Interleukin Research Articles

P-57. Durability of SARS-CoV-2 specific Cellular Immune Response after COVID-19 Vaccination in PLWH

Abstract Background People living with HIV (PLWH) face the dual challenge of human immunodeficiency virus(HIV) infection and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. It is associated with severe outcomes and a higher risk of mortality. In January 2021, the vaccination campaign against SARS-CoV-2 was launched in India and started mass administration of two types of vaccines. The two vaccines used on a large scale were, a recombinant, replication-deficient chimpanzee adenovirus vectored vaccine/ ChAdOx1 nCoV-19 (Covishield) and whole virion inactivated vaccine/ BBV152 (Covaxin).The study of cellular immune responses to COVID-19 vaccination in PLWH was vital for safeguarding this vulnerable population. Methods Study design: Prospective observational study Study population: The individuals recruited in the study were vaccinated with at least 2 doses and/or booster dose of the COVID-19 vaccine (COVISHIELD/COVAXIN). The individuals were segregated into two groups (PLWH and non-HIV-infected healthy individuals). Testing Method: Peripheral blood mononuclear cell (PBMC) were cultured to test the SARS-CoV-2-specific T cell immune response using activation induced marker/ intracellular cytokine staining (AIM/ICS) assay. Distribution of CD4 T lymphocytes count in HIV patients with mean CD4 count 470 cell/µl. Results We conducted did this study in two phases. The first phase was conducted between August 2022 and December 2022, while the second phase, or the follow-up, took place from January 2023 to December 2023. In the first phase/ first visit (V1), we recruited 50 adult PLWH and 40 adult healthy controls. Subsequently, in phase 2/follow-up/ second visit (V2), we achieved a follow-up rate of 100% in each arm. SARS-CoV2-specific cellular immune response after COVID-19 vaccination was comparable and durable IN PLWH as compared to healthy controls. Conclusion Comparable and durable immune response was observed in PLWH population on regular highly active antiretroviral therapy (HAART), with stable viral suppression and good CD4+ T cell counts. This was consistent over time across 1 year gap in both phases of the study and across different vaccine types. This outcome indicates that the cellular immune responses in our PLWH cohort is comparable and durable to that of the general population, which is a reassuring finding for the public health. SARS-CoV-2-specific CD8 T-cell immune response (scatter plots displaying the results of CD8 T cell assays, specifically evaluating the production of various cytokines (IFNγ (Interferon-gamma), TNFα (Tumor Necrosis Factor-alpha), and IL-2 (Interleukin 2), Granzyme B (Gran-B) at two different time points V1 and V2 (visit 1 and visit 2) with p values mentioned at the top. Disclosures All Authors: No reported disclosures

The dynamics of CD4+ T cell proliferation and regulation

We use mathematical modeling to study the proliferation dynamics of CD4+ T cells within an immune response. This proliferation is driven by the autocrine reaction of helper T cells and interleukin-2 (IL-2), and regulated by natural regulatory T cells (nTregs). Previous studies suggested that a fratricidal mechanism is necessary to eliminate helper T cells post-infection. Contrary to this, our mathematical analysis establishes that the depletion of these cells is due to two pivotal factors: the saturation in the proliferation rate of helper CD4+ T cells at high IL-2 concentrations, and the activation rate of nTregs outpacing their death rate. This yields an excitable process, such that the proliferation starts once the helper T cell population passes a threshold. Additionally, we find that when the proliferation of nTregs lags behind their mortality, induced regulatory T cells (iTregs) are crucial to curbing the proliferation of helper CD4+ T cells.

Nephroprotective potential of robinin to counteract aldicarb induced renal dysfunction via modulating TLR4/MyD88, HMGBI/RAGE, NF-κB pathway: A biochemical and pharmacodynamic approach.

The current investigation was conducted to evaluate the nephroprotective potential of robinin (RBN) to avert aldicarb (ALD) induced renal impairments. Thirty-two adult albino rats (Sprague Dawley) were categorized into four groups including control, ALD (15 mgkg-1), ALD (15 mgkg-1) + RBN (6 mgkg-1) and RBN (6 mgkg-1) alone treated group. The results of the current trial demonstrated that ALD intoxication promoted the gene expression of receptor for advanced glycation end products (RAGE), tumor necrosis factor- α (TNF-α), toll-like receptor 4 (TLR4), high mobility group box1 (HMGB1), nuclear factor- kappa B (NF-κB), monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and interleukin-1β (IL-1β). Moreover, activities of HO-1, GSH, GPx, SOD, GSR, and CAT were suppressed while the levels of ROS and MDA were escalated following the ALD exposure. ALD intoxication upregulated the levels of cystatin C, KIM-1, creatinine, NAG, uric acid, urea, NGAL and BUN while reducing the levels of creatinine clearance in renal tissues. The levels of Bax, Caspase-9 and Caspase-3 were promoted while the levels of Bcl-2 were suppressed after ALD administration. Histopathological analysis showed ALD disrupted the normal architecture of renal tissues. However, RBN therapy substantially protected the renal tissues owing to its antioxidative, anti-inflammatory and anti-apoptotic potential.

