Pathogenic role of different phenotypes of immune cells in airway allergic diseases: a study based on Mendelian randomization.

Background: Airway allergic disease (AAD) is a class of autoimmune diseases with predominantly Th2-type inflammation, mainly including allergic rhinitis (AR), allergic asthma (AS), and chronic sinusitis (CRS). There are very complex regulatory mechanisms between immune cells and AAD; however, previous reports found that the functions of the same immune cells in AAD are not identical. Objective: The aim of this study was to explore the causal relationship between different phenotypic immune cells and their association with AAD. Method: Utilizing the publicly available Genome-Wide Association Studies (GWAS) database, this study conducted a bidirectional Mendelian randomization (MR) to assess the causal relationship between immune cells of 731 different immunophenotypes and AAD. The primary assessment methods included inverse variance weighting, weighted median, and MR Egger. Additionally, sensitivity analyses such as MR-PRESSO, leave-one-out, and scatter plots were employed to eliminate the interference of heterogeneity and pleiotropy, ensuring the stability of the causal inference. Result: A total of 38 immune cells with different immunophenotypes were found to be positively and causally associated with AR, of which 26 were protective factors and 12 were risk factors. Positive associations were found between 33 immune cells and AS, of which 14 were protective factors and 19 were risk factors, as well as between 39 immune cells and CRS, of which 22 were protective factors and 17 were risk factors. Finally, the results of all relevant immune cells for the three diseases were taken and intersected, and it was found that CD3 on CD39+-activated Treg (IVWAR = 0.001, IVWCRS = 0.043, IVWAS = 0.027) may be the key immune cell that inhibits the development of AAD (ORAR = 0.940, ORAS = 0.967, ORCRS = 0.976). Conclusion: This study reveals that different immune phenotypes of immune cells are closely related to AAD at the genetic level, which provides a theoretical basis for future clinical studies.

Undersized telomeres in regulatory T cells link to the pathogenesis of allergic rhinitis.

Telomeres are an important biomarker in the cell destiny. The relationship between telomeres and regulatory T cells (Tregs) has not yet been investigated. The objective of this study is to evaluate the link between Tregs' telomere length and allergic rhinitis (AR)'s pathogenesis. Here, we report that low telomerase activity and high endoplasmic reticulum stress status were observed in Tregs from AR patients, as shown in the results. Immune regulatory molecules levels were correlated with the length of Tregs' telomeres. The immune-suppressive functions of Tregs were associated with the telomere length/Telomerase reverse transcriptase/Telomerase protein component 1 status in Tregs. The levels of telomere length/telomerase in airway Tregs were reduced by sensitization. Endoplasmic reticulum stress signaling pathway of proline-rich receptor-like protein kinase-eukaryotic translation initiation factor 2A (eIF2a) was associated with the regulation of telomerase. Inhibiting eIF2a had an effect on upregulating telomerase activity in Tregs and mitigating experimental AR.

[CGRP inhibits proliferation, activation and cytokine secretion of group 2 innate lymphoid cells (ILC2) in peripheral blood from patients with allergic rhinitis].

