Inflammatory Cytokine Gene Research Articles

Cultured fibroblasts of the Okinawa rail present delayed innate immune response compared to that of chicken.

The Okinawa rail is endemic to Okinawa Island and is categorized as an endangered animal. In this study, we focused on innate immunity because it is the first line of host defense. In particular, signals recognizing foreign RNA (e.g., viruses) are important for host defense because they activate the host immune system. The retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) families (RIG-I, MDA5, and LGP2) are sensors that activate innate immunity. Therefore, we analyzed these functions in the Okinawa rail using genomic and cellular analyses of fibroblasts. Fibroblasts can be obtained from dead individuals, allowing these cells to be obtained from dead individuals, which is particularly useful for endangered species. The MDA5 gene of Okinawa rail was sequenced using the Sanger method following PCR amplification and extraction of the amplified sequence from agarose gel. Additionally, mRNA expression analysis of cultured fibroblasts exposed to poly I:C was done. The MDA5 gene was found to be a mutated nonfunctional gene in the Okinawa rail. The mRNA expression rates of inflammatory cytokine genes type I IFN, and Mx1 were slower in Okinawa rail than in chicken cultured fibroblasts. Similar to the mRNA expression results, cell number and live cell ratio also slowly decreased in the Okinawa rail compared with chicken cultured fibroblasts, indicating that the innate immune reaction differs between chicken and the Okinawa rail. To the best of our knowledge, this is the first experimental evaluation of the loss of function of the Okinawa rail innate immune genes. In conclusion, our results provide a basis for conservation strategies for the endangered Okinawa rail.

Iridoid from Eucommia ulmoides Oliv. Exerts Antiarthritis Effects by Inhibiting the JAK2/STAT3 Signaling Pathway In Vivo and In Vitro.

The purpose of this study was to investigate the anti-inflammatory effects of EU-Idd both in vivo and in vitro. In vivo, we used the collagen-induced arthritis (CIA) rat model to investigate the efficacy of EU-Idd on rheumatoid arthritis. Hematoxylin-eosin staining and Safranin O-fast green staining were used to evaluate the pathological status of the ankle joints in CIA rats. Micro-CT scanning was used to investigate bone erosion of the ankle joints. In vitro, the effect of EU-Idd on Th17 cell differentiation was identified by flow cytometry. TRAP staining was used to detect osteoclast cells. HFLS-RA model cells, induced by tumor necrosis factor-α(TNF-α), were used to evaluate the anti-inflammatory effects of EU-Idd while the levels of related inflammatory cytokines and JAK2/STAT3 proteins were detected by RT-qPCR and western blotting. EU-Idd alleviated joint inflammation in CIA rats and exerted protective effects on the ankle joints. EU-Idd also prevented the differentiation of CD4+ T cells into Th17 cells, reduced the number of osteoclasts, and improved the expression levels of bone metabolism-related proteins including OPG and RANKL. Moreover, EU-Idd inhibited the invasion and migration of HFLS-RA cells and downregulated the expression of related inflammatory cytokine genes and the protein expression levels of p-JAK2 and p-STAT3, both in vivo and in vitro. EU-Idd exerts anti-inflammatory and osteoprotective effects by regulating the JAK2/STAT3 pathway in rheumatoid arthritis. These results are beneficial to excavate new pharmaceutical ingredients for rheumatoid arthritis from iridoid.

Генетический полиморфизм воспалительных цитокинов в крови пациентов с инвазией Opisthorchis felineus