Halofuginone prevents inflammation and proliferation of high-altitude pulmonary hypertension by inhibiting the TGF-β1/Smad signaling pathway

The inflammatory response of lung tissue and abnormal proliferation of pulmonary artery smooth muscle cells are involved in the pathogenesis of high-altitude pulmonary hypertension (HAPH). Halofuginone (HF), an active ingredient derivative of Chang Shan (Dichroa febrifuga Lour. [Hydrangeaceae]), has antiproliferative, antihypertrophic, antifibrotic, and other effects, but its protective effects on HAPH remains unclear. In the present study, we evaluated the efficacy of HF on HAPH by establishing a 6000 m HAPH rat model. Male Sprague–Dawley rats were divided into normoxia, normoxia + halofuginone (1 mg/kg), hypoxia, and hypoxia + halofuginone (1 mg/kg) groups. The results showed that HF (1 mg/kg) could prevent hypoxia-induced hemodynamic abnormalities, right ventricular hypertrophy, and pulmonary vascular remodeling in rats. We further detected the expression levels of inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α) and proliferative/antiproliferative indicators proliferating cell nuclear antigen (PCNA), cyclin-dependent kinase 6 (CDK6), Cyclin D1, p21 in lung tissue, and found that HF could attenuate the lung tissue inflammatory response and proliferative response in HAPH rats. In addition, we also examined the expression levels of transforming growth factor-β1 (TGF-β1), Smad2/3 and p-Smad2/3 in lung tissue, and found that HF exerted therapeutic effects by inhibiting the TGF-β1/Smad signaling pathway.

Interleukin-23 versus Interleukin-17 Inhibitors in Preventing Incidental Psoriatic Arthritis in Patients with Psoriasis: A Real-World Comparison From the TriNetX US Collaborative Network.

Psoriatic arthritis (PsA) is a common comorbidity in patients with psoriasis (PsO) that leads to significant disease burden. Biologic therapies targeting the interleukin (IL)-23/IL-17 axis have been widely used for PsO, but their comparative effectiveness in preventing PsA remains unclear. The study objective was to compare the occurrence of developing incidental PsA among PsO patients treated with interleukin-23 inhibitors (IL23is) or interleukin-17 inhibitors (IL17is). A retrospective cohort study was conducted using real-world data from the TriNetX US Collaborative Network, including 53 healthcare organizations. Adult PsO patients treated with IL23is or IL17is between January 2019 and June 2022 were identified. Cox regression analysis was used to assess the risk of PsA incidence, with hazard ratios (HRs) and 95% confidence intervals (CIs) reported. Subgroup analyses were performed based on age, sex, and ethnicity. Sensitivity analyses included comparisons with tumor necrosis factor (TNF) inhibitors (TNFis) to ensure robustness. A total of 4,580 PsO patients were included in the study, with 2,273 receiving IL23is and 2,307 receiving IL17is. Treatment with IL23is was associated with a significantly lower incidence of PsA compared to IL17is (HR = 0.60, 95% CI 0.44-0.82, P = 0.001). This reduction in risk was particularly notable in the 41- to 65-year age group (HR = 0.42, 95% CI 0.27-0.64, P < 0.001) and among females (HR = 0.57, 95% CI 0.38-0.86, P = 0.007). Subgroup analyses based on ethnicity revealed varying outcomes, with White patients showing a significant risk reduction (HR = 0.55, 95% CI 0.38-0.79, P = 0.001) but no significant risk reduction was observed in Black or African American patients (HR = 1.37, 95% CI 0.37-5.13, P = 0.637). Sensitivity analyses comparing IL23is and TNFis confirmed the robustness of the findings. IL23is are associated with a lower risk of PsA incidence compared to IL17is in PsO patients, particularly in specific age, sex, and ethnic groups. These findings suggest that IL23is may be more suitable for PsO patients at high risk of PsA and could inform potential updates to treatment guidelines. Further research should focus on refining therapeutic strategies by incorporating patient-specific factors such as comorbidities, ethnicity, and genetic predispositions, which could optimize biologic selection and enhance PsA prevention efforts in clinical practice.

Increased serum GM-CSF at diagnosis of biliary atresia is associated with improved biliary drainage.

The immune heterogeneity of biliary atresia (BA) presents a challenge for development of prognostic biomarkers. This study aimed to identify early immune signatures associated with biliary drainage after Kasai Portoenterostomy (KPE). Serum samples, liver slides, and clinical data were obtained from patients enrolled in the NIDDK-supported Childhood Liver Disease Research Network. Serum cytokines and hepatic immune cell subsets were measured at diagnosis and compared among 3 groups: 38 infants with BA (20 with evidence of bile flow after KPE; 18 without) and 17 non-BA cholestatic infants. BA participants had lower numbers of lipid associated macrophages (LAM), and increased serum levels of Eotaxin-3, interleukin (IL) 12p70, and IL-8 versus non-BA groups (p < 0.05 for all). Among BA participants, monocyte like macrophages and serum levels of granulocyte-macrophage colony stimulating factor (GM-CSF) were increased in BA participants with good biliary drainage (p = 0.004 and p < 0.001 respectively). Levels of GM-CSF, IL-16, c-reactive protein, TNF-β predicted successful biliary drainage with an area under the receiver operating curve of 0.84 (p < 0.001). These findings suggest that distinct macrophage-associated immune networks at diagnosis may impact biliary drainage after KPE. Identification of early prognostic immune-modulatory markers has potential to improve patient stratification for medical and surgical therapies. We identify serum cytokines, particularly GM-CSF, that are associated with future biliary drainage in patients with biliary atresia. Characterization of macrophage-associated immune networks provides novel insight into early disease mechanism that may impact patient outcomes. Early prognostic biomarkers markers in biliary atresia can help in patient stratification for medical and surgical therapies.

