Excessive Production Of Proinflammatory Cytokines Research Articles

The Therapeutic Effects of SP-8356, a Verbenone Derivative, with Multimodal Cytoprotective Mechanisms in an Ischemic Stroke Rat Model

An ischemic cerebral stroke results from the interruption of blood flow to the brain, triggering rapid and complex cascades of excitotoxicity, oxidative stress, and inflammation. Current reperfusion therapies, including intravenous thrombolysis and mechanical thrombectomy, cause further brain injury due to reperfusion-induced cytotoxicity. To date, novel cytoprotective therapies that could address these challenges have yet to be developed, likely due to the limitations of targeting a single pathologic mechanism. To address these unmet clinical needs, we investigated a synthetic verbenone derivative, SP-8356, as a potential multi-target cytoprotective agent for acute ischemic strokes. In transient middle cerebral artery occlusion (MCAO) rats, SP-8356 significantly reduced brain infarct and edema volumes while improving acute neurological deficits in a dose-dependent manner. Furthermore, SP-8356 improved long-term outcomes, particularly by reducing mortality. These potent cytoprotective effects of SP-8356 were achieved by suppressing the excessive production of free radicals and pro-inflammatory cytokines, reducing the infiltration of inflammatory cells, and mitigating increases in blood–brain barrier permeability. Additional research is needed to determine whether co-administration of SP-8356 can extend the therapeutic time window of reperfusion therapies by mitigating ischemia/reperfusion injury.

P05 The role of JAK inhibitors in difficult-to-treat adult-onset Still’s disease: a case series

Abstract Introduction Adult-onset Still’s Disease (AOSD) is a rare systemic auto-inflammatory disorder characterised by fever, arthritis and rash. Some patients with AOSD exhibit resistance or intolerance to conventional treatments, necessitating alternative therapeutic strategies. This case series aims to explore the efficacy and safety of JAK inhibitors in managing difficult-to-treat AOSD. The driving force behind Still’s disease lies in the excessive and inappropriate production of pro-inflammatory cytokines, notably IL-6 and interferon. Given the pivotal role of JAK inhibitors in mitigating these cytokine pathways, this study highlights their potential therapeutic impact in the management of difficult to treat AOSD. Case description In this retrospective case series, three adult patients with difficult-to-treat AOSD were evaluated for their response to JAK inhibitors, specifically tofacitinib and baricitinib. Case 1: A 32-year-old male presented with a three-week history of fevers, night sweats, weight loss, and lymphadenopathy. Diagnosed with AOSD 18 months later, his treatment began with prednisolone and methotrexate, which was subsequently switched to azathioprine. Tocilizumab was then introduced, alleviating joint pain but causing dermatitis. He was switched to anakinra, allowing a reduction in prednisolone but failing to induce remission and resulting in abnormal liver functions and increased hepatitis B virus titers. Finally, he commenced tofacitinib, achieving symptom resolution, reduced serum ferritin levels, and cessation of steroids. Case 2: A 32-year-old woman presented with a six-month history of joint pain, swelling, stiffness, night sweats, weight loss, and a recurrent rash. Laboratory tests showed anemia, lymphopenia, elevated inflammatory markers, and high ferritin levels. Initial treatment with 60 mg oral prednisolone showed some response, but polyarthritis necessitated further intervention. Anakinra initially managed systemic symptoms, though joint swelling persisted. Tocilizumab was tried but proved ineffective. Subsequently, transitioning to baricitinib at 4 mg daily resulted in good symptomatic control. Case 3: An 18-year-old female presented with recurrent fevers, an intermittent macular rash, joint pains, diarrhoea and splenomegaly. Despite initial antibiotic treatment, persistent symptoms led to an AOSD diagnosis, prompting steroid therapy. Methotrexate failed to control symptoms and caused significant nausea, leading to a switch to tocilizumab. While anakinra initially resolved skin and joint symptoms, recurrent flares necessitated further changes. Azathioprine was trialed but discontinued due to side effects. Colchicine was tried next. Finally, tofacitinib was introduced, and one year later, she achieved significant progress, reducing prednisolone dosage to 5 mg. Discussion The findings from this case series underscore the potential of JAK inhibitors, particularly tofacitinib and baricitinib, as effective treatments for refractory AOSD. In all three cases, patients showed substantial clinical improvement and were able to reduce or discontinue steroids. This suggests a promising role for JAK inhibitors in managing AOSD, especially given the disease’s association with excessive pro-inflammatory cytokine production, including IL-6 and IFN pathways. The transition to JAK inhibitors was often necessitated by inadequate responses or adverse effects from conventional therapies like methotrexate, azathioprine, and biologics such as tocilizumab and anakinra. The successful outcomes with JAK inhibitors highlight their ability to achieve disease control where other treatments failed, demonstrating their efficacy in reducing systemic inflammation and disease activity. Further research is warranted to establish optimal dosing, duration of therapy, and long-term safety profiles of JAK inhibitors in this patient population. These preliminary results are encouraging and suggest a new avenue for treating AOSD. Key learning points • Efficacy of JAK inhibitors: JAK inhibitors, particularly tofacitinib and baricitinib, demonstrated substantial clinical improvement in patients with refractory AOSD. • Steroid reduction: All three patients were able to reduce or discontinue steroid use, highlighting the potential of JAK inhibitors to minimise long-term steroid dependency. • Management of refractory AOSD: JAK inhibitors provided effective disease control where conventional therapies such as methotrexate, azathioprine, and biologics like tocilizumab and anakinra were inadequate or resulted in adverse effects. • Mechanism of action: The efficacy of JAK inhibitors may be attributed to their ability to block pro-inflammatory cytokine pathways, particularly IL-6 and IFN, which are crucial in AOSD pathogenesis. • Safety profile: The treatment was well-tolerated, with significant symptomatic relief and no severe adverse events reported during the study period. • Need for further research: There is a need for further prospective studies to determine the optimal dosing, duration of therapy, and long-term safety profiles of JAK inhibitors in AOSD patients. • Personalised treatment approach: The study supports a personalised treatment approach, considering JAK inhibitors for patients with difficult-to-treat AOSD who do not respond adequately to conventional therapies.

