Nasal Polyps Patients Research Articles

Dysregulated fatty acid metabolism in nasal polyp-derived eosinophils from patients with chronic rhinosinusitis.

Eosinophils are multifunctional granulocytes capable of releasing various cytokines, chemokines, and lipid mediators. We previously reported dysregulated fatty acid metabolism in peripheral blood-derived eosinophils from patients with severe asthma. However, functional characteristics of eosinophils present in allergic inflammatory tissues remain largely uncharacterized. We established a method for isolating CD69hi CCR3low CXCR4- siglec-8int eosinophils from nasal polyps of patients with eosinophilic rhinosinusitis (NP-EOS). Multi-omics analysis including lipidomics, proteomics, and transcriptomics was performed to analyze NP-EOS as compared to peripheral blood-derived eosinophils from healthy subjects (PB-EOS). Lipidomic analysis revealed impaired synthesis of prostaglandins and 15-lipoxygenase (15-LOX)-derived mediators, and selective upregulation of leukotriene D4 production. Furthermore, proteomics and transcriptomics revealed changes in the expression of specific enzymes (GGT5, DPEP2, and 15-LOX) responsible for dysregulated lipid metabolism. Ingenuity pathway analysis indicated the importance of type 2 cytokines and pattern recognition receptor pathways. Stimulation of PB-EOS with eosinophil activators IL-5, GM-CSF, and agonists of TLR2 and NOD2 mimicked the observed changes in lipid metabolism. Inflammatory tissue-derived eosinophils possess a specific phenotype with dysregulated fatty acid metabolism that may be targeted therapeutically to control eosinophilic inflammatory diseases.

Differences in the distribution of CD20, CD3, CD34 and CD45RO in nasal mucosa and polyps from patients with chronic rhinosinusitis.

The present study focused on the assessment of the inflammatory infiltrate that characterizes nasal polyps in patients with chronic rhinosinusitis and nasal polyposis. Inflammatory cell type was determined using specific markers. This evaluation was made possible by determining the expression of the following markers: CD20, a marker of B lymphocytes [using activated T cells (ATC) armed with CD20 antibody]; CD3, a marker of T lymphocytes (using ATC armed with anti-CD3 antibody); CD45, the leukocyte common antigen (using ATC armed with anti-CD45 antibody; and CD34, for the microvasculature of the nasal polyp (using anti-CD34 antibody). The diagnosis of chronic rhinosinusitis with nasal polyps (CRSwNP) was made according to current EPOS guidelines based on patient history, clinical examination and nasal endoscopy. We examined surgically resected nasal polyps from 127 patients diagnosed with CRSwNP, who benefited from surgical procedures at the Department of Otorhinolaryngology of our institution. The polyps were analyzed at the Department of Pathology of our institution utilizing histopathological and immunohistochemical methods as follows: Firstly, the tissues were paraffin-impregnated, sectioned and stained with hematoxylin and eosin. We then examined the expression of CD3, CD20, CD34 and CD45RO by immunohistochemistry with soluble labeled streptavidin biotin (LSAB)/horseradish peroxidase (HRP) complexes. We observed the following histopathological changes: The structure of the epithelium was evidenced by collagenous subjacent stroma with mixed areas, sometimes associated with hyaline zones. In all types of polyps, we also observed a diffuse underlayer or periglandular lymphoplasmacytic in filtrate composed predominantly from T lymphocytes and eosinophils. The histopathological changes suggest the chronic inflammation of the sinus mucosa, which was diffusely distributed in allergic polyps and with nodular distribution in fibro-inflammatory polyps. The number of B lymphocytes was greater in the fibro-inflammatory polyps. On the whole, the findings of this study indicate that the inflammatory infiltrate in nasal polyps from patients with CRSwNP is mainly composed of T cells and eosinophils in all types of polyposis. In addition, a diffuse distribution of allergic polyps and the nodular distribution of fibro-inflammatory polyps, and the hyperplasia of the seromucous glands was observed. The determination of CD20, CD3, CD34 and CD45RO could be used to assess the inflammatory infiltrate of the nasal poplyps in these patients.