Explorative Study of Modulatory Effects of Notochordal Cell-Derived Extracellular Vesicles on the IL-1β-Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants.

Cell-free regenerative strategies, such as notochordal cell (NC)-derived extracellular vesicles (EVs), are an attractive alternative in developing new therapies for intervertebral disc (IVD) degeneration. NC-EVs have been reported to elicit matrix anabolic effects on nucleus pulposus cells from degenerated IVDs cultured under basal conditions. However, the degenerative process is exacerbated by pro-inflammatory cytokines contributing to the vicious degenerative cycle. Therefore, this study explores whether NC-EVs modulate interleukin (IL)-1β-mediated pro-inflammatory responses in the degenerating disc. This study utilized two IL-1β induced pro-catabolic culture models; a dog 3D nucleus pulposus (NP) cell pellet culture and a human patient-derived, exvivo NP tissue culture system. Porcine NC-EVs were generated from NC-conditioned medium by differential centrifugation followed by size exclusion chromatography. Donor matched EV-depleted media were generated by overnight ultracentrifugation, whereafter the EV-depleted NCCM supernatant was subjected to size exclusion chromatography. To investigate whether observed effects were EV-associated, NC-EVs conditions were compared to EV-depleted controls in the absence and presence of IL-1β. The size and concentration of NC-EVs were quantified by nanoparticle tracking analysis, which showed minimal donor variation and confirmed depletion of EVs in the EV-depleted media. In the IL-1β-induced catabolic cascade, the NC-EVs did not elicit anabolic effects at the matrix level nor did they rescue the pro-catabolic phenotype within dog pellets. Modification of the CCL2 secretion seemed to be context dependent in the human explants: where EVs treatment stimulated CCL2 secretion but in the presence of IL-1β this effect was counteracted. Secretion of IL-6 and C-X-C motif chemokine ligand 1 was significantly decreased in NC-EV + IL-1β vs. control+IL-1β but not compared to EV-depleted human explant controls. Altogether, this data provides evidence for a protective modulatory role of NC-EVs. Considering the homeostatic function EVs exert, inherently encompassing subtle biologic modifications, the current study may have lacked sufficient power to demonstrate statistical significance in a sample set with evident donor variation. NC-EVs may modulate the production of specific cytokines and chemokines in human degenerate explants when the key pro-inflammatory cytokine IL-1β is present. Implementation of the technical EV-depleted controls in further studies is essential to robustly demonstrate that these effects are EV-mediated and not associated with other secreted factors co-isolated during EV-isolation.

Evaluation of Plasma Neurodegenerative Biomarkers for Diagnosing Minimal Hepatic Encephalopathy and Predicting Overt Hepatic Encephalopathy in Chinese Patients with Hepatic Cirrhosis.

The performance of neurodegenerative biomarkers-neurofilament light chain (NfL), glial fibrillary acidic protein (GFAP), tau, and ubiquitin carboxy-terminal hydrolase L1 (UCHL1)-in diagnosing minimal hepatic encephalopathy (MHE) has not been systematically evaluated, simultaneously, nor have their associations with the development of overt hepatic encephalopathy (OHE). This study aimed to evaluate the performance of plasma NfL, GFAP, tau, and UCHL1 in diagnosing MHE and predicting the development of OHE in Chinese patients with hepatic cirrhosis. In this prospective study, 124 patients with hepatic cirrhosis were recruited. The Psychometric Hepatic Encephalopathy Score was used to diagnose MHE, and OHE development was observed during a 30-day follow-up period. Plasma levels of NfL, GFAP, tau, and UCHL1 were measured using the highly sensitive single-molecule array when MHE was diagnosed. Additionally, serum interleukin-6 (IL-6) levels and the model for end-stage liver disease (MELD) and MELD-Na scores were also measured. MHE was diagnosed in 57 (46.0%) patients. Patients with MHE had significantly higher plasma levels of NfL and GFAP (34.2 vs. 22.4 pg/mL and 173 vs. 97.6 pg/mL, respectively; both p < 0.001) and lower tau levels (8.4 vs. 11.6 pg/mL, p = 0.048) compared to those without MHE. Plasma NfL (odds ratios = 1.027, 95% confidence interval [CI]: 1.006-1.048; p = 0.013) and serum ammonia levels (odds ratios = 1.021, 95% CI: 1.006-1.036; p = 0.007) were independently associated with MHE occurrence. A combination of NfL, GFAP, tau, and UCHL1 was effective in diagnosing MHE in all cirrhotic patients (area under the receiver operating characteristic curve [hereinafter referred to as AUROC]: 0.748, 95% CI: 0.662-0.821), with an accuracy, sensitivity, and specificity of 71.0%, 71.9%, and 71.6%, respectively. In patients without previous OHE, the combination had an AUROC of 0.764 (95% CI: 0.673-0.840), with an accuracy, sensitivity, and specificity of 72.5%, 71.7%, and 73.0%, respectively. Furthermore, GFAP (hazard ratio (HR) = 1.003, 95% CI: 1.000-1.005; p = 0.044), IL-6 (HR = 1.003, 95% CI: 1.001-1.004; p < 0.001), and MELD score (HR = 1.139, 95% CI: 1.072-1.210; p < 0.001)-but not NfL, tau, and UCHL1-were identified as risk factors for 30-day OHE development. The combination of plasma levels of NfL, GFAP, tau, and UCHL1 performs well in diagnosing MHE. Additionally, MELD score, IL-6, and GFAP appear to be significant predictors of OHE development in patients with hepatic cirrhosis.