Aim: To research the relationship between laboratory and instrumental manifestations of pathology and polymorphisms of inflammatory cytokine genes in patients with Opisthorchis felineus invasion. Design. During 2022, we conducted a comparative study of patients who were hospitalized for etiotropic therapy of O. felineus invasion. Materials and methods. 139 patients with O. felineus invasion were examined. All patients underwent clinical and biochemical blood tests, ultrasound of the abdominal organs and liver elastometry with an assessment of liver fibrosis according to the METAVIR scale. Genotyping of single nucleotide polymorphisms of the IL28b, IL6, TNFA, and IFNG genes was performed using real-time PCR. Results. In patients with opisthorchiasis, liver fibrosis was associated with the presence of the minor allele A of the rs1800630 TNFA polymorphism and the TC genotype of rs2069705 IFNG. Polyps in the gallbladder were associated with the CC genotype of rs1800630 TNFA, while an increase in alkaline phosphatase and an increase in the proportion of eosinophils in the blood were associated with the CC genotype of rs2069705 IFNG. We did not find differences in the studied parameters depending on the genetic polymorphisms rs12979860 IL28b and rs1800795 IL6 in patients with O. felineus invasion. Conclusion. The obtained regularities should be used to increase the efficiency of dispensary observation in order to prevent the development of complications of parasitic invasion. Keywords: Opisthorchis felineus, single nucleotide polymorphism, cytokines, liver fibrosis.

Preventive effect of <i>Enterococcus faecalis</i> and β-glucan on the onset and exacerbation of symptoms of atopic dermatitis in mice

<abstract> <p>Atopic dermatitis (AD) is associated with an immune response caused by the excessive production of IgE antibodies. This is accompanied by excessive itching as a clinical symptom, leading to a great burden and anxiety in daily life. At present, there is no established treatment that can completely cure AD, and symptomatic treatment alleviates symptoms to the extent that they do not interfere with daily life. Recent research has suggested that the ingestion of either various lactic acid bacteria or β-glucan suppressed the production of IgE antibodies and alleviated the symptoms of AD. Therefore, in this study, we used a drug-induced AD mouse model to evaluate the amelioration effect of EC-12 and β-glucan derived from black yeast to investigate the mechanism involved. The oral administration of lactic acid bacteria inhibited the expression of inflammatory cytokine genes at the lesion site, and the administration of β-glucan downregulated serum IgE levels and inhibited the expression of inflammatory cytokine genes at the lesion site. The concomitant administration of EC-12 and β-glucan strongly suppressed AD symptoms, downregulated serum IgE levels, and inhibited the expression of inflammatory cytokines at the lesion site. In summary, the oral administration of EC-12 and β-glucan prevented AD in mice.</p> </abstract>

Glutathione Protects against Paraquat-Induced Oxidative Stress by Regulating Intestinal Barrier, Antioxidant Capacity, and CAR Signaling Pathway in Weaned Piglets.

Endogenous glutathione (GSH) effectively regulates redox homeostasis in the body. This study aimed to investigate the regulatory mechanism of different dietary levels of GSH supplementation on the intestinal barrier and antioxidant function in a paraquat-induced stress-weaned piglet model. Our results showed that dietary 0.06% GSH supplementation improved the growth performance of weaned piglets under normal and stressful conditions to some degree and decreased the diarrhea rate throughout. Exogenous GSH improved paraquat-induced changes in intestinal morphology, organelle, and permeability and reduced intestinal epithelial cell apoptosis. Moreover, GSH treatment alleviated intestinal oxidative stress damage by upregulating antioxidant (GPX4, CnZnSOD, GCLC, and GCLM) and anti-inflammatory (IL-10) gene expression and downregulating inflammatory cytokines (IFN-γ and IL-12) gene expression. Furthermore, GSH significantly reduced the expression levels of constitutive androstane receptor (CAR), RXRα, HSP90, PP2Ac, CYP2B22, and CYP3A29, and increased the expression levels of GSTA1 and GSTA2 in the jejunum and ileum of paraquat-induced piglets. We conclude that exogenous GSH protects against oxidative stress damage by regulating the intestinal barrier, antioxidant capacity, and CAR signaling pathway.