Detection of microparticles in nasal lavage fluid that express both integrin β6 and a marker of epithelial-mesenchymal transition (either alpha-smooth muscle actin or Snail) is useful for the identification of severe chronic rhinosinusitis

Microparticles (MPs) are one type of extracellular vesicle shed from cell membranes. Integrin β6 (ITGB6) is an epithelial marker that is up-regulated during epithelial repair. Snail is a transcription factor that induces epithelial-mesenchymal transition (EMT) during epithelial repair. Alpha-smooth muscle actin (αSMA) is a mesenchymal marker which is also up-regulated during EMT. We hypothesized that abnormal EMT status is associated with polyp recurrence in chronic rhinosinusitis (CRS) and endeavored to establish MP assays that can detect EMT in nasal lavage fluids (NLFs). Snail, αSMA and ITGB6(+)MPs were measured by flow-cytometry in NLFs collected from 196 CRS with nasal polyp patients (CRSwNP) and 47 control subjects whose age, sex and smoking status were matched to the CRSwNP group. Levels of Snail(+)ITGB6(+)MPs (1.6-fold, p<0.0001) and αSMA(+)ITGB6(+)MPs (2.0-fold, p<0.0001) were increased in CRSwNP compared with control. Among CRSwNP subjects, levels of Snail(+)ITGB6(+)MPs (1.7-fold, p<0.0001) and αSMA(+)ITGB6(+)MPs (2.0-fold, p<0.0001) were higher in those with a history of previous polypectomy than in those without, independent of age, atopy, asthma or steroid use. Area under the curve (AUC) analyses showed that levels of αSMA(+)ITGB6(+)MPs (AUC:0.78 [95%CI:0.71-0.84], sensitivity 81%, specificity 61%) and Snail(+)ITGB6(+)MPs (AUC:0.77 [0.69-0.84], sensitivity 58%, specificity 66%) have potential utility to detect EMT and assess the degree of disease severity in CRS patients. Elevated EMT, with up-regulation of ITGB6 on MPs along with EMT markers Snail or αSMA, is associated with severe polyp recurrences which necessitated revision polypectomy. Snail or αSMA(+)ITGB6(+)MPs are potential biomarkers to detect severe CRS patients at high risk for revision polypectomy.

Efficacy and Safety of Dupilumab in Patients with Chronic Rhinosinusitis with Nasal Polyps: Results from the Randomized Phase 3 Sinus-24 Study

Dupilumab, a human IL-4R-α mAb inhibiting IL-4 and IL-13, key drivers of type 2-mediated inflammation, is approved for uncontrolled moderate-to-severe atopic dermatitis in adults and in the USA for patients aged ≥12 years with moderate-to-severe eosinophilic or oral corticosteroid-dependent asthma. In a phase 2a study, dupilumab improved endoscopic, radiologic, clinical, and patient-reported outcomes in chronic rhinosinusitis with nasal polyps (CRSwNP) patients. For the first time we report results from a phase 3 dupilumab study (SINUS-24; NCT02912468) in severe CRSwNP patients (with/without comorbid asthma) on daily intra-nasal mometasone furoate (MFNS). Adults with CRSwNP previously treated with systemic corticosteroids (SCS) and/or surgery (n = 276) were randomized 1:1 to subcutaneous dupilumab 300mg or placebo every 2 weeks for 24 weeks. Outcomes included change from baseline to Week 24 in nasal polyp score (NPS), patient-reported nasal congestion (NC), CT Lund-Mackay (LMK), UPSIT smell test, SNOT-22, FEV1, and ACQ-6 scores. Mean [SD] baseline values were comparable in dupilumab/placebo for NPS (5.64 [1.23] / 5.86 [1.31]), NC (2.26[0.57]/2.45[0.55]), LMK (18.55 [4.55] / 19.55 [4.26]), UPSIT (14.68 [8.66] / 14.44[8.31]), and SNOT-22 (48.00 [20.16] / 50.87[20.22]). Dupilumab significantly (P<0.0001) improved NPS (−2.06), NC (−0.89), LMK (−7.44), UPSIT (+10.56), and SNOT-22 (−21.12) from baseline to Week 24 (LS-mean differences vs placebo). Dupilumab treatment reduced SCS use/NP surgery (P<0.001), and in comorbid asthma patients (58.3%) improved lung function (FEV1; P<0.001) and asthma control (ACQ-6; P<0.0001). The most common adverse event occurring at higher frequency with dupilumab was epistaxis (7.7% vs 3.0% placebo). Add-on dupilumab to MFNS significantly improved endoscopic, radiologic, clinical, and patient-reported sino-nasal outcomes, and asthma outcome measures in severe CRSwNP patients, and was well-tolerated.

Endoscopic Sinus Surgery for Type-2 CRS wNP: An Endotype-Based Retrospective Study.