Immune responses in children with secondary infection of mycoplasma pneumoniae after COVID-19: focus on eosinophils and IgE

BackgroundThe COVID-19 (SARS-CoV-2) epidemic has posed a major challenge to global public health, especially in children. Some children may experience secondary infection with Mycoplasma pneumoniae after SARS-CoV-2 infection, which has attracted widespread attention. Studies have shown that eosinophils play an important role in respiratory tract infections and are involved in regulating immune responses and inflammatory processes. However, there is a lack of systematic research on the specific manifestations and mechanisms of eosinophils in secondary infection with Mycoplasma pneumoniae after SARS-CoV-2 infection.ObjectiveThis study aims to explore the characteristics of immune response in children with SARS-CoV-2 infection and Mycoplasma pneumoniae infection, focusing on the changes in immune indicators such as eosinophils (EOS), immunoglobulin E (IgE), interleukin-6 (IL-6), C-reactive protein (CRP), and procalcitonin (PCT).MethodsThis study is a retrospective observational study, and a total of pediatric patients who were treated in our hospital from January 2023 to December 2023 were included. The study group included children who were diagnosed with SARS-CoV-2 infection and further infected with Mycoplasma pneumoniae, and the control group included children who were only infected with SARS-CoV-2 and had no other pathogens. The clinical data of the two groups of patients, including absolute eosinophil value, IgE quantification, IL-6, CRP and PCT levels, were collected and analyzed, and statistical comparisons were performed.ResultsA total of 134 children were included, including 79 in the study group and 55 in the control group. The absolute eosinophil value [0.17 (0.09, 0.31) vs. 0.09 (0.06, 0.23), P < 0.01] and IgE level [59.28 (37.54, 256.88) vs. 22.00 (11.00, 113.10) P < 0.01] of the children in the study group were significantly higher than those in the control group, while IL-6 [16.81(4.72,31.86) vs. 9.5(3,57.3), P = 0.602], CRP [2.82(1.10,6.13) vs. 1.94(0.50,8.94), P = 0.528] and PCT[0.12(0.08,0.20) vs. 0.12(0.10,0.24), P = 0.329] were no significant difference between the two groups. Binary logistic regression analysis showed that the absolute value of eosinophils and IgE were independent risk factors for secondary infection of Mycoplasma pneumoniae after SARS-CoV-2 infection.ConclusionThis study shows that after SARS-CoV-2 infection, the increase in eosinophils and the increase in related immune indicators IgE may be closely related to secondary infection with Mycoplasma pneumoniae. This study provides an important basis for understanding the immune response of children after SARS-CoV-2 infection and its related clinical management, suggesting that clinicians should closely monitor the eosinophil count and IgE level of children after SARS-CoV-2 infection, especially for children at risk of secondary infection, so as to take timely intervention measures to prevent secondary infection with Mycoplasma pneumoniae and improve the prognosis of children.

Network Pharmacology and Computational Approach to Explore the Potential Underlying Mechanism of Centella asiatica in the Treatment of Diabetes Mellitus

