Inhibition Of Cytokine Release Research Articles

Exosomes of Human Umbilical Cord MSCs Protect Against Hypoxia/Reoxygenation-Induced Pyroptosis of Cardiomyocytes via the miRNA-100-5p/FOXO3/NLRP3 Pathway.

BackgroundAcute myocardial infarction (AMI) is one of the leading causes of morbidity and death worldwide. Studies have indicated that microRNAs in mesenchymal stem cell (MSC)-derived exosomes are crucial for treating various diseases.MethodsHuman umbilical cord MSC (hucMSC)-derived exosomes (hucMSC-exo) were isolated and used to treat cardiomyocytes that underwent hypoxia/reoxygenation (H/R) injury. Bioluminescence assessment was used to study binding of miRNA to its targeting gene.ResultsWe found that H/R decreased the viability of AC16 cells, increased the expression of NLRP3, and activated caspase-1(p20) and GSDMD-N as well as release of IL-1β and IL-18, and such effects were abolished by administration of hucMSC-exo. Administration of exosomes from negative scramble miRNA (NC)-transfected hucMSCs blocked H/R-caused lactate dehydrogenase release, pyroptosis, and over-regulation of NLRP3 and activated caspase-1(p20) and GSDMD-N as well as release of IL-1β and IL-18. More importantly, in comparison to exsomes from NC-transfected hucMSCs, exsomes from miR-100-5p-overexpressing hucMSCs had more obvious effects, and those from miR-100-5p-inhibitor-transfected hucMSCs showed fewer effects. Functional study showed that miR-100-5p bound to the 3’-untranslated region (3’-UTR) of FOXO3 to suppress its transcription. Moreover, overexpression of FOXO3 abolished the protective effects of miR-100-5p.ConclusionEnriched miR-100-5p in hucMSC-exo suppressed FOXO3 expression to inhibit NLRP3 inflammasome activation and suppress cytokine release and, therefore, protected cardiomyocytes from H/R-induced pyroptosis and injury.

Resveratrol alleviates sepsis-induced acute kidney injury by deactivating the lncRNA MALAT1/MiR-205 axis.

IntroductionResveratrol plays a protective role against sepsis development, and the long noncoding RNA (lncRNA) MALAT1 is an inflammation-relevant biomarker. This investigation attempted to reveal whether resveratrol attenuated inflammation of sepsis-induced acute kidney injury (AKI) by regulating MALAT1.Material and methodsIn total 120 rats were divided into a control group (n = 20), a Sham group (n = 20), a sepsis group (n = 40) and a resveratrol group (n = 40), and serum levels of inflammatory cytokines and AKI biomarkers were determined. An equal number of rats under identical treatments were, additionally, tracked for their survival, and the serum level of lncRNA MALAT1 was measured by RT-PCR. Moreover, septic cell models were constructed by treating HK-2 cells with lipopolysaccharide (LPS), and tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 levels released by the cells were determined with ELISA.ResultsResveratrol treatment significantly brought down serum levels of inflammatory cytokines (i.e. TNF-α, IL-1β and IL-6), kidney function indicators (i.e. Scr, blood urea nitrogen [BUN] and Scys C), AKI biomarkers (i.e. NGAL and KIM-1) and MALAT1 in cecal ligation and puncture (CLP)-induced septic model rats (all p < 0.05), and the life span of septic rats was elongated by resveratrol treatment (p < 0.05). Viability and cytokine release of LPS-treated HK2 cells were rescued by resveratrol (p < 0.05), which was accompanied by a marked fall of MALAT1 expression (p < 0.05). In addition, si-MALAT1 diminished viability and suppressed cytokine release of HK2 cells, while pcDNA3.1-MALAT1 hindered the impact of resveratrol on the inflammatory response of HK2 cells (p < 0.05). Ultimately, miR-205, a protective molecule in sepsis-relevant AKI, was down-regulated by resveratrol and si-MALAT1 (p < 0.05).ConclusionsResveratrol relieved sepsis-induced AKI by restraining the lncRNA MALAT1/miR-205 axis.

LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC

BackgroundDysregulation of transcription and cytokine expression has been implicated in the pathogenesis of a variety inflammatory diseases. The resulting imbalance between inflammatory and resolving transcriptional programs can cause an overabundance of pro-inflammatory, classically activated macrophage type 1 (M1) and/or helper T cell type 1 (Th1) products, such as IFNγ, TNFα, IL1-β, and IL12, that prevent immune switching to resolution and healing. The low molecular weight fraction of human serum albumin (LMWF5A) is a novel biologic drug that is currently under clinical investigation for the treatment of osteoarthritis and the hyper-inflammatory response associated with COVID-19. This study aims to elucidate transcriptional mechanisms of action involved with the ability of LMWF5A to reduce pro-inflammatory cytokine release.MethodsELISA arrays were used to identify cytokines and chemokines influenced by LMWF5A treatment of LPS-stimulated peripheral blood mononuclear cells (PBMC). The resulting profiles were analyzed by gene enrichment to gain mechanistic insight into the biologic processes and transcription factors (TFs) underlying the identified differentially expressed cytokines. DNA-binding ELISAs, luciferase reporter assays, and TNFα or IL-1β relative potency were then employed to confirm the involvement of enriched pathways and TFs.ResultsLMWF5A was found to significantly inhibit a distinct set of pro-inflammatory cytokines (TNFα, IL-1β, IL-12, CXCL9, CXCL10, and CXCL11) associated with pro-inflammatory M1/Th1 immune profiles. Gene enrichment analysis also suggests these cytokines are, in part, regulated by NF-κB and STAT transcription factors. Data from DNA-binding and reporter assays support this with LMWF5A inhibition of STAT1α DNA-binding activity as well as a reduction in overall NF-κB-driven luciferase expression. Experiments using antagonists specific for the immunomodulatory and NF-κB/STAT-repressing transcription factors, peroxisome proliferator-activated receptor (PPAR)γ and aryl hydrocarbon receptor (AhR), indicate these pathways are involved in the LMWF5A mechanisms of action by reducing LMWF5A drug potency as measured by TNFα and IL-1β release.ConclusionIn this report, we provide evidence that LMWF5A reduces pro-inflammatory cytokine release by activating the immunoregulatory transcription factors PPARγ and AhR. In addition, our data indicate that LMWF5A suppresses NF-κB and STAT1α pro-inflammatory pathways. This suggests that LMWF5A acts through these mechanisms to decrease pro-inflammatory transcription factor activity and subsequent inflammatory cytokine production.

Evaluation of Anti-asthmatic Activity of Salvadora oleoids Leaves

Salvadora oleoids (Family: Salvadoraceae) is being used in the traditional medicine for the treatment of severe bronchitis and asthma. So, the study aims to evaluate anti-asthmatic activity of S. oleoids leaves. Leaves are reported to contain flavonoids, saponins, steroids, alkaloids, and tannins, known to possess similar effects. Flavonoids have been shown to smooth muscle relaxant, bronchodilator, antioxidant, and anti-inflammatory activity. The flavonoids are also reported to inhibit mast cell degranulation. Saponins are reported to mast cell stabilizing, anti-allergic, anti-inflammatory and antihistaminic activity. Tannins are reported to inhibit cyclooxygenase (COX) enzyme, decrease vascular permeability and antioxidant activity. Steroids are reported to an extraordinarily strong anti-inflammatory activity by inhibiting cytokine release IL-1, 2 and 6. The present investigation was undertaken to evaluate the anti-asthmatic activity of ethanolic extract of S. oleoids leaves in experimental animals. Asthma is a chronic inflammatory disorder of the airways. Limitations of currently available therapies and associated side effects have created an urge to search new and better treatment options. Thus, present study was designed to evaluate anti-asthmatic activity of S. oleoids leaves. Ethanolic extract of S. oleoides was prepared and used in the dose of 200 mg/kg (p.o.). Various in-vivo models like histamine induced bronchoconstriction in guinea pigs and milk induced leukocytosis in mice were used for evaluating anti-asthmatic activity of the drugs. Ethanolic extract of S. oleoides showed a significant broncho-dilatory, anti-histaminic and anti-inflammatory activity. Thus, ethanolic extract of S. oleoides have significant anti-asthmatic activity. Thus, ethanolic extract of S. oleoides leaves have proved to be an effective drug in prevention of asthma.

Tocilizumab: The Key to Stop Coronavirus Disease 2019 (COVID-19)-Induced Cytokine Release Syndrome (CRS)?