Nasal polyps are often characterized by type 2 inflammation and disease recurrence. We developed a new surgical technique, referred to as reboot approach, which aims to maximally remove all sinus mucosa and allow healthy re-epithelialization from the preserved nasal mucosa. We here review type 2 endotype chronic rhinosinusitis with nasal polyps (CRSwNP) patients who underwent classical mucosa-sparing endoscopic sinus surgery (ESS) or the reboot approach. Retrospective case-control study of 50 consecutive CRSwNP patients who underwent endoscopic sinus surgery between 2015 and 2017, either as a classical non-reboot ESS (n = 20); a partial reboot approach removing the mucosa of the ethmoidal, sphenoidal, and maxillary sinuses (n = 18); or a complete reboot approach including Draf III and removal of all frontal sinus mucosa (n = 12). Polyp recurrence over the follow-up period of 2 years served as the primary outcome. All patients demonstrated a type 2 inflammation of the mucosal tissue harvested during surgery. In the classical approach group (n = 20), nine patients relapsed within 2 years (45%); in the partial reboot group, three out of 18 patients (17%) relapsed; and in the full reboot group one out of 12 patients (8%) relapsed. The relapse rates were significantly different between the non-reboot and the reboot groups (P = 0.02) but also between all treatment groups (P = 0.038). Complete removal of diseased mucosa from the paranasal sinuses (reboot approach) significantly reduces the recurrence of nasal polyps for 30 months postoperatively compared to the current mucosa-sparing approach in type 2 inflammatory CRSwNP. 3b Laryngoscope, 129:1286-1292, 2019.

Short-term pre-operative systemic administration with low-dose of steroid does not make a false-negative diagnosis of definite eosinophilic chronic rhinosinusitis after endoscopic sinus surgery.

According to JESREC criteria, definite eosinophilic chronic rhinosinusitis (ECRS) is postoperatively diagnosed based on over 70 eosinophils in high power magnification fields of the resected nasal polyps in patients with probable ECRS. Preoperative systemic administration with steroid is a standard practice, because it reduced intraoperative bleeding during endoscopic sinus surgery (ESS) in patients with ECRS. However, it was recently reported that systemic administration with steroid decreased the number of eosinophils in the nasal polyps, leading to a false negative diagnosis of definite ECRS. To overcome the risk, we have adopted short-term pre-operative systemic administration with low-dose of steroid and examined if our steroid administration makes a false-negative diagnosis. We performed a retrospective chart review of 42 patients with probable ECRS. Eleven patients were administered with 0.5 mg of bethametasone for 7 days before ESS, and other 31 patients were not administered. The average number of eosinophils in nasal polyps in patients who were administered with steroid was 188 ± 167, which was not different from 199 ± 149 in the patients who were not administered. These findings suggest that short-term preoperative administration with low-dose of steroid has few risk of a false-negative diagnosis of definite ECRS. J. Med. Invest. 66 : 233-236, August, 2019.

Increased expression of L‐plastin in nasal polyp of patients with nonsteroidal anti‐inflammatory drug‐exacerbated respiratory disease

Most patients with nonsteroidal anti-inflammatory drug-exacerbated respiratory disease (NERD) suffer from recurrence of nasal polyps. However, little is known about the specific cellular and molecular mechanisms contributing to the pathogenesis of nasal polyp development in patients with NERD in particular, especially at baseline when cyclooxygenase 1 inhibitors are not present. The objectives of this study were to identify proteins involved in the pathogenesis of nasal polyps in patients with NERD. We collected nasal polyp tissue from patients with NERD and from patients with aspirin-tolerant chronic rhinosinusitis with nasal polyps (CRSwNP). Protein profiles were analyzed by 2-dimensional electrophoresis and identified several proteins, including L-plastin, as highly expressed. We examined L-plastin and tissue factor (TF) expression by immunohistochemical and immunofluorescence analyses. To examine the role of L-plastin in eosinophils, we knocked down L-plastin expression in Eol-1 cells by using siRNA transfection. L-plastin protein levels in nasal polyp tissue were increased in patients with NERD relative to those in patients with aspirin tolerant CRSwNP. Immunofluorescence analysis revealed that L-plastin was dominantly expressed in eosinophils and L-plastin and TF were co-expressed in eosinophils in NERD nasal polyp tissue. Knockdown of L-plastin in Eol-1 cells disrupted the cell surface distribution of TF by stimulation with granulocyte macrophage colony-stimulating factor. Increased expression of L-plastin by eosinophils may contribute to abnormal fibrin deposition through TF translocation to the eosinophil cell surface in NERD nasal polyp tissue, which in turn may contribute to the pathogenesis of NERD.

