-13 exposure, suggesting antiviral effects [9]. In our study, cultures exposed to Th2 cytokines unexpectedly exhibited markedly reduced RV-A16 transcripts post-infection compared to unexposed cultures, further supporting the antiviral effect of Th2 cytokines. This finding was somewhat unexpected, as cytokine-induced downregulation of multiple tight junction and junction transcripts might have been anticipated to increase viral transcription by compromising the epithelial barrier. Furthermore, gene expression of rhinoviral receptor ICAM-1 was unchanged, suggesting that this observation was independent of receptor status, in contrast to previous studies [15, 51, 52]. Furthermore, CRS-derived cultures exhibited reduced rhinovirus transcript levels compared to those from healthy controls, suggesting that patient populations may differ in their susceptibility to rhinovirus infection. Taken together, these findings highlight the complexity of epithelial cells responses to rhinovirus, influenced by epithelial maturity, the interplay of pro- and anti-inflammatory cytokines, structural changes caused in the epithelium caused by noxious agents or metaplasia, and disease cohorts. While reduced viral replication in vitro may appear to be a beneficial aspect of chronic Th2 exposure, the concurrent structural and functional impairment of the epithelium likely predisposes patients to prolonged inflammation and secondary infections, ultimately compromising the resilience of the airway barrier to future insults. Notably, this outcome is not consistently observed across studies and may depend on baseline epithelial differences among patient populations.

Several factors should be considered when interpreting these findings. The sample size (3 healthy and 6 CRS donors) limits the generalizability of the results. Furthermore, participants included numerous subtypes of CRS, rather than focusing on CRS with nasal polyps (CRSwNP), a subtype strongly associated with Th2-mediated inflammation [1]. Although CRSwNP is traditionally associated with Th2-mediated inflammation, recent studies indicate variability in immune profiles among populations [53], suggesting the need for tailored approaches to understanding CRS pathogenesis. Future studies would benefit from refined characterization of CRS subtypes, specifically focusing on individuals with eosinophilic CRSwNP, to better explore how Th2-driven inflammation modulates viral susceptibility.

Patient-specific factors, such as a genetic predisposition toward a particular inflammatory milieu, may also influence epithelial immune responses, and contribute to the sex-specific differences observed in this study. Given the limited sample size, the sex-related transcriptomic and cytokine patterns observed in this study should be considered hypothesis-generating and warrant validation in larger, independent cohorts. Although the RUV method was applied to account for unwanted variation, including age, race, and smoking status, the high prevalence of smokers in the CRS cohort (50%) may have influenced epithelial responses. Smoking is known to impair epithelial barrier function, reduce mucociliary clearance, and alter innate immune signaling, which could have affected both baseline gene expression and responses to Th2 cytokines or viral infection [54]. Consequently, some of the observed variability in epithelial antiviral responses may reflect the combined effects of disease status and smoking, highlighting the importance of accounting for smoking and other exposures in future studies.

While ALI cultures more accurately recapitulate epithelial architecture than monolayer cultures, they lack immune cells, which may amplify or modulate epithelial immune responses, limiting the model’s ability to fully capture the interplay between a Th2 environment and host antiviral responses. In addition, functional measures of epithelial barrier integrity such as transepithelial electrical resistance or paracellular permeability were not assessed. While loss of cilia on histology was observed in this study after Th2 cytokine exposure, additional functional studies such as ciliary beat frequency and mucociliary clearance rate were not assessed, which precludes direct assessment of how these molecular and histologic alterations translate into epithelial functional impairment.

Furthermore, the study utilized a single rhinoviral subtype (RV-A16) for infection modeling, limiting the ability to generalize findings to other viral families such as Orthomyxoviridae (e.g., influenza) or Coronaviridae (e.g., coronaviruses). While the presence of rhinoviral RNA detected by bulk RNA-sequencing and RV-A16–specific RT-qPCR reflects viral exposure, productive infection was not definitively confirmed. Unbound virus was removed by PBS washes and samples were collected at 36 h post-infection, but additional approaches such as viral titers or negative-strand RNA detection would strengthen these findings. Future studies should also investigate viral content in CRS patients before and after therapies targeting the Th2 pathway, such as dupilumab, could reveal how in vivo modulation of Th2 inflammation influences viral dynamics.

