IL-8 Levels Research Articles

Augmented IFNγ producing ILC1 and IL 17 producing ILC3 in pemphigus vulgaris: Plausible therapeutic target.

Innate Lymphoid cells (ILCs) are innate counterparts of helper T cells. Although low in number, they have proven to play major roles in many autoimmune diseases. In Pemphigus Vulgaris (PV), the gaps in the knowledge of functional role of ILCs remain. To bridge the gap, our study investigated the phenotype along with the functional determinants of ILCs involved in PV immunopathogenesis. Our data suggested augmentation in overall ILC population in circulation of PV patients. Specifically, ILC1 and ILC3 subtypes were significantly increased in peripheral circulation of PV patients compared to healthy controls. We observed no changes in ILC2 population. mRNAs from ILC enriched population showed significant upregulation in transcription factors- ID2, T bet and RORγt and a downregulation in GATA3 and RORα. The mRNA levels of ILC related cytokines- IFNγ and IL17 were significantly upregulated while no change was observed in the levels of IL13, IL 22, AHR. The levels of autoantibodies against desmoglein (Dsg) 3 which is the characteristic of PV pathogenesis were also checked in the serum which confirmed significant upregulation in PV patients. The levels of proinflammatory- IFNγ, IL 17 and IL 15 were elevated and anti-inflammatory cytokines- IL10 was downregulated in the serum of PV patients. The results of this study offer insights into the functional attributes of ILCs and related cytokines, potentially contributing to the development of future therapeutic interventions.

A multicenter, prospective phase II trial of second-line aflibercept plus FOLFIRI in patients with metastatic colorectal cancer refractory to anti-EGFR agents (HGCSG1801): Updated analyses.

147 Background: The HGCSG1801 trial evaluated the treatment outcomes of aflibercept (AFL) plus FOLFIRI for patients(pts) with metastatic colorectal cancer (mCRC) refractory to an oxaliplatin-based regimen combined with an anti-EGFR agent. Here we report results of the updated efficacy, safety, and a biomarker analysis. Methods: This was a prospective open-label phase II trial. AFL (4 mg/kg iv) followed by FOLFIRI (irinotecan 180 mg/m 2 , leucovorin 200 mg/m 2 iv, bolus 5-fluorouracil [5-FU] 400 mg/m 2 , and infusional 5-FU 2400 mg/m 2 /46 h) was given every 2 weeks until progression or unacceptable toxicities. The primary endpoint was 6-month progression-free survival (PFS) rate, and the secondary endpoints included overall survival (OS), PFS, overall response rate (ORR), disease control rate (DCR), and adverse events. Angiogenic factors were analyzed in plasma sample using a Luminex multiplex assay using the Cox proportional hazards model. This study was sponsored by the Non-Profit Organization Hokkaido Gastrointestinal Cancer Study Group and supported by a grant from Sanofi. Results: Forty-three pts were enrolled between November 2019 and October 2022. The data cut-off date was April 30, 2024. The 6-month PFS rate was 59.0% (90% confidence interval [CI], 45.8–72.1%). Median PFS and OS were 7.3 months (95% CI, 5.5–11.0 months) and 18.8 months (95% CI, 12.9–26.6 months), respectively. The ORR was 20.9% (95% CI, 10.0–36.0%) and DCR was 88.4% (95% CI, 74.9–96.1%). No deaths and no new safety signals with a causal relation to the study treatment were observed. A biomarker analysis using pretreatment plasma samples showed a trend toward better PFS in pts with low VEGF-A (hazard ratio [HR] 0.40 [95% CI, 0.18-0.91]), TSP-2 (HR 0.40 [95% CI, 0.18-0.88]) or IL-8 levels (HR 0.43 [95% CI, 0.20-0.93]). There was also a trend toward better OS in pts with low levels of TSP-2 (HR 0.30 [95% CI, 0.11-0.81]) or TIMP-1 (HR 0.20 [95% CI, 0.06-0.69]). Conclusions: These updated data further support the activity and manageable safety profile of AFL plus FOLFIRI for pts with mCRC who failed an oxaliplatin-based regimen combined with an anti-EGFR agent. It was suggested the association between some angiogenic factors in plasma samples and the activity of AFL plus FOLFIRI in mCRC. Clinical trial information: jRCTs011190006 .

Childhood allergy and anxiety/depression in early adulthood: A longitudinal study in the ALSPAC birth cohort.

