Direct targeting of STAT3 by xanthatin suppresses allergic airway inflammation via inhibition of TSLP release and NK2 cell polarization.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joint inflammation and destruction. Identifying novel therapeutic targets is crucial for improving RA treatment. This study aimed to identify immune-related biomarkers in RA and investigate the potential of CD27 as a therapeutic target. We employed bioinformatics analysis of gene expression data from RA patients and healthy controls (GSE55457), followed by machine learning approaches (LASSO regression and Boruta algorithm) to identify potential biomarkers. Findings were validated in an independent dataset (GSE55235). The therapeutic potential of CD27 neutralization was evaluated in a collagen-induced arthritis (CIA) mouse model. Mechanisms were explored through quantitative real-time PCR, Western blot analysis, ELISA, and flow cytometry to assess T cell subsets, cytokine profiles, and signaling pathways. Bioinformatics analysis identified 714 differentially expressed genes, and machine learning analyses identified CD27, CD24, TNFRSF4, and PDCD1LG2 as potential RA biomarkers, all demonstrating strong diagnostic performance. CD27 showed significant positive correlations with T lymphocyte infiltration. In the CIA model, CD27 neutralization significantly reduced arthritis severity scores. This therapeutic effect was associated with suppression of Th1 responses, evidenced by significantly decreased serum levels of Th1 cytokines (IFN-γ, IL-2, TNF-α) and reduced CD4 + IFN-γ + cell populations, while Th2-related cytokines (IL-4, IL-5) remained largely unaffected. Mechanistically, CD27 neutralization attenuated phosphorylation of AKT and NF-κB p65 in vivo, while p38 MAPK remained unchanged. In vitro, recombinant CD27 protein stimulation of naive CD4 + T cells promoted Th1-biased differentiation, increasing CD4 + IFN-γ + cells and enhancing the phosphorylation of NF-κB p65 and AKT. Our study identifies CD27 as a potential therapeutic target in RA. CD27 neutralization attenuates arthritis severity by suppressing Th1 responses, possibly through modulation of AKT and NF-κB signaling pathways. These findings provide new insights into RA pathogenesis and suggest CD27 as a promising target for RA treatment.

Pythium insidiosum is a fungus-like oomycete capable of causing disease with high mortality rates. Although pulmonary Pythium insidiosum infections are rare, they are associated with an extremely high fatality rate. The mechanisms underlying Pythium insidiosum infection remain unclear and may be associated with immune dysregulation or iron overload. Existing animal models do not adequately replicate the pathophysiological process of pulmonary invasive infection by Pythium insidiosum in humans. Therefore, this study aims to establish a murine model of pulmonary Pythium insidiosum infection and to investigate the role of the Th1/Th2 immune balance in the progression of pulmonary infection. BALB/c mice were divided into the following groups: control group, iron dextran (ID) group, cyclophosphamide (CTX) group, lipopolysaccharide (LPS) group, and groups affected by various factor combinations (e.g., LPS + CTX group, ID + CTX group, ID + LPS group). Mice were inoculated intratracheally with Pythium insidiosum hyphae after pretreatment. Body weight, clinical symptoms, inflammatory cell counts in venous blood and bronchoalveolar lavage fluid (BALF), pulmonary histopathological damage, pathogen burden, as well as serum levels of ferritin (FER), Th1 cytokines (IL-2, IL-12p70, IFN-γ, TNF-α) and Th2 cytokines (IL-4, IL-5, IL-13, IL-10) were assessed in each group. Among all groups, LPS + CTX and ID + CTX groups exhibited severe infection symptoms, significant weight loss, and the highest clinical symptom scores (all P < 0.001). The ID + CTX group showed more pronounced increases in hematoxylin eosin (HE)-stained lung injury scores and Grocott's Methenamine Silver (GMS)-stained hyphal burden scores (all P < 0.001). Compared with the control group, the ID + CTX group demonstrated decreased venous blood leukocyte (WBC), neutrophil (NEUT), and lymphocyte (LYMPH) counts, elevated FER levels, and significantly reduced LYMPH counts in BALF (all P < 0.01). Cytokine analysis revealed that Th1 cytokine (IFN-γ, TNF-α) levels were significantly suppressed in the ID + CTX group after preconditioning (both P < 0.001). Following infection, Th1 responses remained relatively suppressed, while Th2 responses showed an upward trend that did not reach statistical significance (all P > 0.05), more consistent with a relative Th2 shift resulting from Th1 suppression. This study established a murine model of pulmonary Pythium insidiosum infection for the first time through intratracheal hyphae inoculation following ID + CTX preconditioning. The findings suggest that iron overload is associated with Th1 immune response suppression and relative Th2 shift, which may be related to increased susceptibility and pathological damage in pulmonary Pythium insidiosum infection. However, further functional experiments are needed to validate causal relationships.