Conclusions

This study demonstrates that chronic Th2 cytokine exposure drives key molecular and structural changes in sinonasal epithelial cells, impacting mucociliary dynamics and rhinoviral susceptibility, regardless of CRS pathology. These findings underscore the complex interplay between Th2 cytokine exposures and rhinoviral infections and highlight the need for a deeper understanding of how chronic inflammation influences epithelial integrity. Such insights could inform therapeutic strategies to restore mucociliary function and mitigate the broader systemic effects of chronic inflammatory diseases.

Supplementary Information

Supplementary Material 1. (1.4MB, xlsx)
Supplementary Material 2. (2.1MB, pdf)
Supplementary Material 3. (2.6MB, mp4)

Acknowledgements

We would like to thank Dr. Stephanie Joe for her help with patient recruitment in this study and the staff at the Roy J. Carver Biotechnology Center for their technical support.

Abbreviations

ALI

Air-Liquid Interface

ANOVA

Analysis of Variance

BBS

Bardet-Biedl Syndrome

CRS

Chronic Rhinosinusitis

ELISA

Enzyme-Linked Immunosorbent Assay

FDA

Food and Drug Administration

FDR

False Discovery Rate

GATA

3-GATA Binding Protein 3

ICAM

1-Intercellular Adhesion Molecule 1

IFN

Interferon

IL

Interleukin

IPA

Ingenuity Pathway Analysis

IRB

Institutional Review Board

JAK

STAT-Janus Kinase-Signal Transducer and Activator of Transcription

NCBI

National Center for Biotechnology Information

NF

κB-Nuclear Factor Kappa-light-chain-enhancer of activated B cells

PCA

Principal Component Analysis

SOCS

Suppressor of Cytokine Signaling

TCID50

Tissue Culture Infectious Dose 50%

TNF

Tumor Necrosis Factor

Authors' contributions

Juliana Theorell: Investigation, Methodology, Software, Formal analysis, Visualization, Writing – Original Draft, Writing – Review; Jenny Drnevich: Software, Formal analysis, Writing – Review; Sofija Jovanovic Gasovic: Investigation, Methodology; Bridget Lawate: Investigation; Victoria S. Lee: Resources, Writing – Review; Almudena Veiga-Lopez: Conceptualization, Visualization, Writing – Original Draft, Supervision, Project administration, Funding acquisition.

Funding

Research reported in this publication was supported by the Department of Pathology at the University of Illinois Chicago, the National Institutes of Health through a NIEHS P30ES027792 pilot project and K12AR084225 (supporting V.S.L).

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Declarations

Ethics approval and consent to participate

Approval for this study was granted by the University of Illinois Institutional Review Board (STUDY2021-0676).

