Expression Of Interleukin Research Articles

Ningxiang pig-derived Lactobacillus reuteri improves the gut health of weaned piglets by regulating intestinal barrier function and cytokine profiles

Weaning stress in piglets can induce intestinal damage, resulting in impaired growth performance. Probiotics have emerged as significant contributors to enhancing gut health in piglets. This study aimed to evaluate the effects of Lactobacillus reuteri (L. reuteri) supplementation on growth performance, intestinal immunity, and intestinal barrier integrity in weaned piglets. In this investigation, fourteen healthy weaned piglets of similar age and weight, were randomly assigned to two groups (n = 7), receiving either normal saline (Control group) or L. reuteri (L-treatment group) over a 16-day period. The findings revealed no significant impact of L. reuteri on growth performance compared to controls. However, it lowered serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels (P < 0.05) and elevated serum concentrations of Immunoglobulin A (IgA), immunoglobulin G (IgG), and secretory immunoglobulin A (sIgA) (P < 0.05). Additionally, L. reuteri notably enhanced the villus height-to-crypt depth ratio in the ileum (P < 0.05) and increased mRNA expression of zonula occludens-1 (ZO-1), Claudin-1, and ZO-1 in ileal tissue (P < 0.05). Furthermore, L. reuteri reduced mRNA expression of pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) (P < 0.05) in the jejunum and colon while enhancing the expression of the anti-inflammatory cytokine interleukin-10 (IL-10) (P < 0.05) in both the ileum and colon. This study demonstrates that L. reuteri isolated from Ningxiang pigs can improve the intestinal health of weaned piglets by modulating gut barrier function and cytokine levels.

Impact of Bambusa vulgaris-supplemented diet on Nile tilapia challenged with Pseudomonas putida: Hematological, immune, and oxidative responses.

This study investigated the effects of bamboo shoot extract (Bambusa vulgaris) as a feed additive on the health profiles and infection resistance of Nile tilapia (Oreochromis niloticus) against Pseudomonas putida. Bamboo shoot extract was added at levels of 0g, 40g, and 60g per 1000g of diet over a 60-day period. The fish were then challenged with a pathogenic P. putida strain. Chemical analysis of the bamboo shoot extract identified 3,5-dinitrophenol and hydroquinone as the two most abundant compounds. Results showed that fish fed bamboo-enriched diets exhibited significantly enhanced levels of red blood cells, hemoglobin, hematocrit, white blood cells, and platelets, and improved erythrocyte cellular and nuclear morphologies, indicating improved health profiles after the challenge. Liver function indicators, including AST, ALT, and ALP, were notably balanced in fish receiving bamboo shoot extract post-challenge (p<0.05). Blood levels of K+ were lower in the bamboo-fed groups. Additionally, blood levels of Ca++ and Na+ were reduced in fish fed 40g and 60g of bamboo, respectively, compared to the control group (p<0.01). The bamboo extract also enhanced immune and oxidative capacities, as demonstrated by increased catalase, superoxide dismutase, lysozyme activity, and phagocytic activity, along with reduced malondialdehyde levels and elevated serum immunoglobulin M (p<0.01). Gene expression analysis revealed significant effects of Bambusa vulgaris extract, Pseudomonas infection, and their interaction on the expression of interleukin-1β, interleukin-10, and NK-lysin genes, with varying expression levels at 1, 3, and 7 days post-challenge (p<0.05). The liver bacterial load in fish exposed to P. putida significantly decreased in the bamboo-fed groups, with the lowest count observed in the 60g bamboo group. Additionally, survival rates were markedly higher in the bamboo-fed groups compared to the control, with no significant difference between the two bamboo-fed groups. In conclusion, dietary supplementation with bamboo shoot extract enhances hematological parameters, blood cell and nuclear morphology, and increases survival rates in Nile tilapia following infection.

Roles of S100A1 and S100A10 from hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀) in the immune response to Vibrio infection

The S100 proteins are highly conserved EF-hand calcium-binding proteins found only in vertebrates. In the current study, two S100 genes (S100A1 and S100A10) were successfully identified and characterized from hybrid grouper Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀. The deduced S100A10 protein contained two EF-hand domains, and S100A1 only possessed the N-terminal EF-hand. Phylogenetic analysis revealed that S100A1 and S100A10 from hybrid grouper were evolutionarily closely related to their counterparts in other selected vertebrates. Quantitative real-time PCR results revealed that the transcripts of S100A1 and S100A10 mRNA were ubiquitously distributed in all the examined tissues. After Vibrio alginolyticus infection, the expression of S100A1 and S100A10 in the spleen increased significantly. Moreover, overexpression of S100A1 and S100A10 could not only regulate the expression of interleukin 8 (IL-8), IL-10, and IL-16 in the head kidney, liver, and spleen, change the activities of acid phosphatase, catalase, lysozyme, and superoxide dismutase in serum, but also reduce the promoter activities of interferon 3 and nuclear factor kappa-B in vitro. Taken together, this study indicated that S100A1 and S100A10 participate in the immune response of hybrid grouper against bacterial infection.