Objective To investigate the effect of calcitonin gene-related peptide (CGRP) on the regulation of group 2 innate lymphoid cells (ILC2) in the peripheral blood of patients with allergic rhinitis (AR). Methods Peripheral blood mononuclear cells (PBMCs) were extracted from normal healthy individuals and AR patients, then stimulated with CGRP, interleukin 33 (IL-33) and CGRP combined with IL-33 for 3 days, with blank stimulus as control. The percentage of ILC2 in the four groups was measured by flow cytometry. After being sorted, ILC2 was given to CGRP, IL-33 and CGRP combined with IL-33 stimulation for 3 days, with blank stimulus as control. The percentage of IL-5 and IL-13 positive cells in ILC2 was detected by flow cytometry, and the levels of IL-5 and IL-13 in ILC2 supernatant were measured by ELISA. Results The percentage of ILC2 in the peripheral blood of AR patients was significantly higher than that of the control group. The levels of IL-5+ILC2 and IL-13+ILC2 were significantly increased by IL-33 single stimulation after culturing PBMCs. After adding IL-33 combined with CGRP stimulation, the levels of IL-5+ILC2 and IL-13+ILC2 in PBMCs were significantly reduced; after CGRP single stimulation, the levels of IL-5+ILC2 and IL-13+ILC2 in PBMCs were further decreased. After ILC2 was sorted and cultured, the levels of IL-5+ILC2 and IL-13+ILC2 showed significant increase after IL-33 single stimulation. The levels of IL-5+ILC2 and IL-13+ILC2 were decreased by IL-33 and CGRP co-stimulation, and they were further reduced after CGRP single stimulation. Compared to IL-33 single stimulation, IL-5 and IL-13 levels dropped significantly due to the IL-33 and CGRP co-stimulation. The levels of IL-5 and IL-13 were further reduced by CGRP single stimulation. Conclusion CGRP inhibits the proliferation and activation of peripheral blood ILC2 in AR and exert anti-inflammatory effects in AR.

Repair of Oroantral Fistulas Using Free Bone and Mucoperiosteal Flaps After Endoscopic Removal of the Medial Wall of the Maxillary Sinus.

INTRODUCTION: Oroantral fistula (OAF) refers to a pathological connection between the oral cavity and maxillary sinus. It may lead to symptoms that include purulent mucus, nasal congestion, facial swelling, bad breath, and nasal regurgitation of food, all of which negatively impact patients' quality of life. OBJECTIVE: This study presents a novel approach for OAF repair using a 2-layered structure composed of free bone and a mucoperiosteal flap obtained from the medial wall of maxillary sinus. MATERIALS AND METHODS: Ten OAF patients who underwent repair using this method were retrospectively reviewed between August 2015 and June 2022. RESULTS: The extraction of maxillary molars was the most common cause of OAF. The size of the fistulas ranged from 1×2 mm to 5×8 mm. Nine of the 10 patients achieved successful OAF closure following the initial operation using the 2-layer structure composed of free bone and mucoperiosteal flag. One patient was lost to follow-up. The 9 patients were observed for 6 months to 1 year, and they exhibited no obvious complications or recurrence. CONCLUSION: The use of free bone and mucoperiosteal flag from the medial wall of maxillary sinus through an endoscope was effective for OAF repair.

Phosphatidylserine promotes immunotherapy for airway allergy.

Type 1 regulatory T cells (Tr1 cells) play an important role in the maintenance of the immune homeostasis in the body. The induction of Tr1 cell is to be further investigated. The interaction of phosphatidylserine (PS) with TIM3 has immune regulation functions. The objective of this study is to elucidate the role of PS-TIM3 signals in inducing Tr1 cells. In this study, mice were treated using PS or specific immunotherapy by nasal instillation. A murine model of allergic rhinitis was developed using ovalbumin as a specific antigen. We found that PS-containing nasal instillation induced Tr1 cells in the airway tissues. PS promoted gene activities associated with IL-10 through activation of TIM3 in CD4+ T cells. TIM3 mediated the effects of PS on inducing Tr1 cells, in which the TIM3- PI3K-AKT pathway played a critical role. PS boosted allergen-specific immunotherapy by inducing specific antigen Tr1 cell generation. Concomitant administration of PS and SIT resulted in better therapeutic effects on AR. In conclusion, the data demonstrate that PS can promote the specific immunotherapy for AR through inducing antigen specific Tr1 cells in the airway tissues.

Neuromedin U Induces Activation of Peripheral Group 2 Innate Lymphoid Cells through the ERK Pathway in Allergic Rhinitis Patients.

