BackgroundAcute infusion reactions to oxaliplatin, a chemotherapeutic used to treat gastrointestinal cancers, are observed in about 20% of patients. Rapid drug desensitization (RDD) protocols often allow the continuation of oxaliplatin in patients with no alternative options. Break-through symptoms including anaphylaxis can still occur during RDDs.ObjectiveTo evaluate if pretreatment with acalabrutinib, a Bruton’s tyrosine kinase inhibitor, can prevent anaphylaxis during a RDD in a patient sensitized to oxaliplatin.MethodsA 52-year-old male with locally advanced gastric carcinoma developed anaphylaxis during his 5th cycle of oxaliplatin. As he required 6 additional cycles to complete his curative-intent treatment regimen, he underwent RDD to oxaliplatin but still developed severe acute reactions. The risks and benefits of adding acalabrutinib prior to and during a RDD, were reviewed and he elected to proceed.ResultsWith acalabrutinib taken prior to and during the RDD, the patient was able to tolerate oxaliplatin RDDs without complication. Consistent with its mechanism of action, acalabrutinib completely blocked the patient’s positive skin prick response to oxaliplatin. Acalabrutinib did not alter the percentage of circulating basophils (1.24% versus 0.98%) prior to the RDD but protected from basopenia (0.74% versus 0.09%) after the RDD. Acalabrutinib was associated with a drastic reduction in the ability of basophils to upregulate CD63 in vitro following incubation with oxaliplatin (0.11% versus 2.38%) or polyclonal anti-human IgE antibody (0.08% versus 44.2%).ConclusionsFive doses of acalabrutinib 100mg orally twice daily starting the evening 2 days before and continuing through a RDD allowed a sensitized patient to successfully and safely receive oxaliplatin.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by type 2 (T2) inflammation. Recent studies, including our own, suggest that neutrophils are also elevated in eosinophilic T2 nasal polyps (NPs) and that elevated neutrophils display an activated phenotype. However, the actual roles of neutrophils in NP pathogenesis in T2 CRSwNP are still largely unclear. In this study, we aimed to reveal the characteristics of NP neutrophils by single cell RNA-Sequencing (scRNA-Seq). We obtained NP tissues, control sinus (CS) tissues, and matched peripheral blood (PB) from patients with CRSwNP (n=3) and control patients (n=3) and isolated neutrophils. The gene expression profiles were determined by scRNA-Seq, and we performed clustering with UMAP analysis to separate contaminating cells and identify neutrophils. We then examined the differential expression of genes in NP neutrophils by the Benjamini-Hochberg algorithm. We found that 93 and 39 genes were >2-fold significantly elevated in NP neutrophils, compared to PB and CS neutrophils, respectively (p<0.05). Inflammatory response-associated genes including IL1B, CXCL8, C5AR1, and IL1RN were significantly elevated in NP neutrophils. Neutrophil activation-associated genes including PLAUR, CD44, TNFAIP6, FTH1, and LPCAT1 were also elevated in NP neutrophils. Furthermore, ALOX5AP, which encodes an enzyme of the leukotriene synthesis pathway, was elevated in NP neutrophils. Based on these observations, NP neutrophils might affect other NP cells by producing cytokines, chemokines, and leukotrienes. We believe this is the first research to analyze NP neutrophils by scRNA-Seq in CRS. Highly activated neutrophils may play important pathogenic roles in CRSwNP.

