RÉSUMÉ
Objective: To identify potential therapeutic targets of chronic sinusitis with nasal polyps (CRSwNP) through proteomics screening of and verify its effectiveness experimentally. Methods: The nasal tissue samples were collected from patients undergoing surgical treatment in the Department of Otorhinolaryngology, Head and Neck Surgery in Yuhuangding Hospital of Yantai from June 2010 to December 2021, including 69 patients with CRSwNP and 39 patients in the control group. Tissue samples were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in data-independent acquisition (DIA) mode to find differentially expressed proteins. Bioinformatics tools were employed to analyze the functions of differentially expressed proteins. The expression of hematopoietic cell kinase (HCK) in nasal tissues of patients with CRSwNP was further confirmed by qPCR and western blot. The mouse model of CRSwNP was established and treated with HCK inhibitor. The levels of inflammatory factors IgE, IL-4 and IL-5 in serum of CRSwNP mice, both treated and untreated with HCK inhibitors, were detected by enzyme-linked immunosorbent assay (ELISA) across different experimental groups. The experimental data were analyzed by Graphpad Prism 9 software. Results: DIA analysis identified 1 850 differential proteins, including 760 up-regulated proteins and 1 090 down-regulated proteins. Weighted correlation network analysis (WGCNA) correlation analysis of phenotypic data such as cell count and CT score with the results of genomics indemnified 575 proteins of MEBrown module which intersected with 35 kinases further screened from 1 850 differential proteins, yielding eight protein kinases: HCK, SYK, PDK2, FGR, PRKCB, ROR1, CAMK1 and GRK6. qPCR showed that the expression of HCK in CRSwNP was significantly higher than that in the control group (P<0.05). Further experiments in mice confirmed that the secretion of IgE, IL-4 and IL-5 in the serum of CRSwNP group was significantly higher than the control group (all P<0.05), indicating successful model establishment. The intervention of HCK significantly decreased the secretion of IgE, IL-4 and IL-5 in serum of mice (all P<0.05). Conclusion: The HCK inhibitor can reduce the inflammatory index of mice with CRSwNP, and HCK is a potential therapeutic target of CRSwNP.
Sujet(s)
Modèles animaux de maladie humaine , Polypes du nez , Protéomique , Sinusite , Sinusite/métabolisme , Animaux , Souris , Protéomique/méthodes , Maladie chronique , Polypes du nez/métabolisme , Humains , Interleukine-4/métabolisme , Interleukine-5/métabolisme , Spectrométrie de masse en tandem , Immunoglobuline E/sang , Chromatographie en phase liquideRÉSUMÉ
Objective:The purpose of this study is to explore the expression of prostacyclin receptorï¼IPï¼ in patients with chronic rhinosinusitisï¼CRSï¼ and its possible association with type 2 inflammation. Methods:HE staining was used to observe the morphological changes of nasal mucosa, qRT-PCR was used to detect the expression of IP in polyps and nasal mucosa, and IHC was used to detect the expression of IP, IL-4, IL-5 and IL-13 in polyps and nasal mucosa. Results:Compared with the control group, the nasal mucosa of patients with various types of CRS was obviously thickened, accompanied by inflammatory cell infiltration and gland hyperplasia. The statistical results of IHC showed that the expression levels of IL-4, IL-5 and IL-13 in CRS group were significantly higher than those in control groupï¼P<0.05ï¼, and the IP expression in control group was significantly higher than that in ECRS group and non-ECRS groupï¼P<0.05ï¼. The IP expression in ECRS group was negatively correlated with IL-4, IL-5 and IL-13. The results of qRT-PCR showed that the expression of IP mRNA in control group was significantly higher than that in ECRS group and non-ECRS groupï¼P<0.05ï¼. Conclusion:IL-4, IL-5 and IL-13 are highly expressed in the nasal mucosa of CRS patients, while IP is poorly expressed in the nasal mucosa of CRS patients, and IP is negatively correlated with IL-4, IL-5 and IL-13, suggesting that IP is related to the occurrence and development of type 2 inflammation and may be a potential therapeutic target for CRS patients.
Sujet(s)
Inflammation , Muqueuse nasale , Polypes du nez , Récepteurs de l'époprosténol , , Adulte , Femelle , Humains , Mâle , Adulte d'âge moyen , Maladie chronique , Inflammation/métabolisme , Interleukine-13/métabolisme , Interleukine-4/métabolisme , Interleukine-5/métabolisme , Muqueuse nasale/métabolisme , Polypes du nez/métabolisme , Récepteurs de l'époprosténol/métabolisme , /métabolismeRÉSUMÉ
OBJECTIVE: To investigate the levels of 12 kinds of cytokines in seminal plasma and their correlations with routine semen parameters. METHODS: The remaining seminal plasma samples of 134 patients undergoing routine semen examination were collected for detecting cytokines. The parameters for sperm concentration, percentage of progressively motile sperm (PR), and motility were analyzed by a computer-assisted sperm analysis (CASA) system. According to the results of sperm concentration, PR and motility, 134 patients were divided into the normal routine semen parameters group, oligoasthenospermia group and azoospermia group. The levels of 12 kinds of cytokines in seminal plasma, including interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17, interferin (IFN)-α, IFN-γ, and tumor necrosis factor (TNF)-α, were detected by flow cytometry. Two seminal plasma samples were detected for 10 times, respectively, to calculate the coefficients of variation (CV) of each cytokine. The linear range of each cytokine was measured using the standard, and the correlation coefficient (r) was calculated. RESULTS: The r2 of 12 kinds of cytokines detected by flow cytometry were all greater than 0.99. The reproducibility of 2 seminal plasma samples showed that the CVs of all cytokines were lower than 15 % except for TNF-α in sample 1 (15.15 %). Seminal plasma IL-6 levels were negatively correlated with semen volume (P < 0.01). Seminal plasma IL-5 levels were positively correlated with sperm concentration (P < 0.01). Seminal plasma IL-8 levels were negatively correlated with sperm motility (P < 0.01). Seminal plasma IL-8, IL-17 and IL-12P70 levels were negatively correlated with sperm PR (P < 0.05). In addition to the significant negative correlation between IL-5 and IL-17 (P < 0.05), there was a significant positive correlation between the majority of other cytokines. The levels of seminal plasma IL-17 and IL-12P70 in the oligoasthenospermia group and IL-1ß and IL-12P70 in the azoospermia group were significantly higher than those in the normal routine semen parameters group (P ≤ 0.05), while the levels of IL-10 in the azoospermia group were significantly lower than that in the normal routine semen parameters group (P < 0.05). CONCLUSION: There are certain correlations between seminal plasma cytokines and routine semen parameters and strong correlations between different seminal plasma cytokines, suggesting that the imbalance between seminal plasma cytokines may affect sperm quality. However, it still needs to be further confirmed by large samples and multi-center clinical studies and related basic researches.