Allergic disease and common mental disorders frequently co-occur. However, little is known about the longitudinal impact of childhood allergy on the subsequent risk of developing anxiety or depression, and the possible biological mechanisms for this. We performed longitudinal analyses of data from the Avon Longitudinal Study of Parents and Children (ALSPAC) birth cohort. The baseline sample comprised n=5256 children with allergy data available at age 7yrs. We used multivariable regression to test associations between childhood allergy at age 7yrs and: a) four inflammatory markers at age 9yrs; b) depression and anxiety measures between ages 10-24yrs. Allergy measures included biological markers (total serum immunoglobulin E (tIgE), number of positive skin prick tests (SPTs)), and presence of eczema, asthma and/or food allergy (mother reported). Inflammatory markers were interleukin-6 (IL-6), C-reactive protein (CRP), IL-4 and IL-13. We used structural equation modelling to test whether inflammatory markers mediated the association between tIgE and depression/anxiety. tIgE and having≥1 positive SPT at age 7 were associated with IL-6 levels at age 9 (adjusted β=0.09; 95% CI 0.06-0.13; p<0.001 and adjusted β=0.06; 95% CI 0.03-0.09; p<0.001 respectively), but not with CRP, IL-4 or IL13 levels. We found no strong evidence of an association between childhood allergy and subsequent depression/anxiety during adolescence and early adulthood. This finding was consistent across biological and mother-reported allergy measures. Biological markers of childhood allergy are associated with IL-6, a key inflammatory cytokine. However, childhood allergy may not have a large long-term effect on subsequent depression/anxiety.

Impact of microbial diversity on inflammatory cytokines and respiratory pattern measured in whole-body plethysmography in guinea pig models.

This study investigates the breathing patterns and immune status of guinea pigs raised under specific pathogen-free (SPF) conditions compared to conventionally bred (CON). Breathing pattern parameters were assessed using whole-body plethysmography (WBP) during quiet breathing and saline nebulisation. Blood and bronchoalveolar lavage fluid (BALF) were analysed for white blood cell, neutrophil and eosinophil counts, and cytokine levels (TNF-α, IL-1β, IL-4). SPF guinea pigs exhibited higher tidal volume, expired volume, minute volume, and airflow parameters than CON guinea pigs. The immune analysis revealed lower white blood cell counts and IL-4 levels in SPF guinea pigs. These findings indicate that SPF guinea pigs have different respiratory and immune responses than CON guinea pigs. The study highlights that the maturation processes affecting breathing pattern parameters in SPF guinea pigs differ significantly from those in CON guinea pigs. This suggests potential limitations of SPF animals in respiratory physiology research due to their different immune and respiratory responses.

Inflammatory factors predict infection risk during chemotherapy in leukemia patients.

This study explores the novel role of specific inflammatory factors in predicting infection risk among leukemia patients undergoing chemotherapy, providing new insights into managing leukemia-associated infections. This prospective cohort study included 244 leukemia patients suspected of infection, divided into the infected group (n=194) and the uninfected group (n=50) based on etiological testing. The infected group was further categorized into bacterial (n=123), fungal (n=22), and viral (n=49) infection subgroups. Inflammatory cytokines including IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-α, IFN-γ, IL-17A, IL-127P0, and IFN-α were measured using ELISA kits. Additional markers such as hs-CRP, SAA, and PCT were also assessed for the infected group. The levels of IL-1β, IL-4, IL-5, IL-6, IL-8, IL-10, IFN-γ, IL-12p70, and IFN-α significantly differed between the infected and uninfected groups (P<0.05). IL-1β, IL-4, IL-6, and IL-10 were significantly higher in the infected group compared to the uninfected group. Significant differences in PCT, IL-2, IL-4, IL-5, and IL-10 levels were observed among patients with bacterial, fungal, and viral infections (P<0.05). However, none of these inflammatory factors were predictive of infection type. IL-4, IL-6, IL-8, and IL-10 were identified as independent predictors of infection risk. However, no inflammatory factors could be used to distinguish between different types of infections.

Interleukin-13 partly induced by the NLRP3 inflammasome promotes Trichinella spiralis encapsulation in infected mice.

Trichinella spiralis infection is a serious parasitic zoonosis in which a collagenous capsule surrounding the larva is developed in the striated muscle cells. However, the mechanism of T. spiralis encapsulation is currently poorly understood. It has been reported that T. spiralis infection can induce the production of IL-13 via the NLRP3 inflammasome, and it has also been suggested IL-13 thus produced may be involved in T. spiralis encapsulation. This research aimed to clarify the involvement of NLRP3 and IL-13 in the T. spiralis capsule formation process. IL-13 and NLRP3 inhibitors were used in a T. spiralis infected mouse model and in C2C12 cells to analyze the role of IL-13 and NLRP3 in encapsulation. The results showed that T. spiralis infection significantly increased the expression levels of IL-13 and collagen IV and VI. The production of collagen around the T. spiralis encapsulation zone was significantly inhibited when an IL-13 inhibitor was applied. Moreover, the expression levels of IL-13 and collagen IV and VI were significantly decreased by the NLRP3 inhibitor in vitro and in vivo. The above results indicated that NLRP3 can participate in the development of T. spiralis encapsulation by regulating IL-13 expression and stimulating collagen IV and VI synthesis during T. spiralis infection.

