RÉSUMÉ
Fine particulate matter (PM2.5) can damage airway epithelial barriers. The anion transport system plays a crucial role in airway epithelial barriers. However, the detrimental effect and mechanism of PM2.5 on the anion transport system are still unclear. In this study, airway epithelial cells and ovalbumin (OVA)-induced asthmatic mice were used. In transwell model, the adenosine triphosphate (ATP)-induced transepithelial anion short-circuit current (Isc) and airway surface liquid (ASL) significantly decreased after PM2.5 exposure. In addition, PM2.5 exposure decreased the expression levels of P2Y2R, CFTR and cytoplasmic free-calcium, but ATP can increase the expressions of these proteins. PM2.5 exposure increased the levels of Th2-related cytokines of bronchoalveolar lavage fluid, lung inflammation, collagen deposition and hyperplasisa of goblet cells. Interestingly, the administration of ATP showed an inhibitory effect on lung inflammation induced by PM2.5. Together, our study reveals that PM2.5 impairs the ATP-induced transepithelial anion Isc through downregulating P2Y2R/CFTR pathway, and this process may participate in aggravating airway hyperresponsiveness and airway inflammation. These findings may provide important insights on PM2.5-mediated airway epithelial injury.
Sujet(s)
Asthme , Protéine CFTR , Matière particulaire , Récepteurs purinergiques P2Y2 , Animaux , Souris , Récepteurs purinergiques P2Y2/métabolisme , Récepteurs purinergiques P2Y2/génétique , Asthme/métabolisme , Asthme/anatomopathologie , Asthme/traitement médicamenteux , Asthme/induit chimiquement , Asthme/immunologie , Matière particulaire/effets indésirables , Matière particulaire/toxicité , Protéine CFTR/métabolisme , Protéine CFTR/génétique , Humains , Adénosine triphosphate/métabolisme , Ovalbumine/immunologie , Transduction du signal/effets des médicaments et des substances chimiques , Cellules épithéliales/effets des médicaments et des substances chimiques , Cellules épithéliales/métabolisme , Régulation négative/effets des médicaments et des substances chimiques , Muqueuse respiratoire/métabolisme , Muqueuse respiratoire/effets des médicaments et des substances chimiques , Muqueuse respiratoire/anatomopathologie , Liquide de lavage bronchoalvéolaire/cytologie , Liquide de lavage bronchoalvéolaire/composition chimique , Liquide de lavage bronchoalvéolaire/immunologieRÉSUMÉ
Natural antibodies are used to compare immune systems across taxa, to study wildlife disease ecology, and as selection markers in livestock breeding. These immunoglobulins are present prior to immune stimulation. They are described as having low antigen specificity or polyreactive binding and are measured by binding to self-antigens or novel exogenous proteins. Most studies use only one or two antigens to measure natural antibodies and ignore potential effects of antigen specificity in analyses. It remains unclear how different antigen-specific natural antibodies are related or how diversity among natural antibodies may affect analyses of these immunoglobulins. Using genetically distinct lines of chickens as a model system, we tested the hypotheses that (1) antigen-specific natural antibodies are independent of each other and (2) antigen specificity affects the comparison of natural antibodies among animals. We used blood cell agglutination and enzyme-linked immunosorbent assays to measure levels of natural antibodies binding to four antigens: (i) rabbit erythrocytes, (ii) keyhole limpet hemocyanin, (iii) phytohemagglutinin, or (iv) ovalbumin. We observed that levels of antigen specific natural antibodies were not correlated. There were significant differences in levels of natural antibodies among lines of chickens, indicating genetic variation for natural antibody production. However, line distinctions were not consistent among antigen specific natural antibodies. These data show that natural antibodies are a pool of relatively distinct immunoglobulins, and that antigen specificity may affect interpretation of natural antibody function and comparative immunology.
Sujet(s)
Poulets , Animaux , Poulets/immunologie , Lapins , Antigènes/immunologie , Érythrocytes/immunologie , Spécificité des anticorps/immunologie , Ovalbumine/immunologie , Anticorps/immunologie , Hémocyanine/immunologie , Phytohémagglutinine/immunologie , Test ELISARÉSUMÉ
BACKGROUND: The prevalence of food allergies is on the rise, posing a significant challenge to public health. Rodents serve as the predominant animal model in food allergy research; yet, the application of rodent models proves to be a laborious and time-consuming endeavor. It is imperative to develop novel in vivo models. METHODS: Ovalbumin (OVA) was administered as the allergen, following the recommended dosage used in other species. During the sensitization phase, a dosage of 0.25â¯mg per 10 tails per 1â¯L was administered twice daily, and during the challenge phase, the dosage was increased to 3 times the initial level. The study explored two dimensions of sensitization: the mode of exposure, which can be either continuous or intermittent, and the duration of exposure, which includes 3 days, 5 days, and 7 days. We examined midgut pathological changes, immunoglobulins contents, and mRNA expressions associated to T helper cells (Th) 2 cytokines following exposure. RESULTS: A significant 109.3â¯% increase in the number of eosinophils was observed in the midgut histopathology following intermittent 5-day OVA exposure, which emerged as the most effective model. OVA exposure increased concentrations of immunoglobulin M (IgM) (105.2â¯%), IgZ (312.1â¯%), and IgD (304.3â¯%) in this model. The mRNA expressions of Th2-related interleukin (IL)-4 and IL-13 were also elevated by 132.8â¯% and 421.0â¯%, respectively. CONCLUSION: The intermittent 5-day OVA exposure was suggested to be the best constructed zebrafish food allergy model, which may be a potential tool for research into food allergies.