Screening of key genes involved in endometritis in cows and the regulatory role of CD83 in bovine endometrial epithelial cells.

Endometritis in dairy cows involves complex molecular regulatory mechanisms. Therefore, uncovering the molecular regulatory mechanisms of endometritis in dairy cows is crucial to understand its development, prevention, and treatment. This study aimed to screen and validate key genes associated with endometritis using transcriptome sequencing of blood samples and previously obtained metabolomic sequencing data. Based on gain-of-function and loss-of-function experiments on the gene, multiple techniques, including qRT-PCR, western blotting, detection of reactive oxygen species (ROS), measurement of mitochondrial membrane potential, EdU assay, flow cytometry, and CCK-8 assay were used to explore the function of the key gene in lipopolysaccharide (LPS)-stimulated bovine endometrial epithelial cells (BEECs). The results identified 536 differentially expressed genes (DEGs) between healthy cows and those with endometritis. These DEGs were significantly enriched in apoptosis and HIF-1 signaling pathways. Weighted gene co-expression network analysis of transcriptomic and metabolomic data identified CD83, CTNNAL1, LRRC25, and NR1H3 as potential key genes for endometritis in dairy cows, with CD83 being more significantly expressed in LPS-induced BEECs. Consequently, in vitro functional studies were performed on CD83. In overexpression experiments, downregulation of the expression of inflammatory markers interleukin (IL)-1β, IL-6, and IL-8 and reduced ROS release primarily indicated the role of CD83 in attenuating the inflammatory response of BEECs. Furthermore, overexpression of CD83 regulated the S/G2 phase transition of BEECs by affecting the mRNA and protein expression of proliferation marker genes, thereby promoting proliferation of BEECs. The increased EdU positivity and the cell proliferation rate further provided evidence for the promotion of cell proliferation after overexpression of CD83. Additionally, overexpression of CD83 attenuated LPS-stimulated mitochondrial damage in BEECs, as well as the downregulation of apoptosis marker gene expression. In contrast, knockdown of CD83 expression showed the opposite trend. In summary, CD83 attenuated the inflammatory response of BEECs, promoted their proliferation, and inhibited apoptosis. This study provided basic data for understanding the mechanisms of endometritis regulation at the gene level in dairy cows.

Triptolide suppresses IL-1β-induced expression of interleukin-8 by inhibiting ROS-Mediated ERK, AP-1, and NF-κB molecules in human gastric cancer AGS cells

Triptolide suppresses IL-1β-induced expression of interleukin-8 by inhibiting ROS-Mediated ERK, AP-1, and NF-κB molecules in human gastric cancer AGS cells

Hydrogen Sulfide (H2S) Generated in the Colon Induces Neuropathic Pain by Activating Spinal NMDA Receptors in a Rodent Model of Chronic Constriction Injury.

Neuropathic pain (NP) imposes a significant burden on individuals, manifesting as nociceptive anaphylaxis, hypersensitivity, and spontaneous pain. Previous studies have shown that traumatic stress in the nervous system can lead to excessive production of hydrogen sulfide (H2S) in the gut. As a toxic gas, it can damage the nervous system through the gut-brain axis. However, whether traumatic stress in the nervous system leading to excessive production of H2S in the gut can ultimately cause neuropathic pain through the gut-brain axis remains to be investigated. This study established a model of chronic constriction injury (CCI) in mice to determine its effects on gut H2S production, the associated damage via the gut-brain axis, the potential neuropathic pain, as well as the probable mechanism. A CCI mouse model was developed using a spinal nerve ligation approach. Subsequently, the mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were used to determine the mice's pain thresholds. A variety of assays were performed, including immunofluorescence, western blotting, real-time quantitative Polymerase Chain Reaction (PCR), and membrane clamp whole-cell recordings. Mice subjected to CCI showed decreased MWT and TWL, decreased ZO-1 staining, decreased HuD staining, increased Glial fibrillary acidic protein (GFAP) staining, increased expression of tumor necrosis factor-alpha (TNF-α) protein and interleukin-6 (IL-6) protein, increased expression of NMDAR2B (NR2B) protein and NR2B mRNA, increased colocalization of vGlut2- and c-fos-positive cells, and a higher amplitude of evoked excitatory postsynaptic potential (EPSP) compared to Sham group. These changes were significantly reversed by H2S inhibitor treatment, and the specific NMDA receptor inhibitor MK-801 effectively restored the neurotoxicity of H2S. H2S is involved in CCI-induced neuropathic pain in mice, which might be mediated by the activation of the NMDA signaling pathway.