P07 The use of JAK inhibitors in difficult-to-treat adult-onset Still’s disease

Abstract Introduction Adult-onset Still’s Disease (AOSD) is a rare systemic auto-inflammatory disorder characterised by fever, arthritis and rash. Some patients with AOSD exhibit resistance or intolerance to conventional treatments, necessitating alternative therapeutic strategies. This case explores the efficacy and safety of janus kinase (JAK) inhibitors in managing difficult-to-treat AOSD. The driving force behind Still’s disease lies in the excessive and inappropriate production of pro-inflammatory cytokines, notably interleukin-6 (IL-6) and interferon (IFN). Given the pivotal role of JAK inhibitors in mitigating these cytokine pathways, this study highlights their potential therapeutic impact in the management of difficult-to-treat AOSD. Case description A 32-year-old male presented in July 2021 with a three-week history of fevers, night sweats, rigors, and weight loss. A CT scan revealed widespread lymphadenopathy above and below the diaphragm. Blood cultures were negative and, despite antibiotics, the patient continued to experience intermittent fevers. Anemia was asymptomatic, and the patient was transfused with two units of blood. Chronic B infection led to the initiation of entecavir. The patient also experienced palpitations and tachycardia, with an echo revealing a small pericardial effusion, hypertrophied and hyperdynamic left ventricle (LV) with an ejection fraction of 70-75%. A lymph node biopsy was inconclusive, prompting a second biopsy. In September 2021, hyponatremia due to SIADH developed. The patient had ongoing symptoms, including weight loss, and a bone marrow biopsy showed reactive features. CT and PET scans revealed generalised lymphadenopathy. Rheumatology consultation in November 2021 indicated intermittent fever, weight loss, joint pain, and elevated inflammatory markers. The patient had dermatographic lymphadenopathy, and a negative lymph node biopsy lowered the probability of HLH. Methotrexate was initiated, later switched to azathioprine and prednisolone due to ongoing symptoms. In June 2022, the patient started tocilizumab, which resolved joint pain but caused ongoing dermatitis. Tocilizumab was discontinued in January 2023 due to inefficacy, and anakinra was initiated. Prednisolone was gradually tapered off. The patient experienced shortness of breath, and echo revealed pulmonary hypertension. Tocilizumab was discontinued due to injection site reactions and increased hepatitis B virus titers. In March 2023, the decision was made to start the JAK inhibitor tofacitinib. By May 2023, ferritin levels were 1196, and the patient was successfully coming off prednisolone with well-controlled symptoms. Discussion This case underscores the efficacy of JAK inhibitors as a successful therapeutic intervention in challenging cases of AOSD. The 32-year-old male presented with severe symptoms including fevers, night sweats, weight loss, and widespread lymphadenopathy. Initial treatments with antibiotics, methotrexate, azathioprine, prednisolone, and tocilizumab were either ineffective or led to adverse reactions. Despite these setbacks, the patient showed a remarkable clinical response to the JAK inhibitor tofacitinib, initiated in March 2023. The swift and sustained improvement in symptoms, along with a favourable safety profile, suggests that JAK inhibitors can be a promising treatment for refractory AOSD. The ability to taper off steroids while maintaining symptom control is particularly noteworthy, as long-term steroid use is associated with significant side effects. The success of JAK inhibitors in this case is likely attributed to their mechanism of action, which strategically blocks pro-inflammatory cytokine pathways, particularly IL-6 and IFN. This targeted approach addresses the underlying pathophysiology of AOSD, leading to better disease management. However, this case also highlights the importance of monitoring for potential side effects, such as increased hepatitis B virus titres observed in this patient. This necessitated adjustments in therapy and careful monitoring of liver function. The encouraging outcomes in this case prompt the need for further prospective studies to validate these findings. Future research should focus on elucidating the optimal dosing and duration of JAK inhibitor therapy in patients with refractory AOSD. Additionally, long-term follow-up studies are essential to fully understand the safety profile of these agents. Key learning points • Refractory AOSD treatment: JAK inhibitors, such as tofacitinib, can be effective for patients with refractory AOSD who do not respond to conventional therapies. • Steroid sparing: JAK inhibitors may allow for the tapering off of steroids, reducing the risk of long-term steroid-related side effects. • Cytokine pathway targeting: The success of JAK inhibitors highlights the importance of targeting specific pro-inflammatory cytokine pathways, particularly IL-6 and IFN, in managing AOSD. • Monitoring side effects: Continuous monitoring for potential side effects, including increased the risk of thromboembolic events is crucial when using JAK inhibitors. • Need for further research: Prospective studies are needed to validate the efficacy and safety of JAK inhibitors in AOSD and to determine optimal dosing and treatment duration.