Sujet(s)
Modèles animaux de maladie humaine , Hypersensibilité alimentaire , Ovalbumine , Danio zébré , Animaux , Danio zébré/immunologie , Hypersensibilité alimentaire/immunologie , Ovalbumine/immunologie , Lymphocytes auxiliaires Th2/immunologie , Allergènes/immunologie , Cytokines/immunologie , Cytokines/métabolisme , Immunoglobuline E/immunologie , Immunoglobuline E/sangRÉSUMÉ
BACKGROUND: The meticulous selection of appropriate vaccine adjuvants is crucial for optimizing immune responses. Traditionally, pemphigus vulgaris (PV), an autoimmune disorder, has been modelled using complete Freund's adjuvant (CFA). In this study, we aimed to discern potential variations in immune responses elicited by Toll-like receptor (TLR) ligands as compared to CFA. METHODS: A comprehensive investigation was conducted, comparing the effects of these adjuvants in conjunction with ovalbumin or desmoglein-3. Flow cytometry was employed to analyse distinct cell subsets, while enzyme-linked immunosorbent assay quantified antigen-specific antibodies and cytokine levels. Histological examination of harvested skin tissues and transcriptome analysis of skin lesions were performed to identify differentially expressed genes. RESULTS: TLR ligands demonstrated efficacy in inducing PV-like symptoms in wild-type mice, in contrast to CFA. This underscored the substantial impact of the adjuvant on self-antigen tolerance. Furthermore, we proposed an enhanced method for establishing a PV model through adoptive transfer, substituting CFA with TLR ligands. Our results revealed that in contrast to the perception that CFA being the most potent immunopotentiator reported, CFA promoted regulatory T cells (Treg), follicular regulatory T cells and IL-10-producing neutrophils, whereas TLR ligands downregulated CCL17 and IL-10. This suggested potential implications for the recruitment and activation of Treg subsets. While B cell and CD8+ T cell responses exhibited similarity, CFA induced less activation in dendritic cell subsets. A novel mouse model of PV and systemic comparison of immunostimulatory effects of adjuvants were provided by this study. CONCLUSIONS: The systematic comparison of CFA and TLR ligands shed light on the distinctive properties of these adjuvants, presenting innovative mouse models for the investigation of pemphigus. This study significantly contributes to adjuvant research and advances our understanding of PV pathogenesis. KEY POINTS/HIGHLIGHTS: Immunization with desmoglein 3 and Toll-like receptor (TLR) ligands effectively induces pemphigus symptoms in wild-type mice, whereas complete Freund's adjuvant (CFA) fails. TLR ligands heightened the autoreactivity of donor cells in the adoptive transfer pemphigus model. CFA promoted regulatory T cells and IL-10-producing neutrophils, whereas TLR ligands downregulated CCL17 and IL-10, leading to more effective immune responses.
Sujet(s)
Adjuvants immunologiques , Modèles animaux de maladie humaine , Pemphigus , Récepteurs de type Toll , Animaux , Pemphigus/immunologie , Souris , Récepteurs de type Toll/métabolisme , Récepteurs de type Toll/immunologie , Récepteurs de type Toll/agonistes , Adjuvants immunologiques/pharmacologie , Adjuvant Freund/immunologie , Souris de lignée C57BL , Ligands , Ovalbumine/immunologie , FemelleRÉSUMÉ
Yolkin, an egg yolk immunoregulatory protein, stimulates the humoral but inhibits the cellular immune response in adult mice. The aim of this investigation was to evaluate the effects of yolkin administration on the immune response using a model of juvenile, i.e., 28-day- and 37-day-old, mice. We examined the yolkin influence on the magnitude of the cellular immune response, which was determined as contact sensitivity (CS) to oxazolone (OXA), and the humoral immune response, which was determined as the antibody response to ovalbumin (OVA). Yolkin was administered in drinking water, followed by immunization with OXA or OVA. In parallel, the phenotypic changes in the lymphoid organs were determined following yolkin treatment and prior immunization. The results showed that yolkin had a stimulatory effect on CS in the mice treated with yolkin from the 37th day of life but not from the 28th day of life. In contrast, no regulatory effect of yolkin on antibody production was found in 28-day- and 37-day-old mice. Phenotypic studies revealed significant changes in the content of B cells and T cell subpopulations, including CD4+CD25+Foxp3 regulatory T cells. The association between the effects of yolkin on the magnitude of CS and phenotypic changes in main T- and B-cell compartments, as well the importance of changes in T-regulatory and CD8+ cells in the age categories, are discussed. We conclude that the immunoregulatory effects of yolkin on the generation of CS in mice are age dependent and change from stimulation in juvenile to suppression in adult mice.
Sujet(s)
Ovalbumine , Animaux , Souris , Ovalbumine/immunologie , Phénotype , 4-Éthoxyméthylène-2-phényl-oxazol-5(4H)-one , Femelle , Immunité humorale/effets des médicaments et des substances chimiques , Lymphocytes T régulateurs/immunologie , Lymphocytes T régulateurs/effets des médicaments et des substances chimiques , Lymphocytes B/immunologie , Lymphocytes B/effets des médicaments et des substances chimiques , Immunité cellulaire/effets des médicaments et des substances chimiques , Facteurs âges , Eczéma de contact/immunologie , Vieillissement/immunologieRÉSUMÉ
Although oral tolerance is a critical system in regulating allergic disorders, the mechanisms by which dietary factors regulate the induction and maintenance of oral tolerance remain unclear. To address this, we explored the differentiation and function of various immune cells in the intestinal immune system under fasting and ad libitum-fed conditions before oral ovalbumin (OVA) administration. Fasting mitigated OVA-specific Treg expansion, which is essential for oral tolerance induction. This abnormality mainly resulted from functional defects in the CX3CR1+ cells responsible for the uptake of luminal OVA and reduction of tolerogenic CD103+ dendritic cells. Eventually, fasting impaired the preventive effect of oral OVA administration on asthma and allergic rhinitis development. Specific food ingredients, namely carbohydrates and arginine, were indispensable for oral tolerance induction by activating glycolysis and mTOR signaling. Overall, prior food intake and nutritional signals are critical for maintaining immune homeostasis by inducing tolerance to ingested food antigens.