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  • 1.Wise SK, Damask C, Roland LT, Ebert C, Levy JM, Lin S, et al. International consensus statement on allergy and rhinology: Allergic rhinitis – 2023. Int Forum Allergy Rhinol. 2023;13(4):293–859. [DOI] [PubMed] [Google Scholar]
  • 2.Yan B, Lan F, Li J, Wang C, Zhang L. The mucosal concept in chronic rhinosinusitis: Focus on the epithelial barrier. J Allergy Clin Immunol. 2024;153(5):1206–14. [DOI] [PubMed] [Google Scholar]
  • 3.Wynne M, Atkinson C, Schlosser RJ, Mulligan JK. Contribution of Epithelial Cell Dysfunction to the Pathogenesis of Chronic Rhinosinusitis with Nasal Polyps. Am J Rhinol Allergy. 2019;33(6):782–90. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Hewitt RJ, Lloyd CM. Regulation of immune responses by the airway epithelial cell landscape. Nat Rev Immunol. 2021;21(6):347–62. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Ha J-G, Cho H-J. Unraveling the Role of Epithelial Cells in the Development of Chronic Rhinosinusitis. IJMS. 2023;24(18):14229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Zhu Z, Zhao C, Wang M. IL-4/IL‐13 pathway in nasal type 2 inflammation: The central role and targeted therapy. Eye ENT Res. 2024;1(1):39–48. [Google Scholar]
  • 7.Shankar A, McAlees JW, Lewkowich IP. Modulation of IL-4/IL-13 cytokine signaling in the context of allergic disease. J Allergy Clin Immunol. 2022;150(2):266–76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Kotas ME, Patel NN, Cope EK, Gurrola JG, Goldberg AN, Pletcher SD, et al. IL-13–associated epithelial remodeling correlates with clinical severity in nasal polyposis. J Allergy Clin Immunol. 2023;151(5):1277–85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Huang Z-Q, Liu J, Ong HH, Yuan T, Zhou X-M, Wang J, et al. Interleukin-13 Alters Tight Junction Proteins Expression Thereby Compromising Barrier Function and Dampens Rhinovirus Induced Immune Responses in Nasal Epithelium. Front Cell Dev Biol. 2020;8:572749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Lachowicz-Scroggins ME, Boushey HA, Finkbeiner WE, Widdicombe JH. Interleukin-13–Induced Mucous Metaplasia Increases Susceptibility of Human Airway Epithelium to Rhinovirus Infection. Am J Respir Cell Mol Biol. 2010;43(6):652–61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Wise SK, Laury AM, Katz EH, Den Beste KA, Parkos CA, Nusrat A. Interleukin-4 and interleukin‐13 compromise the sinonasal epithelial barrier and perturb intercellular junction protein expression. Int Forum Allergy Rhinol. 2014;4(5):361–70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Zissler UM, Chaker AM, Effner R, Ulrich M, Guerth F, Piontek G, et al. Interleukin-4 and interferon-γ orchestrate an epithelial polarization in the airways. Mucosal Immunol. 2016;9(4):917–26. [DOI] [PubMed] [Google Scholar]
  • 13.Zuyderduyn S, Ninaber DK, Schrumpf JA, Van Sterkenburg MA, Verhoosel RM, Prins FA, et al. IL-4 and IL-13 exposure during mucociliary differentiation of bronchial epithelial cells increases antimicrobial activity and expression of antimicrobial peptides. Respir Res. 2011;12(1):59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Lee HS, Volpe SJ, Chang EH. The Role of Viruses in the Inception of Chronic Rhinosinusitis. Clin Exp Otorhinolaryngol. 2022;15(4):310–8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Contoli M, Ito K, Padovani A, Poletti D, Marku B, Edwards MR, et al. Th2 cytokines impair innate immune responses to rhinovirus in respiratory epithelial cells. Allergy. 2015;70(8):910–20. [DOI] [PubMed] [Google Scholar]
  • 16.Jakiela B, Gielicz A, Plutecka H, Hubalewska-Mazgaj M, Mastalerz L, Bochenek G, et al. Th2-Type Cytokine–Induced Mucus Metaplasia Decreases Susceptibility of Human Bronchial Epithelium to Rhinovirus Infection. Am J Respir Cell Mol Biol. 2014;51(2):229–41. [DOI] [PubMed] [Google Scholar]
  • 17.Ordovas-Montanes J, Dwyer DF, Nyquist SK, Buchheit KM, Vukovic M, Deb C, et al. Allergic inflammatory memory in human respiratory epithelial progenitor cells. Nature. 2018;560(7720):649–54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Geng B, Bachert C, Busse WW, Gevaert P, Lee SE, Niederman MS, et al. Respiratory Infections and Anti-Infective Medication Use From Phase 3 Dupilumab Respiratory Studies. J Allergy Clin Immunology: Pract. 2022;10(3):732–41. [DOI] [PubMed] [Google Scholar]