Delayed macrophage targeting by clodronate liposomes worsens the progression of cytokine storm syndrome.

Excessive macrophage activation and production of pro-inflammatory cytokines are hallmarks of the Cytokine Storm Syndrome (CSS), a lethal condition triggered by sepsis, autoimmune disorders, and cancer immunotherapies. While depletion of macrophages at disease onset protects from lethality in an infection-induced CSS murine model, patients are rarely diagnosed early, hence the need to characterize macrophage populations during CSS progression and assess the therapeutic implications of macrophage targeting after disease onset. In this study, we identified MHCII+F4/80+Tim4- macrophages as the primary contributors to the pro-inflammatory environment in CSS, while CD206+F4/80+Tim4+ macrophages, with an anti-inflammatory profile, become outnumbered. Additionally, we observed an expansion of Tim4- macrophages coinciding with increased hematopoietic stem progenitor cells and reduction of committed myeloid progenitors in bone marrow and spleen. Critically, macrophage targeting with clodronate liposomes at disease onset prolonged survival, while their targeting in mice with established CSS exacerbated disease severity, leading to a more dramatic loss of Tim4+ macrophages and an imbalance in pro- versus anti-inflammatory Tim4- macrophage ratio. Our findings highlight the significance of timing in macrophage-targeted interventions for effective management of CSS and suggest potential therapeutic strategies for diseases characterized by uncontrolled inflammation, such as sepsis.

Mesenchymal stromal cells can block palmitate training of macrophages via cyclooxygenase-2 and interleukin-1 receptor antagonist

Innate training of macrophages can be beneficial for the clearance of pathogens. However, for certain chronic conditions, innate training can have detrimental effects due to an excessive production of pro-inflammatory cytokines. Obesity is a condition that is associated with a range of increased pro-inflammatory training stimuli including the free fatty acid palmitate. Mesenchymal stromal cells (MSCs) are powerful immunomodulators and known to suppress inflammatory macrophages via a range of soluble factors. We show that palmitate training of murine bone-marrow-derived macrophages and human monocyte-derived macrophages (MDMs) results in an increased production of TNFα and IL-6 upon stimulation with lipopolysaccharide and is associated with epigenetic remodeling. Palmitate training led to metabolic changes, however, MSCs did not alter the metabolic profile of human MDMs. Using a transwell system, we demonstrated that human bone marrow MSCs block palmitate training in both murine and human macrophages suggesting the involvement of secreted factors. MSC disruption of the training process occurs through more than one pathway. Suppression of palmitate-enhanced TNFα production is associated with cyclooxygenase-2 activity in MSCs, while secretion of interleukin-1 receptor antagonist by MSCs is required to suppress palmitate-enhanced IL-6 production in MDMs.

The dual role of glucocorticoid regeneration in inflammation at parturition.

Fetal membrane inflammation is an integral event of parturition. However, excessive pro-inflammatory cytokines can impose threats to the fetus. Coincidentally, the fetal membranes express abundant 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1), which generates biologically active cortisol to promote labor through induction of prostaglandin synthesis. Given the well-recognized anti-inflammatory actions of glucocorticoids, we hypothesized that cortisol regenerated in the fetal membranes might be engaged in restraining fetus-hazardous pro-inflammatory cytokine production for the safety of the fetus, while reserving pro-labor effect on prostaglandin synthesis to ensure safe delivery of the fetus. The hypothesis was examined in human amnion tissue and cultured primary human amnion fibroblasts as well as a mouse model. 11β-HSD1 was significantly increased in the human amnion in infection-induced preterm birth. Studies in human amnion fibroblasts showed that lipopolysaccharide (LPS) induced 11β-HSD1 expression synergistically with cortisol. Cortisol completely blocked NF-κB-mediated pro-inflammatory cytokine expression by LPS, but STAT3-mediated cyclooxygenase 2 expression, a crucial prostaglandin synthetic enzyme, remained. Further studies in pregnant mice showed that corticosterone did not delay LPS-induced preterm birth, but alleviated LPS-induced fetal organ damages, along with increased 11β-HSD1, cyclooxygenase 2, and decreased pro-inflammatory cytokine in the fetal membranes. There is a feed-forward cortisol regeneration in the fetal membranes in infection, and cortisol regenerated restrains pro-inflammatory cytokine expression, while reserves pro-labor effect on prostaglandin synthesis. This dual role of cortisol regeneration can prevent excessive pro-inflammatory cytokine production, while ensure in-time delivery for the safety of the fetus.