Sujet(s)
Cytokines , Cytométrie en flux , Analyse du sperme , Sperme , Mobilité des spermatozoïdes , Humains , Mâle , Sperme/métabolisme , Adulte , Cytokines/sang , Cytokines/métabolisme , Cytométrie en flux/méthodes , Analyse du sperme/méthodes , Interleukine-5/métabolisme , Interleukine-5/sang , Interleukine-17/sang , Interleukine-17/métabolisme , Interleukine-17/analyse , Numération des spermatozoïdes , Interleukine-6/sang , Interleukine-6/analyse , Interleukine-6/métabolisme , Interleukine-8/sang , Interleukine-8/métabolisme , Interleukine-8/analyse , Interleukine-12/sang , Interleukine-12/métabolisme , Facteur de nécrose tumorale alpha/métabolisme , Facteur de nécrose tumorale alpha/sang , Facteur de nécrose tumorale alpha/analyse , Interféron gamma/sang , Interféron gamma/métabolisme , Interleukine-1 bêta/métabolisme , Interleukine-1 bêta/sang , Interleukine-1 bêta/analyse , Interleukine-10/sang , Interleukine-10/métabolisme , Interleukine-10/analyse , Azoospermie/métabolisme , Azoospermie/sang , Interleukine-2/sang , Interleukine-2/métabolisme , Interleukine-2/analyse , Interleukine-4/sang , Interleukine-4/métabolisme , Interleukine-4/analyse , Oligospermie/métabolismeRÉSUMÉ
Interleukin-5 (IL-5) has been reported to be involved in cardiovascular diseases, such as atherosclerosis and cardiac injury. This study aimed to investigate the effects of IL-5 on cardiac remodelling. Mice were infused with angiotensin II (Ang II), and the expression and source of cardiac IL-5 were analysed. The results showed that cardiac IL-5 expression was time- and dose-dependently decreased after Ang II infusion, and was mainly derived from cardiac macrophages. Additionally, IL-5-knockout (IL-5-/-) mice were used to observe the effects of IL-5 knockout on Ang II-induced cardiac remodelling. We found knockout of IL-5 significantly increased the expression of cardiac hypertrophy markers, elevated myocardial cell cross-sectional areas and worsened cardiac dysfunction in Ang II-infused mice. IL-5 deletion also promoted M2 macrophage differentiation and exacerbated cardiac fibrosis. Furthermore, the effects of IL-5 deletion on cardiac remodelling was detected after the STAT3 pathway was inhibited by S31-201. The effects of IL-5 on cardiac remodelling and M2 macrophage differentiation were reversed by S31-201. Finally, the effects of IL-5 on macrophage differentiation and macrophage-related cardiac hypertrophy and fibrosis were analysed in vitro. IL-5 knockout significantly increased the Ang II-induced mRNA expression of cardiac hypertrophy markers in myocardial cells that were co-cultured with macrophages, and this effect was reversed by S31-201. Similar trends in the mRNA levels of fibrosis markers were observed when cardiac fibroblasts and macrophages were co-cultured. In conclusions, IL-5 deficiency promote the differentiation of M2 macrophages by activating the STAT3 pathway, thereby exacerbating cardiac remodelling in Ang II-infused mice. IL-5 may be a potential target for the clinical prevention of cardiac remodelling.