Lipocalin 2 inhibits the expressions of interleukin-8 and macrophage inflammatory protein-1α in human neutrophil-like cells.

Lipocalin 2 (LCN2) is a glycoprotein with multiple functions, including antimicrobial activity, inflammatory response modulation, and cell migration. LCN2 is expressed in some cells, such as epithelial cells and neutrophils, and its levels are increased in inflammatory diseases. This study investigated the presence of LCN2 receptor (24p3R) in cells around periodontal tissues and function of LCN2 in cells with a receptor to explore the role of LCN2 in periodontal diseases. The presence of 24p3R was examined in periodontal cells, including human gingival fibroblasts, periodontal ligament fibroblasts, human oral epithelial cells (HOECs), and neutrophil-like cells (HL-60), by Western blotting. Changes in periodontal disease-associated proteins in the presence of recombinant LCN2 (rLCN2) were examined using a protein array in differentiated HL-60 (dHL-60) cells. Interleukin-8 (IL-8) and macrophage inflammatory protein-1α (MIP-1α) mRNA expressions were analyzed by qRT-PCR, and IL-8 and MIP-1α levels in dHL-60 cells treated with rLCN2 or Porphyromonas gingivalis-lipopolysaccaharide (P.g-LPS) were determined by enzyme-linked immunosorbent assay. We detected 24p3R in dHL-60 cells. IL-8 was highly expressed and MIP-1α was weakly expressed in dHL-60 cells using a protein array. rLCN2 significantly decreased IL-8 mRNA and protein levels and suppressed P.g-LPS-induced IL-8 production in dHL-60 cells. As dHL-60 cells were co-cultured with HOECs in which LCN2 was knocked down, IL-8 mRNA expression increased in dHL-60 cells. Furthermore, rLCN2 inhibited MIP-1α production in dHL-60 cells. LCN2 suppresses inflammatory responses by regulating IL-8 and MIP-1α expression in periodontal diseases.

NF-κB oscillation profiles decode response to anti-EGFR monoclonal antibodies.

A direct connection between an inflammatory environment and cancer has been extensively proven over the years. We previously reported that the presence of interleukin 1 (IL-1) is responsible for the lack of response to monoclonal antibody targeting EGFR in colorectal cancer (CRC). Considering the driver role of NF-κB in controlling the expression of IL-1, herein, we investigate the dynamics of the oscillatory profile of the NF-κB response to monoclonal antibody, on the background of an inflammatory environment. NF-kB is a typical transcription factor that displays intrinsic oscillatory behavior, whose biological relevance in term for example of decoding response to monoclonal antibodies, remains unclear. Using live cell luciferase techniques, we recorded NF-κB activity over time in response to cetuximab (CTX) alone or in combination with IL-1 cytokines. Our results revealed an additive effect of these two agents on NF-κB activation, which was specific to CTX responsive cells. In contrast, CTX resistant cells did not display a significant change in the NF-κB profile under the IL-1 plus CTX combination. These results suggest an immediate interactive crosstalk between IL-1 and EGFR in the activation of NF-κB signaling pathways, which may lay the basis for the development of drug tolerant persistent cells (DTP), leading to CTX resistance.

Fluvoxamine may reverse the decrease in filtering capacity that occurs in acute kidney injury by increasing IL-10 and ACE expressions and preserving AQ-2, AQ-4, CL-5, and ZO-1 expressions.