VEGFR1 TK signaling protects the lungs against LPS-induced injury by suppressing the activity of alveolar macrophages and enhancing the anti-inflammatory function of monocyte-derived macrophages

Acute lung injury (ALI) is characterized by hyperinflammation followed by vascular leakage and respiratory failure. Vascular endothelial growth factor (VEGF)-A is critical for capillary permeability; however, the role of VEGF receptor 1 (VEGFR1) signaling in ALI progression remains unclear. Here, we show that deletion of VEGFR1 tyrosine kinase (TK) signaling in mice exacerbates lipopolysaccharide (LPS)-induced ALI as evidenced by excessive pro-inflammatory cytokine production and interleukin(IL)-1β-producing neutrophil recruitment to inflamed lung tissues. ALI development involves reduced alveolar macrophage (AM) levels and recruitment of monocyte-derived macrophages (MDMs) in a VEGFR1 TK-dependent manner. VEGFR1 TK signaling reduced pro-inflammatory cytokine levels in cultured AMs. VEGFR1 TK-expressing MDMs displayed an anti-inflammatory macrophage phenotype. Additionally, the transplantation of VEGFR1 TK-expressing bone marrow (BM)-derived macrophages into VEGFR1 TK-deficient mice reduced lung inflammation. Treatment with placental growth factor (PlGF), an agonist for VEGFR1, protected the lung against LPS-induced ALI associated with increased MDMs. These results suggest that VEGFR1 TK signaling prevents LPS-induced ALI by suppressing the pro-inflammatory activity of AMs and enhancing the anti-inflammatory function of MDMs.

A Randomized Placebo-Controlled Trial of Leronlimab in Mild-To-Moderate COVID-19

PurposeEarly in the course of the SARS-CoV-2 pandemic it was hypothesised that host genetics played a role in the pathophysiology of COVID-19 including a suggestion that the CCR5-Δ32 mutation may be protective in SARS-CoV-2 infection. Leronlimab is an investigational CCR5-specific humanized IgG4 monoclonal antibody currently in development for HIV-1 infection. We aimed to explore the impact of leronlimab on the severity of disease symptoms among participants with mild-to-moderate COVID-19. MethodsThe TEMPEST trial was a randomized, double-blind, placebo-controlled study in participants with mild-to-moderate COVID-19. Participants were randomly assigned in a 2:1 ratio to receive subcutaneous leronlimab (700 mg) or placebo on days 0 and 7. The primary efficacy endpoint was assessed by change in total symptom score based on fever, myalgia, dyspnea, and cough, at end of treatment (day 14). FindingsOverall, 84 participants were randomized and treated with leronlimab (n = 56) or placebo (n = 28). No difference was observed in change in total symptom score (P = 0.8184) or other pre-specified secondary endpoints between treatments. However, in a post hoc analysis, 50.0% of participants treated with leronlimab demonstrated improvements from baseline in National Early Warning Score 2 (NEWS2) at day 14, compared with 20·8% of participants in the placebo group (post hoc; p = 0.0223). Among participants in this trial with mild-to-moderate COVID-19 adverse events rates were numerically but not statistically significantly lower in leronlimab participants (33.9%) compared with placebo participants (50.0%). ImplicationsAt the time the TEMPEST trial was designed although CCR5 was known to be implicated in COVID-19 disease severity the exact pathophysiology of SARS-CoV-2 infection was poorly understood. Today it is well accepted that SARS-CoV-2 infection in asymptomatic-to-mild cases is primarily characterized by viral replication, with a heightened immune response, accompanied by diminished viral replication in moderate-to-severe disease and a peak in inflammatory responses with excessive production of pro-inflammatory cytokines in critical disease. It is therefore perhaps not surprising that no differences between treatments were observed in the primary endpoint or in pre-specified secondary endpoints among participants with mild-to-moderate COVID-19. However, the results of the exploratory post hoc analysis showing that participants in the leronlimab group had greater improvement in NEWS2 assessment compared to placebo provided a suggestion that leronlimab may be associated with a lower likelihood of people with mild-to-moderate COVID-19 progressing to more severe disease and needs to be confirmed in other appropriately designed clinical trials.ClinicalTrials.gov number, NCT04343651 https://classic.clinicaltrials.gov/ct2/show/NCT04343651

TLR4/TNFR1 blockade suppresses STAT1/STAT3 expression and increases SOCS3 expression in modulation of LPS-induced macrophage responses

