Search

Interleukin-27 receptor (IL-27R) is expressed in a variety of immune cells and structural cells, including dendritic cells. The mechanism of IL-27 in asthma has not been fully elucidated. This study aimed to examine whether IL-27 regulated the CD39/ATP axis of dendritic cells in asthma. Our results showed that in ovalbumin (OVA)-induced asthma mouse model, IL-27Rα -/- asthmatic mice showed increased airway resistance, increased infiltration of inflammatory cells in lung tissue, proliferation of goblet cells, enhanced expression of Muc5 AC around airway epithelium, increased total number of cells and eosinophils, increased levels of total IgE, OVA-IgE, IL-4, IL-5, IL-13 and IL-17 A, and increased expression of transcription factors GATA-3 and RORγt in lung tissue. The expression of CD39 mRNA and protein in the lung tissue of IL-27Rα -/- asthmatic mice decreased, and the expression of NLRP3, ASC and Caspase-1 in NLRP3 inflammasome components increased. The concentration of ATP was significantly increased compared with WT asthmatic mice. In vitro experiments showed that the expression of CD39 in lung dendritic cells of IL-27Rα -/- asthmatic mice decreased, while the expression of NLRP3 inflammasome components NLRP3, ASC and Caspase-1 increased. These findings indicate that IL-27 directly and indirectly regulates immunoinflammatory responses in asthma by acting on dendritic cells CD39/ATP Axis., (© 2023. The Author(s).)

Disruption of T-cell differentiation is characteristic of airway inflammation in allergic asthma. How miR-19a works in asthma has not been completely elucidated. This study aimed to examine whether microRNA-19a regulates helper T-cell proliferation and to identify the factors involved and the underlying mechanisms. Our results showed that miR-19a levels were upregulated in parallel with a reduction in RUNX3 expression in a house dust mite (HDM)-induced murine model of asthma. A dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay showed that RUNX3 was a direct target of miR-19a. Inhibiting the expression of miR-19a attenuated inflammation and mucus production, induced Th1 cells, suppressed the Th2 inflammatory response, and repressed dendritic cell (DC) maturation by increasing RUNX3 expression in WT asthmatic mice but not RUNX3 +/- mice. In vitro experiments revealed that miR-19a inhibition could target RUNX3 to induce Th1 polarization and inhibit Th2 polarization by directly acting on naïve CD4 + T cells or indirectly mediating the maturation and antigen-presenting abilities of DCs. These findings indicate that miR-19a directly and indirectly regulates immunoinflammatory responses in asthma by targeting RUNX3., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Eosinophilic asthma accounts for 40% to 50% of asthmatic patients. However, 5% to 10% of patients with asthma need high-dose drug control, which is clinically referred to as severe asthma patients. Interleukin (IL)-5 plays an important role in the proliferation, maturation, and migration of eosinophils. Benralizumab, as an antagonist of the IL-5 receptor, can treat eosinophilic asthma by promoting the apoptosis of eosinophils. The implications for efficacy and/or adverse events are unclear. This article reviews the findings about benralizumab in the treatment of severe eosinophilic asthma in recent years. Results indicated the effectiveness of benralizumab for the treatment of severe asthma., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

The aim of this assessment was to determine the information needs, resources, and resource preferences and self/family-management challenges of parents caring for a child with asthma in China. The acceptance of asthma and consolidated framework for implementation research guided the study. A cross-sectional, descriptive design was used. Seventy-one parents of a child receiving care in an outpatient respiratory clinic completed a three-part, pragmatic, self-report survey. Quantitative data were analyzed using descriptive statistics and qualitative data using thematic analysis. Parents, who needed more knowledge about asthma, sought information from internet sources. However, most parents preferred receiving information during in-person consultation with trained specialists. Management challenges revolved around understanding asthma information, formulating beliefs about asthma, experiencing distressing thoughts and feelings, forming supportive networks, and meeting their child's emotional needs. Evidence supports expanding nursing roles in China to include extended time for initial in-person parental interactions and follow-up using reliable clinic-based internet counseling.

Asthma is a common airway inflammation with an intricate underlying mechanism. The role played by circulating miRNAs in asthma remains unclear. In the present study, we aimed to investigate the role of miR-223-3p in leukocytes of asthma and identify the relationship between miR-223-3p and inflammatory cytokines in asthma. Using real-time polymerase chain reaction (RT-PCR), we detected miR-223-3p expression in peripheral blood leukocytes from 23 asthmatic patients and 20 healthy controls. The levels of IFN-γ (Th1 cytokine), IL-4 (Th2 cytokine), IL-17A (Th17 cytokine) in plasma were examined using enzyme-linked immunosorbent assay (ELISA). Analysis of variance (ANOVA) and Spearman's test was used for statistical analysis. The expression of miR-223-3p in peripheral blood leukocytes was upregulated in the asthmatic patients compared with that in the healthy controls. Increased miR-223-3p expression was associated with forced expiratory volume in 1-second percent predicted (FEV1% predicted). A positive correlation was noted between miR-223-3p and IL-17A. The findings of this study showed that miR-223-3p plays a vital role in the pathogenesis of asthma and can serve as a novel biomarker for asthma.