Background: Centella asiatica is a tropical medicinal herb traditionally used for the treatment of different diseases, such as arthritis, kidney disease, diabetes mellitus, etc. Diabetes mellitus is emerging as a global health concern, demanding research to provide insights into it. Objective: The current research study aimed at employing the Network Pharmacology and Molecular Docking approach to unearth and validate the possible molecular mechanism involved in the treatment of diabetic mellitus with herbal constituents from Centella asiatica. Methods: The phytocompounds and targets of Centella asiatica were screened from different databases. An herb-core-target-ingredient-diabetes mellitus network was established via Cystoscope 3.7.2. Next, Go and KEGG enrichment analysis was performed. Lastly, the interaction between ligands and targets was investigated via molecular docking. Results: According to the results obtained, we identified 49 core targets of diabetes mellitus and 37 active ingredients of Centella asiatica. Next, Go and KEGG resulted in a total of 455 biological processes for the treatment of diabetes mellitus. The KEGG enrichment analysis reported that the targets were related to metabolic pathways, insulin signaling pathways, glycolysis/gluconeogenesis, oxidative stress, insulin resistance, etc. On the basis of KEGG enrichment and protein-protein interaction, we selected Fructose-1-6 bisphosphate1 (FBP1), Glucokinase (GCK), Cytochromes P450 (CYP19A1), fatty acid binding protein 1 (FABP1), Interleukin 2 (IL2) and angiotensin-converting enzyme (ACE), and phytocompounds from Centella asiatica for docking. From the docking study, it was concluded that several targets had a stable binding affinity with Centella asiatica phytocompounds. result: Results: According to the results obtained, we identified 49 core-targets of diabetes mellitus and 37 active ingredients of Centella asiatica. Next, Go and KEGG resulted in total 455 biological process for treatments of diabetes mellitus. The KEGG enrichment analysis reported that the targets were related to metabolic pathways, insulin signaling pathways glycolysis/gluconeogenesis, oxidative stress, insulin-resistance etc. On the basis of KEGG enrichment and protein-protein interaction, we selected Fructose-1-6 bisphosphate1 (FBP1), Glucokinase (GCK), Cytochromes P450 (CYP19A1), fatty acid binding protein 1 (FABP1), Interleukin 2 (IL2) and angiotensin converting enzyme (ACE) and phytocompounds from Centella for Docking. From docking study, it is concluded that several targets have stable binding affinity with Centella phytocompounds Conclusion: We explored the biological mechanism of phytocompounds involved in the treatment of diabetes mellitus through different biological processes and signaling pathways, and lastly, docking provides us commending results that direct for experiments ahead.

Chronic IL-1-Exposed LNCaP Cells Evolve High Basal p62-KEAP1 Complex Accumulation and NRF2/KEAP1-Dependent and -Independent Hypersensitive Nutrient Deprivation Response

Chronic inflammation is a cancer hallmark and chronic exposure to interleukin-1 (IL-1) transforms castration-sensitive prostate cancer (PCa) cells into more fit castration-insensitive PCa cells. p62 is a scaffold protein that protects cells from nutrient deprivation via autophagy and from cytotoxic reactive oxygen via NFκB and NRF2 antioxidant signaling. Herein, we report that the LNCaP PCa cell line acquires high basal accumulation of the p62-KEAP1 complex when chronically exposed to IL-1. p62 promotes non-canonical NRF2 antioxidant signaling by binding and sequestering KEAP1 to the autophagosome for degradation. But despite high basal p62-KEAP1 accumulation, only two of several NRF2-induced genes analyzed, GCLC and HMOX1, showed high basal mRNA levels, suggesting that the high basal p62-KEAP1 accumulation does not result in overall high basal NRF2 activity. Nutrient starvation induces NRF2-dependent GCLC upregulation and HMOX1 repression, and we found that chronic IL-1-exposed LNCaP cells show hypersensitivity to serum starvation-induced GCLC and HMOX1 regulation. Thus, chronic IL-1 exposure affects cell response to nutrient stress. While HMOX1 expression remains NRF2/KEAP1-dependent in chronic IL-1-exposed LNCaP cells, GCLC expression is NRF2/KEAP1-independent. Furthermore, the high basal p62-KEAP1 complex accumulation is not required to regulate GCLC or HMOX1 expression, suggesting cells chronically exposed to IL-1 evolve a novel NRF2-independent role for the p62/KEAP1 axis.

Epigenetic mechanisms underlying endometrial cancer (EC) outcomes: race-specific patterns of DNA methylation associated with molecular subtypes and survival.

Endometrial cancer [EC] is the fourth most common cancer in women in the United States. Stark racial disparities are present in EC outcomes in which Black women have significantly higher EC-related mortality than White women. The social and biologic factors that contribute to these disparities are complex, and may include racial differences in epigenetic landscapes. To investigate race-specific epigenetic differences in EC tumor characteristics and outcomes, we utilized the most recent data within the Cancer Genome Atlas (TCGA). Genome-wide CpG methylation data for more than 850 000 CpG sites were analyzed across 245 tumor samples, including 52 from Black women and 181 from White women. Race-adjusted and race-stratified associations among CpG methylation in ECs and molecular subtypes and disease-free survival (DFS) were examined. Race-specific analysis identified subtype-associated CpGs within 9572 genes in tumors from White women, and only 10 genes in tumors that were from Black women. Race-specific analyses also identified survival-associated CpGs with 1119 unique genes identified in tumors from White women, and none identified in tumors from Black women. Genes identified with differential methylation among subtypes included those involved in oxidative stress (HIF3A), and DNA repair (MLH1). Replication cohort data highlighted genes overlapping with those identified within the TCGA, such as G Protein Subunit Beta 1 (GNB1), involved in G-protein signaling, and Interleukin 37 (IL37), involved in cytokine signaling. Identification of these racial differences in EC tumor epigenetic landscapes and associated changes in gene expression may provide insight into strategies to improve outcomes and reduce disparities.