Due to the urgent need to create appropriate treatment techniques, which are currently unavailable, LPS-induced sepsis has become a serious concern on a global scale. The primary active component in the pathophysiology of inflammatory diseases such as sepsis is the Gram-negative bacterial lipopolysaccharide (LPS). LPS interacts with cell surface TLR4 in macrophages, causing the formation of reactive oxygen species (ROS), TNF-α, IL-1β and oxidative stress. It also significantly activates the MAPKs and NF-κB pathway. Excessive production of pro-inflammatory cytokines is one of the primary characteristic features in the onset and progression of inflammation. Cytokines mainly signal through the JAK/STAT pathway. We hypothesize that blocking of TLR4 along with TNFR1 might be beneficial in suppressing the effects of STAT1/STAT3 due to the stimulation of SOCS3 proteins. Prior to the LPS challenge, the macrophages were treated with antibodies against TLR4 and TNFR1 either individually or in combination. On analysis of the macrophage populations by flowcytometry, it was seen that receptor blockade facilitated the phenotypic shift of the M1 macrophages towards M2 resulting in lowered oxidative stress. Blocking of TLR4/TNFR1 upregulated the SOCS3 and mTOR expressions that enabled the transition of inflammatory M1 macrophages towards the anti-inflammatory M2 phenotype, which might be crucial in curbing the inflammatory responses. Also the reduction in the production of inflammatory cytokines such as IL-6, IL-1β due to the reduction in the activation of the STAT1 and STAT3 molecules was observed in our combination treatment group. All these results indicated that neutralization of both TLR4 and TNFR1 might provide new insights in establishing an alternative therapeutic strategy for LPS-sepsis.

Expression of pro-inflammatory cytokines (IL-18, IL-33) at the level of the mucous membrane, the entry point of infection, in persons who have experienced COVID-19 disease

Introduction. The mucous membrane of the upper respiratory tract is the entrance gate for a large number of infections, including the SARS-CoV-2 virus. That is why the main task of the immune system of the mucous membranes of the entrance gate of infection is to maintain respiratory function. High production of proinflammatory cytokines, which play a key role in the development of severe COVID-19 infection, leads to detrimental consequences for all body systems. Their long-term influence can not only aggravate chronic pathologies, but also significantly increase the recovery period, leading to a decrease in the quality of life of patients. Therefore, the purpose of this work is to study the expression of IL-18 and IL-33 genes at the level of the mucous membranes of the upper respiratory tract in patients who have had COVID-19 disease. Materials and methods. The present study involved patients who had moderate or severe COVID-19. The control group consisted of relatively healthy individuals. The expression levels of IL-18 and IL-33 were detected using RT-PCR. Results. During the entire period of rehabilitation after suffering from COVID-19, patients showed a tendency to increase the level of IL-18 expression at the level of the mucous membranes of the nasopharynx and oropharynx. The level of IL-33 production was also increased, but varied depending on the location and period of sample collection. Conclusion. This increase in the level of IL-18 during the rehabilitation period of patients after COVID-19 may be explained by the fact that the virus, by activating glia through olfactory receptor neurons, triggers a powerful immune response and promotes the production of a large number of pro-inflammatory cytokines. On the contrary, overexpression of IL-33 in the late stages of rehabilitation is most likely associated with its ability to restore barrier tissues of the mucous membranes of the upper respiratory tract. Thus, we can conclude that the virus promotes excessive production of pro-inflammatory cytokines, the amount of which increases maximally at the 6th month of rehabilitation after suffering from COVID-19.

Excessive palmitic acid disturbs macrophage α-ketoglutarate/succinate metabolism and causes adipose tissue insulin resistance associated with gestational diabetes mellitus

Abnormal polarization of adipose tissue macrophages (ATMs) results in low-grade systemic inflammation and insulin resistance (IR), potentially contributing to the development of diabetes. However, the underlying mechanisms that regulate the polarization of ATMs associated with gestational diabetes mellitus (GDM) remain unclear. Thus, we aimed to determine the effects of abnormal fatty acids on macrophage polarization and development of insulin resistance in GDM. Levels of fatty acids and inflammation were assessed in the serum samples and adipose tissues of patients with GDM. An in vitro cell model treated with palmitic acid was established, and the mechanisms of palmitic acid in regulating macrophage polarization was clarified. The effects of excessive palmitic acid on the regulation of histone methylations and IR were also explored in the high-fat diet induced GDM mice model. We found that pregnancies with GDM were associated with increased levels of serum fatty acids, and inflammation and IR in adipose tissues. Increased palmitic acid could induce mitochondrial dysfunction and excessive ROS levels in macrophages, leading to abnormal cytoplasmic and nuclear metabolism of succinate and α-ketoglutarate (αKG). Specifically, a decreased nuclear αKG/succinate ratio could attenuate the enrichment of H3K27me3 at the promoters of pro-inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, leading to cytokine secretion. Importantly, GDM mice treated with GSK-J4, an inhibitor of histone lysine demethylase, were protected from abnormal pro-inflammatory macrophage polarization and excessive production of pro-inflammatory cytokines. Our findings highlight the importance of the metabolism of αKG and succinate as transcriptional modulators in regulating the polarization of ATMs and the insulin sensitivity of adipose tissue, ensuring a normal pregnancy. This novel insight sheds new light on gestational fatty acid metabolism and epigenetic alterations associated with GDM.