Background: Neutrophilic subtype asthma occurs in approximately 15-25% of the asthma cases and is associated with severe airflow obstruction, corticosteroid resistance. MicroRNA plays a vital role in regulating many immune processes, but how miRNA circuits coordinate airway inflammation during neutrophilic asthma is unclear., Methods: To investigate the molecular mechanism of miR-223 in regulation of neutrophilic airway inflammation, miR-223 knockout mice were used to the OVA/CFA-induced neutrophilic asthma or treated with NLRP3 inhibitor and IL-1β receptor antagonist. Based on the results obtained, wide-type mice were subsequently treated with miR-223 agomirs or negative control agomirs, and the effects on airway inflammation were assessed using morphometric techniques, quantitative RT-PCR, western blot, ELISA and other molecular approaches., Results: The expression of miR-223 was upregulated in lung tissues of experimental mice model. Furthermore, miR-223 -/- mice led to aggravated neutrophilic airway inflammation with heightened histopathological, inflammatory cells and cytokines readouts. Moreover, miR-223 -/- mice also presented with enhanced NLRP3 inflammasome level with elevated IL-1β. Blocking NLRP3 or IL-1β diminished this phenotype. Finally, overexpression of miR-223 via treatment with miR-223 agomirs attenuated airway inflammation, NLRP3 levels and IL-1β release., Conclusions: The findings of this study revealed a crucial role for miR-223 in regulating the immunoinflammatory responses by depressing the NLRP3/ IL-1β axis in neutrophilic asthma.

Our previous study showed that sulfatide-activated type II natural killer T (NKT) cells can prevent allergic airway inflammation in an ovalbumin (OVA)-induced murine model of asthma, but the underlying mechanism is unclear. Recently, sulfatide-activated type II NKT cells were shown to modulate the function of dendritic cells in experimental autoimmune encephalomyelitis and nonobese diabetic mice. Thus, it was hypothesized that sulfatide-activated type II NKT cells may modulate the function of lung dendritic cells (LDCs) in asthmatic mice. Our data showed that, in our mouse models, activation of type II NKT cells by sulfatide administration and adoptive transfer of sulfatide-activated type II NKT cells resulted in reduced expression of surface maturation markers and proinflammatory cytokine production of LDCs. LDCs from sulfatide-treated asthmatic mice, in contrast to LDCs from PBS-treated asthmatic mice, significantly reduced allergic airway inflammation in vivo. However, we found no influence of sulfatide-activated type II NKT cells on the phenotypic and functional maturation of bone marrow-derived dendritic cells in vitro. In addition, adoptive transfer of sulfatide-activated type II NKT cells did not influence the phenotypic and functional maturation of LDCs in CD1d -/- mice, which lack both type I and II NKT cells, immunized and challenged with OVA. Our data reveal that sulfatide-activated type II NKT cells can suppress immunogenic maturation of LDCs to reduce allergic airway inflammation in mouse models of asthma, and it is possible that the immunomodulatory effect needs type I NKT cells.

Interleukin (IL)-4-producing-CD8 (cytotoxic T cells, Tc) contribute to lung eosinophilia and airway hyper-responsiveness (AHR) to an antigen. CD4+CD25+ regulatory T cells (Tregs) attenuate airway inflammation and AHR. This study investigated whether Tregs decrease Tc2frequencies in ovalbumin (OVA)-induced asthma model of mice. Female C57BL/6 mice were sensitized with OVA intraperitoneally and challenged with OVA intranasally to induce allergic asthma model. Tregs were sorted by fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS) microbeads. OVA-sensitized mice were injected with Tregs or phosphate buffer saline (PBS) by tail vein ahead of the first challenge. Airway inflammation and airway hyper-responsiveness (AHR)were evaluated by histological analysis and invasive method, respectively. OVA-specific IgE and cytokine levels were detected by ELISA. Flow cytometry was used to detect the percentages of Tc1 and Tc2. Gata3 and T-bet mRNA was determined by quantitative PCR (qPCR). OVA-sensitized and challenged mice displayed typical asthma features, which included eosinophilic airway inflammation, higher levels of Th2 cytokines and AHR. Gata3 mRNA, Tc2 frequencies and OVA-specific IgE levels were significantly increased in OVA-sensitized and challenged mice. Compared to PBS treatment, Tregs decreased Tc2 frequencies, airway inflammation, Th2 cytokine levels and AHR in OVA-sensitized and challenged mice. IL-13 levels were negatively correlated with Tc1 frequencies and with IFNg levels in experimental mice. Our results demonstrated that Tregs could prevent airway inflammation and AHR by decreasing Tc2 frequencies and cytokine levels in OVA-induced asthma model of mice, supporting Tregmight be as a potent therapeutic target for alleviating airway inflammation and AHR.