Elevated expression of IL-17RB and ST2 on myeloid dendritic cells is associated with a Th2-skewed eosinophilic inflammation in nasal polyps

BackgroundInterleukin(IL)-25, IL-33, and thymic stromal lymphopoietin (TSLP) underlie the crosstalk between epithelial cells and dendritic cells (DCs) during the development of Th2 responses. This study aimed to measure the expressions of IL-17RB, ST2 and TSLPR, receptor of IL-25, IL-33, and TSLP respectively, on myeloid DCs in nasal polyps (NP) and evaluate their association with local Th2 inflammation and disease severity in patients with NP.MethodsSamples were collected from 30 NP patients and 16 control subjects recruited prospectively. The mRNA expression of cytokines, including TSLP, IL-25 and IL-33, as well as interferon (IFN)-γ, IL-4, IL-5, IL-13 and IL-17A in NP and control tissues was examined by qualitative polymerase chain reaction (qPCR). The expression of IL-17RB, ST2 and TSLPR as well as other surface markers on myeloid DCs (mDCs) was examined by flow cytometry.ResultsIncreased numbers of total and activated mDCs were found in NP patients. mDCs demonstrated significantly higher expression of IL-17RB, ST2 and TSLPR than those in control tissues. The activated mDCs exhibited up-regulations of OX40L and ICOSL, but down-regulation of PDL1 in NP. Moreover, the IL-17RB, ST2 and TSLPR levels on mDCs were positively correlated with IL-25, IL-33 and TSLP mRNA levels, respectively, in NP. Furthermore, IL-17RB and ST2 expressions on mDCs were correlated with the IL-5 mRNA level as well as eosinophil number in NP. Importantly, the IL-17RB expression on mDCs and the OX40L expression on activated mDCs in NP were positively correlated with CT score and total nasal symptom score.ConclusionsIncreased expressions of IL-17RB and ST2 on mDCs are associated with enhanced local Th2 inflammation in NP, suggesting that mDCs might play a role in IL-25- and IL-33-induced type 2 responses and eosinophilic inflammation in NP.

Interleukin-4-induced posttranscriptional gene regulation of CCL26 by the RNA-binding protein HuR in primary human nasal polyp-derived epithelial cells.

Much attention on the pathophysiology of nasal polyp (NP) has focused on eosinophils. Interleukin (IL)-4 and eotaxin-3 (C-C motif chemokine ligand 26, or CCL26) levels have been reported to be increased in eosinophilic nasal polyps. The aim of this study was to characterize CCL26 posttranscriptional regulation by the RNA-binding protein HuR in primary human nasal polyp-derived epithelial cells (hNPDECs) challenged with IL-4. A prospective, observational study was conducted. Nasal polyp tissues were obtained from eosinophilic (n = 12) and non-eosinophilic (n = 10) NP patients, and inferior turbinate (IT) tissues were taken from control subjects (n = 9) and cultured into hNPDECs. Expression of HuR and CCL26 were measured by immunohistochemistry, Western blot analysis, enzyme-linked immunoassay, and real-time polymerase chain reaction (PCR). The nucleocytoplasmic shuttling of HuR in hNPDECs was detected by immunofluorescence. Posttranscriptional regulation of CCL26 by HuR was tested by ribonucleoprotein immunoprecipitation assay (RIP) and dual-luciferase reporter assay. CCL26 mRNA stabilization was measured by quatititative PCR after treatment with actinomycin D. Student's t test and one-way analysis of variance were used. Immunohistochemical data show that both HuR and CCL26 were highly expressed in NP tissues, especially eosinophilic NP tissues (p < 0.05). IL-4 stimulation increased CCL26 mRNA stability, and overexpression and knockdown of HuR affected CCL26 expression. Immunofluorescence data indicate that IL-4 altered the subcellular distribution of HuR. The RIP and dual-luciferase reporter assay results supply strong evidence for HuR binding to CCL26. Our results provide strong support for the hypothesis that IL-4-induced expression of CCL26 in hNPDECs relies partly on CCL26 mRNA stabilization mediated by the interaction of HuR with CCL26 3'UTR.

Cysteinyl leukotriene 1 receptor expression in nasal polyps.