The kidneys have a regulatory role in many diseases with their diuresis function and capacity to maintain electrolyte balance. In case of extensive damage, the kidney's filtration capacity is impaired and cannot fulfill its functions. Fluvoxamine (FLV), an antidepressant agent, has antioxidant and anti-inflammatory effects. This study aimed to investigate the protective effects of FLV against renal tissue damage in the lipopolysaccharide (LPS) induced systemic inflammation model. Thirty-two Wistar Albino rats were divided into four equal groups: control, LPS, LPS + FLV, and FLV alone. Six hours after LPS administration, rats were sacrificed and kidney tissues were removed. Histopathological examination of kidney tissues, immunohistochemical expression of the ACE-1, Cas-9, and interleukin-10 (IL-10), as well as spectrophotometric assessment of total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), urea, creatinine levels, and zonulin-1 (ZO-1), claudin-5 (CL-5), aquaporins 2 and 4 (AQ-2, AQ-4) were evaluated. TOS, OSI, urea, creatinine levels, and Cas-9 expressions were significantly increased, and TAS levels, ACE-1, IL-10, AQ-2, AQ-4, ZO-1, and CL-5 expressions significantly decreased in the LPS group compared with the control group. In addition, marked hyperemia, slight-to-moderate hemorrhages, and degeneration of the tubule epithelial cells were observed in kidney sections in the LPS group, in addition to the inflammatory cell infiltrations. FLV treatment significantly improved all these findings. FLV treatment ameliorates renal tissues against inflammatory scenes and protects renal function by enhancing IL-10 and ACE-1 expressions and preserving AQ-2, AQ-4, CL-5, and ZO-1 expressions.

Lack of ST2 aggravates glioma invasiveness, vascular abnormality and immune suppression

Abstract Background Glioblastoma (GBM) is the most common primary malignant brain tumor in adults, characterized by aggressive growth and a dismal prognosis. Interleukin-33 (IL-33) and its receptor ST2 have emerged as regulators of glioma growth, but their exact function in tumorigenesis have not been deciphered. Indeed, previous studies on IL-33 in cancer have yielded somewhat opposing results as to whether it is pro- or anti-tumorigenic. Methods IL-33 expression was assessed in a GBM tissue microarray and public databases. As in vivo models we used orthotopic xenografts of patient-derived GBM cells, and syngenic models with grafted mouse glioma cells. Results We analyzed the role for IL-33 and its receptor ST2 in non-malignant cells of the glioma microenvironment and find that IL-33 levels are increased in cells surrounding the tumor. Protein complexes of IL-33 and ST2 are mainly found outside of the tumor core. The IL-33-producing cells consist primarily of oligodendrocytes. To determine the function for IL-33 in the tumor microenvironment, we used mice lacking the ST2 receptor. When glioma cells were grafted to ST2-deficient mouse brains, the resulting tumors exhibit a more invasive growth pattern, and are associated with poorer survival, compared to wild-type mice. Tumors in ST2-deficient hosts are more invasive, with increased expression of extracellular matrix remodeling enzymes and enhanced tumor angiogenesis. Furthermore, the absence of ST2 leads to a more immunosuppressive environment. Conclusions Our findings reveal that glia-derived IL-33 and its receptor ST2 participates in modulating tumor invasiveness, tumor vasculature and immunosuppression in glioma.

Novel treatment strategy targeting interleukin-6 induced by cancer associated fibroblasts for peritoneal metastasis of gastric cancer

Cancer-associated fibroblasts (CAFs) are a crucial component in the tumor microenvironment (TME) of peritoneal metastasis (PM), where they contribute to tumor progression and metastasis via secretion of interleukin-6 (IL-6). Here, we investigated the role of IL-6 in PM of gastric cancer (GC) and assessed whether anti-IL-6 receptor antibody (anti-IL-6R Ab) could inhibit PM of GC. We conducted immunohistochemical analysis of IL-6 and α-smooth muscle (α-SMA) expressions in clinical samples of GC and PM, and investigated the interactions between CAFs and GC cells in vitro. Anti-tumor effects of anti-IL-6R Ab on PM of GC were investigated in an orthotopic murine PM model. IL-6 expression was significantly correlated with α-SMA expression in clinical samples of GC, and higher IL-6 expression in the primary tumor was associated with poor prognosis of GC. Higher IL-6 and α-SMA expressions were also observed in PM of GC. In vitro, differentiation of fibroblasts into CAFs and chemoresistance were observed in GC cells cocultured with fibroblasts. Anti-IL-6R Ab inhibited the progression of PM in GC cells cocultured with fibroblasts in the orthotopic mouse model but could not inhibit the progression of PM consisting of GC cells alone. IL-6 expression in the TME was associated with poor prognosis of GC, and CAFs were associated with establishment and progression of PM via IL-6. Anti-IL-6R Ab could inhibit PM of GC by the blockade of IL-6 secreted by CAFs, which suggests its therapeutic potential for PM of GC.