The severity of COVID-19 upon hospital admission is associated with plasma omega-3 fatty acids

Fatty acids are precursors of inflammatory oxylipins. In the context of COVID-19, an excessive production of pro-inflammatory cytokines is associated with disease severity. The objective was to investigate whether the baseline omega 3/omega 6 fatty acids ratio and the oxylipins were associated with inflammation and oxidative stress in unvaccinated patients with COVID-19, classified according to the severity of the disease during hospitalization. This Prospective population-based cohort study included 180 hospitalized patients with COVID-19. The patients were classified into five groups according to the severity of their disease. Group 1 was the least severe and Group 5 was the most severe. Three specific types of fatty acids—eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA)—as well as their enzymatic and non-enzymatic oxylipins were determined using chromatography coupled mass spectrometry. There was no difference in the ratio of omega-3 to omega-6 fatty acids between the groups (p = 0.276). However, the EPA/AA ratio was lower in Group 4 compared to Group 1 (p = 0.015). This finding was associated with an increase in both C-Reactive Protein (p < 0.001) and Interleukin-6 (p = 0.002). Furthermore, the concentration of F2-Isoprostanes was higher in Group 4 than in Group 1 (p = 0.009), while no significant changes were observed for other oxylipins among groups. Multivariate analysis did not present any standard of biomarkers, suggesting the high complexity of factors involved in the disease severity. Our hypothesis was confirmed in terms of EPA/AA ratio. A higher EPA/AA ratio upon hospital admission was found to be associated with lower concentration of C-Reactive Protein and Interleukin-6, leading to a better prognosis of hospitalized SARS-CoV-2 patients. Importantly, this beneficial outcome was achieved without any form of supplementation. The trial also provides important information that can be further applied to reduce the severity of infections associated with an uncontrolled synthesis of pro-inflammatory cytokines.Trial registration: https://clinicaltrials.gov/study/NCT04449718—01/06/2020. ClinicalTrials.gov Identifier: NCT04449718.

Ser9p-GSK3β Modulation Contributes to the Protective Effects of Vitamin C in Neuroinflammation.

The prolonged activation of microglia and excessive production of pro-inflammatory cytokines can lead to chronic neuroinflammation, which is an important pathological feature of Parkinson's disease (PD). We have previously reported the protective effect of Vitamin C (Vit C) on a mouse model of PD. However, its effect on microglial functions in neuroinflammation remains to be clarified. Glycogen synthase kinase 3β (GSK3β) is a serine/threonine kinase having a role in driving inflammatory responses, making GSK3β inhibitors a promising target for anti-inflammatory research. In this study, we investigated the possible involvement of GSK3β in Vit C neuroprotective effects by using a well-known 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced animal model of PD and a cellular model of neuroinflammation, represented by Lipopolysaccharide (LPS)-activated BV-2 microglial cells. We demonstrated the ability of Vit C to decrease the expression of different mediators involved in the inflammatory responses, such as TLR4, p-IKBα, and the phosphorylated forms of p38 and AKT. In addition, we demonstrated for the first time that Vit C promotes the GSK3β inhibition by stimulating its phosphorylation at Ser9. This study evidenced that Vit C exerts an anti-inflammatory function in microglia, promoting the upregulation of the M2 phenotype through the activation of the Wnt/β-catenin signaling pathway.

Risk Factors for Pulmonary Embolism in Individuals Infected with SARS-CoV2-A Single-Centre Retrospective Study.

The emergence of SARS-CoV2 has presented itself as a significant global health crisis. The prevalence of thrombotic events is known to be high in these patients, affecting various organ systems, sometimes leading to cutaneous thrombosis, pulmonary embolism (PE), stroke, or coronary thrombosis. The available evidence suggests that thromboembolism, hypercoagulability, and the excessive production of proinflammatory cytokines play a significant role in the development of multiorgan failure. Methodology: This retrospective single-centre study was conducted at "Victor Babes" University of Medicine and Pharmacy from Timisoara, Romania, involving a total of 420 patients diagnosed with COVID-19. We separated them into a CONTROL group that included 319 patients, and an intervention group (PE) with 101 patients that, subsequent to infection with the virus, developed pulmonary embolism. The study included the reporting of demographic data, laboratory findings, and comorbidities. Results: Out of a total of 420 patients, 24% experienced pulmonary embolism, while 21.42% died. Arterial thrombotic events were found to be associated with factors such as age, cardiovascular disease, levels of white blood cells, D-dimers, and albumin in the blood. The findings of the study indicate that there is an independent association between pulmonary thrombosis and hypertension (odds ratio (OR): 1.1; 95% confidence interval (CI): 0.7 to 1.7; p = 0.6463), cancer (OR: 1.1; 95% CI: 0.6 to 2.3; p = 0.6014), and COPD (OR: 1.2; 95% CI: 0.6 to 2.3; p = 0.4927). On the other hand, there is a stronger correlation between PE and obesity (OR: 2.8; 95% CI: 1.7 to 4.6; p < 0.0001), diabetes (OR: 3.3; 95% CI: 2 to 5.3; p < 0.0001), and dyslipidemia (OR: 3.6; 95% CI: 2.3 to 5.8; p < 0.0001) in a multivariable regression logistic model. Conclusions: Patients diagnosed with severe forms of COVID-19 display a comparable incidence of arterial thrombotic events, which have been linked to poor survival rates.