Sujet(s)
Arginine , Cellules dendritiques , Tolérance immunitaire , Ovalbumine , Lymphocytes T régulateurs , Sérine-thréonine kinases TOR , Animaux , Arginine/métabolisme , Lymphocytes T régulateurs/immunologie , Ovalbumine/immunologie , Cellules dendritiques/immunologie , Cellules dendritiques/métabolisme , Souris , Sérine-thréonine kinases TOR/métabolisme , Souris de lignée C57BL , Administration par voie orale , Récepteur-1 de la chimiokine CX3C/métabolisme , Intestins/immunologie , Antigènes CD/métabolisme , Intégrines alpha/métabolisme , Sucres/métabolisme , Glycolyse , Jeûne , Transduction du signal , Muqueuse intestinale/immunologie , Muqueuse intestinale/métabolisme , FemelleRÉSUMÉ
Herein, we report a transdermal patch prepared using an ionic liquid-based solid in oil (IL-S/O) nanodispersion and a pressure-sensitive adhesive (PSA) to deliver the macromolecular antigenic protein, ovalbumin (OVA). The IL-S/O nanodispersion and a PSA were first mixed at an equal weight ratio, then coated onto a release liner, and covered with a support film. To evaluate the effect of the PSA, three types of PSAs, DURO-TAK 87-4098, DURO-TAK 87-4287, and DURO-TAK 87-235A, were used to obtain the corresponding IL-S/O patches SP-4098, SP-4287, and SP-235A, respectively. The prepared IL-S/O patches were characterized for surface morphology, viscoelasticity, and moisture content. In vitro skin penetration and in vivo immunization studies of the IL-S/O patches were performed using Yucatan micropig skin and the C57BL/6NJc1 mice model, respectively. The SP-4098 and SP-4287 delivered 5.49-fold and 5.47-fold higher amounts of drug compared with the aqueous formulation. Although both patches delivered a similar amount of drug, SP-4287 was not detached fully from the release liner after 30 days, indicating low stability. Mice immunized with the OVA-containing SP-4098 produced a 10-fold increase in anti-OVA IgG compared with those treated with an aqueous formulation. These findings suggested that the IL-S/O patch may be a good platform for the transdermal delivery of antigen molecules.
Sujet(s)
Administration par voie cutanée , Antigènes , Immunisation , Liquides ioniques , Ovalbumine , Patch transdermique , Liquides ioniques/composition chimique , Animaux , Souris , Ovalbumine/immunologie , Ovalbumine/administration et posologie , Antigènes/immunologie , Antigènes/administration et posologie , Antigènes/composition chimique , Suidae , Peau/métabolisme , Peau/immunologie , Systèmes de délivrance de médicaments , Souris de lignée C57BL , Femelle , Absorption cutanéeRÉSUMÉ
Tripartite motif (Trim) 31 is important for numerous inflammatory diseases. However, whether Trim31 regulates airway inflammation in asthma remains undetermined. The present work explored the role of Trim31 in airway inflammation in asthmatic mice established by ovalbumin (OVA) stimulation. Trim31 expression was markedly downregulated in the lungs of asthmatic mice. Compared with wild-type (WT) mice, Trim31-/- mice showed more severe pathological changes accompanied by increased inflammatory cell infiltration after OVA induction. House dust mite (HDM) stimulation evoked airway epithelial cell injury and inflammation, which were exacerbated by Trim31 silencing or attenuated by Trim31 overexpression. Further examination revealed that Trim31 deficiency exacerbated the activation of the NLRP3 inflammasome in OVA-induced asthmatic mice and HDM-stimulated airway epithelial cells. The inhibition of NLRP3 markedly diminished the Trim31 silencing-mediated enhancement of HDM-induced injury and inflammation in airway epithelial cells. In conclusion, this work demonstrates that Trim31 acts as a crucial mediator of airway inflammation in asthma. Trim31 deficiency may contribute to the progression of asthma by increasing NLRP3 inflammasome activation, suggesting that Trim31 is a potential therapeutic target for asthma.