Sujet(s)
Angiotensine-II , Cardiomégalie , Fibrose , Interleukine-5 , Macrophages , Souris knockout , Facteur de transcription STAT-3 , Transduction du signal , Remodelage ventriculaire , Animaux , Angiotensine-II/pharmacologie , Facteur de transcription STAT-3/métabolisme , Facteur de transcription STAT-3/génétique , Remodelage ventriculaire/effets des médicaments et des substances chimiques , Souris , Macrophages/métabolisme , Macrophages/effets des médicaments et des substances chimiques , Interleukine-5/métabolisme , Interleukine-5/génétique , Cardiomégalie/métabolisme , Cardiomégalie/anatomopathologie , Cardiomégalie/génétique , Cardiomégalie/induit chimiquement , Mâle , Souris de lignée C57BL , Différenciation cellulaire , Myocarde/métabolisme , Myocarde/anatomopathologieRÉSUMÉ
OBJECTIVES: Chronic rhinosinusitis (CRS) endotypes have demonstrated clinical value in guiding treatment decisions. Bacterial lysates are immunomodulators that have shown beneficial effects in various respiratory inflammatory diseases. This study aimed to evaluate the effect of postoperative bacterial lysate therapy on different CRS endotypes. METHODS: Patients diagnosed with CRS who underwent endoscopic sinus surgery were recruited. Bacterial lysates were administered postoperatively for 10 days per month for 3 months to the experimental group comprising patients with a history of frequent upper respiratory infections without adverse reactions. The remaining participants were allocated to the control group. The results of the postoperative 3-, 6-, and 12-month assessments, including the modified Lund-Kennedy (mLK) endoscopic and Sinonasal Outcome Test (SNOT) 22 scores, for the groups were compared. The tissue samples obtained from the participants were evaluated to detect the presence of relevant inflammatory mediators. RESULTS: Among the 92 participants, 47 started bacterial lysate therapy 2 weeks after the surgery. The tissue cytokine profiles and clinical parameters, such as the disease severity and blood eosinophil percentage, of the bacterial lysate and control groups were comparable before treatment. The mLK endoscopic and SNOT-22 scores did not differ after 3, 6, and 12 months of follow-up. The subgroup analysis revealed that the bacterial lysate group had significantly lower mLK endoscopic scores than the control group for CRS without nasal polyps, while there was a tendency toward significance for the interleukin (IL)-5 negative group after 6 months. CONCLUSION: Postoperative bacterial lysate therapy has some beneficial effects on the endoscopic findings of patients with CRS without nasal polyps or those who are negative for IL-5.
Sujet(s)
Endoscopie , Rhinite , Sinusite , Humains , Sinusite/chirurgie , Sinusite/thérapie , Maladie chronique , Rhinite/chirurgie , Rhinite/thérapie , Rhinite/métabolisme , Mâle , Femelle , Adulte d'âge moyen , Adulte , Phénotype , Extrait cellulaire , Polypes du nez/chirurgie , Polypes du nez/métabolisme , Polypes du nez/complications , Test d'impact des symptômes sino-nasaux , Interleukine-5/métabolisme , Soins postopératoires/méthodes , Cytokines/métabolisme , Résultat thérapeutique , ,RÉSUMÉ
BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease resulting in decreased quality of life. Histamine and specifically the H4 receptor play a key role in the inflammatory process in AD and serve as targets for novel therapeutic approaches. OBJECTIVE: In the present study we aimed to elucidate the immunopathological mechanisms with which the H4 receptor impacts TH2 cells and contributes to AD pathophysiology. METHODS: Total CD4+ T cells obtained from healthy or AD individuals and in vitro differentiated TH2 cells were cultured under different conditions and the mRNA expression or protein production of target molecules were determined using quantitative real-time PCR and ELISA. RESULTS: H4 receptor mRNA expression was upregulated concentration dependent upon IL-4 stimulation in in vitro differentiated TH2 cells progressively during the differentiation. Transcriptomic analysis of in vitro differentiated TH2 versus TH1 cells revealed that the H4 receptor among other genes represents one of the highly upregulated genes in TH2 cells. Most importantly, increased amounts of IL-5 and IL-13 mRNA expression were detected in in vitro differentiated TH2 cells as well as protein secretion in the presence of histamine or of the H4 receptor-selective-agonist when compared to the untreated control. CONCLUSION: We show for the first time an H4 receptor dependent upregulation of the pro-inflammatory cytokines IL-5 and IL-13 in human TH2 cells by histamine. This suggests that the blockade of the H4 receptor may lead to downregulation of these cytokines and amelioration of AD symptoms as reported in first clinical studies.
Sujet(s)
Eczéma atopique , Interleukine-13 , Interleukine-5 , Récepteur histaminergique H4 , Lymphocytes auxiliaires Th2 , Humains , Lymphocytes auxiliaires Th2/immunologie , Lymphocytes auxiliaires Th2/métabolisme , Eczéma atopique/immunologie , Eczéma atopique/métabolisme , Interleukine-13/métabolisme , Interleukine-5/métabolisme , Récepteur histaminergique H4/métabolisme , Différenciation cellulaire/immunologie , Activation des lymphocytes/immunologie , Cellules cultivéesRÉSUMÉ
Asthma is a common and burdensome chronic inflammatory airway disease that affects both children and adults. One of the main concerns with asthma is the manifestation of irreversible tissue remodelling of the airways due to the chronic inflammatory environment that eventually disrupts the whole structure of the airways. Most people with troublesome asthma are treated with inhaled corticosteroids. However, the development of steroid resistance is a commonly encountered issue, necessitating other treatment options for these patients. Biological therapies are a promising therapeutic approach for people with steroid-resistant asthma. Interleukin 5 is recently gaining a lot of attention as a biological target relevant to the tissue remodelling process. Since IL-5-neutralizing monoclonal antibodies (mepolizumab, reslizumab and benralizumab) are currently available for clinical use, this review aims to revisit the role of IL-5 in asthma pathogenesis at large and airway remodelling in particular, in addition to exploring its role as a target for biological treatments.