Development of anti-murine IL-18 binding protein antibodies to stimulate IL-18 bioactivity

Abstract Interleukin (IL)-18 is an immunoregulatory cytokine that acts as a potent inducer of T helper 1 and cytotoxic responses. IL-18 activity is regulated by its decoy receptor IL-18 binding protein (IL-18BP), which forms a high-affinity complex with IL-18 to block binding of the cognate receptors. A disbalance between IL-18 and IL-18BP associated with excessive IL-18 signaling can lead to systemic inflammation. Indeed, the severity of CpG-induced macrophage activation syndrome is exacerbated in IL-18BP knockout (KO) mice. On the contrary, targeting IL-18BP can have promising effects to enhance immune responses against pathogens and cancer. We generated monoclonal rabbit anti-mouse IL-18BP antibodies labeled from 441 to 450. All antibodies, except from antibody 443, captured mouse (m)IL-18BP when used in a sandwich enzyme-linked immunosorbent assay. Using an IL-18 bioassay, we showed that antibody 441 did not interfere with the regulatory effect of mIL-18BP, whereas all other antibodies displayed different levels of antagonism. Further experiments were performed using antibody 445 endowed with potent neutralizing activity and antibody 441. Despite binding to IL-18BP with the same affinity, antibody 445, but not antibody 441, was able to release IL-18 from preformed IL-18–IL-18BP complexes. Administration of antibody 445 significantly aggravated the severity of CpG-induced macrophage activation syndrome as compared with antibody 441. Additional experiments using naïve wild-type, IL-18BP KO, and IL-18 KO mice confirmed the specificity of the neutralizing effect of antibody 445 toward IL-18BP. Our studies led to the development of a monoclonal anti-IL-18BP antibody with neutralizing activity that results in the promotion of IL-18 activities.

Deciphering the therapeutic effects of Xiyanping injection: insights into pulmonary and gut microbiome modulation, SerpinB2/PAI-2 targeting, and alleviation of influenza a virus-induced lung injury

Infection with Influenza A virus (IAV) induces severe inflammatory responses and lung injury, contributing significantly to mortality and morbidity rates. Alterations in the microbial composition of the lungs and intestinal tract resulting from infection could influence disease progression and treatment outcomes. Xiyanping (XYP) injection has demonstrated efficacy in clinical treatment across various viral infections. However, its specific effects and mechanisms against IAV remain unclear. In this study, we established an IAV infection mice model, and utilized 16 S rRNA sequencing, RNA sequencing, protein chips, and molecular docking, to investigate the mechanisms of XYP injection on altering pulmonary and gut microbiota, and identifying its target sites. We revealed that XYP injection significantly reduced mortality, weight loss, lung viral titers, and lung pathology in IAV-infected mice. XYP injection down-regulated the activity of malondialdehyde, and the levels of interleukin (IL)-1β, IL-5, IL-6, tumor necrosis factor-α, IL-18, IL-15, granulocyte colony-stimulating factor, IL-9, chemokine (C-C motif) ligand-5, and C-X-C motif chemokine ligand 5, while up-regulated the activities of glutathione peroxidase reactive and superoxide dismutase, and the level of interferon-γ. The diversity of the pulmonary and gut microbiota was altered slightly after XYP injection. The linear discriminant analysis of the gut microbes revealed a higher proportion of potentially beneficial bacteria, including Akkermansia, Parabacteroides goldsteinii, Defluviitaleaceae, Oscillospirales, and Eubacterium_coprostanoligenes_group characterized the XYP group. Peritoneal macrophage RNA sequencing highlighted Serpinb2 as the most significantly regulated gene by XYP injection, along with consistent changes in multiple downstream Th2 structure genes. KEGG pathway analysis indicated significant modifications in genes associated with influenza A, mitogen-activated protein kinase signaling, nuclear factor kappa-B signaling, and apoptosis following XYP injection. Finally, human protein chips and molecular docking were carried out to confirm the binding of the main component of XYP injection, andrographolide, with SERPINB2/PAI-2 protein. Overall, our study provides valuable insights into the therapeutic potential of XYP injection in treating influenza, highlighting its multifaceted effects on host microbiota and immune responses, and pinpointing SerpinB2/PAI-2 as the target for XYP injection in exerting anti-inflammatory and antiviral therapeutic mechanisms.

Hallmarks of Quercetin Benefits as a Functional Supplementary in the Management of Diabetes Mellitus-Related Maladies: From Basic to Clinical Applications.

Quercetin (QE), a particular flavonoid, is well known for its medicinal effects, including anti-oxidant, hypoglycemic, and anti-inflammatory effects. In this review, the findings of QE effects on diabetes STZinduced, alloxan-induced, and its complications have been summarized with a particular focus on in vitro, in vivo, and clinical trials. Consequently, QE mediates several mechanisms, including ameliorating tumor necrosis factor (TNF)-α, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interleukin (IL)-1β, IL-8, and IL-10 expression, increasing insulin glucose uptake to inhibit insulin resistance. Moreover, QE stimulates insulin secretion and attenuates insulin resistance through various pathways, namely transient KATP channel, motivating peroxisome proliferator-activated receptor expression, increasing glucose transporter-4, and decreasing inducible nitric oxide synthase in skeletal muscle. QE has protective effects on the complications caused by diabetes, such as polycystic ovary syndrome, high-fat diet-induced obesity, diabetic-induced hepatic damage, vascular inflammation, nephropathy, and neuropathy.