What are the levels and interactions of neuroligin-1, neuroligin-3, and inflammatory cytokines (IL-6, IL-8) in children diagnosed with autism spectrum disorder?

Autism spectrum disorder (ASD) is characterized by deficits in social interaction, restricted interests, and repetitive behaviors. Several genes, including synaptic proteins and environmental risk factors, play a role in the etiology of autism. We aimed to evaluate the relationship between neuroligin-1 (NLGN-1) and neuroligin-3 (NLGN-3) levels, which are neuronal cell adhesion molecules (CAMs), and inflammatory cytokine (IL-6, IL-8) levels with disease severity and symptom clusters and with each other in children with ASD. Eighty children diagnosed with autism who met the inclusion criteria and sixty-five typically developing children matched for age and sex were included in the study. The children were evaluated psychiatrically through a semi-structured interview, DSM-5 criteria, the Childhood Autism Rating Scale (CARS), and the Social Communication Questionnaire (SCQ). IL-6, IL-8, NLGN-1, and NLGN-3 levels were analyzed in peripheral serum samples using human ELISA kits. IL-8 and NLGN-3 levels were higher in the autism group (p < 0.001, p < 0.001). IL-6 was positively related to CARS and SCQ total scores (p = 0.021, p = 0.040, respectively). IL-8, and NLGN-3 were positively associated with the all subtests of the SCQ and the SCQ total score (all p values <0.001). NLGN-1, NLGN-3, and inflammatory cytokine (IL-6, IL-8) levels were positively correlated (all p values <0.001). Neuroligins play a central role in the brain's ability to process information and maybe a key target in the pathogenesis of ASD. Further research is needed to determine whether, to what extent and how neuronal CAMs and immunity modulate each other and whether this contributes to ASD pathogenesis. Future studies should also be expanded to investigate the influence of variables such as oxidative stress, metalloproteases responsible for ectodomain shedding, or epigenetic regulation.

Impact of Dandelion (Taraxacum officinale) leaf aqueous extract on immunological response of mice after Schistosoma mansoni infection.

This study investigated the effect of dandelion (Taraxacum officinale) leaf aqueous extract (DLE) on the immunological response of mice following infection with Schistosoma mansoni. Mice (in groups of 7) were first experimentally infected with S. mansoni and, 6 weeks later, were treated with praziquantel (PZQ) and/or DLE. Control mice were uninfected. In contrast to the untreated group, animals given PZQ and/or DLE exhibited an enhanced immunological response, as indicated by increased serum IFNγ, TNFα, IL4 and IL10 levels, increased numbers of CD4+ and CD25+ cells in blood and spleen and altered expression of apoptosis-related genes (low Bax and caspase3 and high Bcl2) in the spleen. DLE treatment had a significantly bigger impact in all these parameters compared with PZQ alone and combined DLE/PZQ treatment have the largest effect. While DLE treatment alone significantly decreased parasite burden, it did not improve upon the greater protective effect of PZQ, even when given in combination.

Exploring the therapeutic effect of human recombinant IL11 on lesioned OA human osteochondral explants

ObjectiveTo explore IL11 co-expression profiles in our previously reported RNA-sequencing dataset of OA articular cartilage, in interaction with IL6, and to investigate the effects of hrIL11 administration as potential therapeutic strategy for OA articular cartilage using our biomimetic aged human osteochondral explant model of OA.MethodsWe used RNA-sequencing datasets of macroscopically preserved and lesioned OA articular cartilage (N = 35 patients). Spearman correlations were calculated between IL11 and IL6 expression levels and genes expressed in cartilage (N = 20048 genes). Osteochondral explants were isolated from macroscopically preserved and lesioned areas of the joint and were kept in culture for two weeks, with or without exposure to 200ng/ml hrIL11.ResultsWe found no overlap in correlating genes between IL11 and IL6, indicating their distinct roles in articular cartilage. Moreover, we identified more genes being correlated to IL11 in the lesioned compared to preserved articular cartilage (N = 203 and 106 genes, respectively). Upon treatment of ex vivo OA articular cartilage with hrIL11, we overall observed unbeneficial effects on chondrocyte phenotype, as illustrated by upregulation of MMP13, EPAS1, RUNX2, and POSTN. We did not observe significant differences in Mankin scores upon addition of hrIL11.ConclusionThe current study showed that treatment of OA articular cartilage with hrIL11 is unlikely to be beneficial despite previous indications of hrIL11 as potential druggable target. These findings underscore the importance of functionally investigating OA risk genes. Better understanding of IL11 signaling and the underlying pathways is necessary towards the development of OA treatment strategy.

