Erzhong Fan

Allergen-Dependent Differences in ILC2s Frequencies in Patients With Allergic Rhinitis

Purpose Group 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens. Methods Flow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage- and Lineage+ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage- and Lineage+ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit. Results The percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33±IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage- cells compared to Lineage+ cells. Conclusions AR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.

Association of periostin expression with eosinophilic inflammation in nasal polyps

by GSK-3b leads to the instability of b-catenin, reducing translocation of b-catenin into the nucleus, which can be associated with tumorigenesis. In this study we established the critical link between GSK-3b and Foxp3 by means of tissue examination and in vitro analysis. Previously, Graham et al reported that GSK-3b inhibition is able to potentiate the suppressive activity of Treg cells, but whether other forms of posttranslational modifications, such as phosphorylation, can affect Foxp3 stability and Treg cell function has been largely unknown. In a recent study on rheumatoid arthritis, Nie et al investigated the relationship between Treg cells with pathogenic T cells at the site of chronic inflammation. They identified Ser 418 as a key phosphorylation site of Foxp3 that was responsible for the impaired Treg cell function in response to TNF-a. In this study we found a novel phosphorylation motif (Ser 270/274) of Foxp3, which can be recognized by proinflammatory GSK-3b. Interestingly, when we examined the p-Foxp3 protein using specific antibody against Ser 270 in NP tissues, we found p-Foxp3 (Ser 270) and showed a positive association of GSK-3b and p-Foxp3 (Ser 270) protein levels. Therefore these findings propose a molecular mechanism regulating inhibitory function of Treg cells, which might expand our knowledge on the pathogenesis of CRSwNP. Xingmei Wu, MD* Sihua Wang, PhD* Miaomiao Han, PhD* Bin Song, PhD Ping Ye, MD Shanshan Ma, MD Jun Li, MD Fenghong Chen, MD Geng Xu, PhD Qingqing Ding, PhD Jiahong Xia, PhD Huabin Li, PhD From the Allergy Center, Otorhinolaryngology Hospital, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China; the Department of Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; the Allergy Division, Department of Otolaryngology, Head and Neck Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; the Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases, Shanghai, China; the Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China; and the Department of Pathology, University of Missouri, Columbia, Mo. E-mail: qingqing_ding@ hotmail.com. Or: jiahong.xia@hust.edu.cn. Or: allergyli@163.com. *These authors contributed equally to this work. Supported by grants from the National Natural Science Foundations of China (nos. 81172512, 81271054, 81271056, and 81470673) and the Shanghai Committee of Science and Technology (no. 14DZ2260300). Disclosure of potential conflict of interest: The authors declare that they have no relevant conflicts of interest.

TMEM16A-Mediated Mucin Secretion in IL-13-Induced Nasal Epithelial Cells From Chronic Rhinosinusitis Patients

Purpose Chronic rhinosinusitis with nasal polyps (CRSwNP), a mainly Th2 cytokine-mediated disease, often involves mucus secretion. Recent evidence suggests that transmembrane protein 16A (TMEM16A), a calcium-activated Cl- channel (CaCC), can regulate mucus secretion from airway epithelium by transepithelial electrolyte transport and hydration. However, the role of TMEM16A in mucin production/secretion in the airway epithelium is not clear. This study was conducted to determine the role of TMEM16A in mediating mucin secretion in human nasal polyp epithelial cells (HNPECs) induced by IL-13. Methods Human sinonasal mucosa tissue and dissociated sinonasal epithelium from control subjects and patients with CRSwNP were assessed for the expression of TMEM16A and the secretion of human mucin 5AC (MUC5AC) by immunohistochemistry, Western blot analysis, and enzyme-linked immuno-sorbent assay (ELISA). A model of the Th2 inflammatory environment was created by exposure of primary air-liquid interface (ALI)-cultured HNPECs to interleukin-13 (IL-13) for 14 days, with subsequent assessment of TMEM16A expression in cell lysates by Western blotting and MUC5AC secretion in apical washings of cells by ELISA. Results The expressions of TMEM16A and MUC5AC were increased in human nasal polyp tissue and dissociated nasal polyp epithelium. TMEM16A was detected in IL-13-treated HNPECs, specifically in MUC5AC-positive cells but not in ciliated cells. IL-13 treatment increased percentages of TMEM16A-positive cells, MUC5AC-positive cells, and cells coexpressing TMEM16A/MUC5AC, the expression of TMEM16A protein, and the secretion of MUC5AC. T16Ainh-A01, a TMEM16A inhibitor, attenuated these IL-13-induced effects. Conclusions The expression of TMEM16A and MUC5AC are increased in CRSwNP, which might be a direct effect of Th2 cytokines present in the sinonasal mucosa in CRSwNP. Down-regulation of TMEM16A expression and MUC5AC secretion in HNPECs by T16Ainh-A01 indicates that TMEM16A might play an important role in mucin secretion in upper airway inflammatory diseases.