Background: Oncostatin M (OSM) may promote type 2 inflammation in chronic rhinosinusitis with nasal polyps (CRSwNP) by inducing thymic stromal lymphopoietin (TSLP). Objective: We sought to study the impact of OSM on TSLP synthesis and release from nasal epithelial cells (NECs). Methods: OSM receptors, IL-4 receptors (IL-4R), and TSLP were evaluated in mucosal tissue and primary NECs from patients with CRSwNP by quantitative PCR and immunofluorescence. Air-liquid interface-cultured NECs were stimulated with cytokines, including OSM, and quantitative PCR, ELISA, Western blot, and flow cytometry were used to assess the expression of OSM receptors, IL-4R, and TSLP. Results: Increased levels of OSM receptor 13 chain (OSMR13), IL-4Ra, and TSLP were observed in nasal polyp tissues and primary epithelial cells from nasal polyps of patients with CRSwNP compared with control tissues or cells from control subjects. The level of expression of OSMR13 in tissue was correlated with levels of both IL-4Ra and TSLP. OSM stimulation of NECs increased the expression of OSMR13 and IL-4Ra. Stimulation with IL-4 plus OSM augmented the production of TSLP; the response was suppressed by a signal transducer and activator of transcription 6 inhibitor. Stimulation of NECs with IL-4 plus OSM increased the expression of proprotein convertase subtilisin/kexin 3, an enzyme that truncates and activates TSLP. Conclusions: OSM increases the expression of IL-4Ra and synergizes with IL-4 to induce the synthesis and release of TSLP in NECs. Because the combination of IL-4 and OSM also augmented the expression of proprotein convertase subtilisin/ kexin 3, these results suggest that OSM can induce both synthesis and posttranslational processing/activation of TSLP, promoting type 2 inflammation. (J Allergy Clin Immunol 2023;151:1379-90.)

In the US, chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by elevated levels of IL-4 and IL-13. Several single nucleotide polymorphisms within the gene encoding for the shared IL-4 receptor alpha chain have been identified, including Q576R (c.1727A>G). The frequency of the RR genotype was significantly higher among asthmatics and the variant was associated with more severe lower airway disease. The impact of the RR genotype in CRSwNP is unclear. Genotyping was performed to determine the frequency of the Q576R polymorphism in 226 patients with CRSwNP and 74 with Aspirin Exacerbated Respiratory Disease (AERD). Gene expression levels of various type 2 inflammatory mediators in NP were evaluated by RT-PCR from 14 patients with CRSwNP and 29 with AERD. There was a significantly higher frequency of the RR genotype in patients with AERD compared to those with CRSwNP (18% vs 6%; p=0.007) and compared to the subset of CRSwNP patients with asthma (18% vs 5%; p=0.01). Compared to AERD patients with the QQ genotype, those with at least one R allele (QR/RR) had significantly higher mean tissue expression levels (% expression of GUSB) of CCL26 (74 vs 29; p=0.03), MUC5AC (3,274 vs 1,801; p=0.04), and POSTN (1,505 vs 562; p=0.04). Similar differences were not observed in patients with CRSwNP based on genotype. The frequency of the RR genotype is elevated in AERD patients and is associated with enhanced expression of type 2 inflammatory mediators. Similar findings were not observed in CRSwNP suggesting a fundamental pathophysiological difference between AERD and CRSwNP.

BACKGROUND:Viruses may drive immune mechanisms responsible for chronic rhinosinusitis with nasal polyposis (CRSwNP), but little is known about the underlying molecular mechanisms. OBJECTIVES:To identify epigenetic and transcriptional responses to a common upper respiratory pathogen, rhinovirus (RV), that are specific to patients with CRSwNP using a primary sinonasal epithelial cell culture model. METHODS:Airway epithelial cells were collected at surgery from patients with CRSwNP (cases) and from controls without sinus disease, cultured, and then exposed to RV or vehicle for 48 h. Differential gene expression and DNA methylation (DNAm) between cases and controls in response to RV were determined using linear mixed models. Weighted gene co-expression analysis (WGCNA) was used to identify (a) co-regulated gene expression and DNAm signatures, and (b) genes, pathways, and regulatory mechanisms specific to CRSwNP. RESULTS:We identified 5585 differential transcriptional and 261 DNAm responses (FDR <0.10) to RV between CRSwNP cases and controls. These differential responses formed three co-expression/co-methylation modules that were related to CRSwNP and three that were related to RV (Bonferroni corrected p < .01). Most (95%) of the differentially methylated CpGs (DMCs) were in modules related to CRSwNP, whereas the differentially expressed genes (DEGs) were more equally distributed between the CRSwNP- and RV-related modules. Genes in the CRSwNP-related modules were enriched in known CRS and/or viral response immune pathways. CONCLUSION:RV activates specific epigenetic programs and correlated transcriptional networks in the sinonasal epithelium of individuals with CRSwNP. These novel observations suggest epigenetic signatures specific to patients with CRSwNP modulate response to viral pathogens at the mucosal environmental interface. Determining how viral response pathways are involved in epithelial inflammation in CRSwNP could lead to therapeutic targets for this burdensome airway disorder.