Asthma is a widespread chronic inflammatory condition of the airways, often triggered by allergens such as ovalbumin. This study aimed to evaluate the potential anti-asthmatic effects of a synthetic isoxazolone derivative, (Z)-4-(4-hydroxy-3-methoxybenzylidene)-3-methylisoxazole-5(4H)-one (HMM), in a mouse model of allergic asthma. Molecular docking was carried out to explore the HMM interaction with asthma-associated proteins, providing insights into its potential therapeutic targets. Thirty-six albino mice were randomly divided into six groups: negative control, positive control, standard drug group (methylprednisolone, 15mg/kg), and three groups receiving HMM at doses of 3.2, 6.3, and 12.6mg/kg. Mice were sensitized with ovalbumin on days 0 and 14, followed by intranasal exposure from days 15 to 21 to induce allergic airway inflammation. Blood and bronchoalveolar lavage fluid (BALF) were collected to measure total and differential leukocyte counts. Cytokine expression (IL-4, IL-5) and aquaporins (AQP-1, AQP-5) were quantified via real-time PCR. HMM-treated groups showed reduced inflammation, lowered Th2 cytokine levels, and improved lung water transport markers. Computational studies supported these findings, revealing strong binding affinities between HMM and TNF-α (- 6.07kcal/mol), AKT1 (- 6.32kcal/mol), and COX-2 (- 5.87kcal/mol), forming stable hydrogen-bond interactions with key active-site residues. These results suggest HMM holds promise as a potential anti-asthmatic agent.

Cellular senescence has emerged as a key contributor to the pathogenesis of chronic lung diseases. Peroxisome proliferator-activated receptor gamma (PPAR-γ), a nuclear transcription factor, regulates senescence across multiple cell types. However, the role of PPAR-γ in allergic airway inflammation, particularly through regulation of macrophage senescence, remains poorly defined. Cellular senescence was evaluated in an allergic asthma mouse model using single-cell RNA sequencing (scRNA-seq). Senescent cells were selectively eliminated with dasatinib and quercetin (D&Q) to assess their contribution to disease pathogenesis. Macrophage-lineage-specific PPAR-γ conditional knockout model (PpargΔCD11c) were generated to define the role of macrophage PPAR-γ in senescence and allergic airway inflammation. PPAR-γ activity was further examined in isolated alveolar macrophages and in vivo using rosiglitazone, including macrophage-targeted delivery via phosphatidylserine-modified liposomes (PSL-ROSI). scRNA-seq analysis revealed enhanced senescence signatures in mononuclear phagocytes (MNPs), characterised by increased SenMayo scores and elevated Cdkn2a (p16) expression. Clearance of senescent cells significantly reduced airway inflammation and Th2 cytokine levels (IL-4, IL-5). Correlation analysis identified PPAR-γ as a key transcriptional regulator inversely associated with cellular senescence. Macrophage-lineage-specific deletion of PPAR-γ (PpargΔCD11c) exacerbated airway inflammation and increased cellular senescence. In vitro, rosiglitazone reduced allergen-induced senescence and suppressed proinflammatory mediators (IL-6, ICAM-1, CCL4, CCL5, TIMP-1, TNF-α) in alveolar macrophages. In vivo, rosiglitazone and inhaled PSL-ROSI attenuated cockroach allergen-induced airway inflammation, with PSL-ROSI effectively bypassing the airway mucus barrier to deliver rosiglitazone to lung macrophages. Integrated chromatin binding and transcriptomic analyses demonstrated that PPAR-γ promotes macrophage lipid metabolic programs (e.g., CD36, Fabp4). These findings identify macrophage senescence as a pathogenic driver of allergic airway inflammation and establish PPAR-γ as a critical regulator of macrophage senescence and homoeostasis, highlighting its potential as a therapeutic target for asthma. US National Institutes of Health (NIH) (1R01AI153331 and R01AI141642 to Dr. Gao), Anhui Provincial Key projects of Natural Science Foundation for Colleges and Universities 2025AHGXZK31476 (Dr. Wan).