Cysteinyl leukotrienes (CysLTs) have been implicated in the pathogenesis of chronic rhinosinusitis (CRS). This study was undertaken to better understand the role of CysLTs in the development of CRS through 1). assessment of the pattern of expression of CysLT1 receptor in nasal polyps from patients with CRS and 2). correlation of expression of CysLT1 receptor with clinical features. Expression of CysLT1 receptor was evaluated immunohistochemically in nasal polyps from 20 patients with CRS and nasal mucosa from 10 control subject undergoing plastic surgery of the nose. Patients with CRS showed significantly higher expression of CysLT1 receptor as compared with the control group (p&lt;0.05). The expression of CysLT1 receptor in sub-epithelial inflammatory infiltrates tended to be higher in patients with CRS and allergy as compared with patients with CRS but without allergy (p=0.07). In particular, the expression of CysLT1 receptor in sub-epithelial inflammatory infiltrates was significantly greater in 3 patients with CRS and drug allergy as compared with patients with CRS but without drug allergy (p=0.03). Increased expression of CysLT1 receptor in inflamed mucosa of nasal polyps in patients with allergy might suggest particular role of CysLTs in the pathogenesis of nasal polyps in this group of patients.

Differential effects of postoperative oral corticosteroid on eosinophilic vs. non-eosinophilic CRSwNP subtypes

PurposeThe efficacy of postoperative oral corticosteroids on surgical outcomes in chronic rhinosinusitis with nasal polyps (CRSwNP) patients following endoscopic sinus surgery (ESS) remains controversial. This study evaluated the potential benefits of postoperative oral corticosteroids on surgical outcomes in CRSwNP patients and investigated the differential effects on eosinophilic CRSwNP (ECRSwNP) and noneosinophilic CRSwNP (NECRSwNP). Materials and methodsPatients with bilateral CRSwNP who underwent ESS were enrolled and randomized to receive either oral prednisolone (30 mg/day) or placebo for 2 weeks after surgery. Visual analog scale (VAS) and Sino-Nasal Outcome Test 22 (SNOT-22) scores were chosen as the subjective outcomes, evaluated at preoperative baseline and 1, 3, and 6 months postoperatively. Lund-Kennedy Endoscopic Scores (LKESs) were used as the objective outcome, evaluated at preoperative baseline and at 2 weeks and 2, 3, and 6 months postoperatively. ResultsIn total, 100 patients with bilateral CRSwNP were enrolled, of whom only 82 completed the 6-month follow-up. The subjective outcomes showed no significant difference at each follow-up points. Of the objective outcomes, the corticosteroid group reporting a trend of improvement in LKESs at 6 months postoperatively (p = 0.05). After stratification by tissue eosinophils, only patients with NECRSwNP (<10 eosinophils/HPF) demonstrated a significant improvement in LKESs at 3 months postoperatively (p = 0.03). ConclusionsPostoperative oral corticosteroids did not provide additional improvements in VAS and SNOT-22 scores; nevertheless, a trend of LKES improvement was noted at 6 months postoperatively. After stratification by tissue eosinophils, this effect was significant only among NECRSwNP patients at 3 months follow-up.

Understanding the Propensity for Chronic Sinusitis in Patients on Immunosuppressive Therapy.

Chronic rhinosinusitis (CRS) is a frequently observed condition in patients with immunodeficiency secondary to immunosuppressive medications. The histologic features of CRS among patients undergoing immunosuppressive treatment have yet to be determined and may have important implications on understanding the pathophysiology of the disease process. A structured histopathology report was utilized to analyze sinus tissue removed during functional endoscopic sinus surgery (FESS). Histopathology variables, Lund-Mackay score (LMS), and sinonasal outcome test 22 scores were compared among patients with CRS on immunosuppressive therapy (CRSi), CRS without nasal polyps (CRSsNP) patients, and CRS with nasal polyps (CRSwNP) patients. Fifteen CRSi, 36 CRSwNP, and 56 CRSsNP patients undergoing FESS were analyzed. Compared to CRSsNP, CRSi patients exhibited a trend toward increased moderate-severe inflammation (66.7% vs 42.1%, P < .080), increased neutrophil infiltrate (40.0% vs 24.6%, P < .192), and decreased fibrosis (26.7% vs 43.9%, P < .182). Compared to CRSwNP, CRSi patients demonstrated decreased fibrosis (26.7% vs 66.7%, P < .010), decreased eosinophil aggregates (13.3% vs 44.4%, P < .032), and a trend toward fewer eosinophils per high-power field (46.7% vs 66.7%, P < .154). CRSi cases had significantly lower mean LMS (8.20 ± 4.30 vs 12.78 ± 6.56, P < .017) compared to CRSwNP. CRSi patients exhibit histopathology and disease severity more similar to CRSsNP with trends toward increased neutrophilia and reduced fibrosis. In the appropriate clinical context, discontinuing or changing a patient's immunosuppressive regimen may be a valid treatment option in patients with CRSi. This study provides initial insight into understanding the propensity for chronic sinusitis in patients undergoing immunosuppressive treatment which may have implications on disease management.