Microparticles (MPs) are extracellular vesicles that are released by shedding from plasma membrane of activated cells and new biomarkers to evaluate status of cell activation. MP profilings of activated eosinophils, mast cells and basophils in nasal mucosa in chronic rhinosinusitis with nasal polyps (CRSwNP) remain unclear.

BACKGROUND:Increased activation of the coagulation cascade and diminished fibrinolysis combine to promote fibrin deposition and polyp formation in chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP). More information is needed concerning mechanisms of coagulation in CRSwNP.OBJECTIVE:We investigated the mechanisms as well as the initiation and regulation of coagulation cascade activation in CRS.METHODS:Samples were collected from 135 subjects with CRSwNP, 80 subjects with chronic CRS without nasal polyps (NP), and 65 control subjects. The levels of activated factor X (FXa), prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex, tissue factor (TF), and TF pathway inhibitor (TFPI) were monitored in CRS by real-time PCR, ELISA, immunohistochemistry, or immunofluorescence. Heteromeric complexes of TF with activated factor VII (FVII) and TF with activated FVII and FXa were assessed by coimmunoprecipitation and Western blotting.RESULTS:Increased levels of FXa, F1+2, and thrombin-antithrombin complex were detected in NP tissue compared to uncinate tissue from CRS and control subjects. Although free TF protein levels were not increased in NP, immunoprecipitation of TF in NP tissue revealed increased complexes of TF with FVII. Local expression of FVII was detected in sinonasal mucosa, and the ratio of TFPI to FXa was lower in NP tissue.CONCLUSION:The coagulation cascade is associated with NP compared to control and uncinate tissue from CRS patients, and TF and FVII are produced locally in sinonasal mucosa in patients. TF and FVII can activate the extrinsic coagulation pathway, suggesting that this pathway may activate fibrin deposition in CRSwNP. Reduced formation of the complex of FXa and TFPI in NP may reduce natural suppression of the extrinsic coagulation pathway in CRSwNP.

Background: Patients with aspirin-exacerbated respiratory disease (AERD) regularly exhibit severe nasal polyposis. Studies suggest that chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by excessive fibrin deposition associated with a profound decrease in epithelial tissue plasminogen activator (tPA). Retinoids, including vitamin A and its active metabolite retinoic acid (RA), are necessary for maintaining epithelial function and well-known inducers of tPA in endothelial cells. Objectives: This study sought to determine whether endogenous retinoids are involved in NP pathophysiology and disease severity in patients with CRSwNP and AERD. Methods: NP tissue was collected from patients with AERD or CRSwNP, and concentrations of retinoids and fibrinolysis markers were measured using ELISA. Normal human bronchial epithelial cells were stimulated alone or in combination with RA and IL-13 for 24 hours. Results: This study observed lower retinoid levels in nasal polyps of patients with AERD than those with CRSwNP or healthy controls (P < .01). Levels of the fibrin-breakdown product d-dimer were the lowest in AERD polyps (P < .01), which is consistent with lower tPA expression (P < .01). In vitro, all-trans RA upregulated tPA levels in normal human bronchial epithelial cells by 15-fold and reversed the IL-13-induced attenuation of tPA expression in cultured cells (P < .01). Conclusions: RA, a potent inducer of epithelial tPA in vitro, is reduced in tissue from patients with AERD, a finding that may potentially contribute to decreased levels of tPA and fibrinolysis in AERD. RA can induce tPA in epithelial cells and can reverse IL-13-induced tPA suppression in vitro, suggesting the potential utility of RA in treating patients with CRSwNP and/or AERD.