Expression of interleukin-17 in oral tongue squamous cell carcinoma and its effect on biological behavior

Tongue squamous cell carcinoma (TSCC) is a common malignant oral cancer characterized by substantial invasion, a high rate of lymph node and distant metastasis, and a high recurrence rate. This study aims to provide new ideas for the diagnosis and treatment of TSCC patients by exploring the related mechanisms that affect the migration and invasion of TSCC and inhibit the migration and spread of cancer cells. The results indicated the rate of high expression of IL-17 in cancer tissues was greater than that in tongue tissues, and the expression of IL-17 was related to the TNM stage. The expression of IL-17 in Cal-27 cells was greater than that in HOEC. With increasing IL-17 concentration, cell proliferation, migration, and invasion increased, and the apoptosis rate decreased. After adding the IL-17 inhibitor, the cell proliferation, invasion, and migration abilities decreased, the apoptosis rate increased, and the expression of JAK1and p-STAT3 decreased.IL-17 is highly expressed in oral tongue squamous cell carcinoma and is involved in the occurrence and development of TSCC, possibly through the JAK‒Stat signaling pathway. This study provides a new target and theoretical basis for treating tongue squamous cell carcinoma.

Lactiplantibacillus plantarum 22 A-3 ameliorates leaky gut in mice through its anti-inflammatory effects

There are limited studies on the improvement of leaky gut with minor inflammation associated with various diseases. To explore the therapeutic potential of Lactiplantibacillus plantarum 22 A-3, a member of the Lactobacillus species, in addressing a leaky gut. Lactiplantibacillus plantarum 22 A-3 was administered to a leaky gut mice model with low dextran sulfate sodium concentrations. The Lactiplantibacillus plantarum 22 A-3-treated group exhibited amelioration of increased intestinal permeability, as indicated by lower blood fluorescein isothiocyanate-dextran levels compared with that of the control group. Furthermore, the messenger RNA expression of interleukin-10, an anti-inflammatory cytokine, was upregulated in the small intestine of Lactiplantibacillus plantarum 22 A-3-treated mice. Moreover, forkhead box P3 was upregulated in the small intestine and colon following Lactiplantibacillus plantarum 22 A-3 administration. Flow cytometry showed that forkhead box P3-positive regulatory T cells tended to increase in the small intestine and colon; however, this was not significant. Messenger RNA levels for the pro-inflammatory cytokines, interleukin-1 beta, and tumor necrosis factor-alpha showed no significant changes in the small intestine; however, their expressions significantly decreased in the colon. Blood fluorescein isothiocyanate-dextran levels showed that intestinal permeability also decreased in Lactiplantibacillus plantarum 22 A-3-dead bacteria. The bacterial component of Lactiplantibacillus plantarum 22 A-3 ameliorates increased intestinal permeability through its anti-inflammatory effect in the intestinal tract and may be a novel treatment for leaky gut.

Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitor Roxadustat Accelerates Wound Healing in a Mouse Hind limb Lymphedema Model.

Objective: Drugs regulating hypoxia-inducible factor (HIF)-1α have not been investigated for wound healing in lymphedema. Therefore, we examined the effects of drug modulation of HIF-1α activity for wound healing in our previously developed mouse model of nonirradiated hind limb lymphedema. Approach: Mouse hind limb lymphedema models (n = 17) and a sham group (n = 6) were created using 8- to 10-week-old male C57BL/6N mice. Mice with hind limb lymphedema were randomized into experimental groups receiving roxadustat, 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1), or dimethyl sulfoxide and were given intraperitoneal injections every 2 days for up to 2 weeks. Four days after the surgery, an 8-mm diameter full-thickness skin wound was created in the hind limb. The number of days required for wound closure and the percentage of wounds closed were measured. Skin samples taken at wound creation were evaluated by histological and molecular analysis. Results: Administration of roxadustat accelerated wound healing, whereas YC-1 delayed it, with a significant decrease and increase in skin thickness, respectively. The relative mRNA expression of Hif1α, matrix metalloproteinase-3, and interleukin-6 was significantly higher in the roxadustat group and that of metalloproteinase-9 was significantly lower in the roxadustat group compared with the control group. Innovation: This study is the first to demonstrate delayed wound healing in a mouse model of hind limb lymphedema and the first to demonstrate the promotion of significant wound healing through the use of roxadustat. Conclusion: Roxadustat exerts wound-healing effects and may promote the regulation of extracellular matrix remodeling via gene expression in hind limb lymphedema wound models.