INTRODUCTION: In allergic diseases, group 2 innate lymphoid cells (ILC2s) play critical roles. Neuromedin U (NMU), a highly conserved multifunctional neuropeptide, is secreted by cholinergic neurons and involved in asthma pathogenesis by amplifying lung inflammation driven by ILC2s. However, the precise effects of NMU on ILC2s in allergic rhinitis (AR) and related diseases remain unclear. METHODS: A total of 15 patients with persistent AR and 8 healthy controls (HCs) were enrolled in the study. Visual analog scale (VAS) scores are used to assess the severity of clinical symptoms in AR patients. The percentages of ILC2s in peripheral blood mononuclear cells (PBMCs) were enumerated using flow cytometry. The soluble or intracellular cytokines (IL-5 and IL-13) in PBMCs or sorted ILC2s were assessed in response to various stimuli with IL-33, NMU, IL-33 combined with extracellular signal-related kinase (ERK) inhibitor or NMU combined with ERK inhibitor in the presence of IL-2. RESULTS: We confirmed the proportion of circulating ILC2s was significantly higher in AR patients than in HCs. ILC2s levels were found to be positively related to VAS scores. We also discovered that the release of IL-5 and IL-13 in AR patients' PBMCs stimulated by NMU (p < 0.0001 and p < 0.0001, respectively) or IL-33 (p = 0.002; p = 0.044, respectively) was significantly higher than in HCs. In AR patients, NMU stimulated PBMCs or ILC2s to generate greater inflammatory factors IL-5 and IL-13 compared to IL-33. Furthermore, we observed that NMU-promoted ILC2s activation and proliferation functions were restricted when the ERK pathway was inhibited. CONCLUSION: NMU effectively activated ILC2s in AR patients to produce Th2-type cytokines, and this activation can be prevented by ERK pathway inhibitors. Our findings shed new light on the neuro-immune mechanism of AR and offer new insights into its prevention and treatment.

Characterization of the immune regulatory property of CD22+ CD9+ B cells.

Immunodisruptive homeostasis is recognized in allergic disorders. The mechanism of restoration of immunologic homeostasis in the body is not fully understood. Galectin-9 (Gal9) and CD22 have immune regulatory functions. The goal of this study is to test the role of CD22+ CD9+ B regulatory cells in immune homeostasis the body. A much smaller amount of IL-10 in B10 cells was detected in patients with allergic rhinitis (AR) in contrast to healthy subjects. The IL-10 expression levels in B10 cells were positively correlated with the CD22 expression. CD22 mediated the effects of Gal9 on the enhanced expression of IL-10 in AR B10 cells. Gal9 overcame the refractory induction of IL-10 in B-cells of AR subjects. The immune regulatory ability of AR B10 cells could be restored by Gal9. Combination of Gal9 and SIT induced and activated antigen-specific B10 cells. The B10 cells of Gal9/specific immunotherapy-treated AR mice showed immunosuppressive functions on T-cell activities and induction of type 1 regulatory T cells in an antigen-specific manner. Administration of Gal9 potentiated the effects of specific immunotherapy in mice with AR. In summary, a fraction of regulatory B cells, the CD19+ CD22+ CD9+ B cells, was characterized in the present study. CD22 mediates the effects of Gal9 to promote immunotherapy for allergic diseases by inducing B10 cells. In an antigen-specific manner, the B10 cells suppressed CD4+ T cell activities, and alleviated experimental AR.

Livin promotes Th2-type immune response in airway allergic diseases.

OBJECTIVES: To investigate the effects of livin on the Th2 immune response in airway allergic diseases (AAD) and explore the interaction among livin, GATA3, IL-4 in peripheral blood CD4+ T cells of AAD patients. METHODS: WT mice and livin KO mice were developed for model of AAD. Th2 cell levels in the lung tissues and spleen were assessed by flow cytometry. Also, it was assessed in the culture after exposing to livin inhibitor (Lp-15); the protein and mRNA levels of livin, GATA3 and IL-4 in peripheral blood CD4+ T cells isolated from patients with or without AAD were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting, respectively. Finally, Co-immunoprecipitation (Co-IP) was employed to identify the interaction between livin and GATA3. RESULTS: Compared with WT mouse, Th2 cell frequency in lung tissues and spleen was significantly decreased in livin KO mouse; after adding Lp-15, the differentiation from Naive CD4+T cells in spleen to Th2 cells was blocked; the protein and mRNA levels of livin, GATA3 and IL-4 in AAD group were higher than that in control group. The levels of livin were positively correlated with IL-4, and GATA3 was also positively correlated with IL-4 and livin. GATA3 was detected in the protein complex co-precipitated with livin antibody, and livin was also detected in the protein complex co-precipitated by GATA3 antibody. CONCLUSION: Livin increases the expression of IL-4 and facilitates naive CD4+ T cells to differentiate into Th2 cells, which triggers airway allergy.