Comparative analysis of salivary cytokine profiles in newly diagnosed pediatric patients with cancer and healthy children

Salivary cytokines have the potential to serve as biomarkers for evaluating cancer progression and treatment response in specific cancer types. This study explored salivary cytokine profiles in pediatric cancer patients and healthy controls, examining changes during chemotherapy. We conducted a prospective study involving newly diagnosed cancer patients and healthy controls under 19 years old. Saliva samples were collected at diagnosis, and three and six months post-diagnosis for cancer patients, while healthy controls provided samples at a single time point. Cytokine levels were analyzed using Luminex technology. Our study included 19 cancer patients (10 with leukemia, 5 with lymphoma, and 4 with solid tumors) and 128 healthy controls aged 4 to 18 years. At diagnosis, patients with leukemia and solid tumors showed elevated levels of interferon-γ, interleukin (IL)-1α, IL-1β, IL-4, IL-5, IL-8, IL-10, and tumor necrosis factor. After three months, IL-6, IL-10, and inducible protein-10 levels significantly increased, while IL-1α, IL-1β, and IL-8 rose by six months. These findings indicate that salivary cytokines are elevated at diagnosis and during initial treatment phases in pediatric cancer patients, highlighting saliva’s potential as a noninvasive medium for early detection of systemic diseases in children.

Ginsenoside Rd protects against acute liver injury by regulating the autophagy NLRP3 inflammasome pathway

Ginsenoside Rd (Rd) is a bioactive compound predominantly found in Panax ginseng C.A. Meyer and Panax notoginseng (Burkill) F.H. Chen ex C.H. Chow, both species belonging to genus Panax in the Araliaceae family. However, its hepatic protective effect against acute liver injury and related mechanistic action remain unexplored. To investigate the protective effect of Rd against thioacetamide (TAA)-induced acute liver injury and assess its underlying regulatory mechanisms related to autophagy and inflammation. Forty-eight 8 weeks old C57BL/6 mice were treated with saline (control or model group), Rd (12.5 mg/kg, 25 mg/kg or 50 mg/kg), and diammonium glycyrrhizinate (DG, 30 mg/kg) for three days. Then the mice were stimulated with TAA to establish acute liver injury model, excluding the control group. HSC-T6 cells were treated with Rd at concentrations of 2.5, 5, or 10 µM, for 12 h with or without Lipopolysaccharide (LPS) stimulation at 100 ng/mL. Immunofluorescence staining, qPCR and Western blot were employed to analyze the expressions of genes and proteins associated with inflammation and autophagy. To validate the role of Rd in regulating autophagy and inflammation, the autophagy inducers, rapamycin and GSK621, were utilised in reverse validation experiments in cells. Rd exhibited significant hepatic protective effects in mice by reducing the serum levels of Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Glutathione S-transferase (GST) and Lactate dehydrogenase (LDH) with acute liver injury. It exhibited strong anti-inflammatory effect by reducing inflammation associated protein, such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), nod-like receptor protein 3 (NLRP3), associated speck-like protein containing a CARD (ASC), interleukin-18 (IL-18) and interleukin-1β(IL-1β) proteins and the mRNA expression levels of COX-2, Tumor Necrosis Factor α (TNF α), interleukin-6 (IL-6) and iNOS were decreased in liver tissue. And Rd inhibited LPS-induced inflammation by reducing the expression of COX-2 and NLRP3 in HSC-T6 cells. Moreover, not only in vivo but also in vitro, Rd downregulated the expression of LC3II, Beclin1, phosphorylation-AMP-activated protein kinase (p-AMPK), phosphorylation-ULK1 (p-ULK1) and upregulated the expression of p62 and phosphorylation-mechanistic target of rapamycin (p-mTOR) to suppress autophagy via the AMPK/mTOR/ULK1 pathway. Finally, the inhibitory effects of Rd on autophagy and inflammation in HSC-T6 cells were partially blocked by rapamycin and GSK621. Rd is a promising therapeutic agent to protect liver against TAA-induced acute liver injury by regulating the autophagy-NLRP3 inflammasome pathway.

Biomarkers associated with progression of infantile hemangioma: Exploratory study.

To identify patients with infantile hemangioma (IH) in need of early-stage treatment in this multicenter, prospective, observational study, we investigated the potential of plasma cytokines as a clinically useful marker. Plasma samples were collected at three time points from 41 patients with infantile hemangioma: baseline (days 14-60 after delivery), visit 1 (days 61-150, the proliferative phase), and visit 2 (days 151-395, the involuting phase). With a twofold or more increase in tumor volume during the baseline-visit 1 period regarded as progression, progression was seen in 15 cases (36.6%). In the first step, cytokine arrays were performed using plasma samples in five progressive and five non-progressive cases. Plasma levels of six cytokines at baseline were selected for a prediction marker of change in tumor volume during baseline-visit 1. Validation enzyme-linked immunosorbent assay indicated that the baseline growth differentiation factor 1 (GDF1) concentration tended to correlate with the proliferation ratio of total lesions or target lesion during baseline-visit 1, although without statistical significance. However, the plasma GDF1 concentrations were significantly lower in patients with a fourfold or more increase in total volume (p = 0.013). Furthermore, changes in plasma interleukin (IL)-7Rα levels showed a statistically significant inverse correlation between volume change of the target lesion during baseline-visit 1 (p = 0.0069). Our results suggest that plasma GDF1 measurement after birth is a useful marker for progression of IH. Additionally, the downregulation of IL-7Rα that begins several months after birth may also contribute to tumor growth. (UMIN-CTR: UMIN000038574).