Sujet(s)
Asthme , Inflammasomes , Protéine-3 de la famille des NLR contenant un domaine pyrine , Ovalbumine , Protéines à motif tripartite , Ubiquitin-protein ligases , Animaux , Femelle , Humains , Souris , Asthme/immunologie , Asthme/métabolisme , Modèles animaux de maladie humaine , Cellules épithéliales/immunologie , Cellules épithéliales/métabolisme , Inflammasomes/métabolisme , Poumon/anatomopathologie , Poumon/immunologie , Souris de lignée C57BL , Souris knockout , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Ovalbumine/immunologie , Pyroglyphidae/immunologie , Protéines à motif tripartite/génétique , Protéines à motif tripartite/métabolisme , Ubiquitin-protein ligases/génétique , Ubiquitin-protein ligases/déficit , Ubiquitin-protein ligases/métabolismeRÉSUMÉ
Asthma is a widely prevalent chronic disease that brings great suffering to patients and may result in death if it turns severe. Jolkinolide B (JB) is one diterpenoid component separated from the dried roots of Euphorbia fischeriana Steud (Euphorbiaceae), and has anti--inflammatory, antioxidative, and antitumor properties. However, the detailed regulatory role and associated regulatory mechanism in the progression of asthma remain elusive. In this work, it was demonstrated that the extensive infiltration of bronchial inflammatory cells and the thickening of airway wall were observed in ovalbumin (OVA)-induced mice, but these impacts were reversed by JB (10 mg/kg) treatment, indicating that JB relieved the provocative symptoms in OVA-induced asthma mice. In addition, JB can control OVA-triggered lung function and pulmonary resistance. Moreover, JB attenuated OVA-evoked inflammation by lowering the levels of interleukin (IL)-4, IL-5, and IL-13. Besides, the activated nuclear factor kappa B (NF-κB) and transforming growth factor-beta-mothers against decapentaplegic homolog 3 (TGFß/smad3) pathways in OVA-induced mice are rescued by JB treatment. In conclusion, it was disclosed that JB reduced allergic airway inflammation and airway remodeling in asthmatic mice by modulating the NF-κB and TGFß/smad3 pathways. This work could offer new opinions on JB for lessening progression of asthma.
Sujet(s)
Remodelage des voies aériennes , Asthme , Modèles animaux de maladie humaine , Diterpènes , Souris de lignée BALB C , Facteur de transcription NF-kappa B , Ovalbumine , Animaux , Asthme/traitement médicamenteux , Asthme/immunologie , Remodelage des voies aériennes/effets des médicaments et des substances chimiques , Souris , Diterpènes/pharmacologie , Diterpènes/administration et posologie , Diterpènes/usage thérapeutique , Ovalbumine/immunologie , Facteur de transcription NF-kappa B/métabolisme , Femelle , Facteur de croissance transformant bêta/métabolisme , Cytokines/métabolisme , Protéine Smad-3/métabolisme , Transduction du signal/effets des médicaments et des substances chimiques , Humains , Anti-inflammatoires/pharmacologie , Anti-inflammatoires/usage thérapeutique , Antiasthmatiques/pharmacologie , Antiasthmatiques/usage thérapeutique , Euphorbia/composition chimiqueRÉSUMÉ
Studies have demonstrated that prior to puberty, girls have a lower incidence and severity of asthma symptoms compared to boys. This study aimed to explore the role of progesterone (P4), a sex hormone, in reducing inflammation and altering the immune microenvironment in a mouse model of allergic asthma induced by OVA. Female BALB/c mice with or without ovariectomy to remove the influence of sex hormones were used for the investigations. Serum, bronchoalveolar lavage fluid (BALF), and lung tissue samples were collected for analysis. The results indicated that P4 treatment was effective in decreasing inflammation and mucus secretion in the lungs of OVA-induced allergic asthma mice. P4 treatment also reduced the influx of inflammatory cells into the BALF and increased the levels of Th1 and Th17 cytokines while decreasing the levels of Th2 and Treg cytokines in both BALF and lung microenvironment CD45+ T cells. Furthermore, P4 inhibited the infiltration of inflammatory cells into the lungs, suppressed NETosis, and reduced the number of pulmonary CD4+ T cells while increasing the number of regulatory T cells. The neutrophil elastase inhibitor GW311616A also suppressed airway inflammation and mucus production and modified the secretion of immune Th1, Th2, Th17, and Treg cytokines in lung CD45+ immune cells. These changes led to an alteration of the immunological milieu with increased Th1 and Th17 cells, accompanied by decreased Th2, Treg, and CD44+ T cells, similar to the effects of P4 treatment. Treatment with P4 inhibited NETosis by suppressing the p38 pathway activation, leading to reduced reactive oxygen species production. Moreover, P4 treatment hindered the release of double-stranded DNA during NETosis, thereby influencing the immune microenvironment in the lungs. These findings suggest that P4 treatment may be beneficial in reducing inflammation associated with allergic asthma by modulating the immune microenvironment. In conclusion, this research indicates the potential of P4 as a therapeutic agent for ameliorating inflammation in OVA-induced allergic asthma mice.
Sujet(s)
Asthme , Ovalbumine , Progestérone , Animaux , Femelle , Souris , Asthme/immunologie , Asthme/traitement médicamenteux , Asthme/métabolisme , Liquide de lavage bronchoalvéolaire , Microenvironnement cellulaire/effets des médicaments et des substances chimiques , Cytokines/métabolisme , Modèles animaux de maladie humaine , Pièges extracellulaires/effets des médicaments et des substances chimiques , Pièges extracellulaires/métabolisme , Pièges extracellulaires/immunologie , Inflammation/traitement médicamenteux , Inflammation/immunologie , Inflammation/métabolisme , Poumon/immunologie , Poumon/anatomopathologie , Poumon/effets des médicaments et des substances chimiques , Poumon/métabolisme , Souris de lignée BALB C , Ovalbumine/immunologie , Progestérone/pharmacologie , Cellules Th17/immunologie , Cellules Th17/effets des médicaments et des substances chimiques , Cellules Th17/métabolismeRÉSUMÉ
Airway epithelial-mesenchymal transition (EMT) is the important pathological feature of airway remodeling in asthma. While macrolides are not commonly used to treat asthma, they have been shown to have protective effects on the airways, in which mechanisms are not yet fully understood. This study aims to investigate the impact of clarithromycin on airway EMT in asthma and its potential mechanism. The results revealed an increase in Kv1.3 expression in the airways of ovalbumin (OVA)-induced asthmatic mice, with symptoms and pathological changes being alleviated after treatment with the Kv1.3 inhibitor 5-(4-phenoxybutoxy)psoralen (PAP-1). Clarithromycin was found to attenuate airway epithelial-mesenchymal transition through the inhibition of Kv1.3 and PI3K/Akt signaling. Further experiments in vitro confirmed that PAP-1 could mitigate EMT by modulating the PI3K/Akt signaling in airway epithelial cells undergoing transformation into mesenchymal cells. These findings confirmed that clarithromycin might have a certain protective effect on asthma-related airway remodeling and represent a promising treatment strategy.