Indoleamine 2,3-Dioxygenase 1/Aryl Hydrocarbon Receptor Feedback Loop Mediates Anti-inflammation in lipopolysaccharide-stimulated Astrocytes to Dampen Inflammatory Neurotoxicity.

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates cell immune responses in a cell type-specific and ligand-dependent manner. In the central nervous system, astrocytic AhR plays important roles in regulating neuroinflammation by mediating responses to endogenous ligands generated from the inflammation-induced indoleamine 2,3-dioxygenase 1 (IDO1)/kynurenine (KYN) pathway. We previously demonstrated that reduction of AhR expression decreases lipopolysaccharide (LPS)-induced pro-inflammatory responses in microglia. However, the role of AhR in the astrocytic immune responses and its subsequent effects on microglial activation and neurotoxicity remain unclear. In this study, we used LPS-induced neuroinflammation in rat cortical glia-neuron (GN) mix cultures, which increased the expression of tumor necrosis factor-α and interleukin-6 and microglial activation. These proinflammatory responses were attenuated by a specific AhR agonist 6-formylindolo [3,2-b] carbazole (FICZ), but not by the AhR antagonist CH223191. CH223191, which inhibits LPS- and FICZ-induced AhR activation, enhanced neurotoxicity induced by LPS-glutamate co-treatment in GN mix cultures. Furthermore, inhibition of AhR expression and activation enhanced LPS-induced proinflammatory responses, and LPS-induced AhR activation was abrogated by the inhibition of IDO1 expression in astrocytes. Notably, AhR knockdown inhibited the anti-inflammatory effects of KYN while enhancing LPS-induced IDO1 expression in astrocytes, suggesting that AhR mediates the anti-inflammatory effect of KYN and the negative feedback regulation of IDO1 expression. Finally, we examined the role of astrocytic AhR in inflammatory astrogliosis-induced neurotoxicity by treating primary cortical neurons with LPS-treated astrocyte-conditioned medium (ACM). The results revealed that ACM derived from siAhR-transfected astrocytes increased neurotoxicity. In conclusion, inflammation-activated AhR mediates the anti-inflammatory effects and negative feedback regulation of the IDO1/KYN pathway in astrocytes, thereby dampening inflammatory astrogliosis-induced neurotoxicity.

Diagnostic and prognostic biomarkers at idiopathic pulmonary arterial hypertension

Introduction. Idiopathic pulmonary arterial hypertension (IPAH) is a subtype of pulmonary arterial hypertension (PAH). IPAH is characterized by a progressive increase in pulmonary vascular resistance (PVR), which can lead to right ventricular heart failure and even mortality. The aim was to study the levels of C-reactive protein (CRP), endothelin-1 (ET-1) and interleukin-6 (IL-6) in patients with IPAH, as well as to assess their relationship with clinical and hemodynamic parameters.Materials and methods. Expression of CRP, ET-1 and IL-6 in blood serum samples of patients with IPAH (n=53) and control group (n=52) of the appropriate age/gender was determined using enzyme immunoassay (ELISA). Demographic, clinical characteristics and hemodynamic parameters were studied in patients with IPAH according to catheterization of the right heart (CRH).Result and discussion. The average age range of patients with IPAH was 35.0-51.0 years, there were 46 (86.8%) women in this group. The average age of the participants in the control group was 31.0-42.0 years, there were 46 (88.5%) women. The levels of ET-1 (p &lt; .016) and IL-6 (p &lt; .001) were higher in patients with IPAH compared to patients in the control group, whereas the level of CRP (p = .270) was no different. Meanwhile, the level of ET-1 positively correlated with the average pressure in the right atrium (r = .728, p &lt; .001) and pulmonary vascular resistance (r = .360, p = .008), while IL-6 positively correlated with the functional class of heart failure according to the classification of the New York Heart Association (NYHA FC).Conclusion. Levels of IL-6 and ET-1 can be included in the diagnostic algorithm for assessing the severity and prognosis of the disease.