An animal model of severe acute respiratory distress syndrome for translational research

BackgroundDespite the fact that an increasing number of studies have focused on developing therapies for acute lung injury, managing acute respiratory distress syndrome (ARDS) remains a challenge in intensive care medicine. Whether the pathology of animal models with acute lung injury in prior studies differed from clinical symptoms of ARDS, resulting in questionable management for human ARDS. To evaluate precisely the therapeutic effect of transplanted stem cells or medications on acute lung injury, we developed an animal model of severe ARDS with lower lung function, capable of keeping the experimental animals survive with consistent reproducibility. Establishing this animal model could help develop the treatment of ARDS with higher efficiency.ResultsIn this approach, we intratracheally delivered bleomycin (BLM, 5 mg/rat) into rats’ left trachea via a needle connected with polyethylene tube, and simultaneously rotated the rats to the left side by 60 degrees. Within seven days after the injury, we found that arterial blood oxygen saturation (SpO2) significantly decreased to 83.7%, partial pressure of arterial oxygen (PaO2) markedly reduced to 65.3 mmHg, partial pressure of arterial carbon dioxide (PaCO2) amplified to 49.2 mmHg, and the respiratory rate increased over time. Morphologically, the surface of the left lung appeared uneven on Day 1, the alveoli of the left lung disappeared on Day 2, and the left lung shrank on Day 7. A histological examination revealed that considerable cell infiltration began on Day 1 and lasted until Day 7, with a larger area of cell infiltration. Serum levels of IL-5, IL-6, IFN-γ, MCP-1, MIP-2, G-CSF, and TNF-α substantially rose on Day 7.ConclusionsThis modified approach for BLM-induced lung injury provided a severe, stable, and one-sided (left-lobe) ARDS animal model with consistent reproducibility. The physiological symptoms observed in this severe ARDS animal model are entirely consistent with the characteristics of clinical ARDS. The establishment of this ARDS animal model could help develop treatment for ARDS.

Association between serum levels of 12 different cytokines and short-term efficacy of chemoradiotherapy in esophageal squamous cell carcinoma

BackgroundEsophageal squamous cell carcinoma (ESCC) has a poor prognosis, with chemoradiotherapy (CRT) being a key treatment method. This study focused on circulating cytokines as potential predictors of treatment response and prognosis in patients with ESCC.Materials and methodsSerum samples were collected from 36 ESCC patients, and 12 different cytokines were quantified using a multiplex immunofluorescence assay. We used non-parametric Wilcoxon unpaired rank tests to examine the relationship between cytokine concentrations and clinical outcomes. The duration of progression-free survival was assessed through imaging studies and telephone follow-ups. Kaplan–Meier survival plots, analyzed with the log-rank test, were utilized to depict survival trends.ResultsPre-treatment serum IL-8 levels were significantly elevated in patients with lymphoid metastases (p = 0.036). Lower initial levels of IL-8 and IL-1β were observed in patients with partial response group compared to those with stable disease (p = 0.002, p = 0.01). Elevated baseline levels of IL-8 and interferon-gamma (IFN-γ) were correlated with a poorer prognosis. Higher levels of IL-5 and IFN-γ levels following therapy were associated with worse outcomes.ConclusionsOur findings indicate that IL-8, IL-1β, IL-5, and IFN-γ may serve as potential biomarkers for treatment efficacy and prognosis in ESCC. Patients with low levels of IL-8 and IL-1β demonstrate a favorable response to CRT. Elevated serum levels of IL-8, IL-1β, IFN-γ, and IL-5 may predict poorer clinical outcomes.

Sargassum fusiforme polysaccharides protect mice against Citrobacter rodentium infection via intestinal microbiota-driven microRNA-92a-3p-induced Muc2 production.