Role of IFN-γ, IL-13, and IL-17 on mucociliary differentiation of nasal epithelial cells in chronic rhinosinusitis with nasal polyps.

Mucociliary dysfunction is a prominent pathophysiological feature of chronic rhinosinusitis with nasal polyps (CRSwNP); however, the precise mechanisms underlying mucociliary dysfunction are still unclear.

Frequency, Suppressive Capacity, Recruitment and Induction Mechanisms of Regulatory T Cells in Sinonasal Squamous Cell Carcinoma and Nasal Inverted Papilloma

Background Sinonasal squamous cell carcinoma (SSCC) and nasal inverted papilloma (NIP) represent the predominant type of malignant and benign tumors in sinonasal tract, respectively. CD4+CD25+Foxp3+ natural regulatory T (Treg) cells might play critical role(s) in the suppression of anti-tumor immune response and thus shed light on tumor progression from benign to malignant. Objective This study aimed to evaluate the frequency and suppressive capacity of Treg cells in SSCC compared to NIP and further to explore the underlying mechanisms. Patients and Methods Frequencies of Treg, Th1 and Th2 cells were evaluated by flow cytometry in tissue homogenate and peripheral blood from 31 SSCC patients, 32 NIP patients and 35 normal controls. Treg cells were tested for regulatory function by co-culture with effector T cells. CCR4 and its ligands, CCL22 and CCL17, were analyzed by flow cytometry and Luminex, respectively. The chemoattractant properties of CCR4/CCL22 and CCR4/CCL17 for Treg cells were assessed using the Boyden chamber technique, to elucidate the potential mechanisms of Treg recruitment in tumor microenvironment. Treg cells induction via TGF-β was assessed with transwells after local CD4+Foxp3+ T cells were assessed by immunohistochemistry and TGF-β concentration was measured by Luminex. Results Tumor-infiltrating Treg cells increased significantly from normal to NIP to SSCC (P ≤ 0.001 for normal vs. NIP and P = 0.004 for NIP vs. SSCC). Significantly elevated frequency and enhanced suppression capacity of circulating Treg cells in SSCC were detected compared to NIP and healthy controls, concomitant with Th1 decrease and Th2 increase. Apparently increased CCL22 attracted CCR4-expressing Treg cells to tumor microenvironment in SSCC, compared to NIP. SSCC produced significantly more TGF-β than NIP and thus possessed greater potential for Treg cell induction. Conclusion Frequency and suppressive capacity of Treg cells enhanced with progression of malignancy from NIP to SSCC. Circulating Treg cells were recruited to tumor tissue via CCR4/CCL22 signalling, whereas tumor-synthesised TGF-β contributed to induction of peripheral Treg cells.

Matrix metalloproteinases, tissue inhibitor of metalloproteinase and transforming growth factor A1 in the remodeling of chronic rhinosinusitis in North China

Results MMP-8,9 were the highest among all above MMPs. MMP-8 of NP tissue was much higher than that of CRS and control tissue (P<0.001). MMP-9 of NP tissue was also significantly higher than that of CRSsNP and control tissue (P<0.05). MMP-7 of NP tissue and CRSsNP were much higher than that of control tissue (P<0.001). MMP2 of CRSsNP tissue was much higher than that of NP and control tissue (P<0.001). TIMP-1,2 were the highest among all the four TIMPs. TIMP-1 of CRSsNP tissue was much higher than that of NP and control tissue (P<0.001 and P<0.05 respectively). TIMP-2 of NP and CRSsNP tissue were much higher than that of control tissue (P<0.01). TIMP-3 of CRSsNP tissue was much higher than that of NP and control tissue (P<0.01 and P<0.05 respectively). There were no significantly difference of TGF-¦Â1 among the three groups.