Emerging evidence suggests that dysregulated immune homeostasis may play a role in the pathophysiology of ADHD; however, comprehensive profiling of T-helper (Th) cell subsets and microglial regulatory cytokines remains limited. This cross-sectional study included adolescents aged 12 to 18 years with ADHD (n = 45) and healthy controls (n = 41). Serum levels of Th1 (IL-12 and IFN-γ), Th2 (IL-4), Th17 (IL-17 and IL-23), Treg (IL-10 and TGF-β), and microglia regulatory cytokines (IL-34 and CSF-1) were measured by ELISA. ROC curve and logistic regression analyses were performed to determine the diagnostic and predictive value of the parameters in distinguishing ADHD. Significant increases in IL-12, IL-17, IL-23, IL-4, and IL-10 levels were observed in the ADHD group compared to the control group. IL-12 and IL-17 showed the highest diagnostic yield (AUC = 0.77). IL-12 remained a strong independent predictor in multivariate analysis (OR = 55.88, P = .015). IFN-γ and TGF-β did not differ significantly between the groups. CSF-1 levels did not differ between the groups, while IL-34 showed an increase that approached statistical significance in the ADHD group (P = .052; AUC = 0.622). Our results reveal a profile of Th1/Th17-dominated proinflammatory activation, partially compensatory Th2/Treg modulation, and potentially IL-34-mediated microglial divergence in ADHD. IL-12 and IL-17 showed the highest discriminatory performance; however, their AUC values (~0.77) remained at a moderate level. Our findings highlight the need for multidimensional immunophenotyping and point toward novel immune-targeted therapeutic strategies.

Umbilical cord mesenchymal stem cell-derived exosomes delivered via a thermosensitive hydrogel ameliorate atopic dermatitis by mitigating keratinocyte mitochondrial damage through activating the Wnt/β-catenin pathway.

PM2.5 exposure exacerbates airway pyroptosis related inflammatory response in asthmatic mice by activating NLRP3 inflammasome.

Boehmeria nivea (L.) Gaud. ameliorates airway inflammation by inhibiting Th2 cell differentiation in allergic asthma.

OPTN knockout alleviates OVA-induced airway inflammation in a mouse model of asthma.

Mulberry leaf polyphenols alleviate food allergy and moderate intestinal flora and lipid metabolism in allergic mice

CD70, a member of the tumor necrosis factor receptor superfamily, is expressed in glioblastoma (GB), where it promotes tumor growth, migration, and immunosuppression. Accordingly, it has emerged as a target for chimeric antigen receptor (CAR)-T cell therapy. Despite the influence of CAR structure on therapeutic efficacy, no comparative studies have evaluated different CD70-directed CAR designs in GB. Our study addressed this gap. We first validated CD70 expression in transcriptomic datasets, patient tissue, and GB cell lines. We then generated CD70-specific CAR-T cells featuring distinct target recognition and co-stimulatory domains (CD27z, LF28z, and LFBBz) and performed phenotypic characterization. Using co-culture systems and 3D cerebral organoids, we showed that all constructs eliminated target cells in a CD70-dependent manner, with CD27z secreting the highest levels of Th1 cytokines. This functional advantage translated into superior survival in an orthotopic GB mouse model. Based on these findings, we developed a panel of murine CD27-based constructs, all of which demonstrated potent antitumor activity in vitro and in immunocompetent GB mouse models, further underscoring the therapeutic promise of CD27 integration into the CAR design. Collectively, our comparative analysis highlights the superior efficacy of the ligand-based construct, supporting its incorporation into a clinical trial targeting CD70 in recurrent GB.

Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity disease triggered by inhaled Aspergillus -fumigatus (Af) spores (i.e., conidia), in persons with cystic fibrosis (CF). High iron levels have been commonly described in CF respiratory secretions. Af proteases cause a severe allergic immune response, leading to pulmonary exacerbations and progressive lung function decline. The stimuli that promote Af proteases in ABPA are poorly understood. Determine if airway iron is a critical factor in CF-ABPA by regulating fungal protease production. We used transcriptomics and proteomics to investigate the role of iron in stimulating the production of Af proteases and evaluated the impact of these iron-induced Af proteases on the Th2 immune response in CF bronchial epithelial (CFBE) cells and in a CF-ABPA mouse model. Transcriptional analysis showed a significant upregulation of Af proteases with exposure to iron and a downregulation with iron chelation. This finding was validated in secretome studies showing that Af-culture filtrates (Af-cf) from iron-exposed conidia contained significantly higher levels of 14 proteases compared to controls. Exposure to iron-primed Af-cf increased the production of Th2 promoting cytokines (IL-33, IL-25 and TSLP) in CFBE cells compared to the wild type cells. In vivo, inoculation of CF mice with iron-primed Af conidia led to higher levels of serum IgE and Th2 cytokines (IL-4, IL-5, IL-9, IL-13) in lung tissue. These findings suggest that airway iron is a potent stimulus for Af proteases and represents a modifiable risk factor and potential therapeutic target for CF-ABPA.