Sujet(s)
Remodelage des voies aériennes , Asthme , Interleukine-5 , Humains , Asthme/traitement médicamenteux , Asthme/immunologie , Asthme/métabolisme , Remodelage des voies aériennes/effets des médicaments et des substances chimiques , Interleukine-5/antagonistes et inhibiteurs , Interleukine-5/immunologie , Interleukine-5/métabolisme , Antiasthmatiques/usage thérapeutique , Antiasthmatiques/pharmacologie , AnimauxRÉSUMÉ
Asthma, the most prevalent respiratory disease, affects more than 300 million people and causes more than 250,000 deaths annually. Type 2-high asthma is characterized by interleukin (IL)-5-driven eosinophilia, along with airway inflammation and remodeling caused by IL-4 and IL-13. Here we utilize IL-5 as the targeting domain and deplete BCOR and ZC3H12A to engineer long-lived chimeric antigen receptor (CAR) T cells that can eradicate eosinophils. We call these cells immortal-like and functional IL-5 CAR T cells (5TIF) cells. 5TIF cells were further modified to secrete an IL-4 mutein that blocks IL-4 and IL-13 signaling, designated as 5TIF4 cells. In asthma models, a single infusion of 5TIF4 cells in fully immunocompetent mice, without any conditioning regimen, led to sustained repression of lung inflammation and alleviation of asthmatic symptoms. These data show that asthma, a common chronic disease, can be pushed into long-term remission with a single dose of long-lived CAR T cells.
Sujet(s)
Asthme , Récepteurs chimériques pour l'antigène , Animaux , Asthme/immunologie , Asthme/thérapie , Souris , Récepteurs chimériques pour l'antigène/immunologie , Récepteurs chimériques pour l'antigène/génétique , Récepteurs chimériques pour l'antigène/métabolisme , Immunothérapie adoptive/méthodes , Lymphocytes T/immunologie , Interleukine-5/immunologie , Interleukine-5/métabolisme , Modèles animaux de maladie humaine , Humains , Interleukine-4/immunologie , Interleukine-4/métabolisme , Souris de lignée C57BL , Granulocytes éosinophiles/immunologie , Femelle , Interleukine-13/métabolisme , Interleukine-13/immunologieRÉSUMÉ
Chronic obstructive pulmonary disease (COPD) is a condition characterized by chronic airway inflammation and obstruction, primarily caused by tobacco smoking. Although the involvement of immune cells in COPD pathogenesis is well established, the contribution of innate lymphoid cells (ILCs) remains poorly understood. ILCs are a type of innate immune cells that participate in tissue remodeling processes, but their specific role in COPD has not been fully elucidated. During COPD, the breakdown of pulmonary elastin generates elastin peptides that elicit biological activities on immune cells. This study aimed to investigate the presence of ILC in patients with COPD and examine the impact of elastin peptides on their functionality. Our findings revealed an elevated proportion of ILC2 in the peripheral blood of patients with COPD, and a general activation of ILC as indicated by an increase in their cytokine secretion capacity. Notably, our study demonstrated that serum from patients with COPD promotes ILC2 phenotype, likely due to the elevated concentration of IL-5, a cytokine known to favor ILC2 activation. Furthermore, we uncovered that this increase in IL-5 secretion is partially attributed to its secretion by macrophages upon stimulation by elastin peptides, suggesting an indirect role of elastin peptides on ILC in COPD. These findings shed light on the involvement of ILC in COPD and provide insights into the potential interplay between elastin breakdown, immune cells, and disease progression. Further understanding of the mechanisms underlying ILC activation and their interaction with elastin peptides could contribute to the development of novel therapeutic strategies for COPD management.NEW & NOTEWORTHY Elastin-derived peptides, generated following alveolar degradation during emphysema in patients with COPD, are able to influence the response of type 2 innate lymphoid cells. We show that the orientation of innate lymphoid cells in patients with COPD is shifted toward a type 2 profile and that elastin peptides are indirectly participating in that shift through their influence of macrophages, which in turn impact innate lymphoid cells.
Sujet(s)
Élastine , Immunité innée , Lymphocytes , Broncho-pneumopathie chronique obstructive , Humains , Broncho-pneumopathie chronique obstructive/immunologie , Broncho-pneumopathie chronique obstructive/anatomopathologie , Élastine/métabolisme , Élastine/immunologie , Lymphocytes/immunologie , Lymphocytes/métabolisme , Lymphocytes/effets des médicaments et des substances chimiques , Femelle , Mâle , Sujet âgé , Adulte d'âge moyen , Interleukine-5/métabolisme , Interleukine-5/immunologie , Macrophages/immunologie , Macrophages/métabolisme , Peptides/pharmacologie , Peptides/immunologieRÉSUMÉ
Objective: To investigate the effects of the epidermal growth factor receptor(EGFR) inhibitor Gefitinib on airway inflammation and airway remodelling in asthmatic C57BL/6 mice, and to analyze its possible mechanisms. Methods: Male C57BL/6 mice, aged 6-8 weeks, were randomly assigned into five groups: Group A (control group), Group B (asthma group), Group C (asthma+20 mg/kg gefitinib group), Group D (asthma+40 mg/kg gefitinib group), and Group E (40 mg/kg gefitinib group), with seven mice per group. Mice were sensitized by intraperitoneal injection of a mixture of 0.2 ml solution containing OVA and Al(OH)3 [20 µg OVA+2 mg Al(OH)3 dissolved in 0.2 ml of physiological saline] at Day 0 and 14. Starting from Day 25 to 31, Group B, C, and D were challenged with nebulization of 1% OVA solution (8 ml) to induce asthma, once a day for approximately 40 minutes, with continuous aerosolization for 7 days. Group C and D were given 0.2 ml of Gefitinib dissolved in 0.5% carboxymethylcellulose sodium (CMCNa) by gavage half an hour before challenging, and Group E was simultaneously given with 0.2 ml of Gefitinib dissolved in 0.5% CMCNa only. Group A and B were given an equivalent volume