The COVID-19 disease is an unprecedented international public health emergency and considerably impacts the global economy and health service system. While awaiting the development of an effective vaccine, searching for the therapy for severe or critical COVID-19 patients is essential for reducing the mortality and alleviating the tension of the health service system. Cytokine release syndrome (CRS) induced by elevated interleukin-6 was recognized to underscore the pathology of severe COVID-19 patients. Inhibiting CRS by agents suppressing IL-6 may relieve symptoms, shorten the hospital stay and reduce the need for oxygen therapy. Although evidence from randomized, double-blinded clinical trials is still lacking, the IL-6R inhibitor tocilizumab (TCZ) has shown some clinical benefits in the treatment of severe COVID-19 patients and have been included in clinical guidelines. In this review, we focused on the possible mechanisms of TCZ in the treatment of CRS and highlighted some significant considerations in the use of TCZ to treat COVID-19 patients.

Sesamin attenuates carrageenan-induced lung inflammation through upregulation of A20 and TAX1BP1 in rats

Sesamin is a major component in lignans of sesame seeds, has been described to possess a lot of biological activity. The main objective of our study was to investigate the inhibitory effect and novel molecular mechanisms of sesamin on carrageenan-induced lung inflammation in rats. Here we showed that sesamin can obviously reduce polymorphonuclear neutrophils infiltration and exudate volume. Further studies exhibited sesamin can inhibit cytokines release, polymorphonuclear neutrophils markers production and the degree of lung tissues injury. Western blot analysis revealed that sesamin can inhibit the TRAF6 expression and NF-κB pathway activation in lung tissue. We found that sesamin can increase the expression of A20 and TAX1BP1 in lung tissues, and the interaction between the two molecules. In conclusion, all these results demonstrated that sesamin can attenuate carrageenan-induced lung inflammation, the mechanisms that may be related to upregulation of the novel target A20 and TAX1BP1 which can negative regulation for NF-κB pathway. Importantly, this is the first evidence showing that TAX1BP1 can be as a novel regulatory target to attenuate the lung inflammation.

In Vivo Anti-inflammatory Potential of Viscozyme®-Treated Jujube Fruit.

The fruit of Ziziphus jujuba, commonly called jujube, has long been consumed for its health benefits. The aim of this study was to examine the protective effect of dietary supplementation of enzymatically hydrolyzed jujube against lung inflammation in mice. The macerated flesh of jujube was extracted with aqueous ethanol before and after Viscozyme treatment. The extract of enzyme-treated jujube, called herein hydrolyzed jujube extract (HJE), contained higher levels of quercetin, total phenolics, and flavonoids, and exhibited more effective radical-scavenging abilities in comparison to non-hydrolyzed jujube extract (NHJE). HJE treatment decreased production of inflammation-associated molecules, including nitric oxide and pro-inflammatory cytokines from activated Raw 264.7 or differentiated THP-1 cells. HJE treatment also reduced expression of nuclear factor-κB and its downstream proteins in A549 human lung epithelial cells. Moreover, oral supplementation of 1.5 g of HJE per kg of body weight (BW) attenuated histological lung damage, decreased plasma cytokines, and inhibited expression of inflammatory proteins and oxidative stress mediators in the lungs of mice exposed to benzo(a)pyrene at 50 mg/kg BW. Expression levels of antioxidant and cytoprotective factors, such as nuclear factor erythroid-derived 2-related factor 2 and heme oxygenase-1, were increased in lung and liver tissues from mice treated with HJE, compared to mice fed NHJE. These findings indicate that dietary HJE can reduce benzo(a)pyrene-induced lung inflammation by inhibiting cytokine release from macrophages and promoting antioxidant defenses in vivo.

Allicin Inhibits Proliferation by Decreasing IL-6 and IFN-β in HCMV-Infected Glioma Cells.

PurposeAllicin, an extract of garlic, has antitumor effects in multiple tumor types. However, the efficacy of allicin for treating glioblastoma has not yet been examined. This study examined the antitumor effect of allicin on human cytomegalovirus (HCMV)-infected glioblastoma multiforme (GBM) and its role in cytokine signaling.Materials and MethodsHCMV-infected glioblastoma was modeled by transfection of U87MG glioblastoma cells with HMCV proteins. MTT assay was used to assess the effect of allicin on the proliferation of glioma cells. Western blot analysis was used to detect the effect of allicin on the expression of intermediate-early gene 2 (IE2) and p53. Reverse transcription-quantitative polymerase chain reaction was used to assess and the levels of interleukin (IL)-6 and interferon (IFN)-β. Single cell gel electrophoresis was used to analyze changes in radiotherapy-induced DNA damage.ResultsTransfection of the IE2 protein led to decreased p53 expression and increased glioblastoma cell proliferation. Allicin inhibited this proliferation in a dose- and time-dependent manner. An inhibitory effect on cytokine release was observed in GBM cells treated with allicin. After treatment with allicin, p53 levels increased significantly, whereas expression of the inflammatory factors such as IL-6 and IFN-β decreased. U87MG cells treated with allicin and 10 Gy irradiation had increased intracellular DNA damage compared to either treatment alone.ConclusionAllicin inhibited proliferation of glioblastoma cells in vitro. Allicin also inhibited cytokine release, upregulated p53 activity, and increased the sensitivity of glioblastoma to radiotherapy. These results suggest that allicin is effective against HCMV-infected glioblastomas.