  • 19.Müller L, Brighton LE, Carson JL, Fischer WA II, Jaspers I. Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface. JoVE. 2013;80:50646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Patro R, Duggal G, Love MI, Irizarry RA, Kingsford C. Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods. 2017;14(4):417–9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Risso D, Ngai J, Speed TP, Dudoit S. Normalization of RNA-seq data using factor analysis of control genes or samples. Nat Biotechnol. 2014;32(9):896–902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Peixoto L, Risso D, Poplawski SG, Wimmer ME, Speed TP, Wood MA, et al. How data analysis affects power, reproducibility and biological insight of RNA-seq studies in complex datasets. Nucleic Acids Res. 2015;43(16):7664–74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Wingfield Jenna L, Lechtreck K-F, Lorentzen E. Trafficking of ciliary membrane proteins by the intraflagellar transport/BBSome machinery. Essays Biochem. 2018;62(6):753–63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Nakayama K, Katoh Y. Ciliary protein trafficking mediated by IFT and BBSome complexes with the aid of kinesin-2 and dynein-2 motors. J Biochem. 2018;163(3):155–64. [DOI] [PubMed] [Google Scholar]
  • 25.Avasthi P, Marshall WF. Stages of ciliogenesis and regulation of ciliary length. Differentiation. 2012;83(2):S30–42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Gudis D, Zhao K-q, Cohen NA. Acquired cilia dysfunction in chronic rhinosinusitis. Am J Rhinol Allergy. 2012;26(1):1–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Laoukili J, Perret E, Willems T, Minty A, Parthoens E, Houcine O, et al. IL-13 alters mucociliary differentiation and ciliary beating of human respiratory epithelial cells. J Clin Invest. 2001;108(12):1817–24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Turner J, Roger J, Fitau J, Combe D, Giddings J, Heeke GV, et al. Goblet Cells Are Derived from a < i>FOXJ1 -Expressing Progenitor in a Human Airway Epithelium. Am J Respir Cell Mol Biol. 2011;44(3):276–84. [DOI] [PubMed] [Google Scholar]
  • 29.Ma J, Rubin BK, Voynow JA. Mucins, Mucus, and Goblet Cells. Chest. 2018;154(1):169–76. [DOI] [PubMed] [Google Scholar]
  • 30.Chen G, Korfhagen TR, Karp CL, Impey S, Xu Y, Randell SH, et al. Foxa3 induces goblet cell metaplasia and inhibits innate antiviral immunity. Am J Respir Crit Care Med. 2014;189(3):301–13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Chen G, Korfhagen TR, Xu Y, Kitzmiller J, Wert SE, Maeda Y, et al. SPDEF is required for mouse pulmonary goblet cell differentiation and regulates a network of genes associated with mucus production. J Clin Invest. 2009;119(10):2914–24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Seibold MA. Interleukin-13 Stimulation Reveals the Cellular and Functional Plasticity of the Airway Epithelium. Ann Am Thorac Soc. 2018;15(Suppl 2):S98–102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Hallstrand TS, Hackett TL, Altemeier WA, Matute-Bello G, Hansbro PM, Knight DA. Airway epithelial regulation of pulmonary immune homeostasis and inflammation. Clin Immunol. 2014;151(1):1–15. [DOI] [PubMed] [Google Scholar]
  • 34.Gour N, Wills-Karp M. IL-4 and IL-13 signaling in allergic airway disease. Cytokine. 2015;75(1):68–78. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Junttila IS. Tuning the Cytokine Responses: An Update on Interleukin (IL)-4 and IL-13 Receptor Complexes. Front Immunol. 2018;9:888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.White SR, Martin LD, Stern R, Laxman B, Marroquin BA. Expression of IL-4/IL-13 receptors in differentiating human airway epithelial cells. Am J Physiol Lung Cell Mol Physiol. 