of 0.5% CMCNa by gavage. After 24 h of final challenge, the bronchoalveolar lavage fluid (BALF) was prepared for the determination of total cell count and eosinophil count. The levels of total immune globulin E (IgE) in serum and interleukin (IL)-4, IL-5 and IL-13 in BALF and lung tissue homogenates were measured by ELISA. The mRNA expression levels of IL-4, IL-5, IL-13 in lung were measured. Immunohistochemistry and Western blot experiments were used to detect the expression levels of EGFR in lung tissues. Results: In Group B, the level of total IgE in serum, total cell count, eosinophil count, the levels of IL-4, IL-5, IL-13 in BALF and the phosphorylation of EGFR and its downstream activation in lung were higher than those in Group A (all P<0.05). The levels of total IgE in serum [(261.32±44.38) ng/ml, (194.09±52.39) ng/ml vs (1 023.70±105.51) ng/ml], total cell count [(23.70±4.08)×105/ml, (14.92±4.06)×105/ml vs (35.36±6.30)×105/ml], eosinophil count [(108.00±13.69)×104/ml, (67.00±17.28)×104/ml vs (147.86±20.06)×104/ml], IL-4 [(36.42±4.48) pg/ml, (30.45±8.12) pg/ml vs (58.72±7.17) pg/ml], IL-5 [(16.20±4.62) pg/ml, (13.38±5.14) pg/ml vs (23.46±5.38) pg/ml], IL-13 [(18.45±7.28) pg/ml, (14.33±7.70) pg/ml vs (104.12±24.66) pg/ml] in BALF of Group C and D were lower than those in Group B (all P<0.05). The levels of IL-4, IL-5, and IL-13 as well as their mRNA levels in the lung tissue of Group C and D were lower than those in Group B (all P<0.05). In Group C and D, the positive expression rate of phosphorylated epidermal growth factor receptor (p-EGFR) in lung tissue [(40.53±6.80)%, (23.60±4.42)% vs (70.78±5.36)%], p-EGFR/EGFR (61.68±7.48, 51.13±5.19 vs 105.90±11.66), phosphorylated extracellular regulated protein kinase (p-Erk)/extracellular regulated protein kinase (Erk) (75.28±7.11, 47.54±4.83 vs 98.76±4.71), and phosphorylated protein kinase B (p-Akt)/protein kinase B (Akt) (96.24±5.40, 68.52±2.73 vs 103.30±4.52) was lower than those of Group B (all P<0.05). There was no statistically significant difference in the relevant indicators between Group A and E (all P>0.05). Conclusion: Gefitinib may alleviate airway inflammation and airway remodeling in asthmatic mice by inhibiting EGFR phosphorylation and affecting the activation of downstream Erk and Akt.
Sujet(s)
Remodelage des voies aériennes , Asthme , Géfitinib , Souris de lignée C57BL , Animaux , Asthme/traitement médicamenteux , Asthme/métabolisme , Souris , Géfitinib/pharmacologie , Remodelage des voies aériennes/effets des médicaments et des substances chimiques , Mâle , Liquide de lavage bronchoalvéolaire , Inflammation , Interleukine-4/métabolisme , Quinazolines/pharmacologie , Récepteurs ErbB/métabolisme , Ovalbumine , Poumon/métabolisme , Poumon/anatomopathologie , Interleukine-5/métabolisme , Interleukine-13/métabolisme , Granulocytes éosinophiles , Modèles animaux de maladie humaineRÉSUMÉ
The activities, ontogeny, and mechanisms of lineage expansion of eosinophils are less well resolved than those of other immune cells, despite the use of biological therapies targeting the eosinophilia-promoting cytokine interleukin (IL)-5 or its receptor, IL-5Rα. We combined single-cell proteomics and transcriptomics and generated transgenic IL-5Rα reporter mice to revisit eosinophilopoiesis. We reconciled human and murine eosinophilopoiesis and provided extensive cell-surface immunophenotyping and transcriptomes at different stages along the continuum of eosinophil maturation. We used these resources to show that IL-5 promoted eosinophil-lineage expansion via transit amplification, while its deletion or neutralization did not compromise eosinophil maturation. Informed from our resources, we also showed that interferon response factor-8, considered an essential promoter of myelopoiesis, was not intrinsically required for eosinophilopoiesis. This work hence provides resources, methods, and insights for understanding eosinophil ontogeny, the effects of current precision therapeutics, and the regulation of eosinophil development and numbers in health and disease.
Sujet(s)
Lignage cellulaire , Granulocytes éosinophiles , Interleukine-5 , Souris transgéniques , Protéomique , Analyse sur cellule unique , Transcriptome , Granulocytes éosinophiles/immunologie , Granulocytes éosinophiles/métabolisme , Animaux , Interleukine-5/métabolisme , Interleukine-5/génétique , Humains , Souris , Protéomique/méthodes , Analyse sur cellule unique/méthodes , Différenciation cellulaire/immunologie , Souris de lignée C57BL , Analyse de profil d'expression de gènes/méthodes , Sous-unité alpha du récepteur à l'interleukine-5/métabolisme , Sous-unité alpha du récepteur à l'interleukine-5/génétique , Myélopoïèse/génétique , Facteurs de régulation d'interféron/métabolisme , Facteurs de régulation d'interféron/génétique , Souris knockoutRÉSUMÉ
Asthma is a common airway disease associated with allergic inflammation. Environmental factors, such as pollens, pollution, insect-borne antigens, or commercial chemicals, cause this disease. The common symptoms of this airway allergic reaction are increasing mucus, narrowing of the airway wall, coughing, and chest tightness. Medications, such as steroids, alleviate the disease but with severe side effects. Several studies have reported the anti-inflammatory effects of tree-based essential oil components, particularly 3-carene. Therefore, this study used 3-carene to determine if it alleviates asthmatic symptoms in the murine model. First, BALB/c mice were sensitized to an ovalbumin and aluminium hydroxide mixture on day 7th and 14th. From days 21st to 23rd, the mice were challenged with 3-carene and budesonide. The lung trachea, plasma, and bronchiolar lavage fluid (BAL fluid) were collected on day 24. The 3-carene treatment suppressed the cytokine gene expression, such as interleukin-4 (IL-4), IL-5, and IL-13, reducing the lung epithelial cell thickness in the asthmatic model. These results suggest that essential oil 3-carene has an anti-asthmatic effect.