HIV-1 subtype C transmitted founders modulate dendritic cell inflammatory responses

BackgroundHeterosexual transmission remains the main route of HIV-1 transmission and female genital tract (FGT) inflammation increases the risk of infection. However, the mechanism(s) by which inflammation facilitates infection is not fully understood. In rhesus macaques challenged with simian immunodeficiency virus, dendritic cell (DC) mediated recruitment of CD4+ T cells to the FGT was critical for infection. The aim of this study was to delineate the mechanisms underlying DC-mediated HIV infection by comparing chemokine and pro-inflammatory cytokine production in response to transmitted founder (TF) and chronic infection (CI) Envelope (Env) pseudotyped viruses (PSV).ResultsMonocyte-derived DCs (MDDCs) were stimulated with PSV and recombinant gp140 representing matched TF and CI pairs of four individuals and cytokine secretion measured by multiplex immuno-assay. We found that 4/9 Env induced robust MDDC inflammatory responses and of those, three were cloned from TFs. Overall, TF Env induced MDDCs from healthy donors to secrete higher concentrations of inflammatory cytokines and chemokines than those from CI, suggesting TF Env were better inducers of inflammation. Assessing the signalling pathway associated with inflammatory cytokines, we found that PSV of matched TF and CI variants and a gp140 clone activated ERK and JNK to similar levels. Recombinant soluble DC-SIGN inhibited cytokine release and activation of ERK by PSV, suggesting that Env-DC-SIGN binding was partly involved in MDDC stimulation. Therefore, Env clones might differentially stimulate MDDC immune responses via alternative, yet unidentified signalling pathways.ConclusionOverall, this could suggest that the genetics of the virus itself influences inflammatory responses during HIV infection. In the absence of pre-existing infections, induction of greater inflammatory response by TFs might favour virus survival within the healthy FGT by driving an influx of target cells to sites of infection while suppressing immune responses via IL-10.

Vidofludimus calcium, a next generation DHODH inhibitor for the Treatment of relapsing-remitting multiple sclerosis

BackgroundInhibition of dihydroorotate dehydrogenase (DHODH) is an established mechanism for the treatment of relapsing-remitting multiple sclerosis (RRMS). Currently approved treatments have several shortcomings. Consequently, new and effective treatments with improved safety and convenience profiles are sought after by patients. ObjectiveTo explore the overall profile of vidofludimus for the treatment of RRMS. MethodsPreclinical investigations were done exploring the species-dependency of DHODH inhibition of vidofludimus. In addition, the preclinical efficacy in a rat experimental autoimmune encephalomyelitis (EAE) model and the inhibition of cytokine release from activated PBMC were investigated. Pharmacokinetic data were also obtained in a Phase 1 multiple ascending dose trial of the formulation IMU–838 (vidofludimus calcium). ResultsIt was shown that vidofludimus is 2.6 times more potent in inhibiting DHO oxidation by human DHODH compared to teriflunomide. Although both compounds increased cell apoptosis, vidofludimus was more efficacious in the inhibition of T-lymphocyte proliferation compared to teriflunomide. The same was also observed for the secretion of IL–17 and IFN–γ. Interestingly, the potency or vidofludimus to inhibit rat or mouse DHODH is 7.5 and 64.4 time lower than the for the human DHODH, respectively. The rat EAE study clearly exhibited a dose-dependent inhibition of cumulative disease scores by vidofludimus. In the multiple ascending dose Phase 1 clinical trial, the serum half-life of about 30 h provides a favorable profile for once daily dosing of IMU–838, with quick dosing to steady state through levels within 5 days and the ability to wash out drug quickly, if required. ConclusionsThe investigations highlighted that the desired selective immunomodulatory properties can be separated from general antiproliferative effects seen and related adverse events in first-generation DHODH inhibitors. Based on data obtained from a series of pre-clinical as well as phase 1 and phase 2 studies, IMU–838 is a promising next-generation candidate for the oral treatment of RRMS. However, this will need to be confirmed in the currently ongoing Phase 2 study in RRMS patients.