The ATP/P2X7 axis of dendritic cells (DCs) mediates the activation of NLRP3 inflammasome and promotes secretion of interleukin (IL)-1β and IL-18 to induce T helper (Th) 2, Th17 differentiation in the pathogenesis of asthma. NLRP3 inflammasome also regulates high mobility protein 1 (HMGB1) release in DCs. Recent studies demonstrated the correlation between HMGB1 expression and airway inflammation and hyper-responsiveness (AHR) in asthma. However, the relationship between the ATP/P2X7-NLRP3 axis and HMGB1 in DCs in asthma is still unclear. ATP, apyrase, Brilliant Blue G, BzATP, glibenclamide, and Z-YVAD-FMK were administered to ovalbumin (OVA)-induced murine asthmatic model. For in vitro studies, bone marrow-derived mononuclear cells (BMDCs) were primed with LPS and stimulated with the same reagents. Activation of the ATP/P2X7 axis aggravated airway inflammation and AHR in the lung and induced Th2, Th17 polarization in asthmatic mice. Inhibition of NLRP3 inflammasome weakened cardinal features of asthma and blocked Th2, Th17 polarization. In vitro and vivo, ATP/P2X7 axis activated NLRP3 inflammasome and induced HMGB1 expression and release from DCs. Inhibition of NLRP3 inflammasome reduced HMGB1 expression and release. The ATP/P2X7-NLRP3 axis of DCs participates in mediating airway inflammation, AHR, and promoting Th2, Th17 inflammatory responses in asthmatic mice by inducing HMGB1 expression and secretion., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The Th (T helper) 2 response is characteristic of allergic asthma, and Th17 cells are involved in more severe asthma. Recent studies demonstrated that HMGB1 (High mobility group box 1 protein) regulates airway inflammation and the Th2, Th17 inflammatory response in asthma. HMGB1 can interact with Toll-like receptors (TLR) 2 and 4, and the receptor for advanced glycation end products (RAGE), activating the NF-κB (nuclear factor kappa B) signaling pathway and inducing the release of downstream inflammatory mediators. Both Th cells and dendritic cells express TLR2, TLR4, and RAGE receptors. Therefore, we speculate that HMGB1 could regulate the differentiation of Th2, Th17 cells in asthma through direct and indirect mechanisms. An ovalbumin (OVA)-induced mouse asthmatic model was established. Anti-HMGB1 antibody or rHMGB1 was administered to OVA-sensitized mice 30 min prior to each challenge. For in vitro studies, magnetically separated CD4 + naive T cells were stimulated with or without rHMGB1 and/or anti-HMGB1 antibody. BMDCs (bone marrow-derived dendritic cells)-stimulated with or without rHMGB1 and/or anti-HMGB1 antibody were cocultured with CD4 + naive T cells. Our study showed that administration of rHMGB1 aggravated airway inflammation and mucus production, and induced Th2, Th17 polarization in asthmatic mice, and that anti-HMGB1 antibody weakened characteristic features of asthma and blocked the Th2, Th17 inflammatory responses. HMGB1 could directly act on naive T cells to induce differentiation of Th2, Th17 cells in vitro through activating the TLR2, TLR4, RAGE-NF-κB signal pathway in CD4 + naive T cells. HMGB1 could also indirectly promote Th2, Th17 differentiation via activating the TLR2, TLR4, RAGE-NF-κB signal pathway in DCs to mediate their maturation and antigen-presenting ability in vitro., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Objective: In allergic asthma, regulatory T cell (Treg) number and function are decreased. Antigen-primed CD8 + T cells play an indispensable role in the full development of airway inflammation and airway hyper-responsiveness (AHR) occurring in asthma. In this study, we investigated the relationship between subpopulations of CD8 + T cells and CD39 + Tregs., Methods: Female C57BL/6 mice were used to develop the model of allergic asthma. Experimental mice were immunized with ovalbumin (OVA) by intra-peritoneal (i.p) injection and then challenged with OVA by intra-tracheal administration. Control mice were immunized with vehicle by i.p injection and challenged with OVA. Airway inflammation was determined by histology and AHR was measured by an invasive method. Levels of interferon (IFN)-γ, IL-4, and IL-17 in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The frequencies of CD8 + IFN-γ + cells (Tc1), CD8 + IL-4 + cells (Tc2), CD8 + IL-17 + cells (Tc17), and CD39 + Tregs were measured by flow cytometry. The correlation between CD39 + Tregs and Tc subsets was analyzed by Pearson's test., Results: Experimental mice displayed phenotypes of allergic asthma, including inflammatory cell infiltration into the lungs, goblet cell hyperplasia, increased airway resistance, and increased IL-4 and IL-17 in BALF. Compared to control mice, experimental mice displayed lower CD39 + Tregs and Tc1 but higher Tc2 and Tc17. There was a negative correlation between CD39 + Tregs and Tc2 or Tc17., Conclusion: In allergic asthma, increased Tc2 and Tc17 are possibly related to insufficient CD39 + Tregs.

Objective: Allergic asthma is a chronic airway inflammation resulting from an imbalance of T helper (Th) cell responses to allergens. Interleukin (IL)-35 has been shown to have potent immunoregulatory properties. Whether IL-35 participates in the immunopathogenesis of allergic asthma patients is still unknown., Methods: CD4+ T cells and CD4+ CD25- T cells were obtained from peripheral blood mononuclear cells (PBMCs) using magnetic separation. The concentration of IL-35 in plasma was measured by enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of the IL-35 subunits, EBI3 and IL-12p35, were detected by quantitative real-time PCR (qPCR). The proliferative responses of CFSE-labeled CD4+ CD25- T cells in the presence or absence of rhIL-35 were evaluated by flow cytometry. Cytokine production of activated CD4+ CD25- T cells was examined by flow cytometry and ELISA., Results: IL-35 protein and mRNA levels were decreased in allergic asthmatics. The frequencies of CD4+ CD25+ Foxp3+ Tregs and CD4+ IL-12p35+ T cells in allergic asthma patients were lower than in healthy controls. Moreover, the addition of rhIL-35 suppressed CFSE+ CD4+ CD25- T cell proliferation in vitro in a dose-dependent manner, and the suppression induced by rhIL-35 was associated with decreases in IL-4 but not IFN-γ and IL-17 production of activated CD4+ CD25- T cells. The increased level of Th1/Th2 was observed in allergic asthmatics in the presence of rhIL-35., Conclusions: Our data suggest that IL-35 can effectively suppress the proliferation and IL-4 production of activated CD4+ CD25- T cells in allergic asthma, and that IL-35 may be a new immunotherapy for asthma patients.