Sujet(s)
Remodelage des voies aériennes , Asthme , Clarithromycine , Transition épithélio-mésenchymateuse , Canal potassique Kv1.3 , Souris de lignée BALB C , Ovalbumine , Phosphatidylinositol 3-kinases , Protéines proto-oncogènes c-akt , Transduction du signal , Animaux , Transition épithélio-mésenchymateuse/effets des médicaments et des substances chimiques , Asthme/traitement médicamenteux , Asthme/métabolisme , Asthme/induit chimiquement , Asthme/anatomopathologie , Ovalbumine/immunologie , Clarithromycine/pharmacologie , Clarithromycine/usage thérapeutique , Protéines proto-oncogènes c-akt/métabolisme , Transduction du signal/effets des médicaments et des substances chimiques , Phosphatidylinositol 3-kinases/métabolisme , Souris , Canal potassique Kv1.3/métabolisme , Canal potassique Kv1.3/antagonistes et inhibiteurs , Remodelage des voies aériennes/effets des médicaments et des substances chimiques , Humains , Modèles animaux de maladie humaine , Femelle , Psoralène/pharmacologie , Psoralène/usage thérapeutique , Cellules épithéliales/effets des médicaments et des substances chimiquesRÉSUMÉ
PURPOSE: The etiology of asthma remains elusive, with no known cure. Based on accumulating evidence, autophagy, a self-degradation process that maintains cellular metabolism and homeostasis, participates in the development of asthma. Mycobacterium vaccae vaccine (M. vaccae), an immunomodulatory agent, has previously been shown to effectively alleviate airway inflammation and airway remodeling. However, its therapeutic effect on asthma via the regulation of autophagy remains unknown. Therefore, this study aimed to investigate the impact of M. vaccae in attenuating asthma airway inflammation via autophagy-mediated pathways. METHODS: Balb/c mice were used to generate an ovalbumin (OVA)-immunized allergic airway model and were subsequently administered either M. vaccae or M. vaccae + rapamycin (an autophagy activator) prior to each challenge. Next, airway inflammation, mucus secretion, and airway remodeling in mouse lung tissue were assessed via histological analyses. Lastly, the expression level of autophagy proteins LC3B, Beclin1, p62, and autolysosome was determined both in vivo and in vitro, along with the expression level of p-PI3K, PI3K, p-Akt, and Akt in mouse lung tissue. RESULTS: The findings indicated that aerosol inhalation of M. vaccae in an asthma mouse model has the potential to decrease eosinophil counts, alleviate airway inflammation, mucus secretion, and airway remodeling through the inhibition of autophagy. Likewise, M. vaccae could reduce the levels of OVA-specific lgE, IL-5, IL-13, and TNF-α in asthma mouse models by inhibiting autophagy. Furthermore, this study revealed that M. vaccae also suppressed autophagy in IL-13-stimulated BEAS-2B cells. Moreover, M. vaccae may activate the PI3K/Akt signaling pathway in the lung tissue of asthmatic mice. CONCLUSION: In summary, the present study suggests that M. vaccae may contribute to alleviating airway inflammation and remodeling in allergic asthma by potentially modulating autophagy and the PI3K/Akt signaling pathway. These discoveries offer a promising avenue for the development of therapeutic interventions targeting allergic airway inflammation.
Sujet(s)
Asthme , Autophagie , Modèles animaux de maladie humaine , Inflammation , Souris de lignée BALB C , Mycobacteriaceae , Ovalbumine , Phosphatidylinositol 3-kinases , Protéines proto-oncogènes c-akt , Transduction du signal , Animaux , Protéines proto-oncogènes c-akt/métabolisme , Protéines proto-oncogènes c-akt/immunologie , Phosphatidylinositol 3-kinases/métabolisme , Ovalbumine/immunologie , Asthme/immunologie , Asthme/thérapie , Souris , Inflammation/immunologie , Mycobacteriaceae/immunologie , Femelle , Remodelage des voies aériennes/immunologie , Poumon/anatomopathologie , Poumon/immunologie , Vaccins antibactériens/immunologieRÉSUMÉ
BACKGROUND: The treatment of food allergy (FA) needs improvement. The treatment of immune disorders can be improved by regulating epigenetic marks, which is a promising method. The objective of this research is to alleviate experimental FA by employing an inhibitor of DNA methyltransferase-1 (DNMT1). METHODS: Ovalbumin was used as the specific antigen to establish a mouse model of FA. Intestinal IL-35+ regulatory B cells (Breg cells) were isolated from FA mice, and characterized using immunological approaches. RESULTS: FA mice had a lower frequency of IL-35+ Breg cells, which was inversely correlated with their FA response. The quantity of IL-35 was lower in intestinal Breg cells from FA mice. Hypermethylation status was detected in the Il35 promoter, which was accompanied with high levels of H3K9me3. Enforced expression of DNMT1 hindered the promoter activity of the IL35 gene. Administration of an inhibitor of DNMT1 (RG108) restored the immune regulatory capacity of FA intestinal Bregs, and effectively suppressed the expression of DNMT1, and attenuated experimental FA. CONCLUSIONS: The elevated quantity of DNMT1 in intestinal Breg cells compromises the expression of IL-35 and affects the immune regulatory functions, which facilitates the development of FA. The immune regulatory functions of intestinal Breg cells are restored and experimental FA is attenuated by inhibiting DNMT1.