Effects of corticosteroids on COPD lung macrophage phenotype and function.

The numbers of macrophages are increased in the lungs of chronic obstructive pulmonary disease (COPD) patients. COPD lung macrophages have reduced ability to phagocytose microbes and efferocytose apoptotic cells. Inhaled corticosteroids (ICSs) are widely used anti-inflammatory drugs in COPD; however, their role beyond suppression of cytokine release has not been explored in COPD macrophages. We have examined the effects of corticosteroids on COPD lung macrophage phenotype and function. Lung macrophages from controls and COPD patients were treated with corticosteroids; effects on gene and protein expression of CD163, CD164, CD206, MERTK, CD64, CD80 and CD86 were studied. We also examined the effect of corticosteroids on the function of CD163, MERTK and cluster of differentiation 64 (CD64). Corticosteroid increased CD163, CD164, CD206 and MERTK expression and reduced CD64, CD80 and CD86 expression. We also observed an increase in the uptake of the haemoglobin-haptoglobin complex (CD163) from 59 up to 81% and an increase in efferocytosis of apoptotic neutrophils (MERTK) from 15 up to 28% following corticosteroid treatment. We observed no effect on bacterial phagocytosis. Corticosteroids alter the phenotype and function of COPD lung macrophages. Our findings suggest mechanisms by which corticosteroids exert therapeutic benefit in COPD, reducing iron available for bacterial growth and enhancing efferocytosis.

Brominated Flame Retardants alter Cytokine Production in Mast Cells

Polybrominated diphenyl ethers (PBDEs), extensively used as flame‐retardants have been classified as persistent environmental pollutants. PBDEs have also been shown to cause neurotoxicity, immunotoxicity and endocrine disruption; however, the exact mechanisms are largely unknown. In this study, we propose to investigate PBDE‐induced immunotoxicity in mast cells. We examined the effects of two of the most commonly occurring brominated diphenyl ether (BDE) congeners, BDE‐47 and −99, and a metabolite of BDE‐47, 6‐OH‐BDE‐47 on MC/9 mast cells. MC/9 cells treated with BDE‐47, −99 and 6‐OH‐BDE‐47 (0.25, 0.5, 5, 20 and 40μM, 24h) were stimulated with IgE/antigen (1μg/ml), and levels of IL‐6 and TNF‐α were measured by ELISA. Our results indicate that PBDEs inhibit the IgE‐mediated cytokine release in MC/9 cells in a concentration‐dependent manner. Interestingly, exposure to BDE‐47 and 6‐OH‐BDE47 decreased IL‐6 release in a concentration‐dependent manner; however, BDE‐99 decreased IL‐6 release only at concentrations above 20 μM. These results demonstrate that tetra‐brominated BDE‐47 and its 6‐OH metabolite can suppress cytokine release with increasing concentrations, while penta‐brominated BDE‐99 congener inhibits IL‐6 release only at higher concentrations. Mast cell degranulation measured by β‐hexoaminidase also showed that BDE congeners decreased degranulation in a concentration‐dependent manner in IgE stimulated mast cells. In order to test effects on cell viability, we exposed the MC/9 cells to increasing concentrations of the BDE congeners and measured cytotoxicity at 24 and 48h by MTS assay. We found that tetra‐brominated BDE‐47 exhibited more potent cytotoxicity (EC50 μM =42.38 at 24h and 32.33 at 48h) in comparison to the penta‐brominated BDE‐99 (EC50 μM =48.41 and 35.04 at 48h) in the MC/9 cells. However, the 6‐OH metabolite of BDE‐47 was more cytotoxic in comparison to its parent compound BDE‐47 as well as penta‐brominated BDE‐99 (EC50 μM =25.82 at 24h and 20.16 at 48h). We are further characterizing development of apoptosis in the MC9 cells in the presence of BDE congeners. Our results suggest that PBDE congeners can modulate cytokine release in mast cells and the extent of inhibition is dependent on the bromination of PBDEs. These results address the need to better understand the molecular and cellular mechanisms by which PBDE exposure can mediate immunotoxicity.