Zhichuanling oral liquid (ZCLOL) is a multi-component traditional Chinese medicine formulation that has been widely used in the clinical management of asthma. However, the molecular mechanisms underlying its clinical efficacy remain largely undefined, particularly regarding calcium homeostasis and endoplasmic reticulum stress (ERS) regulation. This study aimed to elucidate the mechanisms by which ZCLOL mitigates asthma-related airway inflammation through the integration of in vivo and in vitro models with proteomic profiling. In this study, an OVA-induced asthma model was established to evaluate the therapeutic effects of ZCLOL by assessing pulmonary function, serum Th2 cytokine levels, and macrophage polarization. To further elucidate the underlying mechanisms of ZCLOL in asthma treatment, we conducted proteomic analysis, measured calcium levels in lung tissue, and performed Western blotting. Additionally, an inflammatory BEAS-2B cell model induced by LPS and IL-13, combined with BAPTA-AM treatment, was used to explore the molecular pathways involved. ZCLOL effectively alleviated airway hyperresponsiveness, reduced lung tissue injury, inhibited M1 macrophage polarization and Th2 cytokine secretion, decreased intracellular calcium ion levels, mitigated endoplasmic reticulum stress, and downregulated the expression of STIM1, Bip, Ero1-Lα, PDI, CHOP, and phosphorylated JNK (p-JNK). ZCLOL alleviates airway inflammation and hyperresponsiveness in asthma by restoring Ca2+ homeostasis and suppressing ERS through inhibition of the STIM1/CHOP/JNK axis. These findings provide mechanistic evidence supporting ZCLOL as a potential multi-target therapeutic strategy for asthma.

IntroductionBrucella is a Gram-negative facultative intracellular bacterium that can cause fever, abortion, and other symptoms in humans and various mammals. btpB, a type IV secretion system (T4SS) effector of Brucella, plays a critical role in regulating Brucella infection and inhibiting the host's innate immune response.MethodsIn this study, a btpB mutant strain of Brucella A19 (ΔbtpB) was constructed using homologous recombination, and its biological characteristics, virulence, and immunogenicity were systematically investigated.ResultsThe results showed that ΔbtpB exhibited weakened resistance to in vitro stress, while its growth characteristics did not differ significantly from the wild-type strain A19. In the mouse immunization model, ΔbtpB induced weaker splenic pathological damage, and the splenic bacterial load was significantly lower than that of A19, indicating its reduced virulence. Additionally, ΔbtpB infection elicited stronger humoral and cellular immune responses in mice, including higher antibody levels, increased levels of Th1 cytokines (such as IFN-γ and IL-2), and enhanced proliferation and activity of CD8+ cells. Detection of Th1 and Th2 cells revealed that ΔbtpB induced stronger Th1 and Th2 responses in the spleen in the early stage, but the Th2 response weakened in the middle and late stages of infection. Notably, ΔbtpB infection did not suppress natural killer (NK) cell activity and even significantly enhanced its cytotoxic activity compared to the A19 strain.ConclusionOur research demonstrates that ΔbtpB leads to a reduced survival capacity of Brucella, while enhancing its immunogenicity. This suggests btpB is an important target for the prevention of Brucella.

Ischemia-free liver transplant (IFLT) has been developed to reduce ischemia-reperfusion injury (IRI). This study aims to investigate how this procedure impacts local and systemic immunity compared to conventional liver transplantation (CLT). Immunohistochemistry, immunofluorescence staining, single-cell RNA sequencing (scRNA-seq), and multiplex cytokine are used to illustrate distinct local and systemic immunity. In contrast to CLT, IFLT reduces neutrophil infiltration and neutrophil extracellular trap formation in grafts. By constructing an immune cell chimerism atlas, we reveal that IFLT reduces recipient-derived monocyte infiltration by suppressing ANXA1-FPR1 signaling through the STAT3-HIF-1α pathway, thereby attenuating inflammatory responses in graft monocytes. Additionally, IFLT confers graft protection by upregulating HMOX1 expression in monocytes and macrophages. Peripherally, IFLT significantly reduces the expression of MHC II molecules in circulating monocytes. Accordingly, CD8+ effector T cell composition, T helper 1 (Th1) and Th17 cytokine levels are reduced, while regulatory T cell (Treg) composition and Th2 cytokine levels are increased in IFLT versus CLT recipients. These results show that IFLT profoundly affects local and systemic immunity in liver transplantation. Recipient-circulating monocytes might play a key role in the interaction between graft IRI and allograft rejection.