Clinical Efficacy of Single Vidian Neurectomy in Chronic Rhinosinusitis with Nasal Polyps and Allergic Rhinitis.

OBJECTIVE: To investigate the clinical efficacy of single Vidian neurectomy (sVN) in the treatment of chronic rhinosinusitis with nasal polyps and allergic rhinitis (CRSwNP &AR). STUDY DESIGN: Descriptive study.   Place and Duration of Study: Otolaryngology-Head &Neck Surgery, Shanxi Medical University Second Affiliated Hospital, Taiyuan, China, February 2016 to February 2019. METHODOLOGY: Patients meeting the diagnostic criteria for AR and CRSwNP confirmed after assessment by an ENT physician; moderately severe and persistent AR, aged ≥18 years to ≤70 years; and testing positive for sIgE and were regularly treated with medications for three months or more before surgery with unsatisfactory symptom control. Exclusion criteria were patients with acute exacerbations of sinusitis or fungal sinusitis combined with nasal polyps, intolerant to aspirin, acute infection or sinus tumours; contraindications to general anaesthetic surgery or oral corticosteroids; and those who have received allergen immunotherapy, corticosteroids and antihistamines within one year. The relevant epidemiological data were collected, including IgE level, VAS, RQLQ, and the Lund-Kennedy scores, to assess patients' clinical symptoms and quality of life before and after surgery. RESULTS: Fifty-five patients were followed up for two years after surgery, and statistical analysis was performed using SPSS version 25.0. It was found that VAS scores, RQLQ scores, and Lund-Kennedy scores of the patients who underwent sVN were significantly lower at six months (all p <0.01), one year (all p <0.01, and two years all p <0.01) after surgery compared with those before surgery. CONCLUSION: sVN has better efficacy in patients with CRSwNP&AR, has the potential to reduce its recurrence rate, and seems to be a safe and effective treatment. KEY WORDS: Chronic rhinosinusitis with nasal polyps, Allergic rhinitis, Eosinophilic chronic rhinosinusitis with nasal polyps, Single Vidian neurectomy, Clinical efficacy.

An eosinophil-Sos1-RAS axis licenses corticosteroid resistance in patients with allergic rhinitis.

BACKGROUND: Corticosteroid resistance (CR) is a serious disadvantage in treating many chronic inflammatory conditions. Eosinophils are the main inflammation cells in allergic reactions. Environmental pollution, such as PM2.5, is associated with the pathogenesis of allergic disorders. The objective of this study is to elucidate the mechanism by which the exposure to PM2.5 confers eosinophil CR status. METHODS: Patients with allergic rhinitis were recruited and assigned to corticosteroid sensitive (CS) and CR groups. Eosinophils were purified from nasal lavage fluids collected from patients with allergic rhinitis. A murine AR mouse model was developed with dust mite allergens and PM2.5 as the sensitization reagents. RESULTS: CR status was detected in about 60% eosinophil collected in patients with AR. Upon exposure to eosinophil activators, CS eosinophils released a large quantity of mediators, which was suppressed by the presence of steroids in the culture. CR eosinophils demonstrated resistance to steroidal therapy. RAS activation levels in eosinophils were higher in CR eosinophils than in CS eosinophils. Higher expression of the Son of sevenless-1 (Sos1) was detected in CR eosinophils, which formed a complex with RAS and glucocorticoidreceptor-α in CR eosinophils to prevent the binding between steroids and glucocorticoidreceptor-α. The presence of an Sos1 inhibitor dissociated glucocorticoid receptor-α from RAS/Sos1 complex, that restored the sensitivity to steroids in eosinophils. Administering the Sos1 inhibitor effectively attenuated the experimental allergic rhinitis. CONCLUSIONS: CR status was detected in approximately 1/3 eosinophils sampled from patients with allergic rhinitis. Sos1 was instrumental in the development and perseverance of CR in eosinophils. Sos1 inhibition restored sensitivity to steroids in CR eosinophils, which effectively reduced experimental allergic rhinitis.