Prednisolone encapsulated PLGA nanoparticles: Characterization, cytotoxicity, and anti-inflammatory activity on C6 glial cells

Prednisolone, a popular glucocorticoid (GC) known for its ability to inhibit cytokine release, is also employed extensively in cancer therapy. GCs have been used to treat brain tumors to reduce tumor-associated edema. They are known to exhibit different effects on different cell lines and in some cases are known to be neurotoxic. In this study, we have investigated and compared the cytotoxic and anti-inflammatory effects of prednisolone and prednisolone encapsulated Poly Lactic-co-Glycolic acid (PLGA) nanoparticles (NPs) on C6 cells that are cancerous but are known for their similarity to astrocyte cells. Design expert software was used to analyze the effect of different variables for NP formulation. By varying different parameters, NPs were synthesized and characterized using particle size analyzer and zeta potential. The surface morphology of the NPs was analyzed using scanning electron microscopy. Lipopolysaccharide activated C6 glial cells witnessed significantly lower cell proliferation in the presence of the drug during the 48 hours of incubation and the prednisolone encapsulated NPs were able to attenuate pro-inflammatory cytokines like Tumor necrosis factor alpha (TNF-α) and nitric oxide substantially even after the 72 hours of incubation when compared with the free drug. The results suggested that prednisolone was more effective as an anti-inflammatory drug in the PLGA nanoformulation

P156 ALPN-101, A FIRST-IN-CLASS DUAL ICOS/CD28 ANTAGONIST, DEMONSTRATES EFFICACY IN PATIENT-DERIVED PBMC IN VITRO AND IN AN IN VIVO T CELL TRANSFER MODEL OF CHRONIC INFLAMMATORY BOWEL DISEASE (IBD)

Abstract Background T cell costimulation has been strongly implicated in the pathogenesis of IBD, yet CD28 costimulatory pathway inhibitors (e.g. abatacept, CTLA4-Fc) have not proven clinically efficacious, implicating an alternative costimulatory pathway. ICOS is a costimulatory receptor highly related to CD28, upregulated upon T cell activation and mediating costimulatory signals in post-activation T cells - suggesting ICOS may be more relevant in active disease. In contrast, CD28 predominates in naïve T cells and is less critical in activated, effector and/or memory T cells. ALPN-101 is an Fc fusion protein of a human inducible T cell costimulator ligand (ICOSL) variant immunoglobulin domain (vIgDTM) engineered to inhibit simultaneously the CD28 and ICOS pathways. It has been shown to have potent in vitro immunosuppressive activity and in vivo efficacy in models of disease for which implication of CD28 and ICOS has been reported (e.g. aGvHD, inflammatory arthritis, Sjögren’s, lupus, MS). Its safety, tolerability, and dose-dependent pharmacokinetics/dynamics are under study in a Ph1 healthy volunteer study. Here, we evaluate ALPN-101 in vitro using PBMC from Crohn’s and ulcerative colitis patients demonstrating superior suppression of T cell activation and cytokine release and show its efficacy to both prevent and treat disease in a mouse T cell transfer model of chronic colitis. Methods Primary cell assays were performed with PBMC stimulated with K562 cells (CD80+, CD86+, ICOSL+, anti-CD3 (OKT3) +) to evaluate suppression of cytokine release and compare to single pathway inhibition. ALPN-101 was assessed in the CD4+CD45RBhigh T cell-induced colitis model either singly dosed on Day 0 or 14 or repeat dosed 2x/week starting at Day 0 or 14 through Day 41, respectively. Serum cytokine and flow analysis of blood was performed throughout the study. Clinical presence of colitis was assessed using a disease activity index based on weight loss and stool consistency. At end of study, colons were measured and assessed histologically. Results ALPN-101 suppressed cytokine release (IFNγ, IL-2) from healthy or IBD patient PBMCs superior to single pathway inhibitors. In vivo, preventively or therapeutically, a single dose of ALPN-101 was efficacious to significantly improve multiple colitis readouts. Repeat dosing completely prevented onset of colitis. ALPN-101-treated mice gained weight and had colon weight-to-length ratios similar to the no-colitis cohort and demonstrated significant suppression of T cells and pro-inflammatory cytokines (e.g. TNFα, IL-12/23, IL-6). Conclusion Dual pathway inhibitor ALPN-101 is superior to single pathway inhibition in human in vitro and mouse in vivo translational studies and may be a novel therapeutic candidate for the treatment of IBD. Clinical trials for ALPN-101 in multiple inflammatory diseases are planned and underway. Consistent with clinical findings, histological analysis confirms efficacy of ALPN-101 in reducing colitis. All ALPN-101 treated groups had lower histological scores compared to the Fc control treated group (B). Mice treated from Day 0 to the end of the study had no pathological colon findings (C), 4/12 mice treated from Day 14 to end of study had no signs of colitis (D). None of the mice in these groups had their muscularis layer of colon affected by inflammation. Mice treated with a single dose at day 0 or day 14 had milder colitis than Fc control-treated mice, although the differences were not statistically significant.