Sujet(s)
DNA (Cytosine-5-)-methyltransferase 1 , Méthylation de l'ADN , Hypersensibilité alimentaire , Interleukines , Régions promotrices (génétique) , Animaux , DNA (Cytosine-5-)-methyltransferase 1/métabolisme , DNA (Cytosine-5-)-methyltransferase 1/génétique , Hypersensibilité alimentaire/immunologie , Souris , Interleukines/immunologie , Interleukines/métabolisme , Interleukines/génétique , Régions promotrices (génétique)/génétique , Lymphocytes B régulateurs/immunologie , Modèles animaux de maladie humaine , Souris de lignée BALB C , Femelle , Ovalbumine/immunologie , Phtalimides/pharmacologie , Intestins/immunologie , Tryptophane/analogues et dérivésRÉSUMÉ
Micro- and nanoparticles delivery systems have been widely studied as vaccine adjuvants to enhance immunogenicity and sustain long-term immune responses. Polygonatum sibiricum polysaccharide (PSP) has been widely studied as an immunoregulator in improving immune responses. In this study, we synthesized and characterized cationic modified calcium carbonate (CaCO3) microparticles loaded with PSP (PEI-PSP-CaCO3, CTAB-PSP-CaCO3), studied the immune responses elicited by PEI-PSP-CaCO3 and CTAB-PSP-CaCO3 carrying ovalbumin (OVA). Our results demonstrated that PEI-PSP-CaCO3 significantly enhanced the secretion of IgG and cytokines (IL-4, IL-6, IFN-γ, and TNF-α) in vaccinated mice. Additionally, PEI-PSP-CaCO3 induced the activation of dendritic cells (DCs), T cells, and germinal center (GC) B cells in draining lymph nodes (dLNs). It also enhanced lymphocyte proliferation, increased the ratio of CD4+/CD8+ T cells, and elevated the frequency of CD3+ CD69+ T cells in spleen lymphocytes. Therefore, PEI-PSP-CaCO3 microparticles induced a stronger cellular and humoral immune response and could be potentially useful as a vaccine delivery and adjuvant system.
Sujet(s)
Carbonate de calcium , Cellules dendritiques , Polygonatum , Polyosides , Animaux , Souris , Carbonate de calcium/composition chimique , Polygonatum/composition chimique , Polyosides/composition chimique , Cellules dendritiques/immunologie , Cellules dendritiques/effets des médicaments et des substances chimiques , Femelle , Adjuvants vaccinaux/composition chimique , Ovalbumine/immunologie , Ovalbumine/administration et posologie , Cytokines/métabolisme , Souris de lignée BALB C , Adjuvants immunologiques/composition chimique , Adjuvants immunologiques/pharmacologie , Adjuvants immunologiques/administration et posologie , Immunoglobuline G/immunologie , Immunoglobuline G/sang , Nanoparticules/composition chimiqueRÉSUMÉ
BACKGROUND: The incidence rate of allergic rhinitis (AR) is on the rise, which seriously affects the quality of life, work efficiency, mental state of patients, and sleeping in AR sleep. This experiment aimed to investigate the changes in Treg/Th17 cells in the nasal mucosa of an AR mouse model and the intervention effect of an Anti-IL-17 antibody. METHODS: A mouse model of AR was induced by intraperitoneal ovalbumin (OVA) injection for sensitization and stimulation with nasal drops. The times of rubbing, sneezing, and symptomatology scores were counted and analyzed. Pathological damage to the nasal mucosa was observed by Hematoxylin-Eosin (HE) staining. Peripheral blood CD4+CD25+CD127- Treg cell levels and the content of Th17 cells were measured by flow cytometry (FCM). ELISA kits were used to detect the levels of relevant cytokines (IL-10 and TGF-ß1) secreted by Treg cells. The intervention effect of Anti-IL-17 antibody was observed by giving Anti-IL-17 antibody treatment. RESULTS: The times of rubbing, sneezing, and symptomatology scores were significantly higher in mice in the OVA group than in the Control group (p < 0.001). The percentage of CD4+CD25+CD127- Treg cells in CD4+CD25+ T cells (p < 0.05) and the levels of IL-10 (p < 0.001) and TGF-ß1 (p < 0.001) were significantly decreased. After OVA induction, the continuity of the nasal mucosa of mice was interrupted, the percentage of Th17 cells, IL-17, and IL-4 levels were increased, and IFN-γ levels were significantly reduced (p < 0.001). And protein expression of RORγt was significantly upregulated (p < 0.001). In addition, all of these results were reversed by Anti-IL-17 antibody treatment, significantly improving AR-related symptoms (p < 0.05). CONCLUSION: Anti-IL-17 antibodies may regulate the body's immune response by promoting CD4+CD25+CD127- Treg cell differentiation, thereby ameliorating the symptoms associated with AR.