Interleukin (IL)-35 is a newly discovered suppressive cytokine and has been shown to alleviate inflammatory and autoimmune diseases. The purpose of this study was to investigate immunomodulatory capacity of IL-35 in patients with allergic asthma. IL-35 mRNA expression levels in peripheral blood mononuclear cells (PBMCs) were detected by quantitative real-time PCR (qPCR). The frequencies of cytotoxic T cells (Tc)1, Tc2 and Tc17 cells were measured by flow cytometry. Plasma levels of IL-35, interferon (IFN)-γ, IL-4, and IL-17 were examined by enzyme-linked immunosorbent assay (ELISA). The correlations between plasma IL-35 levels and Tc1, Tc2, and Tc17 cytokine production in allergic asthmatics (n = 25) and healthy controls (n = 12) were analyzed by Pearson's test. IL-35 protein and mRNA expression levels were down-regulated in allergic asthmatics compared with healthy controls. The frequencies of Tc2 and Tc17 cells were significantly increased in patients with asthma, and the frequency of Tc1 cells did not differ between asthmatic patients and healthy controls. Similarly, plasma levels of IL-4 and IL-17 were significantly increased in asthmatic patients, while there was no difference in IFN-γ levels between allergic asthma patients and healthy controls. More importantly, plasma IL-35 protein levels were negatively correlated with the frequency of IL-4-producing CD8+ T (Tc2) cells and with the IL-4 level in patients with allergic asthma. Our results suggest that decreased circulating IL-35 levels could contribute to the pathogenesis of allergic asthma by regulating CD8+ T cells.

Mesenchymal stem cells (MSCs) have been investigated in the treatment of numerous autoimmune diseases. However, the immune properties of MSCs on the development of asthma have remained to be fully elucidated. Airway dendritic cells (DCs) have an important role in the pathogenesis of allergic asthma, and disrupting their function may be a novel therapeutic approach. The present study used a mouse model of asthma to demonstrate that transplantation of MSCs suppressed features of asthma by targeting the function of lung myeloid DCs. MSCs suppressed the maturation and migration of lung DCs to the mediastinal lymph nodes, and thereby reducing the allergen-specific T helper type 2 (Th2) response in the nodes. In addition, MSC-treated DCs were less potent in activating naive and effector Th2 cells and the capacity of producing chemokine (C-C motif) ligand 17 (CCL17) and CCL22, which are chemokines attracting Th2 cells, to the airways was reduced. These results supported that MSCs may be used as a potential treatment for asthma.

Background: Invariant natural killer T cells (iNKT cells) are a unique subset of T lymphocytes and are considered to play an important role in the development of allergic bronchial asthma. Recently, iNKT cells were shown to play an immunoregulatory role in CD4+ and CD8+ T cell-mediated adaptive immune response. Allergen-specific Th2 inflammatory responses are an important part of the adaptive immune response in asthma. However, the regulatory functions of the Th2 inflammatory response in asthma have not been studied in detail., Method: In this study, we have investigated the regulatory functions of iNKT cells on the Th2 inflammatory response in an ovalbumin (OVA)-induced murine model of asthma., Results: Our results demonstrate that α-Galactosylceramide (α-GalCer) administration activated iNKT cells but could not induce the Th2 inflammatory response in wild-type (WT) mice. In the OVA-induced asthma model, α-GalCer administration and adoptive transfer of iNKT cells significantly augmented the Th2 inflammatory responses, including elevated inflammatory cell infiltration in the lung and bronchoalveolar lavage fluid (BALF); increased levels of IL-4, IL-5, and IL-13 in the BALF and splenocyte culture supernatant; and increased serum levels of OVA-specific IgE and IgG1. In addition, the Th2 inflammatory response was reduced, but not completely abrogated in CD1d-/- mice immunized and challenged with OVA, compared with WT mice., Conclusion: These results suggest that iNKT cells may serve as an adjuvant to enhance Th2 inflammatory response in an OVA-induced murine model of asthma.

Objective: Mast cell infiltration into airway smooth muscle (ASM) bundle is an important feature of asthma. Extracellular adenosine triphosphate (eATP) contributes to the initiation of airway inflammation. eATP induces mast cells migration by acting through purinergic receptors. CD39 is an ectonucleotidase that degrades ATP to ADP and AMP. Whether eATP participates in the migration of mast cell towards ASM cells is still unknown., Methods: Airway smooth muscle cells (ASMCs) were isolated from C57/BL6J mice sensitized and challenged with OVA. ASMCs were in vitro cultured and stimulated with IL-4 + IL-13 in the presence or absence of exogenous CD39 or CD39 inhibitor ARL67156. ATP level in the supernatants was measured with ATP content determination kit. CXCL10 concentration in the ASMCs supernatants was measured by ELISA, the mRNA expression of CXCL10 in ASMCs was determined with real-time PCR. Human mast cell line HMC-1 was cultured in Iscove's-Modified Dubecco's Medium. The expression of CXCR3 in HMC-1 cells was determined with flow cytometry and real-time PCR, respectively. HMC-1 migration rates were determined with transwell system., Results: In the supernatants of Th2 cytokine-stimulated ASMCs, ATP level was higher than that without stimulation. CD39 decreased, whereas ARL67156 increased ATP level in the supernatants. Both ATP and the supernatants of Th2 cytokine-stimulated ASMCs induced migration of HMC-1 cells. The surface and mRNA expression of CXCR3 in HMC-1 cells, and the mRNA expression and secretion of CXCL10 in ASMCs were increased after stimulation with ATP or Th2 cytokines. All these effects were partially inhibited by CD39., Conclusion: Our data suggested ASMCs in the asthma microenvironment promoted the migration of mast cells via secretion of ATP and the expression of CXCL10/CXCR3 axis. CD39 could reverse this effect and may be a new target for the treatment of asthma.