Insights Into the Research Status of Neuromedin U: A Bibliometric and Visual Analysis From 1987 to 2021.

Neuromedin U (NMU) is a regulatory peptide that is widely distributed throughout the body and performs a variety of physiological functions through its corresponding receptors. In recent years, NMU has become the focus of attention in various fields of research as its diverse and essential functions have gradually been elucidated. However, there have been no bibliometrics studies on the development trend and knowledge structure of NMU research. Therefore, in this study, we used VOSviewer software to statistically analyze scientific data from articles related to NMU to track the developmental footprint of this research field, including relevant countries, institutions, authors, and keywords. We retrieved a total of 338 papers related to NMU, written by 1,661 authors from 438 organizations of 41 countries that were published in 332 journals. The first study on NMU was reported by a group in Japan in 1985. Subsequently, nine articles on NMU were published from 1987 to 2006. A small leap in this field could be detected in 2009, with 30 articles published worldwide. Among the various countries in which this research has been performed, Japan and the United States have made the most outstanding contributions. Miyazato M, Kangawa K, and Mori K from the Department of Biochemistry, National Retrain and Cardiovascular Center Research Institute in Japan were the most productive authors who have the highest number of citations. Keyword analysis showed six clusters: central-nervous-system, homeostasis, energy metabolism, cancer, immune inflammation, and food intake. The three most highly cited articles were associated with inflammation. Overall, this study demonstrates the research trends and future directions of NMU, providing an objective description of the contributions in this field along with reference value for future research.

Circadian rhythm disruption exacerbates Th2-like immune response in murine allergic airway inflammation.

BACKGROUND: Chronic jet lag (CJL)-induced circadian rhythm disruption (CRD) is positively correlated with an increased risk of allergic diseases. However, little is known about the mechanism involved in allergic rhinitis (AR). METHODS: Aberrant light/dark cycles-induced CRD mice were randomly divided into negative control (NC) group, AR group, CRD+NC group, and CRD+AR group (n = 8/group). After ovalbumin (OVA) challenge, nasal symptom scores were recorded. The expression of Occludin and ZO-1 in both nasal mucosa and lung tissues was detected by reverse transcription-quantitative polymerase chain reaction (RT-PCR) and immunohistochemical staining. The level of OVA-specific immunoglobulin E (sIgE) and T-helper (Th)-related cytokines in the plasma was measured by enzyme-linked immunosorbent assay (ELISA), and the proportion of Th1, Th2, Th17, and regulatory T cell (Treg) in splenocytes was evaluated by flow cytometry. RESULTS: The nasal symptom score in the CRD+AR group was significantly higher than those in the AR group with respect to eosinophil infiltration, mast cell degranulation, and goblet cell hyperplasia. The expression of ZO-1 and Occludin in the nasal mucosa and lung tissues in the CRD+AR group were significantly lower than those in the AR group. Furthermore, Th2 and Th17 cell counts from splenocytes and OVA-sIgE, interleukin 4 (IL-4), IL-6, IL-13, and IL-17A levels in plasma were significantly increased in the CRD+AR group than in the AR group, whereas Th1 and Treg cell count and interferon γ (IFN-γ) level were significantly decreased in the CRD+AR group. CONCLUSION: CRD experimentally mimicked CJL in human activities, could exacerbate local and systemic allergic reactions in AR mice, partially through decreasing Occludin and ZO-1 level in the respiratory mucosa and increasing Th2-like immune response in splenocytes.

[Progress of IgG4 related sino-nasal diseases].