BackgroundAspirin-exacerbated respiratory disease (AERD) is characterized by asthma, chronic rhinosinusitis with nasal polyps (CRSwNP), and an intolerance of medications that inhibit cyclooxygenase-1. Patients with AERD have more severe upper and lower respiratory tract disease than do aspirin-tolerant patients with CRSwNP. A dysregulation in arachidonic acid metabolism is thought to contribute to the enhanced sinonasal inflammation in AERD.ObjectiveOur aim was to utilize an unbiased approach investigating arachidonic acid metabolic pathways in AERD.MethodsSingle-cell RNA sequencing (10× Genomics, Pleasanton, Calif) was utilized to compare the transcriptional profile of nasal polyp (NP) cells from patients with AERD and patients with CRSwNP and map differences in the expression of select genes among identified cell types. Findings were confirmed by traditional real-time PCR. Lipid mediators in sinonasal tissue were measured by mass spectrometry. Localization of various proteins within NPs was assessed by immunofluorescence.ResultsThe gene encoding for 15-lipooxygenase (15-LO), ALOX15, was significantly elevated in NPs of patients with AERD compared to NPs of patients with CRSwNP (P < .05) or controls (P < .001). ALOX15 was predominantly expressed by epithelial cells. Expression levels significantly correlated with radiographic sinus disease severity (r = 0.56; P < .001) and were associated with asthma. The level of 15-oxo-eicosatetraenoic acid (15-Oxo-ETE), a downstream product of 15-LO, was significantly elevated in NPs from patients with CRSwNP (27.93 pg/mg of tissue) and NPs from patients with AERD (61.03 pg/mg of tissue) compared to inferior turbinate tissue from controls (7.17 pg/mg of tissue [P < .001]). Hydroxyprostaglandin dehydrogenase, an enzyme required for 15-Oxo-ETE synthesis, was predominantly expressed in mast cells and localized near 15-LO+ epithelium in NPs from patients with AERD.ConclusionsEpithelial and mast cell interactions, leading to the synthesis of 15-Oxo-ETE, may contribute to the dysregulation of arachidonic acid metabolism via the 15-LO pathway and to the enhanced sinonasal disease severity observed in AERD.

Additional file 10: Tables showing enrichment and colocalization results from this study. Table S1: Interaction model results for genotype x atopy and genotype x steroid use for eQTLs and meQTLs. Table S2: Enrichment estimates of eQTLs for TAGC asthma GWAS SNPs from six tissues. No P-values were significant after FDR correction. Table S3: Enrichment estimates of eQTLs for adult onset asthma GWAS SNPs from six tissues. No P-values were significant after FDR correction. Table S4: moloc results for molecular QTL-GWAS pairs and triplets. Table S5: Asthma GWAS risk allele effects on gene expression and DNA methylation.