Sargassum fusiforme, widely consumed in Asian countries, has been proven to have various biological activities. However, the impacts and mechanisms of Sargassum fusiforme polysaccharides (SFPs) on intestinal bacterial infection are not yet fully understood. Our findings indicate that SFPs pretreatment ameliorates intestinal inflammation by reducing C. rodentium colonization, increasing colon length and levels of IL-10 and IL-22, decreasing IL-1β, IL-6, TNF-α, and IL-17 levels, inhibiting colonic crypt elongation and hyperplasia, and enhancing the intestinal mucosal barrier. The protective effects against intestinal bacterial infection are linked to enhanced clearance of C. rodentium and improvements in the intestinal mucosal barrier and C. rodentium-induced intestinal microbiota dysbiosis. Fecal microbiota transplantation experiments were conducted to evaluate the functional impact of microbiota induced by SFPs. The results suggest that intestinal microbiota modified by SFPs effectively countered C. rodentium infection. In addition, our study identified that miRNA-92a-3p is partially complementary to the 3'-UTR of the Notch1 gene, thereby repressing the Notch1-Hes1 signaling pathway and enhancing Muc2 secretion. Taken together, these findings reveal that SFPs protect mice from C. rodentium infection by activating the miR-92a-3p/Notch1-Hes1 regulatory axis driven by the intestinal microbiota, which stimulates Muc2 production to maintain intestinal barrier homeostasis.

P0131 Characterisation of a human multi-organ-on-chip model to unravel gut-blood-brain communication

Abstract Background Persistent fatigue is a frequently reported symptom among patients with Inflammatory Bowel Disease (IBD), significantly diminishing quality of life. Due to the lack of understanding how IBD-associated fatigue originates, currently available treatments do not efficiently tackle this symptom. Developing targeted therapies requires tools to unravel the mechanisms behind central nervous system dysfunction in response to chronic intestinal inflammation. We are therefore establishing and characterising a human multi-organ-on-chip model to recapitulate key physiological elements of gut-brain communication via the systemic circulation. Methods In our model, a gut-blood barrier, comprised of Caco-2 (enterocytes) and HT29-MTX cells (goblet cells), is connected to a blood-cerebrospinal fluid (CSF) barrier, composed of HIBCPP and iHCPEnC cells (choroid plexus epithelial and endothelial cells respectively), and to a brain compartment (SH-SY5Y, neurons) via recirculation of cell culture medium. Gut inflammation was mimicked by the addition of the pro-inflammatory cytokines TNFα and IFNγ to the intestinal epithelial cells. Barrier integrity was measured by transepithelial/endothelial resistance (TEER) and by paracellular permeability assays with 4 kDa FITC-dextran. In addition, we performed a bead-based immunoassay to measure secreted IL-8 levels and RT-qPCR to analyse differential gene expression. Results The gut-blood barrier under low perfusion (3,6*10-4 mPa) exhibited increased expression of differentiation marker genes compared to static culture, such as LYZ, SOX9 and CYP3A4, indicating an improvement of cell differentiation. Moreover, low perfusion induced the formation of 3D villi-like structures, mucus production and the reduction of barrier strength towards more physiological relevant levels (TEER: 81,6±44,8 Ω.cm2; Papp: 1.10-5±4.10-6 cm/s). The gut-blood barrier was responsive to the addition of TNFα and IFNγ and high IL-8 levels could be measured in the circulatory blood flow. The barrier properties of the blood-CSF barrier under low perfusion (endothelial: 2,52 mPa, epithelial: 4,6*10-4 mPa) were not significantly different from a static setup, but less variance was observed in TEER. Neurons differentiated with BDNF and retinoic acid in our model displayed long projections and expressed mature neuronal marker genes (SYP, SNAP25, TUBB3). Conclusion We have successfully established and characterised each organ compartment of the gut-blood-brain model. Moreover, we have proven that low perfusion improves the physiological relevance of our model, especially of the gut-blood barrier. We are now exploring phenotypical and transcriptional alterations across the blood-CSF barrier and the brain compartment when gut inflammation is triggered.

The Impact of Inflammatory Cytokines on Recurrent Pregnancy Loss: A Preliminary Investigation.