Sujet(s)
Asthme , Monoterpènes bicycliques , Interleukine-13 , Interleukine-4 , Interleukine-5 , Animaux , Femelle , Souris , Antiasthmatiques/pharmacologie , Antiasthmatiques/usage thérapeutique , Anti-inflammatoires/pharmacologie , Anti-inflammatoires/usage thérapeutique , Asthme/traitement médicamenteux , Asthme/anatomopathologie , Liquide de lavage bronchoalvéolaire/cytologie , Liquide de lavage bronchoalvéolaire/immunologie , Modèles animaux de maladie humaine , Interleukine-13/métabolisme , Interleukine-4/métabolisme , Interleukine-5/métabolisme , Poumon/effets des médicaments et des substances chimiques , Poumon/anatomopathologie , Souris de lignée BALB C , Ovalbumine , Monoterpènes bicycliques/pharmacologieRÉSUMÉ
Cytokines regulate immune responses by binding to cell surface receptors, including the common subunit beta (ßc), which mediates signaling for GM-CSF, IL-3, and IL-5. Despite known roles in inflammation, the structural basis of IL-5 receptor activation remains unclear. We present the cryo-EM structure of the human IL-5 ternary receptor complex, revealing architectural principles for IL-5, GM-CSF, and IL-3. In mammalian cell culture, single-molecule imaging confirms hexameric IL-5 complex formation on cell surfaces. Engineered chimeric receptors show that IL-5 signaling, as well as IL-3 and GM-CSF, can occur through receptor heterodimerization, obviating the need for higher-order assemblies of ßc dimers. These findings provide insights into IL-5 and ßc receptor family signaling mechanisms, aiding in the development of therapies for diseases involving deranged ßc signaling.
Sujet(s)
Cryomicroscopie électronique , Facteur de stimulation des colonies de granulocytes et de macrophages , Interleukine-3 , Multimérisation de protéines , Récepteurs à l'interleukine-5 , Transduction du signal , Humains , Sites de fixation , Chaine bêta commune aux récepteurs des cytokines/métabolisme , Chaine bêta commune aux récepteurs des cytokines/génétique , Chaine bêta commune aux récepteurs des cytokines/composition chimique , Facteur de stimulation des colonies de granulocytes et de macrophages/métabolisme , Facteur de stimulation des colonies de granulocytes et de macrophages/composition chimique , Facteur de stimulation des colonies de granulocytes et de macrophages/génétique , Cellules HEK293 , Interleukine-3/métabolisme , Interleukine-3/composition chimique , Interleukine-3/génétique , Interleukine-5/métabolisme , Modèles moléculaires , Liaison aux protéines , Récepteurs à l'interleukine-5/métabolisme , Récepteurs à l'interleukine-5/génétique , Récepteurs à l'interleukine-5/composition chimique , Imagerie de molécules uniques , Relation structure-activitéRÉSUMÉ
Asthma is a heterogeneous disease characterized by chronic airway inflammation, reversible airflow limitation, and airway remodeling. Eosinophil peroxidase (EPX) is the most abundant secondary granule protein unique to activated eosinophils. In this study, we aimed to illustrate the effect of EPX on the epithelial-mesenchymal transition (EMT) in BEAS-2B cells. Our research found that both EPX and ADAM33 were negatively correlated with FEV1/FVC and FEV1%pred, and positively correlated with IL-5 levels. Asthma patients had relatively higher levels of ADAM33 and EPX compared to the healthy control group. The expression of TSLP, TGF-ß1 and ADAM33 in the EPX intervention group was significantly higher. Moreover, EPX could promote the proliferation, migration and EMT of BEAS-2B cells, and the effect of EPX on various factors was significantly improved by the PI3K inhibitor LY294002. The findings from this study could potentially offer a novel therapeutic target for addressing airway remodeling in bronchial asthma, particularly focusing on EMT.
Sujet(s)
Remodelage des voies aériennes , Asthme , Bronches , Eosinophil Peroxidase , Cellules épithéliales , Transition épithélio-mésenchymateuse , Facteur de croissance transformant bêta-1 , Humains , Asthme/métabolisme , Asthme/anatomopathologie , Asthme/physiopathologie , Asthme/immunologie , Mâle , Femelle , Cellules épithéliales/métabolisme , Eosinophil Peroxidase/métabolisme , Facteur de croissance transformant bêta-1/métabolisme , Adulte d'âge moyen , Adulte , Bronches/anatomopathologie , Interleukine-5/métabolisme , 4H-1-Benzopyran-4-ones/pharmacologie , Cytokines/métabolisme , Lignée cellulaire , Lymphopoïétine stromale thymique , Prolifération cellulaire , Mouvement cellulaire , Morpholines/pharmacologie , Protéines ADAMRÉSUMÉ
Two recent articles by the same research group documented that patients with severe eosinophilic asthma exhibit an increased proportion of a subtype of eosinophils, namely CD62Llow inflammatory eosinophils (iEos) and identified an intriguing correlation between such iEos and asthma control scores. Moreover, CD62Llow iEos were reduced after treatment with the anti-IL-5 monoclonal antibody mepolizumab. In the future, we believe that eosinophil subtypes could represent a useful biomarker in severe eosinophilic asthma, helping clinicians characterize patient endotypes and monitoring the response to biological drugs.