Sujet(s)
Modèles animaux de maladie humaine , Interleukine-17 , Souris de lignée BALB C , Muqueuse nasale , Rhinite allergique , Lymphocytes T régulateurs , Cellules Th17 , Animaux , Lymphocytes T régulateurs/immunologie , Muqueuse nasale/immunologie , Muqueuse nasale/anatomopathologie , Rhinite allergique/immunologie , Rhinite allergique/anatomopathologie , Rhinite allergique/sang , Cellules Th17/immunologie , Souris , Interleukine-17/immunologie , Interleukine-17/métabolisme , Femelle , Ovalbumine/immunologie , Facteur de croissance transformant bêta-1/métabolisme , Facteur de croissance transformant bêta-1/immunologie , Interleukine-10/immunologie , Interleukine-10/métabolisme , Anticorps/immunologieRÉSUMÉ
Lipid vesicles are widely used for drug and gene delivery, but their structural instability reduces in vivo efficacy and requires specialized handling. To address these limitations, strategies like lipid cross-linking and polymer-lipid conjugation are suggested to enhance stability and biological efficacy. However, the in vivo metabolism of these altered lipids remains unclear, necessitating further studies. A new stabilization technique without chemical modification is urgently needed. Here, a bio-mimetic approach for fabricating robust multilamellar lipid vesicles to enhance in vivo delivery and stabilization of protein antigens is presented. This method leverages 1-O-acylceramide, a natural skin lipid, to facilitate the self-assembly of lipid nanovesicles. Incorporating 1-O-acylceramide, anchoring lipid bilayers akin to its role in the stratum corneum, provides excellent stability under environmental stresses, including freeze-thaw cycles. Encapsulating ovalbumin as a model antigen and the adjuvant monophosphoryl lipid A demonstrates the vesicle's potential as a nanovaccine platform. In vitro studies show enhanced immune responses with both unilamellar and multilamellar vesicles, but in vivo analyses highlight the superior efficiency of multilamellar vesicles in inducing higher antibody and cytokine levels. This work suggests ceramide-induced multilamellar lipid vesicles as an effective nanovaccine platform for enhanced antigen delivery and stability.
Sujet(s)
Ovalbumine , Animaux , Souris , Ovalbumine/composition chimique , Ovalbumine/immunologie , Matériaux biomimétiques/composition chimique , Matériaux biomimétiques/pharmacologie , Vaccination/méthodes , Lipide A/composition chimique , Lipide A/analogues et dérivés , Vaccins/composition chimique , Vaccins/immunologie , Céramides/composition chimique , Lipides/composition chimique , Nanoparticules/composition chimique , Femelle , Souris de lignée C57BLRÉSUMÉ
BACKGROUND: For decades, studies have demonstrated the anti-inflammatory potential of proteins secreted by helminths in allergies and asthma. Previous studies have demonstrated the immunomodulatory capabilities of Succinate Coenzyme A ligase beta-like protein (SUCLA-ß) derived from Trichinella spiralis, a crucial excretory product of this parasite. OBJECTIVE: To explore the therapeutic potential of SUCLA-ß in alleviating and controlling ovalbumin (OVA)-induced allergic asthma, as well as its influence on host immune modulation. METHODS: In this research, we utilized the rTs-SUCLA-ß protein derived from T. spiralis to investigate its potential in mitigating airway inflammation in a murine model of asthma induced by OVA sensitization/stimulation, both pre- and post-challenge. The treatment's efficacy was assessed by quantifying the extent of inflammation in the lungs. RESULTS: Treatment with rTs-SUCLA-ß demonstrated efficacy in ameliorating OVA-induced airway inflammation, as evidenced by a reduction in eosinophil infiltration, levels of OVA-specific Immunoglobulin E, interferon-γ, interleukin (IL)-9, and IL-17A, along with an elevation in IL-10. The equilibrium between Th17 and Treg cells plays a pivotal role in modulating the abundance of inflammatory cells within the organism, thereby ameliorating inflammation and alleviating symptoms associated with allergic asthma. CONCLUSIONS AND CLINICAL RELEVANCE: Our data revealed that T. spiralis-derived Ts-SUCLA-ß protein may inhibit the allergic airway inflammation by regulating host immune responses.
Sujet(s)
Asthme , Protéines d'helminthes , Ovalbumine , Trichinella spiralis , Trichinella spiralis/immunologie , Animaux , Asthme/immunologie , Asthme/traitement médicamenteux , Souris , Ovalbumine/immunologie , Protéines d'helminthes/immunologie , Protéines d'helminthes/pharmacologie , Souris de lignée BALB C , Modèles animaux de maladie humaine , Femelle , Cytokines/métabolisme , Cytokines/immunologie , Immunoglobuline E/immunologie , Poumon/immunologie , Poumon/parasitologie , Poumon/anatomopathologie , Lymphocytes T régulateurs/immunologie , Hypersensibilité/immunologie , Hypersensibilité/traitement médicamenteux , Cellules Th17/immunologieRÉSUMÉ
Epithelial barrier dysfunction plays an important role in the pathogenesis of Th2 bias. The mechanism requires further clarification. NEMO is associated with regulating apoptotic activities in the cell. The purpose of this study is to investigate the role of insufficient Nemo signals in developing Th2 bias in the respiratory tract. Nemof/fEpcam-Cre mice (A mouse strain carrying NEMO-deficient epithelial cells. NemoKO mice, in short) was generated. An airway Th2 bias mouse model was established with the ovalbumin/alum protocol. The NemoKO mice exhibited spontaneous airway Th2 bias. Respiratory tract epithelial barrier integrity was compromised in NemoKO mice. Apoptosis was found in approximately 10% of the epithelial cells of the respiratory tract in NemoKO mice. The reconstruction of the Nemo expression restored homeostasis within the epithelial barrier of the airways. Restoration of Nemo gene expression in epithelial cells by Nemo mRNA vaccination alleviated Th2 bias in mice with airway allergy. To sum up, NEMO plays an important role in maintaining the integrity of the epithelial barrier in the respiratory tract. Administration of NEMO mRNA vaccines can restore epithelial barrier functions and alleviate Th2 bias in the airways.