IgG4 related disease（IgG4-RD）is a newly recognized chronic fibroinflammatory disease in recent years.It is often accompanied by the significant elevation of serum IgG4 level，but the diagnostic specificity of the elevation is not high.The diagnosis is mainly based on histopathology，which is characterized by dense IgG4 positive plasma cell infiltration，storiform fibrosis and obliteran phlebitis.IgG4-RDcan involve various organs of the body，but less involve the nasal cavity and paranasal sinuses.The nasal symptoms of IgG4-RD are lack of specificity，so it is easier to be missed and misdiagnosed.PET/CT has important value in the diagnosis and differential diagnosis of the disease.We reviewed the relevant studies of IgG4-RD in nasal cavity and paranasal sinus in recent years,in order to improve the understanding of IgG4-RD in nasal cavity and paranasal sinuses and to improve the diagnosis and treatment rate.

Immunotherapy with toripalimab for lung adenocarcinoma in a real-world patient with an Eastern Cooperative Oncology Group performance status (ECOG PS) score of 4: a case report.

Multidisciplinary treatment, mainly chemotherapy combined with immunotherapy, is preferred in patients with advanced lung cancer who are negative for driver genes and have an Eastern Cooperative Oncology Group performance status (ECOG PS) score of 0-1; in contrast, patients with an ECOG PS score of 3-4 should be managed with supportive treatment and palliative care rather than chemotherapy or other antitumor treatments. In the real-world settings, however, in the Chinese population, a large proportion of patients and their families are willing to take risks to receive benefit from oncological treatments. We encountered a patient who had definite advanced lung adenocarcinoma with multiple metastases and fusion in abdominal lymph nodes. Intestinal obstruction and obstructive jaundice were also observed. Despite the application of gastrointestinal decompression, continuous parenteral nutrition support, common bile duct stenting, and pancreatic duct stenting, no effective antitumor therapy (except in the case of immunotherapy) was attempted due to the high tumor burden and poor ECOG PS score in this patient. After effective communication with the family members, toripalimab was attempted in this patient with great success (the patient did not receive chemotherapy and only received immunotherapy). In the spring of 2019, imported programmed death receptor-1 (PD-1) antibodies (pabolizumab and navulizumab) listed in the Chinese market were expensive, and the people could not afford the cost. However, treprizumab was just on the market, with low price and high pharmacoeconomics. Therefore, the drug was selected for treatment.) The patient achieved almost complete remission, with a progression-free survival (PFS) of about 2.5 years. The expression of programmed death-ligand 1 (PD-L1) was high in this patient. Toripalimab monotherapy may be applied in patients with high PD-L1 expression although any potential adverse effects should be closely monitored. To our knowledge, no other data from clinical trials and real-world cases have thus far been reported concerning the application of immunotherapy in ECOG PS 4 patients with advanced lung cancer. The innovation of this article is that although the patient has advanced cancer and ECOG PS 4 is an end-stage patient, immunotherapy is still given and achieved great success.

Livin in synergy with Ras induces and sustains corticosteroid resistance in the airway mucosa.

Rationale: Corticosteroid resistance (CR) seriously affects the therapeutic effects of steroids on many chronic inflammatory disorders, including airway allergy. The mechanism of CR development is unclear. Recent research indicates that livin, an apoptosis inhibitor, is associated with the regulation in cell activities. This study investigates the role of livin in the inducing and sustaining CR in the airway mucosa. Methods: Nasal epithelial cells (NECs) were isolated from surgically removed nasal mucosal tissues of patients with allergic rhinitis (AR) and nasal polyps with or without CR. Differentially expressed genes in NECs were analyzed by the RNA sequencing. A CR mouse model was developed to test the role of livin in CR development. Results: The results showed that NECs of AR patients with CR expressed high levels of livin, that was positively correlated with the thymic stromal lymphopoietin (TSLP) expression and the high Ras activation status in NECs. Livin and Ras activation mutually potentiating each other in the inducing and sustaining the TSLP expression in NECs. TSLP induced eosinophils and neutrophils to express glucocorticoid receptor-ß (GRß). Eosinophils and neutrophils with high CRß expression were resistant to corticosteroids. Depletion of livin or inhibition of TSLP markedly attenuated CR and airway allergy. Conclusions: Livin facilitates CR development in the airways by promoting TSLP expression in epithelial cells and the GRß expression in eosinophils and neutrophils. Depletion of livin or inhibiting TSLP attenuates CR development and inhibits airway allergy, this has the translational potential to be used in the treatment of airway allergy.