Patients with severe eosinophilic asthma (SEA) have an increased proportion of a subtype of eosinophils, CD62Llow inflammatory eosinophils (iEos), which are reduced after mepolizumab treatment. iEos might represent a novel useful biomarker in SEA.
Sujet(s)
Asthme , Granulocytes éosinophiles , Inflammation , Humains , Asthme/traitement médicamenteux , Asthme/immunologie , Asthme/anatomopathologie , Asthme/métabolisme , Granulocytes éosinophiles/métabolisme , Granulocytes éosinophiles/immunologie , Granulocytes éosinophiles/anatomopathologie , Inflammation/anatomopathologie , Marqueurs biologiques/métabolisme , Anticorps monoclonaux humanisés/usage thérapeutique , Éosinophilie/immunologie , Éosinophilie/anatomopathologie , Interleukine-5/métabolisme , Indice de gravité de la maladieRÉSUMÉ
The relationship between allergic inflammation and gut microbiota has been elucidated, and the effect of probiotics on immune disorders has been studied as well. Identifying the role of probiotics in individual diseases and immune responses and selecting and applying specific microorganisms based on these findings can be an effective strategy for using probiotics. Herein, lactobacilli isolated from kimchi were investigated in depth, focusing on their immune regulatory effects and the mechanisms involved. Lactic acid bacteria (LAB) effectively diminished the increased secretion of T helper 2 cytokines, such as IL-4, IL-5, and IL-13, from ovalbumin (OVA)-sensitized mouse splenocytes. The gene expression of GATA3, IL-4, IL-5, IL-9, and IL-13 was confirmed to be regulated by LAB. LAB also suppressed IL-2 production and STAT5 phosphorylation. An IL-10-neutralizing antibody attenuated these effects, indicating that LAB-induced upregulation of IL-10 in antigen-presenting cells was responsible at least partially for the increased IL-2 production and STAT5 phosphorylation in CD4+ T cells. In conclusion, the current study identified one immunomodulatory mechanism that allows LAB to regulate allergic immune reactions and the potential of LAB from kimchi to modulate various immune reactions.
Sujet(s)
Cellules présentatrices d'antigène , Interleukine-10 , Lactobacillus plantarum , Facteur de transcription STAT-5 , Lymphocytes auxiliaires Th2 , Facteur de transcription STAT-5/métabolisme , Animaux , Interleukine-10/métabolisme , Phosphorylation , Souris , Lymphocytes auxiliaires Th2/immunologie , Cellules présentatrices d'antigène/immunologie , Cellules présentatrices d'antigène/métabolisme , Inflammation , Probiotiques/pharmacologie , Souris de lignée BALB C , Aliments fermentés/microbiologie , Interleukine-4/métabolisme , Femelle , Ovalbumine , Rate/immunologie , Rate/métabolisme , Interleukine-5/métabolisme , Cytokines/métabolisme , Interleukine-2/métabolismeRÉSUMÉ
Eosinophil extracellular traps (EETs) are implicated in various eosinophil-associated diseases; however, their role in chronic rhinosinusitis (CRS) remains unclear. In the present study, 57 CRS patients were enrolled, and immunofluorescence was used to analyze EETs in eosinophilic (eCRS) and non-eosinophilic (Non-eCRS) tissues. MSD was used to examine IL-4, IL-5, and IL-13 concentrations in tissue homogenates. Charcot-Leyden crystals (CLCs) protein expression was detected in PMA, PMA+DNase I, and blank control eosinophils using ELISA. Eotaxin-3 mRNA and protein levels were measured in human nasal epithelial cells (HNECs) cultured with EETs, EETs+DNase I, DNase I, and unstimulated eosinophils using PCR and ELISA. EETs were significantly increased in eCRS tissues compared with Non-eCRS (P<0.001), and correlated with VAS and Lund-Mackay CT scores. IL-5 expression was related to EETs formation (r = 0.738, P<0.001). PMA-stimulated eosinophils exhibited higher CLCs protein levels (P<0.01). Co-culturing HNECs with EETs significantly increased eotaxin-3 mRNA and protein levels (P<0.0001, P<0.001) compared with other groups. The study suggests EETs formation is elevated in eCRS patients and is involved in CLCs formation and chemokine secretion, promoting eosinophilic inflammation.
Sujet(s)
Pièges extracellulaires , Rhinite , , Sinusite , Humains , Granulocytes éosinophiles , Chimiokine CCL26/métabolisme , Interleukine-5/génétique , Interleukine-5/métabolisme , Deoxyribonuclease I/métabolisme , ARN messager/métabolismeRÉSUMÉ
AIM: We investigated the relationship between the group 2 innate lymphoid cells (ILC2s)-myeloid-derived suppressor cells (MDSCs) axis and obesity-related breast cancer. METHODS: Fifty-eight patients with breast cancer who had first relapse and metastasis between January 2019 and August 2021 were enrolled. The proportions of ILC2s and MDSCs in blood and the levels of cytokines in serum were detected with flow cytometry. Correlation analysis among clinical characteristics (such as body mass index [BMI]), cytokines, ILC2s, and MDSCs was conducted. RESULTS: There was a significant difference in the proportions of ILC2s and MDSCs between the high BMI group and the normal BMI group (p < .05). In the triple-negative breast cancer (TNBC) patients, the proportions of ILC2s and MDSCs in the obese group were significantly higher than those in the nonobese group (p < .05). In all breast cancer patients, there was a positive correlation between BMI and the ILC2s-MDSCs axis (p < .05). However, there was no correlation observed between the number of metastases, progression-free survival, and the ILC2s-MDSCs axis (p > .05). Additionally, ILC2s showed positive correlations with MDSCs, interleukin-5 (IL-5), IL-10, IL-17A, (PD-L1), programmed cell death 2 ligand 2 (PD-L2), and molecular typing (p < .05). Similarly, MDSCs exhibited positive correlations with IL-5, IL-8, IL-9, IL-17A, PD-L1, and PD-L2 (p < .05). In patients with TNBC, there was a positive correlation between BMI and IL-5 (p < .05). CONCLUSION: Conclusively, obesity may enhance the immunosuppressive effect of the ILC2-MDSC axis in advanced breast cancer. IL-5 may play a vital role in the ILC2-MDSC axis and obesity in TNBC.