1,8-cineol inhibits the Wnt/β-catenin signaling pathway through GSK-3 dephosphorylation in nasal polyps of chronic rhinosinusitis patients

Chronic rhinosinusitis with nasal polyps (CRSwNP) represents a benign neoplasm of the nasal mucosa, which leads to a decreased breathing capacity and reduced olfaction. The pathogenesis and the molecular mechanisms driving nasal polyps are not very well known. GSK-3 is involved in the regulation of various biosynthetic pathways and various kinases are able to regulate the GSK-3. Therefore, we investigated the effect of the monoterpene oxide 1,8-cineol on the regulation of the Wnt/β-catenin signaling pathway with its central regulator protein GSK-3 in vitro.We determined GSK-3 expression and phosphorylation as well as the expression of negative regulators (Akt and SGK) and downstream activation of β-catenin in nasal polyps of patients with CRSwNP by immunohistochemistry and Western blot experiments. In this study we demonstrated for the first time, that 1,8-cineol acts as a potential inhibitor of the Wnt/β-catenin signaling pathway, by affecting the inhibitory phosphorylation of GSK-3, which is the key regulator of the β-catenin activity. Our data provide novel insights in the regulatory networks responsible for the progression of CRSwNP and furthermore represent a new mechanism of 1,8-cineol activity, which may lead to novel treatment approaches to this natural drug.

Olfactory Change Pattern after Endoscopic Sinus Surgery in Chronic Rhinosinusitis with Olfactory Dysfunction.

To determine the change pattern of olfactory function in chronic rhinosinusitis (CRS) with olfactory dysfunction after endoscopic sinus surgery (ESS), and its association with inspection scores. Observational study. Second Hospital of Shandong University, China, from December 2014 to January 2016. Thirty-four CRS with nasal polyps (CRSwNP) patients and 14 CRS without nasal polyps (CRSsNP) patients were evaluated preoperatively by T&T olfactory test, olfactory VAS test, SNOT-20score and Lund-Mackay CT score. Outcomes were re-evaluated at 1 month, 3 months and 6 months postoperatively. Both olfactory and SNOT-20 scores showed significant improvement within 6 months in both CRSwNP and CRSsNP groups. Patients with anosmia in T&T test showed the largest degree of improvement. No significant recovery of olfactory dysfunction was observed at 1 month and 3 months in CRSsNP groups postoperative. In CRSwNP, the olfactory scores were correlated with the CT scores significantly (r=0.649, p<0.001; r=0.625, p<0.001). However, no correlation was found between the SNOT-20 score and olfactory score preoperatively. Our study has confirmed that the therapeutic effects of ESS on olfactory function last for up to 6 months, particularly in patients with CRSwNP. Although the therapeutic effects plateaued at 3 months postoperatively, the olfactory function continues to recover between 3 and 6 months.

Is there any effect of neurotrophin-3 on the pathogenesis of non-allergic nasal polyps?

Although the role of neurotrophins such as nerve growth factor and brain-derived neurotrophic factor in nasal polyps development has been studied, the contribution of neurotrophin-3 has not been evaluated yet. This study aimed to investigate the possible role of neurotrophin-3 in nasal polyps pathogenesis. The study group comprised 70 non-allergic nasal polyps patients and the control group consisted of 53 patients with middle turbinate concha bullosa. Specimens were taken, during surgery, from the ethmoid sinus nasal polyps in the nasal polyps group and from the lateral part of the middle turbinate concha bullosa in the control group. Tissue and serum levels of neurotrophin-3 were assessed by immunohistochemistry and enzyme-linked immunosorbent assay, respectively. Nasal polyps patients had higher tissue neurotrophin-3 scores (p < 0.001). There was no statistically significant difference between groups regarding serum neurotrophin-3 levels (p = 0.417). Tissue neurotrophin-3 staining scores in the nasal polyps group had no statistically significant correlation with Lund-Mackay scores (p = 0.792). Neurotrophin-3 may have a local effect in nasal polyps pathogenesis, without joining systemic circulation.