Recent studies have demonstrated that extracellular adenosine 5'-triphosphate (eATP) is involved in allergic airway inflammation by activating purinergic receptors. eATP can be hydrolyzed by ectonucleotidases, such as CD39. In this study, we investigated the expression and distribution of CD39 in the lungs of mice, as well as the effects of apyrase on airway inflammation and the chemotactic migration of dendritic cells (DCs). A mouse model of asthma was developed with chicken ovalbumin (OVA)/aluminum hydroxide using female C57BL/6 mice. Apyrase was administered to OVA-sensitized mice prior to each challenge by intraperitoneal injection. The distribution of CD39 was detected by immunofluorescence. The mRNA and protein expression of CD39 was determined by quantitative PCR and western blot analysis, respectively. The levels of Th2 cytokines in the bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay (ELISA). The effect of apyrase on the chemotactic migration of DCs towards ATP was explored by migration assay in vitro. In the lungs, CD39 was primarily located in the cytoplasm and cytomembrane of bronchial epithelial cells and CD39 expression was reduced in mice with allergic asthma. Treatment with apyrase markedly attenuated OVA-induced airway inflammation, including peribronchial eosinophilic inflammation and reduced the number of inflammatory cells, as well as the levels of cytokines in BALF. Furthermore, apyrase also markedly reduced the expression of GATA binding protein 3 (GATA3) and decreased the chemotactic migration of DCs towards ATP.Our data demonstrate that a reduction in CD39 expression may be associated with the development of allergic airway inflammation and that apyrase alleviates airway inflammation by decreasing the chemotactic migration of DCs towards eATP. Therefore, targeting at eATP or ectonucleotidases may provide a novel therapeutic approach for allergic asthma.

Objective: Allergic asthma is a common inflammatory disease regulated by the T helper (Th) cells. Interferon regulatory factor 4 (IRF4) plays an important role in the differentiation of Th cells. This study investigated whether IRF4 is involved in the systemic immune responses in allergic asthma patients., Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from allergic asthmatics (n = 29) and healthy controls (n = 12). The mRNA and protein levels of IRF4 in PBMCs were measured by quantitative RT-PCR and western blotting. The frequencies of Th1, Th2, and Th17 cells were analyzed by flow cytometry. The related cytokine (interferon-γ, interleukin-4 (IL-4), and IL-17) concentrations in plasma and culture supernatants were measured by ELISA. The levels of GATA binding protein 3 (GATA3), retinoic acid-related orphan receptor γt (RORγt), and forkhead box P3 (FOXP3), the master transcription factors controlling Th2, Th17, and T regulatory cells differentiation, respectively, were examined by quantitative RT-PCR., Results: The levels of IRF4 were elevated in allergic asthmatics compared with those in healthy controls. The frequencies of Th2 and Th17 cells as well as the concentrations of Th2- and Th17-related cytokines were higher in plasma from asthma patients than those from healthy controls; similar results were observed in culture supernatants. IRF4 mRNA levels were positively correlated with GATA3, RORγt, and FOXP3 mRNA levels in allergic asthmatics but not in healthy controls., Conclusions: Our results suggested that IRF4 may play a role in the systemic inflammation of allergic asthma patients by facilitating the differentiation of Th2 and Th17 cells at least at the transcriptional level.

Asthma is a common chronic inflammatory disease involving many different cell types. Recently, type I natural killer T (NKT) cells have been demonstrated to play a crucial role in the development of asthma. However, the roles of type II NKT cells in asthma have not been investigated before. Interestingly, type I and type II NKT cells have been shown to have opposing roles in antitumor immunity, antiparasite immunity, and autoimmunity. We hypothesized that sulfatide-activated type II NKT cells could prevent allergic airway inflammation by inhibiting type I NKT cell function in asthma. Strikingly, in our mouse model, activation of type II NKT cells by sulfatide administration and adoptive transfer of sulfatide-activated type II NKT cells result in reduced-inflammation cell infiltration in the lung and bronchoalveolar lavage fluid, decreased levels of IL-4 and IL-5 in the BALF; and decreased serum levels of ovalbumin-specific IgE and IgG1. Furthermore, it is found that the activation of sulfatide-reactive type II NKT cells leads to the functional inactivation of type I NKT cells, including the proliferation and cytokine secretion. Our data reveal that type II NKT cells activated by glycolipids, such as sulfatide, may serve as a novel approach to treat allergic diseases and other disorders characterized by inappropriate type I NKT cell activation.

Background: T cells play an important role in orchestrating allergic asthma by producing various cytokines; however, little information is available on the phenotype of cytokine-producing T cells, especially CD8(+) T and γδ T cells in humans., Objective: The aim of this study was to investigate the role of cytokine expressions from circulating CD4(+), CD8(+), and γδ T cells in patients with allergic asthma., Methods: Peripheral blood mononuclear cells were isolated from patients with allergic asthma (n = 29) and healthy controls (n = 12). The percentage of helper T cell (Th)1-, Th2-, and Th17-type cytokines interferon-γ (IFN-γ), interleukin (IL)-4, and IL-17, respectively, of CD4(+), CD8(+), and γδ T cells was analyzed by flow cytometry., Results: The percentages of IL-4(+) and IL-17(+) CD4(+) T cells increased, whereas IFN-γ(+) CD4(+) T cells decreased in patients with allergic asthma compared with healthy controls. The frequency of IL-17(+) CD4(+) T cells tended to increase with the severity of allergic asthma. Higher frequency of IL-4-producing and lower frequency of IFN-γ-producing CD8(+) and γδ T cells were found in patients with allergic asthma compared with healthy controls. The IFN-γ(+)/IL-17(+) ratio among CD4(+), CD8(+), and γδ T cells was significantly decreased in patients with allergic asthma compared with healthy controls., Conclusion: CD8(+) and γδ T cells might be involved in the pathogenesis of allergic asthma through the release of Th1-, Th2-, and Th17-type cytokines.