Recurrent pregnancy loss (RPL), defined as two or more consecutive miscarriages before 20weeks of gestation, affects 1-2% of couples worldwide. Pro-inflammatory cytokines, such as TNF-α, IL-1β and IL-6 play critical roles in early pregnancy, while anti-inflammatory cytokines like TGF-β and IL-10 promote immune tolerance to prevent harmful inflammatory responses that play important role in placental and fetal development. This aim of the study is to analyse the levels of inflammatory cytokines in blood serum from RPL patients and healthy women (control). The measured cytokines included TNF-α, IL-6, IL-10, TGF-β, CRP, ferritin, IL-1β and IL-4, IFN-γ and IL-17. Using an unpaired t-test and Pearson correlation, significant difference observed between the groups. The results had significantly elevated CRP levels with decreased levels of TGF-β and ferritin (p < 0.05), whereas, IL-1β and IL-4 also found raised indicating a link between systemic inflammation and recurrent miscarriages. IL-4 and CRP increase further suggest potential oxidative stress role in RPL cases. However, no significant differences observed in IL-10, IL-6, or TNF-α level between the groups. This study highlights immune dysregulation as possible contributors to early pregnancy loss, with significant increases in CRP, IL-1β, and IL-4 levels indicating an imbalanced immune response at the maternal-fetal interface. These cytokine elevations may disrupt immune tolerance, suggesting the need for further exploration into cytokine interactions in pregnancy and their potential as an investigatory biomarker and therapeutic target in RPL.

P0122 The Potential Role of Circulating Cytokines and Chemokines To Predict Intestinal Failure in Patients Affected by Short Bowel Syndrome

Abstract Background Short bowel syndrome (SBS) is a life-altering and life-threatening disease resulting from massive small bowel resections, often occurring in patients with Crohn's disease (CD), a severe chronic gastrointestinal disorder characterized by the infiltration of inflammatory cells into the intestinal mucosa and submucosa. Several cytokines and chemokines play a crucial role in the development and progression of inflammatory bowel disease by recruiting inflammatory cells1,2. Currently, no biomarkers indicate a higher risk of developing SBS or intestinal failure (IF); thus, the present study aims to evaluate the circulating cytokine/chemokine profile in patients affected by IF/SBS and CD patients at high risk of developing IF, to identify disease severity-related biomarkers. Methods A total of 150 CD patients were enrolled in this study, and they were stratified into four subgroups based on the disease severity: SBS patients with IF and parenteral nutrition (A1 group) or without IF and inactive disease (A3 group); CD patients with high (B group) or low risk of IF (C2 group). Circulating cytokines and chemokines levels were determined using the human 27-PLEX assay on sera collected from patients at the moment of the enrollment (T0, n=70) and after 6 months (T6M, n=10). Results Among inflammatory cytokines, high levels of RANTES, PDGF-BB, IP-10, MIP-1β, IL-9, and EOTAXIN were detected in all groups, while the levels of anti-inflammatory interleukins such as IL-4 and IL-13 resulted low in all patients at T0 (fig.1). At T6M, a general reduction of all the enhanced inflammatory markers was detected in all the groups except for RANTES, whose levels were almost unchanged and even increased in the A3 group. On the contrary, IP-10, which appears explicitly increased in the A1 group, is the most significantly reduced in all groups at T6M (fig.2). Conclusion Our results show a difference in the levels of circulating cytokines/chemokines profiles among the different groups of patients. In particular, IP-10, EOTAXIN, and PDGF-BB are significantly higher in the A1, A3, and C2 groups, respectively, while IL-9, RANTES, and MIP-1β levels present a similar profile at T0. After 6 months, a general reduction of all the analyzed inflammatory markers was recorded in all the groups. In particular, IP-10 shows the most relevant reduction, while RANTES levels remain almost unaltered at T6M. Literature data confirmed a role for these cytokines/chemokines in CD severity and progression3,4. However, to validate these preliminary data and potentially find a biomarker for IF prediction, more samples will be collected and analyzed.

Inula japonica Thunb. and its active compounds ameliorate airway inflammation by suppressing JAK-STAT signaling.

Asthma, a chronic inflammatory disease, remains a global health challenge due to its complex pathophysiology and the limited treatment efficacy. This study explored the effect of Inula japonica Thunb. water extract (IJW) on asthma and its protective mechanisms. To assess the effects of IJW, we established an experimental asthma model in BALB/c mice using ovalbumin (OVA). Airway hyper-responsiveness (AHR) in response to methacholine was measured. We quantified inflammatory cell infiltration and cytokine and chemokine levels in bronchoalveolar lavage fluid (BALF), as well as total IgE levels in serum. Staining with hematoxylin and eosin and periodic acid-Schiff was used to examine the impact of IJW on lung pathology. We performed RNA sequencing to identify differentially expressed genes, which were subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses. We used interleukin (IL)-4/IL-13-treated human bronchial epithelial (HBE) cells to explore the associated mechanisms. IJW showed therapeutic effects against OVA-induced asthma by alleviating AHR, peribronchial inflammation, mucus hypersecretion, and collagen fiber deposition. It reduced total IgE levels in the serum and IL-4, IL-5, IL-13, eotaxin, macrophage-derived chemokines, and periostin levels in BALF. In IL-4/IL-13-treated HBE cells, IJW and its components suppressed the Janus kinase-signal transducer and activator of the transcription (JAK-STAT) signaling. These findings support IJW's potential as a pharmacological agent for allergic airway inflammation and asthma.