Microparticles (MP) are extracellular vesicles that are released by shedding from the membrane of activated or injured cells. We previously reported that eosinophil MP (EosMP), mast cell MP (MCMP) and basophil MP (BasoMP) were increased in nasal lavage fluids (NLF) collected from chronic rhinosunisitis with polyps (CRSwNP). NLF were collected from 154 CRSwNP cases. MP were measured using flow cytometry. ECP levels were measured by ELISA. Hierarchical cluster analysis of CRSwNP cases based on levels of EosMP (EMR1[+]MP), MCMP (FcεRI[+]c-kit[+]MP) and BasoMP (CD203c[+]c-kit[-]MP) resulted in emergence of 4 main clusters; Cluster[1] (null, n=36): EosMPs(1.0-fold, vs. controls)/MCMPs(0.6-fold)/BasoMPs(1.0-fold), Cluster[2] (BasoMPhigh, n=49): EosMP(1.2-fold)/MCMP(0.4-fold)/BasoMP(3.0-fold), Cluster[3] (EosMPhigh/MCMPhigh, n=44): EosMP(1.7-fold)/MCMP(1.8-fold)/BasoMPs(1.2-fold), Cluster[4] (EosMPhigh/MCMPhigh/BasoMPhigh, n=25): EosMPs(1.9-fold)/MCMPs(2.8-fold)/BasoMP(3.4-fold). Levels of ECP (778±3009 ng/μL vs. 217±1061, p

Background Genome-wide association studies (GWASs) have identified thousands of variants associated with asthma and other complex diseases. However, the functional effects of most of these variants are unknown. Moreover, GWASs do not provide context-specific information on cell types or environmental factors that affect specific disease risks and outcomes. To address these limitations, we used an upper airway epithelial cell (AEC) culture model to assess transcriptional and epigenetic responses to rhinovirus (RV), an asthma-promoting pathogen, and provide context-specific functional annotations to variants discovered in GWASs of asthma. Methods Genome-wide genetic, gene expression, and DNA methylation data in vehicle- and RV-treated upper AECs were collected from 104 individuals who had a diagnosis of airway disease (n=66) or were healthy participants (n=38). We mapped cis expression and methylation quantitative trait loci (cis-eQTLs and cis-meQTLs, respectively) in each treatment condition (RV and vehicle) in AECs from these individuals. A Bayesian test for colocalization between AEC molecular QTLs and adult onset asthma and childhood onset asthma GWAS SNPs, and a multi-ethnic GWAS of asthma, was used to assign the function to variants associated with asthma. We used Mendelian randomization to demonstrate DNA methylation effects on gene expression at asthma colocalized loci. Results Asthma and allergic disease-associated GWAS SNPs were specifically enriched among molecular QTLs in AECs, but not in GWASs from non-immune diseases, and in AEC eQTLs, but not among eQTLs from other tissues. Colocalization analyses of AEC QTLs with asthma GWAS variants revealed potential molecular mechanisms of asthma, including QTLs at the TSLP locus that were common to both the RV and vehicle treatments and to both childhood onset and adult onset asthma, as well as QTLs at the 17q12-21 asthma locus that were specific to RV exposure and childhood onset asthma, consistent with clinical and epidemiological studies of these loci. Conclusions This study provides evidence of functional effects for asthma risk variants in AECs and insight into RV-mediated transcriptional and epigenetic response mechanisms that modulate genetic effects in the airway and risk for asthma.

Background: Aspirin-exacerbated respiratory disease (AERD) is characterized by the triad of chronic rhinosinusitis with nasal polyps (CRSwNP), asthma, and intolerance to cyclooxygenase-1 enzyme inhibitors. The underlying mechanisms contributing to AERD pathogenesis are not fully understood, but AERD is characterized by an enhanced type 2 inflammatory phenotype. Basophils are potent type 2 effector cells, but their involvement in AERD pathophysiology remains unclear. Objective: We sought to characterize the systemic and local basophil responses in patients with AERD compared with patients with CRSwNP. Methods: Sinonasal tissues including inferior turbinate and/or nasal polyps (NPs) and peripheral blood were collected from controls, patients with AERD, and patients with CRSwNP. Expression of cell surface (CD45, Fc epsilon RI, CD203c), activation (CD63), and intracellular (2D7) markers associated with basophils was characterized using flow cytometry. Clinical data including Lund-Mackay scores and pulmonary function were obtained. Results: The mean number of basophils (CD45+CD203c+Fc epsilon RI+CD117-) detected in AERD NPs (147 +/- 28 cells/mg tissue) was significantly elevated compared with that detected in CRSwNP NPs (69 +/- 20 cells/mg tissue; P = .01). The number of circulating basophils was significantly elevated in patients with AERD (P = .04). Basophils in NPs had significantly higher CD203c and CD63 mean fluorescence intensity compared with blood in both conditions (P < .01). Basophils from AERD NPs had lower expression of the granule content marker 2D7 compared with those from matched blood (P <.01) or NPs of patients with CRSwNP (P = .06), suggesting ongoing degranulation. Basophil 2D7 mean fluorescence intensity significantly correlated with pulmonary function (r = 0.62; P = .02) and inversely correlated with sinonasal inflammation (r = -0.56; P = .004). Conclusions: Increased basophil numbers and extent of ongoing degranulation in NPs of patients with AERD compared with patients with CRSwNP may contribute to the exaggerated disease pathogenesis and severity unique to AERD.