Levels of Th1 and Th2 Cytokines in Patients with Nasal Polyps

Objective: We aimed to investigate Th1 / Th2 (T helper) cell balance by examining Interleukin-4 (IL-4), Interleukin-5 (IL-5), Interleukin-10 (IL-10), Interleukin-17 (IL-17) and Interferon gamma (IFN γ) levels in serum in patients with nasal polyps (NPs). Patients and Methods: This study included 45 patients (mean age: 34.8 years) with NPs and a control group (mean age: 37.9) consisting of 45 healthy volunteers. Results: The mean IL-17 level in the NPs group (4.096 pg/ml) was significantly higher than the control group (2.219 pg/ml) (p<0.001). IL-5 levels were significantly higher in the control group. NPs group had significantly lower IL-5 level compared with the control group (1.981 pg/ml vs. 2.720 pg/ml, respectively: p<0.001). The mean IFN γ concentration was significantly lower in the NPs group (0.379 pg/ml) compared to the control group (0.439 pg/ml) (p=0.011). No significant differences were found in the IL-4 (3.25 pg/ml vs. 3.38 pg/ml) and IL-10 (11.249 pg/ml vs. 12.156 pg/ml) concentrations between NPs and the control groups (p=0.799 and p=0.255, respectively). Conclusions: Increased IL-17 levels were found in NPs patients. There was no significant differences in IL-4 and IL-10 values of the Th1 cytokines among the groups, whereas IL-5 was found to be low in the NPs group. IFN gamma values of Th2 cytokines were found to be low in the NPs group. According to these results, low IFN gamma levels and regulatory cytokine IL-7 activation may be effective in the etiopathogenesis of NPs.

Mucus T helper 2 biomarkers predict chronic rhinosinusitis disease severity and prior surgical intervention.

Chronic rhinosinusitis (CRS) is a diverse clinical syndrome with a heterogeneous pathophysiology. Early attempts to identify CRS endotypes and biomarkers have largely relied on analysis of surgically obtained tissue, thus limiting their practical utility. This study examined the ability of mucus T helper 2 (Th2) biomarkers to predict CRS disease severity and clinical characteristics. CRS (n = 90) and healthy control subjects (n = 17) were prospectively enrolled prior to surgical intervention and mucus levels of interleukin (IL)-4, IL-5, and IL-13 were determined using a multiplex cytometric bead assay. Data for relevant cytokines was then scaled, normalized, and later combined to develop standardized metrics indicative of Th2-associated inflammation. Th2-high and Th2-low subgroups were consequently identified and validated against factors associated with disease severity and clinical outcomes. Mucus levels of IL-5 and IL-13 were elevated in CRS subjects compared to controls, while no significant difference was noted for IL-4. IL-5 and IL-13 high CRS were associated with worse objective measures of disease severity and greater rates of revision surgery. Similar relationships were noted for both cytokines when CRS with nasal polyps (CRSwNP) patients were analyzed separately. Th2-high CRS and Th2-low CRS were then categorized using a scaled IL-5/IL-13 metric. Th2-high CRS was characterized by an increased number of subjects with nasal polyps and comorbid asthma, and worse symptom and computed tomography (CT) scores. The Th2-associated cytokines, IL-5 and IL-13, are detectable in sinonasal mucus and their levels can be used to define Th2-high and Th2-low CRS. Identification of Th2-high and Th2-low endotypes using mucus-based biomarkers could facilitate stratification of CRS subgroups and guide personalized therapies.

Chronic rhinosinusitis with nasal polyps is characterized by dysbacteriosis of the nasal microbiota

Chronic rhinosinusitis with nasal polyp (CRSwNP) patients are often characterized by asthma comorbidity and a type-2 inflammation of the sinonasal mucosa. The mucosal microbiota has been suggested to be implicated in the persistence of inflammation, but associations have not been well defined. To compare the bacterial communities of healthy subjects with CRSwNP patients, we collected nasal swabs from 17 healthy subjects, 21 CRSwNP patients without asthma (CRSwNP−A), and 20 CRSwNP patients with co-morbid asthma (CRSwNP+A). We analysed the microbiota using high-throughput sequencing of the bacterial 16S rRNA. Bacterial communities were different between the three groups. Haemophilus influenzae was significantly enriched in CRSwNP patients, Propionibacterium acnes in the healthy group; Staphylococcus aureus was abundant in the CRSwNP−A group, even though present in 57% of patients. Escherichia coli was found in high amounts in CRSwNP+A patients. Nasal tissues of CRSwNP+A patients expressed significantly higher concentrations of IgE, SE-IgE, and IL-5 compared to those of CRSwNP−A patients. Co-cultivation demonstrated that P. acnes growth was inhibited by H. influenzae, E. coli and S. aureus. The nasal microbiota of healthy subjects are different from those of CRSwNP−A and CRSwNP+A patients. However, the most abundant species in healthy status could not inhibit those in CRSwNP disease.