Background: The suppressive cytokine interleukin-10 (IL-10) plays a central role in disease control and clinical therapies of asthma. CD46 was recently identified as costimulatory molecule in inducing IL-10-producing regulatory T cells type 1 (Tr1) from CD4(+) T cells. Alterations in CD46 costimulation pathway were shown to be associated with multiple sclerosis and hemodialysis. In this study, the authors investigated alterations in CD46 costimulatory pathway in asthma., Methods: CD4(+)CD25(+) regulatory T cells (Tregs) and CD4(+)CD25(-) T cells were isolated from peripheral blood mononuclear cells (PBMCs) of asthma patients (n = 13) and healthy subjects (n = 17). Both subsets of CD4(+) T cells were cultured alone or cocultured at a ratio of 1:10 under stimulation with CD3/CD28 or CD3/CD46, and the production of IL-10 in the supernatants was assessed by enzyme-linked immunosorbent assay (ELISA), the proliferation rates of the cells were determined with thymidine incorporation assay., Results: Levels of IL-10 in the supernatants were higher in undivided CD4(+) T cells and 1:10 cocultured CD4(+)CD25(+) Tregs/CD4(+)CD25(-) T cells than in CD4(+)CD25(+) Tregs or CD4(+)CD25(-) T cells alone, either under CD46 or under CD28 costimulation, both in healthy controls (n = 9) and in asthma patients (n = 7). Under anti-CD3/CD46 stimulation, IL-10 production in undivided CD4(+) T cells and cocultured T cells from asthma patients was lower than that in healthy controls. When treated with glucocorticoids, IL-10 production in undivided CD4(+) T cells and 1:10 cocultured CD4(+) T cells was not different between asthma patients (n = 6) and healthy controls (n = 8). The proliferation rates and the surface expression of CD46 were not different in T cells from both groups., Conclusions: This study identified a new functional defect of CD4(+)CD25(+) Tregs in inducing IL-10 production from CD4(+)CD25(-) T cells under CD46 costimulation in asthma patients, which may be involved in the pathogenesis of allergic asthma.

IL-23/IL-17 axis is an important regulator in various inflammatory diseases. However, the role of IL-23 in allergic airway inflammation is not well understood. In this study, we show that in an allergen-induced asthma model, mice with transgenic overexpression of IL-23R exhibited increased airway infiltration of eosinophils and Th2 cytokine production, whereas those deficient in IL-23 displayed reduced airway inflammation. In vitro, IL-23-IL-23R signaling promoted GATA-3 expression and enhanced Th2 cytokine expression. Conversely, in the absence of this signal, Th2 cell differentiation was partially inhibited. Therefore, IL-23 signaling may regulate allergic asthma through modulation of Th2 cell differentiation.

Background: Allergic asthma is an inflammatory disease regulated by the T helper (Th) cells. The Th1/Th2 imbalance has been well documented in the pathogenesis of allergic asthma. Recently, Th17 cells have been found to participate in the development of allergic asthma in animals. However, whether Th17 immunity contributes to the systemic immune responses in allergic asthmatic patients is unclear., Methods: Peripheral blood mononuclear cells were isolated from allergic asthmatics (n = 29) and healthy controls (n = 12). The frequencies of Th1, Th2 and Th17 cells were analyzed by flow cytometry. The related cytokine (IFN-gamma, IL-4, IL-17, IL-22, IL-23 and IL-25) concentrations in plasma and culture supernatants were measured by enzyme-linked immunosorbent assay and Luminex. The level of retinoic acid-related orphan receptor gammat (RORgammat), a key transcription factor controlling Th17 differentiation, was examined by real-time quantitative polymerase chain reaction., Results: The percentages of Th2 and Th17 cells as well as the concentrations of Th2- and Th17-related cytokines were higher in allergic asthmatics than those in healthy controls; some patients were even treated with inhaled glucocorticoid. The percentages of Th17 cells as well as the plasma concentrations of IL-17 and IL-22 tended to increase with the severity of the disease, while the IL-25 level was elevated in mild patients. A parallel elevation of IL-17 and IL-23 concentrations and an increase in RORgammat level were found in allergic asthmatics., Conclusion: Our results suggest that besides predominant Th2 immunity, abnormal Th17 immunity may be also involved in the pathogenesis of allergic asthma., (2009 S. Karger AG, Basel.)

Background: Allergen exposure can lead to the development of allergic asthma, sensitized asymptomatic or nonallergic responses in different conditions. Regulatory T (T(reg)) cells are involved in controlling the immune response direction. The aim of this study was to determine whether allergic asthmatics had defects in CD4+CD25+ T(reg) cells and CD4+IL (interleukin)-10+ T(reg) cells after specific allergen exposure., Methods: Peripheral-blood mononuclear cells (PBMCs) were isolated from 20 patients with asthma sensitized to house dust mite, 24 asymptomatic subjects sensitized to house dust mite and 22 nonallergic subjects. Cells were cultured without stimulation and with rDer p 1. Frequencies and proliferations of CD4+CD25+ T cells and CD4+IL-10+ T cells were measured by flow cytometry. Concentrations of IL-10, IL-4, interferon (IFN)-gamma and tumor growth factor beta(1) in culture supernatants and sera were determined by ELISA., Results: PBMCs derived from sensitized asthmatic patients had the lowest percentage of CD4+CD25+ T(reg) cells and the highest IL-4 level in response to allergen among the 3 groups. PBMCs derived from sensitized asymptomatic subjects had higher percentages of both CD4+CD25+ T(reg) cells and CD4+IL-10+ T cells than those from sensitized asthmatic patients in rDer-p-1-stimulated cultures, and they had the highest levels of IL-10 and IFN-gamma responses to allergen among the 3 groups., Conclusions: Patients with allergic asthma had an insufficient CD4+CD25+ T(reg) cell response and an excessive Th2 response to specific allergen. The high levels of T(reg) cell response and IL-10 and IFN- gamma responses to specific allergen can partly explain the immunological characteristics associated with sensitized asymptomatic subjects., ((c) 2008 S. Karger AG, Basel.)