P1019 Role of Oncostatin M (Osm)-Osmr signaling in colonic inflammation and host defense during citrobacter rodentium infection

Abstract Background Elevated Oncostatin M (OSM) and its receptor (OSMR) are linked to the pathology of inflammatory bowel disease (IBD), particularly in severe, treatment-refractory cases. While anti-OSM therapies hold potential for IBD management, their impact on susceptibility to acute enteric infections remains unclear. This study aims to clarify the role of Osm-Osmr signaling in the context of Citrobacter rodentium infection, a murine model mimicking acute colonic inflammation. Methods Temporal studies were conducted on C57BL/6J mice infected with C. rodentium, assessing gene expression, bacterial load, and immune dynamics at days 3, 7, 14, and 21 post-infection. Histological analysis and flow cytometry were performed to evaluate inflammation and immune infiltration. Additionally, Osm knockout (KO) and Osmr conditional KO mice were utilized to investigate the role of OSM signaling in pathogen control and immune responses. Results Colonic expression of Osm and Osmr peaked at day 14, concurrent with increased levels of Il-22, IFN-g, and antimicrobial peptides (Reg3b, Reg3g), indicating robust immune activation and epithelial repair. ELISA confirmed elevated OSM protein levels. Histology at day 14 revealed severe colonic inflammation, with high Osm/Osmr expression correlating with inflammation scores (p &amp;lt; 0.0001). Flow cytometry showed increased CD45⁺ immune cells, including neutrophils, monocytes, and memory T cells. Notably, OSM signaling was dispensable for bacterial clearance, as Osm knockout mice displayed comparable bacterial loads to wild-type controls despite similar inflammatory responses, suggesting that OSM’s role may be more focused on modulating inflammation and tissue repair rather than direct pathogen control. Conclusion Osm-Osmr signaling plays a key role in modulating inflammation and epithelial repair during C. rodentium infection. The coordinated upregulation of OSM and OSMR enhances immune responses and supports epithelial integrity, highlighting potential therapeutic implications for IBD. Further research is needed to assess the impact of anti-OSM therapies on balancing inflammation control and host defense against infections.

DOP117 Innate lymphoid cells control colitis and intestinal fibrosis via cellular stress response in mice and human

Abstract Background Group 2 innate lymphoid cells (ILC2s) are increased in peripheral blood and intestinal samples from IBD patients and mice with colitis. ILC2s are major producers of type 2 cytokines including IL-5 and IL-13 and growth factor AREG that induce goblet cell hyperplasia and mitigate acute inflammation in the gut. On the other hand, prolonged or uncontrolled ILC2 activation may contribute to chronic inflammation and tissue fibrosis, although the role of ILC2s in stricturing/fibrostenotic Crohn’s disease (CD) has been unknown. In this study, we investigated the function of IRE1a-XBP1 pathway, a regulatory hub of cellular stress response, in colitis and fibrosis in mice and CD patients. Methods We generated conditional knockout Ire1aflo/floxIl5-Cre (Ire1aΔIl5) and Rag-/-Ire1aΔIl5 mice with Ire1a deleted in ILC2s. Dextran sodium sulfate (DSS)-induced and T-cell transfer colitis models were used. Human peripheral ILC2s were collected for flow cytometry and ex vivo stimulations. Results Mouse intestinal ILC2s express high level of Ire1a. The number of ILC2s increased in colon, small intestine (SI), and esophagus of Ire1aΔIl5 mice with elevated proliferation marker Ki67. Additionally, SI and colonic Ire1aΔIl5 ILC2s produced more IL-5, IL-13, and AREG after ex vivo stimulation with IL-33 or IL-25. Consistently, selective IRE1 inhibitor 4u8C enhanced production of IL-13, while selective IRE1 activator IXA4 suppressed IL-13 in sort-purified human ILC2s ex vivo (Figure 1). Bulk RNA-seq of SI Ire1aΔIl5 ILC2s showed reduced Rxrg and elevated fatty acid transport by pathway analysis, which may lead to the increased proliferation and cytokine production in Ire1aΔIl5 ILC2s. Upon DSS challenge, Ire1aΔIl5 mice exhibited milder signs of disease including weight loss, clinical scores as well as colon shortening and histology, which was accompanied by elevations in ILC2-derived cytokines and AREG as well as heightened goblet cell differentiation and mucin production. In contrast, upon adoptive T-cell transfer, Rag-/-Ire1aΔIl5 mice developed worsened chronic colitis and intestinal fibrosis, which was reversed by intraperitoneal injection of neutralizing antibody against IL-13 or AREG. Furthermore, CD patients with intestinal strictures expressed higher level of IL-13 in ILC2s in the blood (Figure 2). Conclusion IRE1a-XBP1 constrains cytokine production by intestinal ILC2s during inflammation. IRE1a deficiency in ILC2s protected against acute colitis with heightened type 2 cytokines and AREG; while loss of IRE1a exacerbated chronic colitis and fibrosis. The presence of CD strictures was associated with elevated IL-13 in peripheral ILC2s . We identified IRE1a and ILC2s as potential biomarkers and therapeutic targets for stricturing CD.