P0135 Effect of human recombinant IL-33 on fibrosis markers in colonic fibroblasts

Abstract Background IBD is frequently complicated by intestinal fibrosis and strictures formation, occurring in more than 50% of Crohn’s disease (CD) and 5% of ulcerative colitis. Intestinal fibrosis leads to surgery in CD and strictures frequently recur leading to repeated surgeries1. There is currently no specific therapy to prevent or inhibit intestinal fibrosis and this therefore constitutes a major treatment challenge. Intestinal fibrosis is characterized by excessive deposition of extracellular matrix (ECM) synthesized by intestinal myofibroblasts1. Interleukin-33 (IL-33) is a pleiotropic nuclear cytokine involved in immunoregulation and tissue repair2. Following cellular injury, IL-33 is released by epithelial, endothelial or fibroblast cells. Binding to the ST2 (suppressor of tumorigenicity 2) receptor, it induces translocation of the transcription factor NF-□□□□B, which regulates the expression of pro-inflammatory genes2. IBD patients have increased colonic expression of IL-33 and ST2 in epithelial cells compared to control patients3. Neutralization of IL-33 enables the repair of lung epithelial cells by limiting the release of pro-inflammatory cytokines such as IL-6, IL-8 and TNF-α in a mouse model of pulmonary epithelial lesions4. We hypothesized that IL-33 may be involved in IBD-associated intestinal fibrosis. We therefore aimed to understand the role of IL-33 in a human colonic fibroblast cell line (CCD-18Co). Methods First, we measured IL-33 and ST2 mRNA expression in CCD-18Co in response to TGF-ß1 (10 ng/mL) and/or TNF-ɑ (100 ng/mL) for 24h. Then, CCD-18Co were induced by increasing doses of IL-33 treatment (10, 50, 100 ng/mL) for 24h and inflammatory and fibrosis markers were measured. Results TGF-ß1 significantly induced mRNA levels encoding ECM-associated proteins such as α-SMA (***p&amp;lt;0.001) or COL1A1 (*p&amp;lt;0.05, Figure 1).Treatment with TGF-ß1 and/or TNF-α had no effect on the expression of mRNAs encoding IL-33 and ST2 in CCD-18Co in our experimental conditions. IL-33 treatment significantly increased mRNA levels for IL-1ß and ECM-associated proteins (COL3A1, CTGF and VIM) (*p&amp;lt;0.05, Figure 1) in CCD-18Co but these effects are abolished in presence of TGF-ß1 and/or TNF-α treatment. Conclusion Treatment with TGF-ß1 and/or TNF-α had no significative impact on IL-33 mRNA levels in the CCD-18Co in our experimental conditions. IL-33 treatment increased ECM-associated proteins in CCD-18Co line under basal conditions but not in TGF-ß1 and/or TNF-α conditions.

P0055 Impact of Indole-3-Propionic Acid on Bacterial Invasion Potential of Klebsiella Strains Isolated from Pediatric Ulcerative Colitis Patients

Abstract Background Indole-3-propionic acid (IPA), a tryptophan metabolite produced by microbes, has demonstrated beneficial effects, including reversing dysbiosis, balancing bacterial populations, and reducing inflammation.1,2,3 However, the mechanisms behind these effects and their impact on pathogenic microbes remain unclear. This study aimed to evaluate the effects of IPA on bacterial invasion in vitro, using potential pathobionts (Klebsiella variicola and Klebsiella quasipneumoniae) isolated from non-inflamed sections of the colon in pediatric ulcerative colitis (UC) patients. We hypothesized that IPA could reduce virulence traits, promoting a more homeostatic environment, which could lower inflammation in pediatric UC patients. Methods Caco-2 cells (colonic epithelial cell line) were infected with Klebsiella strains, with or without 0.5 mM IPA. Bacterial invasion was assessed via gentamicin protection assay, and qPCR measured Interleukin-8, IL-10, and TNF-α expression in infected cells. Transepithelial electrical resistance (TEER) assay evaluated barrier integrity, and qPCR analyzed the expression of bacterial virulence genes, including entB (siderophores), manC (capsule), and hcp1 (type VI secretion) in IPA-treated and untreated cultures. Additionally, siderophore production was quantified using a Chrome-Azurol Sulfonate (CAS) assay, and Anthony’s stain visualized the bacterial capsule. Results IPA-treated Caco-2 cells showed significantly reduced bacterial invasion, with further reductions when bacteria were also exposed to IPA. qPCR revealed a significant decrease in IL-8 expression in IPA-treated cells during infection. IL-10 levels decreased non-significantly, and TNF-α expression showed variable results. A trend toward improved tight junction integrity was observed in the TEER assay with IPA treatment, although no significant differences were detected. IPA treatment increased the expression of entB, manC, and hcp1 genes, except for K. variicola, where manC expression remained unchanged. IPA significantly reduced the capsule thickness of K. variicola and resulted in a significant reduction in K. quasipneumoniae siderophore production. Conclusion IPA reduces bacterial invasion and modulates immune responses in Caco-2 cells infected with Klebsiella pathobiont strains. It reduces the virulence of K. quasipneumoniae by targeting siderophore production and diminishes the virulence of K. variicola by reducing capsule formation, suggesting its potential as an anti-virulence agent against these pathobionts. These findings highlight IPA’s potential as an immunomodulatory agent for managing IBD. However, further research is needed into other Klebsiella virulence pathways and host-microbiota interactions.