Role of FcγRI in Antigen-Dependent Eosinophil Activation in Patients With Allergic Rhinitis.

BACKGROUND: The eosinophil (Eo) activation is a crucial factor evoking allergic rhinitis (AR) attacks; factors; the mechanism of triggering Eo activation remains to be further investigated. The interaction of antigen (Ag) and antibody plays a critical role in evoking allergy attacks. This study aims to elucidate the role of FcγRI, the high affinity receptor of IgG, in the Ag-mediated Eo activation. METHODS: Nasal lavage fluids (NLF) were collected from AR patients and healthy control (HC) subjects. Eos were isolated by flow cytometry cell sorting and analyzed by pertinent immunological approaches. RESULTS: Eos composed more than 60% of the cellular components in AR NLF. Exposure to specific Ags (sAgs) in the culture triggered Eos to release inflammatory mediators. High levels of FcγRI were detected on the surface of AR NLF Eos. Exposure to lipopolysaccharide markedly increased the FcγRI expression in naive Eos, which could be bound by Ag-specific IgG (sIgG) to form complexes on the surface of Eos; this made Eos at the sensitized status. Eos bore with the sIgG/FcγRI complexes could be activated upon exposure to sIgG in the culture; these Eos can be designated as Ag-specific Eos. Passive transfer of Ag-specific Eos resulted in profound AR response in mice upon sAg challenge. Depletion of FcγRI on Eos efficiently abolished AR response in mice. CONCLUSIONS: AR Eos express high levels FcγRI, that can be bound by sIgG to make Eos sensitized. Re-exposure to specific Ags can activate the sensitized Eos.

Quantitative determination of human serum testosterone via isotope dilution ultra­performance liquid chromatography tandem mass spectrometry.

Quantification of testosterone serves an important role in the differential diagnosis of androgen­related endocrine diseases. Mass spectrometry exhibits higher accuracy and lower variability than immunoassays, especially at low testosterone concentrations. The present study developed and validated an isotope dilution ultra­performance liquid chromatography tandem mass spectrometry method for determination of human serum testosterone. The serum was equilibrated with an isotopic internal standard and treated with acidic buffer to release hormones from their binding proteins. Testosterone was extracted via two serial liquid­liquid extractions. In the first stage, the lipid fractions from an acidic buffer solution were isolated using ethyl acetate and n­hexane. The organic phase was evaporated and reconstituted in a basic buffer solution. In the second stage, the polar impurities of n­hexane extraction were removed. Total testosterone in serum was quantified via ultra­performance liquid chromatography tandem mass spectrometry in multiple reaction monitoring mode with positive electrospray ionization. The coefficient of variation of the method for intra­ and inter­assay was 2.13% (1.40­2.77%) and 3.44% (3.06­3.66%), respectively. The recovery ranged from 94.32 to 108.60% for different samples. The limit of detection was 0.50 ng/dl and the linear range was from 1.00 to 1,000.00 ng/dl. In addition, the extraction efficiency in three different levels of quality control of the serum ranged from 85.02 to 93.29%. Moreover, structural analogues were investigated and were not indicated to affect the quantification of testosterone. The present method may enable quantification of testosterone in a clinical setting with high precision and accuracy.

Chinese Society of Allergy and Chinese Society of Otorhinolaryngology-Head and Neck Surgery Guideline for Chronic Rhinosinusitis.