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Cellules myéloïdes suppressives , Tumeurs du sein triple-négatives , Humains , Cellules myéloïdes suppressives/métabolisme , Immunité innée , Antigène CD274/métabolisme , Interleukine-17/métabolisme , Interleukine-5/métabolisme , Lymphocytes/métabolisme , Tumeurs du sein triple-négatives/métabolisme , Cytokines/métabolismeRÉSUMÉ
Asthma is recognized as a chronic respiratory illness characterized by airway inflammation and airway hyperresponsiveness. Wogonoside, a flavonoid glycoside, is reported to significantly alleviate the inflammation response and oxidative stress. Herein, this study aimed to investigate the therapeutic effect and underlying mechanism of wogonoside on airway inflammation and mucus hypersecretion in a murine asthma model and in human bronchial epithelial cells (16HBE). BALB/c mice were sensitized and challenged with ovalbumin (OVA). Pulmonary function and the number of cells in the bronchoalveolar lavage fluid (BALF) were examined. Pathological changes in lung tissue in each group were evaluated via hematoxylin and eosin and periodic acid-Schiff staining, and changes in levels of cytokines in BALF and of immunoglobulin E in serum were determined via an enzyme-linked immunosorbent assay. The expression of relevant genes in lung tissue was analyzed via real-time PCR. Western blotting and immunofluorescence were employed to detect the expression of relevant proteins in lung tissue and 16HBE cells. Treatment with 10 and 20 mg/kg wogonoside significantly attenuated the OVA-induced increase of inflammatory cell infiltration, mucus secretion, and goblet cell percentage and improved pulmonary function. Wogonoside treatment reduced the level of T-helper 2 cytokines including interleukin (IL)-4, IL-5, and IL-13 in BALF and of IgE in serum and decreased the mRNA levels of cytokines (IL-4, IL-5, IL-6, IL-13, and IL-1ß and tumor necrosis factor-α), chemokines (CCL-2, CCL-11, and CCL-24), and mucoproteins (MUC5AC, MUC5B, and GOB5) in lung tissues. The expression of MUC5AC and the phosphorylation of STAT6 and NF-κB p65 in lung tissues and 16HBE cells were significantly downregulated after wogonoside treatment. Thus, wogonoside treatment may effectively decrease airway inflammation, airway remodeling, and mucus hypersecretion via blocking NF-κB/STAT6 activation.
Sujet(s)
Asthme , Flavanones , Glucosides , Facteur de transcription NF-kappa B , Humains , Animaux , Souris , Facteur de transcription NF-kappa B/métabolisme , Ovalbumine/effets indésirables , Ovalbumine/métabolisme , Interleukine-13 , Interleukine-5/métabolisme , Interleukine-5/pharmacologie , Interleukine-5/usage thérapeutique , Asthme/induit chimiquement , Asthme/traitement médicamenteux , Asthme/génétique , Poumon/métabolisme , Inflammation/métabolisme , Mucus/métabolisme , Cytokines/génétique , Cytokines/métabolisme , Liquide de lavage bronchoalvéolaire , Souris de lignée BALB C , Modèles animaux de maladie humaine , Facteur de transcription STAT-6/génétique , Facteur de transcription STAT-6/métabolisme , Facteur de transcription STAT-6/pharmacologieRÉSUMÉ
Eosinophils are key effector cells mediating airway inflammation and exacerbation in patients with severe eosinophilic asthma. They are present in increased numbers and activation states in the airway mucosa and lumen. Interleukin-5 (IL-5) is the key eosinophil growth factor that is thought to play a role in eosinophil priming and activation. However, the mechanism of these effects is still not fully understood. The anti-IL-5 antibody mepolizumab reduces eosinophil counts in the airway modestly but has a large beneficial effect on the frequency of exacerbations of severe eosinophilic asthma, suggesting that reduction in eosinophil priming and activation is of central mechanistic importance. In this study, we used the therapeutic effect of mepolizumab and single-cell ribonucleic acid sequencing to investigate the mechanism of eosinophil priming and activation by IL-5. We demonstrated that IL-5 is a dominant driver of eosinophil priming and plays multifaceted roles in eosinophil function. It enhances eosinophil responses to other stimulators of migration, survival, and activation by activating phosphatidylinositol-3-kinases, extracellular signal-regulated kinases, and p38 mitogen-activated protein kinases signaling pathways. It also enhances the pro-fibrotic roles of eosinophils in airway remodeling via transforming growth factor-ß pathway. These findings provide a mechanistic understanding of eosinophil priming in severe eosinophilic asthma and the therapeutic effect of anti-IL-5 approaches in the disease.