The aim of this study was to investigate and identify the relationship between urinary cysteinyl leukotriene E(4) levels and clinical response to antileukotriene treatment in patients with asthma. Forty-eight patients with stable mild to moderate asthma were treated with montelukast in a four-week trail. Asthmatic symptom score, beta(2)-agonist usage, percentage of eosinophil, total serum IgE concentration, forced expiratory volume in the first second (FEV(1)), peak expiratory flow rate (PEFR), and urinary leukotriene E(4) (uLTE(4)) were measured before and after treatment. Clinical response was assessed by the improvement of asthma symptom scores, beta(2)-agonist usage, and FEV(1). Responders were defined as patients who had to fit the following three criteria: a reduction of more than 20% in mean symptom score; a reduction of more than 20% in beta(2)-agonist usage, and a mean improvement of FEV(1) of more than 10% from baseline value. Others were classified as nonresponders. Logistic analysis was used to access the various clinical factors correlated with the clinical response. There were 25 responders and 23 nonresponders. The mean uLTE(4) level from the responders was higher than that from the nonresponders (224.5 +/- 34.4 vs. 175.3 +/- 37.1 pg/mg creatinine, p < 0.05). There was a significant correlation between the clinical response and the uLTE(4) level but not demographic features, percentage of eosinophils, serum IgE concentration, or spirometry (p > 0.05). Subjects with a uLTE(4) level of >/= 200 pg/mg creatinine were 3.5 times more likely to respond to montelukast than those with less than 200 pg/mg creatinine (95% confidence interval [CI] = 1.7-15.8). The uLTE(4) level is closely correlated with antileukotriene treatment. uLTE(4) is a good biomarker for selecting this drug to treat asthma.

Background: In allergic asthma, allergen-specific T cells have a Th2-biased phenotype, and it is thought that dendritic cells (DCs) contribute to the induction of allergic immune responses. Therefore, we hypothesized that DCs from allergic asthmatics and healthy donors differ with regard to their preference to induce Th1 or Th2 immune responses., Objectives: To investigate differences in DC-expressed costimulatory molecules and DC-secreted cytokines between allergic asthmatics and healthy donors, and their influence on the Th1- and Th2-type cytokine balance., Methods: Circulating monocytes from patients with allergic asthma and healthy donors were cultured with GM-CSF and IL-4, respectively, for 5 days and subsequently with lipopolysaccharide for 2 days to create mature DCs (mDCs). CD1a, CD83, CD40 and CD86 expression on mDCs was examined using a fluorescence-activated cell sorter. IL-12 and IL-10 secreted by mDCs were measured by ELISA. Naïve cord blood T cells were primed by mDCs from two groups, and IL-4 and IFN-gamma production by polarized T-helper cells (Th) was measured by ELISA., Results: (1) CD86 expression on mDCs from allergic asthmatics was higher than that from healthy donors. (2) IL-12, IL-12p40 and IL-10 production by mDCs from allergic asthmatics was significantly lower than that from healthy donors, respectively. (3) IL-4 production by Th cells primed by mDCs from allergic asthmatics was increased compared with that from healthy donors., Conclusions: mDCs from allergic asthmatics preferentially priming naïve T cells towards Th2-cell development might be due to increased expression of CD86 and reduced production of IL-12 and IL-10., (Copyright 2006 S. Karger AG, Basel.)

Objective: To investigate the deficiency of the expression of the phenotypes (CD(1a), CD(83), CD(40), CD(86)) and cytokines (IL-12 and IL-10) by human peripheral blood monocyte (PBMC)-derived dendritic cell (DCs) from asthmatic subjects, and their influence on naive T cell polarization., Methods: Adherent cells were isolated from peripheral blood samples in asthmatic patients and in healthy volunteers, and were cultured with granulocyte-macrophage colony-stimulating factor and IL-4 as immature DC (iDC). iDCs were stimulated with lipopolysaccharide as mature DC (mDCs). Nonadherent cells were obtained from umbilical cord blood by idem methods, and naïve T cells were sorted by adding anti-CD(4) and anti-CD(45RA) in nonadherent cells respectively and magnetic microbeads. Naïve T cells and mDCs from two groups were co-cultured in complete RPMI1640 media respectively, and naive T cells polarized as T helper cells 1 (Th1) and Th2. The expression of the CD(1a), CD(83), CD(40) and CD(86) on mature DCs were examined by fluorescent activated cell sorter. IL-12 and IL-10 released by mDCs and IL-4 and IFNgamma produced by Th cells were measured by ELISA., Results: (1) The expression of CD(86) on dendritic cells from atopic asthmatics was higher than that from healthy control subjects (40.75 +/- 3.99 vs 29.88 +/- 1.25, P < 0.01). (2) The levels of IL-12, IL-12p40 and IL-10 produced by DCs from asthmatic subjects were all significantly lower than those from healthy control group (217.79 +/- 118.65 vs 905.66 +/- 495.32, P < 0.01; 2072.22 +/- 1496.37 vs 5569.43 +/- 2922.75, P < 0.01; 336.89 +/- 261.52 vs 1425.00 +/- 1148.87, P < 0.05, respectively). (3) IL-4 production by Th2 cells which were primed by DCs from asthmatics was significantly increased as compared to that from control group (368.56 +/- 190.72 vs 584.91 +/- 290.13, P < 0.01); On the contrary, IFNgamma in the patient group was reduced as compared to that in the control group (425.33 +/- 164.94 vs 49.86 +/- 18.14, P < 0.05). (4) In the patient group, the level of IL-12 was positively correlated to that of IFNgamma (P < 0.05), negatively correlated to that of IL-4 (P < 0.05); IL-10 was negatively correlated to IL-4 (P < 0.05). (5) There was a positive correlation between IL-12 and IL-10 in the two groups (P < 0.01)., Conclusion: Because of DC deficiency, naïve T cells preferentially polarize to Th2 which synthesize more Th2-type cytokine (i.e. IL-4) and T cell tolerance cannot be induced, which may be one of the important pathogenic mechanisms for allergic asthma.