Objective: This study aimed to investigate the efficacy of acupuncture and moxibustion in the treatment of chronic urticaria and its effect on cellular immune indexes and the imbalance of T helper 1 (Th1)/T helper 2 (Th2) cell function. Methods: A total of 70 patients with chronic urticaria who were diagnosed and treated at Wuhan NO.1 Hospital from January 2020 to April 2022 were selected and divided into a control group and an observation group using the random number table method, with 35 cases in each group. The control group was treated with loratadine, whereas the observation group was treated with loratadine combined with acupuncture. The treatment effects, quality-of-life scores, IgE levels, Th1 and Th2 cytokine levels, and adverse reactions were compared between the two groups. Results: The total effective rate of treatment in the observation group was significantly higher than that in the control group ( P &lt; 0.05). After treatment, the dermatology life quality index score and serum IgE level of the observation group were significantly lower than those before treatment and those in the control group ( P &lt; 0.05). After treatment, the levels of IL-2, IFN-γ, and TNF-α in both groups were significantly higher than those before treatment; however, the levels in the observation group were significantly lower than those in the control group ( P &lt; 0.05). After treatment, the levels of IL-6 and IL-8 in the two groups were significantly higher than those before treatment, and the levels in the observation group were significantly higher than those in the control group ( P &lt; 0.05). The IL-2 and IL-4 levels were significantly lower than before treatment, and the levels in the observation group were significantly lower than those in the control group ( P &lt; 0.05). There was no significant difference in the total incidence of adverse reactions between the two groups ( P &gt; 0.05). Conclusion: Acupuncture and moxibustion in the treatment of chronic urticaria has good clinical efficacy, which is beneficial to improve the patient's cellular immune function, balance the function of Th1/Th2 cells, and improve the clinical symptoms and quality of life of the patient. Moreover, the therapeutic effect is safe and effective.

Anti-airway inflammatory effects of a complex from <i>Scutellaria baicalensis</i> , <i>Nelumbinis semen,</i> and <i>Mori cortex</i> extracts in an ovalbumin-induced allergic asthma model

Level of Th-2 cytokines (IL-4, IL-5, and IL-13) in serum and relative expression of associated cytokine genes (IL-4, IL-5, and IL-13) in peripheral blood along with other parameters of host resistance were compared between the resistant and susceptible Garole sheep infected with Haemonchus contortus. Resistant (n = 12) and susceptible Garole sheep (n = 12) were selected based on faecal egg count (FEC) and packed cell volume (PCV) and were divided into three equal groups: healthy control, infected resistant, and infected susceptible. Sheep of infected groups were orally challenged with infective larvae of H. contortus and then FEC, body weight, haemoglobin (Hb), PCV, peripheral eosinophil counts (PEC), and serum Th-2 cytokines concentration and associated gene (IL-4, IL-5, and IL-13) expressions were measured. Partial sequences of all the three cytokine genes were also analysed. Significantly (p < 0.05) lower FEC was observed in resistant sheep with no eggs in faeces from 31 to 35 DPI while the susceptible sheep continued to pass Haemonchus eggs through faeces till end of the study. Body weight, Hb, PCV, and PEC of resistant sheep were higher (p < 0.05) compared with susceptible sheep. Concentrations of Th-2 cytokines (IL-4, IL-5, and IL-13) as well as expression all the genes (IL-4, IL-5, and IL-13) were found to be greater (p < 0.05) in resistant sheep on different days post infection. No single nucleotide variation was observed in either of the three genes that could be suggested to be responsible host resistance. In resistant sheep, increased eosinophilia, Th-2 cytokine concentration, and relative expression Th-2 cytokine genes in peripheral blood were observed synchronously with that of the elimination of parasites indicating the role of those components in resistance against H. contortus and this needs to be explored by examining local reaction in the abomasum of infected host.

Brucella secreted effector BspH stimulates intracellular ion flux and confers anti-Brucella immunity.