Sujet(s)
ARN messager , Lymphocytes auxiliaires Th2 , Animaux , Souris , ARN messager/métabolisme , ARN messager/génétique , Lymphocytes auxiliaires Th2/immunologie , Lymphocytes auxiliaires Th2/métabolisme , Vaccination , Protéines et peptides de signalisation intracellulaire/métabolisme , Protéines et peptides de signalisation intracellulaire/génétique , Ovalbumine/immunologie , Cellules épithéliales/métabolisme , Apoptose , Souris de lignée C57BL , Hypersensibilité respiratoire , Hypersensibilité , Muqueuse respiratoire/métabolisme , Muqueuse respiratoire/immunologie , Modèles animaux de maladie humaine , Souris knockoutRÉSUMÉ
BACKGROUND: Allergic Rhinitis (AR) is a prevalent chronic non-infectious inflammation affecting the nasal mucosa. NLRP3-mediated pyroptosis of epithelial cells plays a pivotal role in AR pathogenesis. Herein, we evaluated the impact of the long non-coding RNA nuclear paraspeckle assembly transcript 1 (lncRNA NEAT1) on NLR family pyrin domain containing 3 (NLRP3)-mediated pyroptosis in AR. METHODS: Nasal inflammation levels in ovalbumin (OVA)-induced AR mice were assessed using HE staining, and NLRP3 expression was evaluated through immunohistochemistry. ELISA was utilized to detect OVA-specific IgE, IL-6, IL-5, and inflammatory cytokines (IL-1ß, IL-18). Human nasal epithelial cells (HNEpCs) stimulated with IL4/IL13 were used to analyze the mRNA and protein levels of associated genes utilizing RT-qPCR and western blot, respectively. Cell viability and pyroptosis were assessed by CCK-8 and flow cytometry. The targeting relationship between NEAT1, PTBP1 and FOXP1 were analyzed by RIP and RNA pull down assays. FISH and IF analysis were performed to assess the co-localization of NEAT1 and PTBP1. RESULTS: In both the AR mouse and cellular models, increased levels of NEAT1, PTBP1 and FOXP1 were observed. AR mice exhibited elevated inflammatory infiltration and pyroptosis, evidenced by enhanced expressions of OVA-specific IgE, IL-6, and IL-5, NLRP3, Cleaved-caspase 1, GSDMD-N, IL-1ß and IL-18. Functional assays revealed that knockdown of PTBP1 or NEAT1 inhibited pyroptosis while promoting the proliferation of IL4/IL13-treated HNEpCs. Mechanistically, NEAT1 directly interacted with PTBP1, thereby maintaining FOXP1 mRNA stability. Rescue assays demonstrated that FOXP1 upregulation reversed the inhibitory effects of silencing NEAT1 or PTBP1 on IL4/IL13-stimulated pyroptosis activation in HNEpCs. CONCLUSION: NEAT1 acts as a RNA scaffold for PTBP1, activating the PTBP1/FOXP1 signaling cascade, subsequently triggering NLRP3-mediated pyroptosis in HNEpCs, and ultimately promoting AR progression. These findings highlight some new insights into the pathogenesis of AR.
Sujet(s)
Facteurs de transcription Forkhead , Protéine-3 de la famille des NLR contenant un domaine pyrine , Muqueuse nasale , Pyroptose , ARN long non codant , Rhinite allergique , Animaux , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , ARN long non codant/génétique , ARN long non codant/métabolisme , Rhinite allergique/immunologie , Rhinite allergique/anatomopathologie , Rhinite allergique/génétique , Rhinite allergique/métabolisme , Humains , Souris , Facteurs de transcription Forkhead/métabolisme , Facteurs de transcription Forkhead/génétique , Muqueuse nasale/immunologie , Muqueuse nasale/anatomopathologie , Muqueuse nasale/métabolisme , Souris de lignée BALB C , Ovalbumine/immunologie , Ribonucléoprotéines nucléaires hétérogènes/métabolisme , Ribonucléoprotéines nucléaires hétérogènes/génétique , Transduction du signal , Modèles animaux de maladie humaine , Femelle , Cytokines/métabolismeRÉSUMÉ
Self-assembling protein nanoparticles are beneficial platforms for enhancing the often weak and short-lived immune responses elicited by subunit vaccines. Their benefits include multivalency, similar sizes as pathogens and control of antigen orientation. Previously, the design, preparation, and characterization of self-assembling protein vesicles presenting fluorescent proteins and enzymes on the outer vesicle surface have been reported. Here, a full-size model antigen protein, ovalbumin (OVA), was genetically fused to the recombinant vesicle building blocks and incorporated into protein vesicles via self-assembly. Characterization of OVA protein vesicles showed room temperature stability and tunable size. Immunization of mice with OVA protein vesicles induced strong antigen-specific humoral and cellular immune responses. This work demonstrates the potential of protein vesicles as a modular platform for delivering full-size antigen proteins that can be extended to pathogen antigens to induce antigen specific immune responses.