2010;299(5):L681–93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Palikhe NS, Kim S-H, Cho B-Y, Choi G-S, Kim J-H, Ye Y-M, et al. IL-13 Gene Polymorphisms are Associated With Rhinosinusitis and Eosinophilic Inflammation in Aspirin Intolerant Asthma. Allergy Asthma Immunol Res. 2010;2(2):134–40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Hariri BM, Cohen NA. New Insights into Upper Airway Innate Immunity. Am J Rhinol Allergy. 2016;30(5):319–23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Mettelman RC, Allen EK, Thomas PG. Mucosal immune responses to infection and vaccination in the respiratory tract. Immunity. 2022;55(5):749–80. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Liu T, Zhang L, Joo D, Sun S-C. NF-κB signaling in inflammation. Sig Transduct Target Ther. 2017;2(1):17023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Iyer SS, Cheng G. Role of interleukin 10 transcriptional regulation in inflammation and autoimmune disease. Crit Rev Immunol. 2012;32(1):23–63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Hebenstreit D, Luft P, Schmiedlechner A, Regl G, Frischauf A-M, Aberger F, et al. IL-4 and IL-13 Induce < i>SOCS-1 Gene Expression in A549 Cells by Three Functional STAT6-Binding Motifs Located Upstream of the Transcription Initiation Site. J Immunol. 2003;171(11):5901–7. [DOI] [PubMed] [Google Scholar]
  • 43.Ghyselinck NB, Duester G. Retinoic acid signaling pathways. Development. 2019;146(13):dev167502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Garcia E, Lacasa D, Giudicelli Y. Estradiol Stimulation of c-fos and c-jun Expressions and Activator Protein-1 Deoxyribonucleic Acid Binding Activity in Rat White Adipocyte*. Endocrinology. 2000;141(8):2837–46. [DOI] [PubMed] [Google Scholar]
  • 45.Ruh MF, Bi Y, D’Alonzo R, Bellone CJ. Effect of estrogens on IL-1β promoter activity. J Steroid Biochem Mol Biol. 1998;66(4):203–10. [DOI] [PubMed] [Google Scholar]
  • 46.Mahonen A, Jukkola A, Risteli L, Risteli J, Mäenpää PH. Type I procollagen synthesis is regulated by steroids and related hormones in human osteosarcoma cells. J Cell Biochem. 1998;68(2):151–63. [PubMed] [Google Scholar]
  • 47.Zhang Y, Goleva E, Leung DYM. Estrogen Inhibits Corticosteroid Induction of the Mitogen-Induced Kinase Phosphatase Gene. J Allergy Clin Immunol. 2008;121(3):792. [Google Scholar]
  • 48.Smith LC, Moreno S, Robertson L, Robinson S, Gant K, Bryant AJ, et al. Transforming growth factor beta1 targets estrogen receptor signaling in bronchial epithelial cells. Respir Res. 2018;19(1):160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 49.An J, Ribeiro RCJ, Webb P, Gustafsson J-Å, Kushner PJ, Baxter JD, et al. Estradiol repression of tumor necrosis factor-α transcription requires estrogen receptor activation function-2 and is enhanced by coactivators. Proc Natl Acad Sci USA. 1999;96(26):15161–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50.Espersen J, Weber U, Römer-Franz A, Lenarz T, Stolle SRO, Warnecke A. Level of sex hormones and their association with acetylsalicylic acid intolerance and nasal polyposis. PLoS ONE. 2020;15(12):e0243732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 51.Bianco A, Sethi S, Allen J, Knight R, Spiteri M. Th2 cytokines exert a dominant influence on epithelial cell expression of the major group human rhinovirus receptor, ICAM-1. Eur Respir J. 1998;12(3):619–26. [DOI] [PubMed] [Google Scholar]
  • 52.Papi A, Epithelial. ICAM-1 regulation and its role in allergy. Clin Experimental Allergy. 1997;27(7):721–4. [DOI] [PubMed] [Google Scholar]
  • 53.Lin Y-T, Tsai M-H, Su Y-Y, Huang S-C. Comparison of cytokine expression and disease severity between plasma cell-dominant and eosinophil-dominant patients in chronic rhinosinusitis with nasal polyps. Allergy Asthma Clin Immunol. 2024;20(1):34. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Nwozor KO, Hackett TL, Chen Q, Yang CX, Aguilar Lozano SP, Zheng X, et al. Effect of age, COPD severity, and cigarette smoke exposure on bronchial epithelial barrier function. Am J Physiol Lung Cell Mol Physiol. 2025;328(5):L724–37. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Material 1. (1.4MB, xlsx)
Supplementary Material 2. (2.1MB, pdf)
Supplementary Material 3. (2.6MB, mp4)

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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