Canakinumab in the treatment of systemic juvenile idiopathic arthritis: a retrospective single center study in China.

Systemic juvenile idiopathic arthritis (sJIA) is characterized by excessive production of proinflammatory cytokines. As an anti-IL-1 agent, canakinumab has been approved in the USA and Europe for the treatment of sJIA patients aged ≥2 years. However, the use of canakinumab has never been reported in China. In this study, we aimed to assess the efficacy and safety of canakinumab in Chinese patients with sJIA. A total of 11 patients with sJIA who were treated with canakinumab were included in this study. Clinical data were collected retrospectively from medical records. Efficacy was evaluated by the systemic juvenile arthritis disease activity score (sJADAS). The follow-up was performed at canakinumab initiation, at months 1, 3, 6, 9 and 12, or at the last follow-up. Of the 11 patients enrolled, 91.0% (10/11) had previously received treatment with tocilizumab. The mean duration of canakinumab was 9 (3-18) months. 45.5% (5/11) of patients showed complete response, 45.5% (5/11) showed partial response, and 9.0% (1/11) showed no response. 18.2% (2/11) experienced disease flare during the treatment with canakinumab. 81.8% (9/11) of patients successfully reduced the dose of corticosteroids, with six discontinuing corticosteroids. 45.6% (5/11) of patients experienced infection. No serious adverse events occurred during the treatment with canakinumab. Canakinumab may be effective and tolerable for Chinese sJIA patients, helping to reduce the dosage of corticosteroids. However, additional researches on large samples are required to evaluate its efficacy and safety.

Epigenetic and transcriptional regulation of cytokine production by Plasmodium falciparum-exposed monocytes.

Plasmodium falciparum infection causes the most severe form of malaria, where excessive production of proinflammatory cytokines can drive the pathogenesis of the disease. Monocytes play key roles in host defense against malaria through cytokine production and phagocytosis; however, they are also implicated in pathogenesis through excessive proinflammatory cytokine production. Understanding the underlying molecular mechanisms that contribute to inflammatory cytokine production in P. falciparum-exposed monocytes is key towards developing better treatments. Here, we provide molecular evidence that histone 3 lysine 4 (H3K4) methylation is key for inflammatory cytokine production in P. falciparum-exposed monocytes. In an established in vitro system that mimics blood stage infection, elevated proinflammatory TNF and IL-6 cytokine production is correlated with increased mono- and tri-methylated H3K4 levels. Significantly, we demonstrate through utilizing a pharmacological inhibitor of H3K4 methylation that TNF and IL-6 expression can be suppressed in P. falciparum-exposed monocytes. This elucidated epigenetic regulatory mechanism, controlling inflammatory cytokine production, potentially provides new therapeutic options for future malaria treatment.

Inflammation-Responsive Nanoagents for Activatable Photoacoustic Molecular Imaging and Tandem Therapies in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) severely lowers the life quality by progressively destructing joint functions and eventually causing permanent disability, representing a pressing public health concern. The pathogenesis of RA includes the excessive production of proinflammatory cytokines and harmful oxygen-derived free radicals, such as nitric oxide (NO), which constitute vital targets for precise diagnosis and effective treatment of RA. In this study, we introduce an advanced nanoagent that integrates the RA microenvironment-activatable photoacoustic (PA) imaging with multitarget synergistic treatment for RA. A highly sensitive organic probe with NO-tunable energy transformation and molecular geometry is developed, which enables strong near-infrared absorption with a turn-on PA signal, and the active intramolecular motion could further boost PA conversion. The probe is coassembled with an inflammation-responsive prodrug to construct the theranostic nanoagent, on which a macrophage-derived cell membrane with natural tropism to the inflammatory sites is camouflaged to improve the targeting ability to inflamed joints. The nanoagent could not only sensitively detect RA and differentiate the severity but also efficiently alleviate RA symptoms and improve joint function. The combination of activatable probe-mediated NO scavenging and on-demand activation of anti-inflammatory prodrug significantly inhibits the proinflammatory factors and promotes macrophage repolarization from M1 to M2 phenotype. This meticulously designed nanoagent ingeniously integrates RA-specific PA molecular imaging with synergistic multitarget therapy, rendering tremendous promise for precise intervention of RA-related diseases.