Comparative analysis of inflammatory biomarkers for the diagnosis of neonatal sepsis: IL-6, IL-8, SAA, CRP, and PCT.

Neonatal sepsis (NS) is highly likely to cause death; however, early diagnosis of NS is still a great challenge. This study aimed to determine the diagnostic values of IL-6, IL-8, and serum amyloid A (SAA) in NS patients. C-Reactive protein (CRP), procalcitonin (PCT), interleukin (IL)-6, IL-8, and SAA were detected in 120 infants with NS (60 premature infants [NS-PIs] and 60 term infants [NS-TIs]). Sixty noninfected premature infants and 60 noninfected term infants composed the control group. Receiver operating characteristic (ROC) curves were used to determine the sensitivity and specificity of IL-6, IL-8, and SAA alone and in combination with CRP and PCT. The area under the curve (AUC) was calculated to evaluate the diagnostic value. The clinical characteristics of the subjects were recorded. The expression of CRP, PCT, IL-6, IL-8, and SAA was upregulated in patients with NS compared with control subjects. When the SAA cut-off value was 10.18 mg/L, the greatest AUC for the diagnosis of NS-PIs was for SAA (AUC = 0.833, 95% CI 0.762-0.905, P < 0.001). When the CRP cut-off value was 9.562 mg/L, the smallest AUC for the diagnosis of NS-PIs was for CRP (AUC = 0.776, 95% CI 0.684-0.867, P < 0.001). When the IL-8 cut-off value was 52.03 pg/mL, the greatest AUC for the diagnosis of NS-TIs was for IL-8 (0.821). When the IL-8 cut-off value was 52.03 pg/mL, the greatest AUC for the diagnosis of NS-TIs was for IL-8 (AUC = 0.821, 95% CI 0.745-0.898, P < 0.001). When the CRP cut-off value was 13.18 mg/L, the smallest AUC for the diagnosis of NS-TIs was for CRP (AUC = 0.762, 95% CI 0.667-0.857; P < 0.001). Additionally, according to the AUC value, the best combination was SAA and PCT for NS-PI diagnosis, and the best combination was PCT and IL-6 for NS-TI. In conclusion, compared with PCT and CRP, IL-6, IL-8, and SAA are better diagnostic biomarkers. Moreover, PCT combined with SAA is more suitable for diagnosing NS-PIs, and PCT combined with IL-6 is more suitable for diagnosing NS-TIs.

Mechanism of Ginkgo Diterpene Lactone Meglumine Injection Promoting Angiogenesis and Improving Cerebral Perfusion in Cerebral Infarction Rats

Background In recent years, traditional Chinese medicine has shown advantages in treating ischemic cerebrovascular diseases, including multi-component, multi-target, and low toxicity effects. Purpose: The aim of this study was to clarify the impact of ginkgo diterpene lactone meglumine injection (GDLMI) on promoting microvascular neovascularization and improving cerebral perfusion after cerebral infarction (CIF) in rats and to explore its underlying mechanisms in promoting angiogenesis. Methods Seventy-five rats were randomly rolled into Sham (sham surgery) group, Model (acute CIF model) group, low-dose (LD) (1 g/kg bw GDLMI) group, medium-dose (MD) (3 g/kg bw GDLMI) group, and high-dose (HD) (5 g/kg bw GDLMI) group. The neurological function of rats was evaluated using the Longa method. Micro-PET was performed to evaluate brain metabolism status at 7 days post-reperfusion. Serum expression levels (ELs) of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α were measured. Brain tissue samples were collected to assess cerebral infarct volume and to measure oxidative stress (OS) markers, including superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA). TdT-mediated dUTP nick end labeling (TUNEL) was performed to detect the apoptosis rate of cortical neurons. Western blotting was conducted to determine brain tissue’s ELs of vascular endothelial growth factor (VEGF), angiopoietins-1 (Ang-1), p-PI3K, and p-AKT proteins. Results Rats in Model group exhibited higher Longa scores and increased cerebral infarct volume, decreased brain 18F-FDG content, elevated serum ELs of IL-1β, IL-6, and TNF-α, reduced SOD and CAT activities, increased MDA content in brain tissue, elevated apoptosis rate of cortical neurons, and elevated protein ELs of VEGF and Ang-1 in brain tissue, accompanied by decreased ELs of p-PI3K and p-AKT proteins compared to the Sham group ( p &lt; 0.05). Rats in the LD, MD, and HD groups showed decreased Longa scores and cerebral infarct volume, increased brain 18F-FDG content, decreased ELs of IL-1β, IL-6, and TNF-α, increased SOD and CAT activities, decreased MDA content in brain tissue, decreased apoptosis rate of cortical neurons, and increased protein ELs of VEGF, Ang-1, p-PI3K, and p-AKT in brain tissue versus Model group ( p &lt; 0.05). The effects on all indicators in rats showed a dose-dependent trend. Conclusion GDLMI has a protective effect against acute CIF/reperfusion injury (RI), which may be attributed to its ability to suppress inflammation and OS damage by activating PI3K/AKT signaling.