Gold Nanoparticles inhibits cytokine release and alleviates sepsis through of MAPK and NFκB activation

Gold Nanoparticles inhibits cytokine release and alleviates sepsis through of MAPK and NFκB activation

Macrolides and COVID-19: An optimum premise

The epidemic of coronavirus infection disease 19 (COVID-19), which started in Wuhan City, is caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2) which binds angiotensin-converting enzyme 2 (ACE2) receptor, which is highly expressed by the lung epithelial cells. In COVID-19-induced acute respiratory distress syndrome, a hyperinflammatory syndrome with hypercytokinemia leads to acute lung injury and the development of respiratory failure. Macrolides are broad-spectrum, bacteriostatic antibiotics with significant anti-inflammatory and immunomodulatory effects. Different preclinical and clinical studies have shown that macrolides inhibit cytokine release, attenuate the inflammatory response, and improve immunoglobulin response. Azithromycin potentiates the anti-SARS-CoV-2 activity of chloroquine in COVID-19. However, azithromycin alone is effective initially in the management of COVID-19 due to its antiviral and anti-inflammatory activity. The antiviral potential of azithromycin is linked to different mechanisms, including modulation of lysosomal activity and the interaction points between SARS-CoV-2 and ACE2 receptor. Therefore, macrolides, chiefly azithromycin, are an effective drug against COVID-19 through direct antiviral effect or via the modulation of hyperinflammatory status.

JAK2/STAT1-mediated HMGB1 translocation increases inflammation and cell death in a ventilator-induced lung injury model

Janus kinase 2/signal transducer and activators of transcription 1 (JAK2/STAT1) signaling is a common pathway that contributes to numerous inflammatory disorders, including different forms of acute lung injury (ALI). However, the role of JAK2/STAT1 in ventilator-induced lung injury (VILI) and its underlying mechanism remain unclear. In this study, using lipopolysaccharide (LPS) inhalation plus mechanical ventilation as VILI mouse model, we found that the administration of JAK2 inhibitor AZD1480 markedly attenuated lung destruction, diminished protein leakage, and inhibited cytokine release. In addition, when mouse macrophage-like RAW 264.7 cells were exposed to LPS and cyclic stretch (CS), AZD1480 prevented cell autophagy, reduced apoptosis, and suppressed lactate dehydrogenase release by downregulating JAK2/STAT1 phosphorylation levels and inducing HMGB1 translocation from the nucleus to the cytoplasm. Furthermore, HMGB1 and STAT1 knockdown attenuated LPS+CS-induced autophagy and apoptosis in RAW 264.7 cells. In conclusion, these findings reveal the connection between the JAK2/STAT1 pathway and HMGB1 translocation in mediating lung inflammation and cell death in VILI, suggesting that these molecules may serve as novel therapeutic targets for VILI.

Bioactive Plant Compounds in Coffee Charcoal (Coffeae carbo) Extract Inhibit Cytokine Release from Activated Human THP-1 Macrophages.