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Aspirin Exacerbated Respiratory Disease (AERD) is defined by the triad of asthma, chronic rhinosinusitis with nasal polyps (CRSwNP), and an intolerance to cyclooxygenase-1 inhibitors. AERD patients tend to have more severe respiratory disease than aspirin-tolerant patients with CRSwNP. The underlying mechanisms contributing to enhanced disease in AERD are not understood, but a dysregulation in arachidonic acid metabolism is important. We investigated the 15-lipoxygenase (15-LO) pathway in AERD pathogenesis. We examined expression levels of genes important for the synthesis and function of arachidonic acid metabolites in AERD and CRSwNP NP by single-cell RNA sequencing using the 10X Genomics platform and by traditional RT-PCR. Lipid mediators were measured in sinonasal tissue by mass spectrometry. Immunofluorescence was used to examine localization of specific enzymes within NP. ALOX15 expression was significantly elevated in AERD compared to CRSwNP NP (p<0.05) and control tissue (p<0.001) and was predominantly expressed in epithelial cells. ALOX15expression significantly correlated with radiographic sinus disease severity (r=0.56, p<0.001) and was associated with comorbid asthma in CRSwNP. 15-oxo-ETE, a downstream product of 15-LO, was significantly elevated in CRSwNP (27.93 pg/mg tissue) and AERD (61.03 pg/mg tissue) NP compared to control (7.17 pg/mg tissue, p<0.001). Hydroxyprostaglandin dehydrogenase, an enzyme required for 15-oxo-ETE synthesis, was predominantly expressed in mast cells and, by immunofluorescence, localized near 15-LO+epithelium in AERD NP. Epithelial and mast cell interactions, leading to the synthesis of 15-oxo-ETE, may contribute to the dysregulation of arachidonic acid metabolism via the 15-LO pathway as well as to the enhanced sinonasal disease severity observed in AERD.

Alloantigen-specific hyporesponsiveness can be induced in alloreactive T cells contained within the whole peripheral blood mononuclear cell (PBMC) population by stimulating these responder cells ex vivo with HLA-mismatched stimulator PBMC as the antigen presenting cell (APC) source, in the presence of a CD28 costimulation blocking agent. As a result of this approach, specific alloreactivity is markedly decreased (by 1-2 logs), but third-party alloresponses and in vitro responses relying on the activation of pathogen- and tumor-associated antigen T-cell functional activities are not globally impinged upon (Guinan et al. N Engl J Med 340(22):1704-1714, 1999, Davies et al. Transplantation 86(6):854-864, 2008, Davies et al. Cell Transplant 21(9):2047-61, 2012). This method has been used clinically to alloanergize bone marrow and PBMC allografts, creating ex vivo cell therapies for adoptive transfer to blood cancer patients at high risk of disease relapse whose best option was to receive haploidentical hematopoietic cell transplants. These early phase trials consisting of, or containing, alloanergized T-cell infusions show promise in reducing graft-versus-host disease (GvHD), providing more rapid immune reconstitution, and decreasing severe post-transplant infectious complications and disease relapse. Herein, we describe this straightforward technique for generating alloanergized PBMC as it is performed in the research lab setting using belatacept for CD28-mediated costimulatory blockade (CSB) and PBMC isolated by Ficoll Hypaque gradient centrifugation as responders and APC. We also describe methods for evaluating subsequent alloproliferation to first and third party stimulation as well as assessment of cell division, pathogen-specific immunity, or allosuppression. The technique has successfully been transferred to collaborating labs, largely owing to the flexibility of using fresh or frozen PBMC, the lack of a requirement for specially isolated APC populations, and the ability to scale up or scale down the cell numbers that are to be anergized.