Aim: To investigate the mechanism of anti-inflammatory action of theophylline on asthma., Methods: Nineteen asthmatic subjects were administered 200 mg sustained-release theophylline twice daily for 4 weeks. The number of CD4+ T lymphocytes, eosinophils, and the levels of interleukin (IL)-5 and interferon gamma (IFN-gamma) in sputum pre- and post-administration were measured by direct immunofluorescene technique, Wright's stain and enzyme-linked immunosorbent assay, respectively. The symptom scores and lung function were also evaluated., Results: Theophylline treatment significantly improved symptom scores and FEV1.0, FEV1.0 % (P < 0.05) and reduced sputum eosinophils (P < 0.01) in asthmatic subjects. These were accompanied by a decrease in sputum IL-5 level (P < 0.01), but sputum CD4+ T lymphocytes and IFN-gamma had no significant change (P >0.05). The mean (range) serum theophylline concentration in final steady state was 7.9 (3.9 - 12.9) mg/L., Conclusion: The anti-inflammatory action of theophylline in asthma may result from reduction of IL-5 production in the airways.

Objective: In allergic asthma, regulatory T cell (Treg) number and function are decreased. Antigen-primed CD8+ T cells play an indispensable role in the full development of airway inflammation and airway hyper-responsiveness (AHR) occurring in asthma. In this study, we investigated the relationship between subpopulations of CD8+ T cells and CD39+ Tregs. Methods: Female C57BL/6 mice were used to develop the model of allergic asthma. Experimental mice were immunized with ovalbumin (OVA) by intra-peritoneal (i.p) injection and then challenged with OVA by intra-tracheal administration. Control mice were immunized with vehicle by i.p injection and challenged with OVA. Airway inflammation was determined by histology and AHR was measured by an invasive method. Levels of interferon (IFN)-γ, IL-4, and IL-17 in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The frequencies of CD8+IFN-γ+ cells (Tc1), CD8+IL-4+ cells (Tc2), CD8+IL-17+cells (Tc17), and CD39+Tregs were measured by flow cytometry. The correlation between CD39+Tregs and Tc subsets was analyzed by Pearson's test. Results: Experimental mice displayed phenotypes of allergic asthma, including inflammatory cell infiltration into the lungs, goblet cell hyperplasia, increased airway resistance, and increased IL-4 and IL-17 in BALF. Compared to control mice, experimental mice displayed lower CD39+Tregs and Tc1 but higher Tc2 and Tc17. There was a negative correlation between CD39+Tregs and Tc2 or Tc17. Conclusion: In allergic asthma, increased Tc2 and Tc17 are possibly related to insufficient CD39+Tregs. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate and identify the relationship between urinary cysteinyl leukotriene E4 levels and clinical response to antileukotriene treatment in patients with asthma. Forty-eight patients with stable mild to moderate asthma were treated with montelukast in a four-week trail. Asthmatic symptom score, β2-agonist usage, percentage of eosinophil, total serum IgE concentration, forced expiratory volume in the first second (FEV1), peak expiratory flow rate (PEFR), and urinary leukotriene E4 (uLTE4) were measured before and after treatment. Clinical response was assessed by the improvement of asthma symptom scores, β2-agonist usage, and FEV1. Responders were defined as patients who had to fit the following three criteria: a reduction of more than 20% in mean symptom score; a reduction of more than 20% in β2-agonist usage, and a mean improvement of FEV1 of more than 10% from baseline value. Others were classified as nonresponders. Logistic analysis was used to access the various clinical factors correlated with the clinical response. There were 25 responders and 23 nonresponders. The mean uLTE4 level from the responders was higher than that from the nonresponders (224.5 ± 34.4 vs. 175.3 ± 37.1 pg/mg creatinine, p < 0.05). There was a significant correlation between the clinical response and the uLTE4 level but not demographic features, percentage of eosinophils, serum IgE concentration, or spirometry ( p > 0.05). Subjects with a uLTE4 level of ≥ 200 pg/mg creatinine were 3.5 times more likely to respond to montelukast than those with less than 200 pg/mg creatinine (95% confidence interval [CI] = 1.7–15.8). The uLTE4 level is closely correlated with antileukotriene treatment. uLTE4 is a good biomarker for selecting this drug to treat asthma. [ABSTRACT FROM AUTHOR]