Accumulation of CD69+ tissue‑resident memory T cells in the nasal polyps of patients with chronic rhinosinusitis.

In patients with chronic rhinosinusitis with nasal polyps (CRSwNP), a relative accumulation of cluster of differentiation (CD)8+ Tcells over CD4+ Tcells occurs in nasal polyps compared with the peripheral blood. Nasal CD8+ Tcells and CD4+ Tcells predominantly present an effector memory phenotype. Immunological studies have reported that memory Tcells recirculate from the tissues to the peripheral blood and a high percentage of these Tcells persist within the tissue. The aim of the present study was to characterize CD69+ sphingosine‑1‑phosphate receptor1 (S1PR1)‑ tissue resident memory Tcells (Trm) in the polyps of patients with CRSwNP. Tissue and blood samples were collected from 10patients undergoing nasal sinus surgery. Expression of specific extra‑ and intracellular molecules were analyzed using multicolor flow cytometry. A significantly higher level of CD8+ Tcells than CD4+ Tcells was present in nasal polyps, while significantly more CD4+ Tcells than CD8+ Tcells were detected in the peripheral blood of patients with CRSwNP. The frequency of CD69+ Tcells was significantly higher in CD8+ and CD4+ Tcells in nasal polyps compared with the peripheral blood. The frequency of CD69+ S1PR1‑ Trm was also significantly higher in CD4+ and CD8+ Tcells from nasal polyps compared with the peripheral blood. Within polyps, the frequency of CD69+ S1PR1‑ Trm was again significantly higher in CD8+ compared with CD4+ Tcells. In summary, a significantly higher frequency of CD69+ S1PR1‑ Tcells was observed in the nasal polyps compared with the peripheral blood in patients with CRSwNP. The results of the present study suggest that local regulation of the immune response occurs within nasal polyps. As such, Trm should be considered a potential stimulus in the pathogenesis of nasal polyps. However, the role of Trm in nasal polyps as a pathogenic trigger of the local inflammatory reaction requires further investigation.

The expression of CD22 and CD72 on B cells negatively correlate with IgE level in eosinophilic chronic rhinosinusitis with nasal polyps

Abstract Background CD22 and CD72 have been demonstrated to act as B cell inhibitory receptors, which suppress B cells activation. However, the roles of CD22 and CD72 on B cells in nasal polyps (NPs) are largely unknown. Objective To explore the expression of CD22 and CD72 on B cells and the association with immunoglobulins (Igs) in NPs. Methods The expression levels of CD22 and CD72 was measured by RT-PCR, immunohistochemistry, immunofluorescence and flow cytometry. Igs levels in NPs were quantified by Bio-Plex assay or the UniCAP system. Computed tomography scan and symptom severity were graded. Results The mRNA expression of CD22 and CD72 were elevated in eosinophilic chronic rhinosinusitis with nasal polyps (Eos CRSwNP) patients compared with non-eosinophilic chronic rhinosinusitis with nasal polyps (non-Eos CRSwNP) and controls. CD22 and CD72 were mainly expressed on CD19+ B cells. The numbers of CD22+ and CD72+ B cells and the percentage of CD22+ B cells and CD72+ B cells in B cells were increased in Eos CRSwNP compared with non-Eos CRSwNP and controls. IgA, IgG and IgM were elevated in both Eos CRSwNP and non-Eos CRSwNP, while IgD and IgE were increased in Eos CRSwNP. CD22 mRNA and CD72 mRNA expression level were negatively correlated with IgE, but not IgA, IgG, IgM and IgD in Eos CRSwNP. The percentage of CD22+ B cells and CD72+ B cells in B cells negatively correlate with the total visual analog scale symptom scores and total endoscope scores. Conclusion CD22 and CD72 were significantly increased in Eos CRSwNP patients and negatively correlated with IgE level, indicating the down-regulation role of CD22 and CD72 in B cell activation and a suppressive role in the synthesis of IgE in Eos CRSwNP.