BACKGROUND:Chronic rhinosinusitis (CRS) is a heterogeneous chronic inflammatory disease subdivided based on the presence or absence of nasal polyps (NPs). Histologic features of chronic rhinosinusitis with nasal polyps (CRSwNP) include inflammatory cell infiltration and excessive fibrin deposition in NPs. Thrombin-activatable fibrinolysis inhibitor (TAFI) is an enzyme that plays an antifibrinolytic role in the body. The significance of TAFI has been documented in patients with chronic inflammatory diseases, including chronic lung disease; however, it has not been evaluated in the pathogenesis of NPs. OBJECTIVE:The objective of this study was to evaluate the potential role of TAFI in the pathogenesis of NPs. METHODS:Nasal lavage fluid was collected from control subjects and patients with CRS. We measured levels of thrombin/anti-thrombin complex (TATc) and TAFI protein using an ELISA. RESULTS:TATc levels in nasal lavage fluid were significantly increased in patients with CRSwNP and patients with chronic rhinosinusitis without nasal polyps (CRSsNP) compared with control subjects, and TAFI levels in nasal lavage fluid were also significantly increased in patients with CRSwNP compared with those in control subjects and patients with CRSsNP. There was a significant correlation between TATc and TAFI levels in nasal lavage fluid. Interestingly, patients with CRS and asthma showed increased TATc and TAFI levels in nasal lavage fluid compared with those in patients with CRS without asthma, especially patients with CRSwNP. CONCLUSIONS:Increased TATc and TAFI levels in nasal passages of patients with CRSwNP might participate in fibrin deposition in NPs and might play a role in the pathogenesis of CRSwNP and asthma.

B cell activation markers are upregulated in nasal polyps (NP). However, age-related B cell inflammatory changes in NP are not well understood. Sinonasal tissues and nasal lavage fluids (NLF) from young (18-49), mature (50-64), and elderly subjects (≥65) with NP, and age-matched healthy controls were collected. Affymetrix microarray assays were performed using NP and uncinate tissues, and the expression of B cell activation markers was examined. A murine model of NP was generated in 3 age groups: young (2 months), middle-aged (12 months) and old-aged (20 months). Levels of B cell activating factor (BAFF) and anti-dsDNA antibody were measured by ELISA and CD138 (plasma cell marker) was examined using immunohistochemistry. There was increased BAFF protein levels in NLF in mature adults (50-64) with NP compared to controls (75.5 vs 17.2 pg/ml, p=0.0198). Levels of anti-dsDNA antibody were significantly increased in older subjects with NP compared to controls (68.8 vs 7.9 IU/ml, p=0.003). CD138 staining was greater in NP subjects than controls without age differences. The microarray analysis revealed that gene expressions of TNFRSF13C(BAFF receptor), CD19(B-cell marker), MS4A1(CD20; B-cell marker), SDC1(CD138; plasma cell marker) and TPSB2(MCP-6; mast cell marker) were significantly higher in elderly NP subjects vs elderly controls. In the murine model of NP, BAFF protein levels were significantly increased in NLF compared to controls in the middle-aged NP group (299.2 vs 29.4 pg/ml, p=0.007), demonstrating a similar pattern in human NP. This study demonstrates age-related differences of B-cell inflammatory responses in human NP and a murine NP model.