Peripheral blood age-sensitive immune markers in multiple sclerosis: relation to sex, cytomegalovirus status, and treatment.

Immunosenescence is accelerated by chronic infectious and autoimmune diseases and could contribute to the pathobiology of multiple sclerosis (MS). How MS and disease-modifying therapies (DMTs) impact age-sensitive immune biomarkers is only partially understood. We analyzed 771 serum samples from 147 healthy controls and 289 people with MS (PwMS) by multiplex immunoassays. We determined cytomegalovirus (CMV) serostatus and collected retrospective clinical information. We performed unsupervised and multivariable analyses. Unsupervised analyses revealed that MS immune profile was characterized by low relative levels of anti-inflammatory/neuroprotective factors IL-4, IL-10, TNF, and β-NGF but high levels of growth factors EGF and bFGF. Serum levels of IL-4, β-NGF, IL-27, BDNF, and leptin were significantly influenced by sex and/or CMV status. IL-4 and β-NGF levels were lower in untreated PwMS compared to controls, while EGF and bFGF levels were influenced by age and markedly elevated in PwMS in multivariable analysis. Samples from treated PwMS, but not untreated PwMS, showed lower levels of BDNF and TNF than controls. Initiation of high efficacy DMTs, but not low efficacy DMTs, was associated with reduced levels of bFGF and EGF. Samples associated with distinct DMTs exhibited specific profiles for age-sensitive immune markers. Finally, lower levels of IL-6, TNF, IL-10, and β-NGF were observed at baseline in PwMS who subsequently experienced clinical failure after DMTs initiation. Age, sex, CMV status, and specific DMTs significantly influence levels of age-sensitive immune biomarkers associated with MS and must be considered when investigating inflammation-related biomarkers. This work was supported by a Grant for Multiple Sclerosis Innovation by Merck KGaA (ID: 10.12039/100009945).

Inhibiting miR-155-5p promotes proliferation of human submandibular gland epithelial cells in primary Sjogren's syndrome by negatively regulating the PI3K/AKT signaling pathway via PIK3R1.

To investigate the mechanism mediating the regulatory effect of miR-155-5p on proliferation of human submandibular gland epithelial cells (HSGECs) in primary Sjogren's syndrome (pSS). Dual luciferase reporter assay was used to verify the targeting relationship between miR-155-5p and the PI3K/AKT pathway. In a HSGEC model of pSS induced by simulation with TRAIL and INF-γ, the effects of miR-155-inhibitor-NC or miR-155 inhibitor on cell viability, cell cycle, apoptosis and proliferation were evaluated using CKK8 assay, flow cytometry and colony formation assay. ELISA and RT-PCR were used to detect the expressions of inflammatory cytokines and miR-155-5p mRNA in the cells; Western blotting was performed to detect the expressions of proteins in the PI3K/AKT signaling pathway. Dual luciferase assay showed that miR-155-5p targets the PI3K/AKT pathway via PIK3R1 mRNA. The HSGEC model of pSS showed significantly decreased cell viability, cell clone formation ability and expressions IL-10 and IL-4 and increased cell apoptosis, cell percentage in G2 phase, expressions of TNF‑α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-Akt/AKT ratio, and PIK3R1 protein expression. Treatment of the cell models with miR-155 inhibitor significantly increased the cell viability, G1 phase cell percentage, colony formation ability, and expressions of IL-10 and IL-4 levels, and obviously reduced cell apoptosis rate, G2 phase cell percentage, expressions of TNF-α, IL-6, miR-155-5p and PIK3R1 mRNA, p-PI3K/PI3K ratio, p-AKT/AKT ratio, and PIK3R1 protein expression. In HSGEC model of pSS, inhibition of miR-155-5p can promote cell proliferation and reduced cell apoptosis by targeting PI3K1 mRNA to negatively regulate the overexpression of PI3K/AKT signaling pathway.