The current document is based on a consensus reached by a panel of experts from the Chinese Society of Allergy and the Chinese Society of Otorhinolaryngology-Head and Neck Surgery, Rhinology Group. Chronic rhinosinusitis (CRS) affects approximately 8% of Chinese adults. The inflammatory and remodeling mechanisms of CRS in the Chinese population differ from those observed in the populations of European descent. Recently, precision medicine has been used to treat inflammation by targeting key biomarkers that are involved in the process. However, there are no CRS guidelines or a consensus available from China that can be shared with the international academia. The guidelines presented in this paper cover the epidemiology, economic burden, genetics and epigenetics, mechanisms, phenotypes and endotypes, diagnosis and differential diagnosis, management, and the current status of CRS in China. These guidelines-with a focus on China-will improve the abilities of clinical and medical staff during the treatment of CRS. Additionally, they will help international agencies in improving the verification of CRS endotypes, mapping of eosinophilic shifts, the identification of suitable biomarkers for endotyping, and predicting responses to therapies. In conclusion, these guidelines will help select therapies, such as pharmacotherapy, surgical approaches and innovative biotherapeutics, which are tailored to each of the individual CRS endotypes.

Probiotic extracts ameliorate nasal allergy by inducing interleukin-35-producing dendritic cells in mice.

BACKGROUND: The therapeutic efficacy of allergic rhinitis (AR) needs to be improved. Probiotics have immunoregulatory functions. In this study we evaluated the effects of protein extracts of probiotics in the amelioration of AR. METHODS: Extracts of Bifidobacterium infantis (EBI) were prepared by lysing the live probiotics. AR mice were developed to be used to evaluate the therapeutic efficacy of EBI. RESULTS: The results show that EBI induced interleukin (IL)-10-producing dendritic cells (DCs) via increasing IL-35 and signal transducer and activator of transcription 3 (STAT3) phosphorylation. IL-10-expressing DCs induced IL-10-producing B cells (B10 cells), with the latter showing immunosuppressive functions. After challenge with specific antigens, AR mice showed sneezing, nasal itch, and increases in serum-specific immunoglobulin E (IgE) and mouse mast cell protease-1; higher levels of T helper 2 (Th2) cytokines (IL-4, 67.17 ± 10.66; IL-5, 62.83 ± 9.70; IL-13, 51.00 ± 6.69, before treatment) in nasal mucosal protein extracts, which were significantly suppressed (IL-4, 27.00 ± 6.66; IL-5, 23.86 ± 4.53; IL-13, 25.67 ± 4.93, after treatment (p < 0.001) by administration with EBI nasal drops. CONCLUSION: EBI can suppress AR via inducing B10 cells. Thus, after carrying out required preclinical experiments and tests, EBI has the translational potential to be used in the treatment of AR and other allergic diseases.

Survivin facilitates T-helper 2-biased inflammation in the airway.

BACKGROUND: The biased T helper 2 (Th2) responses play a critical role in the pathogenesis of allergy. The underlying mechanism is not fully understood yet. Survivin can regulate multiple cellular activities. This study aims to elucidate the role of survivin in the development and maintenance of Th2 polarization. METHODS: CD4+ T cells were isolated from blood samples collected from patients with allergic asthma (AS) and HS control (HS) subjects. Mice carrying CD4+ T cells with survivin knockout (KO mice) were employed to test the role of survivin in the development of the biased Th2 responses. RESULTS: KO mice failed to induce airway allergy. Peripheral CD4+ T cells expressed survivin, which was higher in the AS group than that in the HS group. Naive CD4+ T cells with higher expression of survivin were prone to differentiating into Th2 cells. Survivin bound to the Il4 promoter in CD4+ T cells to enhance Il4 gene transcription. The expression of Fas was lower in CD4+ T cells of the AS group than that in the HS group. Overexpression of survivin suppressed the expression of Fas and impaired the activation-induced cell death (AICD) of CD4+ T cells. CONCLUSION: Survivin facilitates the development of biased Th2 polarization through promoting expression of interleukin 4 (IL-4) and impairing the AICD machinery of CD4+ T cells. To modulate the expression of survivin in CD4+ T cells has the translational potential in the treatment of allergic diseases.