Curdione combined with borneol treats bacterial mixed HPV infection by regulating the crosstalk among immune cells

BackgroundHuman papillomavirus (HPV) infection is a worldwide reproductive system disease. Baofukang suppository, a traditional herbal preparation that includes curdione and borneol, has been reported to treat bacterial vaginosis (BV) and HPV infection in China. However, the therapeutic mechanism is still unknown. This study aims to explore the molecular mechanisms of curdione and borneol in treating HPV infection.MethodsWe conducted a retrospective cohort analysis of medical records from a single-center study involving 205 HPV patients, focusing on the correlation between HPV clearance and co-infection with other pathogens, confirming the efficacy of Baofukang suppository. Bioinformatics and network pharmacology approaches were employed to identify therapeutic targets of Baofukang suppository for BV/HPV co-infections. qRT-PCR, Western blot, immunofluorescence staining, and flow cytometry were utilized to validate the therapeutic targets of curdione and borneol, along with the associated immune molecular changes. Finally, the molecular mechanisms and therapeutic efficacy of curdione and borneol were confirmed in vivo using an LPS/TC-1 cervical orthotopic injection model.ResultsCurdione and borneol selectively inhibit the secretion of interleukin-6 (IL-6) and interleukin-1β (IL-1β) by macrophages. The reduction in IL-6 and IL-1β levels effectively inhibits the expression of CD274 (Programmed death ligand 1, PD-L1) in infected epithelial cells by inhibiting STAT3 phosphorylation, thereby suppressing their immune evasion capabilities. Furthermore, curdione and borneol enhance the expression of tumor necrosis factor α (TNF-α) and caspase 1 (CASP1) in macrophages, as well as the expression of interleukin 12 (IL-12) and interleukin 23 (IL-23) in dendritic cells (DCs). The expression of these inflammatory factors effectively promotes the migration and differentiation of T cells to the site of infection, completing the clearance of infected epithelial cells.ConclusionThe main components of Baofukang suppository, curdione and borneol, inhibit the progression of HPV infection and the occurrence of cervical cancer by modulating the communication between innate and adaptive immunity, promoting the recruitment and recognition of CD8+ T cells to eliminate HPV-infected epithelial cells.

IL-21 and IL-33 May Be Effective Biomarkers to Predict the Efficacy of PD-1 Monoclonal Antibody for Advanced Cholangiocarcinoma.

Background and Objective: Treatment options for patients with advanced biliary tract cancer (BTC) are limited. The programmed cell death protein-1 (PD-1) inhibitors may have synergistic effects with chemotherapy. Therefore, the aim of our study was to provide real-world data on treatment outcomes in BTC patients receiving chemotherapy alone versus a combination of chemotherapy and PD-1 inhibitors. Additionally, we explored potential markers predictive of PD-1 inhibitor efficacy in this combined therapy. Methods: We conducted a review of patients at Changzhou First People's Hospital who received PD-1 inhibitors in combination with chemotherapy or chemotherapy alone as first-line treatment for advanced BTC. The primary endpoints of the study were progression-free survival (PFS) and overall survival (OS). Kaplan-Meier survival curves and the log-rank test were used to analyze the data. Immunohistochemistry showed the expression of interleukin-21 (IL-21), interleukin-33 (IL-33), and Eomes in the tumor tissue of patients who received PD-1 inhibitors in combination with chemotherapy. Results: The study enrolled 61 patients receiving PD-1 inhibitors combined with chemotherapy and 65 receiving chemotherapy alone. The median OS and PFS for patients receiving PD-1 inhibitors in combination with chemotherapy were 11.7 and 6.7 months, respectively. These durations were significantly longer than those for chemotherapy alone: OS of 10.3 months (95% CI: 0.16-0.21, p = 0.031) and PFS of 5.3 months (95% Confidence interval (CI) 0.25-0.32, p = 0.018). High IL-21 expression or low IL-33 expression in tumor tissue correlated with better response rates to chemotherapy combined with PD-1 inhibitors. Conclusions: Combining PD-1 inhibitors with chemotherapy shows good antitumor activity, making it an effective way to treat BTC. The expression profiles of IL-21 and IL-33 hold promise as potential markers for guiding the chemotherapy combined with immunotherapy in BTC patients.