Deficiency of Trex1 leads to spontaneous development of type 1 diabetes

BackgroundType 1 diabetes is believed to be an autoimmune condition, characterized by destruction of insulin-producing cells, due to the detrimental inflammation in pancreas. Growing evidences have indicated the important role of type I interferon in the development of type 1 diabetes.MethodsTrex1-deficient rats were generated by using CRISPR-Cas9. The fasting blood glucose level of rat was measured by a Roche Accuchek blood glucose monitor. The levels of insulin, islet autoantibodies, and interferon-β were measured using enzyme-linked immunosorbent assay. The inflammatory genes were detected by quantitative PCR and RNA-seq. Hematein-eosin staining was used to detect the pathological changes in pancreas, eye and kidney. The pathological features of kidney were also detected by Masson trichrome and periodic acid-Schiff staining. The distribution of islet cells, immune cells or ssDNA in pancreas was analyzed by immunofluorescent staining.ResultsIn this study, we established a Trex1-deletion Sprague Dawley rat model, and unexpectedly, we found that the Trex1−/− rats spontaneously develop type 1 diabetes. Similar to human diabetes, the hyperglycemia in rats is accompanied by diabetic complications such as diabetic nephropathy and cataract. Mechanistical investigation revealed the accumulation of ssDNA and the excessive production of proinflammatory cytokines, including IFN-β, in Trex1 null pancreas. These are likely contributing to the inflammation in pancreas and eventually leading to the decline of pancreatic β cells.ConclusionsOur study links the DNA-induced chronic inflammation to the pathogenesis of type 1 diabetes, and also provides an animal model for type 1 diabetes studies.

Safflower extract (Carthamus tinctorius Linn.) suppresses proinflammatory cytokines level in rheumatoid arthritis mice model stimulated by complete Freund’s adjuvants

Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammation of the synovial membrane, and swelling, associated with excessive production of proinflammatory cytokines. In addition, anti-citrullinated protein antibodies is excessively high in RA. Carthamus tinctorius Linn. (C. tinctorius), commonly known as Safflower, is widely used as traditional medicine in Indonesia and has anti-inflammatory properties. This study aims to determine cell viability on HEK293 cells and the anti-RA activity of the ethanol extract of C. tinctorius on reducing edema in animal models of RA mice and to determine the anti-RA activity ethanol extract C. tinctorius against rheumatoid factor (RF) in RA mice. In this study, HEK293 cell line was treated with C. tinctorius extract at various concentrations. In addition, mice’s paws were examined after being injected with complete Freund’s adjuvant (CFA) as an intraplantar RA inducer and treated with 2.5 mg/kg body weight (BW) of methotrexate, and extract C. tinctorius at various concentrations orally once a day for 14 days. Then, the paw’s thickness was measured using a gauge meter, arthritis index scoring was observed daily, and agglutination in the RF test was observed on the 17th and 31st days. The collected data was analyzed using one-way analysis of variance. Moreover, an In-Silico test was carried out by measuring the affinity and stability of C.tintorius extract as an anti-RA agent against the cytokines IL-1β and tumor necrosis factor-alpha. The test results showed C. tinctorius at various concentrations were safe to HEK293 cell lines and the administration of C. tintorius extract at 100, 200, and 400 mg/kg BW doses could reduce paw thickness, arthritis index scoring &gt;1, and no agglutination occurred in the RF test. It can concluded that the C. tintorius extract is safe to HEK293 and has activity against CFA-induced RA.

Immunomodulatory effect of propolis Trigona spp. in mice infected with Toxoplasma gondii.

Toxoplasmosis is a zoonotic disease caused by Toxoplasma gondii. Infection with virulent strains of T. gondii induces excessive production of pro-inflammatory cytokines and increases fatality. Propolis has potential as an anti-inflammatory and immunomodulator. This study aimed to evaluate the pro-inflammatory cytokines response of Interleukin-2 (IL-2) and IL-17, and the anti-inflammatory cytokine IL-10 in mice infected with T. gondii RH strain and treated with ethanol extract of Trigona bee propolis (EEP) from Lombok, West Nusa Tenggara (NTB), Indonesia. A total of 36 mice were divided into six groups with six mice per group. Group 1 was considered as a negative control; mice were not infected and not treated while in groups 2 to 6, mice were infected with 100 T. gondii (tachyzoites). Mice in group 2 were infected but not treated whereas in groups 3, 4, 5, and 6, respectively, were treated orally with Cotrimoxazole 40 mg/kg body weight, EEP 1 mg, EEP 2 mg, and EEP 4 mg, the day after infection for 3 days. All mice under study were euthanized following ethical considerations with prior approvals, on the fifth day post-infection and spleens were taken to observe cytokines using flow cytometry. Statistical analysis was performed with one-way ANOVA and Duncan Multiple Range Test (you may add any statistical tool/software if used?? Like R version?, SPSS, version? If you used any of these/others)? EEP (1-4 mg) increased CD4+ T cells, CD8+ T cells, and CD11b+ cells in infected mice, though not significantly (p > 0.05), unlike cotrimoxazole, which showed significant increases (p < 0.05). EEP at 2 mg and 4 mg (G5 and G6) reduced IL-2 and IL-17 in CD4+ T cells, and at 4 mg (G6) in CD8+ T cells. IL-10 expression increased in CD11b+ cells at 1 mg and in CD4+ T cells at 4 mg (G6). Propolis ethanol extract from Trigona spp. bees acts as an immunomodulator and anti-inflammatory in T. gondii-infected mice, supporting its potential as an anti-toxoplasmosis agent.