The herbal preparation coffee charcoal is produced by over-roasting and milling green dried Coffea arabica L. seeds, and has a long-standing tradition in the treatment of inflammatory and gastrointestinal disorders. Its therapeutic properties are commonly attributed to adsorptive and astringent effects. This insufficiently explains its mode of action, especially when used in the treatment of inflammatory diseases in lower dosages. Our investigations aimed to identify bioactive secondary plant metabolites affecting cytokine-signaling. Thus, a phytochemical analysis of coffee charcoal extract was conducted using HPLC and LC/MS. Trigonelline, neochlorogenic acid, chlorogenic acid, caffeine, cryptochlorogenic acid, feruloylquinic acid isomers, and a caffeoylquinolacton were identified in the extract. Subsequently, the effects of coffee charcoal extract, chlorogenic acid isomers, their metabolite caffeic acid, caffeine, and trigonelline on cytokine (TNF, IL-6, MCP-1) release from LPS-challenged human THP-1 macrophages were examined to evaluate anti-inflammatory activity. Coffee charcoal showed concentration-dependent mild-to-medium inhibitory effects. The chlorogenic acid isomers and caffeic acid inhibited the TNF release, with cryptochlorogenic acid exerting the most distinct effects, as well as decreasing the release of IL-6 and MCP-1. In addition, scanning electron microscopic images provided an impression of the particle constitution, indicating a larger particle size and less structured surface of coffee charcoal in comparison to activated charcoal. In conclusion, our findings underline that beyond adsorptive effects, coffee charcoal exhibits pharmacological properties, which derive from a spectrum of secondary plant metabolites and support the therapeutic use in inflammatory diseases. Chlorogenic acids, particularly cryptochlorogenic acid, appear as pivotal bioactive compounds.

4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to exert anti-inflammatory effects

Activated macrophages switch from oxidative phosphorylation to aerobic glycolysis, similar to the Warburg effect, presenting a potential therapeutic target in inflammatory disease. The endogenous metabolite itaconate has been reported to regulate macrophage function, but its precise mechanism is not clear. Here, we show that 4-octyl itaconate (4-OI, a cell-permeable itaconate derivative) directly alkylates cysteine residue 22 on the glycolytic enzyme GAPDH and decreases its enzyme activity. Glycolytic flux analysis by U13C glucose tracing provides evidence that 4-OI blocks glycolytic flux at GAPDH. 4-OI thereby downregulates aerobic glycolysis in activated macrophages, which is required for its anti-inflammatory effects. The anti-inflammatory effects of 4-OI are replicated by heptelidic acid, 2-DG and reversed by increasing wild-type (but not C22A mutant) GAPDH expression. 4-OI protects against lipopolysaccharide-induced lethality in vivo and inhibits cytokine release. These findings show that 4-OI has anti-inflammatory effects by targeting GAPDH to decrease aerobic glycolysis in macrophages.

Green Tea Polyphenol Induced Mg2+-rich Multilayer Conversion Coating: Toward Enhanced Corrosion Resistance and Promoted in Situ Endothelialization of AZ31 for Potential Cardiovascular Applications.

As a promising biodegradable metallic material, magnesium (Mg) and its alloys have attracted special attention in the recent decade. However, challenges still remain due to its high corrosion rate and insufficient biocompatibility after implantation. In this work, we prepare a simple and versatile green tea phenol-metal induced multilayer conversion coating (Mg2+ incorporated epigallocatechin gallate (EGCG) coating) on magnesium alloys' (AZ31) substrate by layer-by-layer (LBL) method. The surface morphology results revealed that, with the incorporation of Mg2+, the as-formed EGCG/Mg coating was rich in phenol-Mg complex and presented more homogeneous and dense morphology, with far less cracks than the pure EGCG coating. The in vitro degradation rate and corrosion resistance were studied by electrochemical corrosion tests and monitoring of the changed pH value and hydrogen evolution, respectively, which revealed that the corrosion rate was effectively decreased compared to that of bare AZ31 after it was protected by EGCG/Mg coating. In vitro and ex vivo thrombogenicity test demonstrated the EGCG/Mg coatings presented an impressive improvement in decreasing the adhesion and activation of platelets and erythrocytes, in activated partial thromboplastin time (APTT), and in antithrombogenicity compared to those of bare AZ31. Owing to the mild degradation rate, in combination with the biological function of EGCG, enhanced endothelial cells' (ECs') adhesion and proliferation, suppressed smooth muscle cells' (SMCs') adhesion/proliferation, and inhibited cytokine release were observed on EGCG/Mg coated AZ31 alloy. Besides, the in vivo subcutaneous embedding experiment suggested that the EGCG/Mg coating performed more mild tissue response due to the improved corrosion resistance to the surrounding microenvironment. Moreover, for in vivo abdominal aorta assay, the EGCG/Mg coated AZ31 wire presented better corrosion resistance and enhanced re-endothelialization compared to bare AZ31 wire. These results suggested the potential of using green tea polyphenol induced Mg2+-rich multilayer conversion coating for enhanced corrosion protection and desired biocompatibility of biodegradable cardiovascular implants.