Polymorphism of the proinflammatory and anti–inflammatory cytokines genes in patients with an acute pancreatitis in the Northern–Eastern region of Ukraine

Objective.To determine the influence of the genes polymorphism of proinflammatory – interleukin–8 A–251T (rs 4073) and antiinflammatory – interleukin–10 C–592A (rs 1800872) cytokines on course of an acute pancreatitis.
 Materials and methods. The genes polymorphism of proinflammatory – interleukin–8 A–251T (rs 4073) and antiinflammatory – interleukin–10 C–592A (rs 1800872) cytokines were studied in 83 patients with an acute pancreatitis, using the method of polymerase chain reaction (PCR). The control group have consisted of 60 healthy persons without an acute pancreatitis in anamnesis.
 Results. The rate of the genes polymorphism of interleukin–8 A–251T (rs 4073) and interleukin–10 C–592A (rs 1800872) was determined in oedematious and destructive forms of an acute pancreatitis, as well as in patients operated on.
 Conclusion. The acute pancreatitis transition into necrotic form was not connected with the kind of the genes polymorphism of proinflammatory – interleukin–8 A–251T (rs 4073) and antiinflammatory – interleukin–10 C–592A (rs 1800872) cytokines. Operative interventions for an acute pancreatitis complications were performed statistically significantlymore frequently in carriers of the T/T gene genotype of interleukin–8 A–251T (rs 4073). The favorable, noncomplicated course of an acute pancreatitis was associated with polymorphism of the A/T gene interleukin–8 A–251T (rs 4073) and, possibly, – with polymorphism of the A/T gene interleukin–10 C–592A (rs 1800872).

Gene polymorphisms and risk of head and neck squamous cell carcinoma: a systematic review.

Exposure to the same environmental factors in different people have resulted in different susceptibility to head and neck squamous cell carcinoma (HNSCC), which suggests genetic variation may be a risk factor for the development of HNSCC. So, the aim was to review literatures on the association between gene polymorphisms and risk of HNSCCs. This systematic review included all articles on the impact of gene polymorphisms on risk and susceptibility to HNSCC published till September 2021 using PubMed, Web of science, SCOPUS, Google Scholar and Cochrane library databases. Of 1163 initial searched articles, 77 articles were eligible to include in this review. Studies were categorized based on gene functions. In each category, studied gene polymorphisms related to growth control genes, cell cycle control, apoptosis, DNA repair genes, carcinogen-metabolizing enzymes, alcohol-metabolizing genes, antioxidant gene, inflammatory cytokine, transcription factor, tumor immunity, folate metabolism, and tumor suppressor gene were discussed separately. Among the polymorphisms that are often significantly associated with HNSCC risk are: GSTM1 null, GSTT1 null, CYP2D6 *4, XRCC1 Arg194Trp and Arg399Gln, ERCC1 C8092A, XPD Lys751Gln, XRCC3 Thr241Met, P53 codon 72 and MTHFR C677T polymorphisms. Varied and contradictory results have been reported in different studies regarding the association of gene polymorphisms with HNSCC risk. To conclude about this association and to overcome these contradictions, it is necessary to use the results of existing meta-analyses or to perform new or updated meta-analyses.

6‐Gingerdione Reduces Apoptotic Conditions in HepG2 Cells and Inhibits Inflammatory Cytokine Gene Expression in Alcoholic Liver Injured Zebrafish Larvae

Antioxidant natural products and their analogs especially phenolic compounds, exhibit diverse biological properties, including anti-inflammatory, antioxidant, and anticancer activities. Ginger which is widely used worldwide for various beneficial effects also contains several phenolic antioxidants, and 6-gingerol is one of the natural products studied extensively. However, the molecular mechanism of synthetically synthesized 6-gingerdione (compound 1) from 6-gingerol was not known. In this study, compound 1 and methylated 6-gingerdione (compound 2) were obtained semi synthetically from 6-gingerol. Compound 1 and 2 are subjected to SwissADME prediction. Then the protective effect of compound 1 was analyzed in 2 % EtOH induced HepG2 cells and zebrafish larvae. Hydroxyl and nitric oxide scavenging assays reveal that compound 1 showed more antioxidant activity than compound 2 at 50 μM. Moreover, compound 1 exhibited good anti-inflammatory activity via lipoxygenase inhibition and proteinase inhibition. Apoptosis and oxidative stress in HepG2 cells were induced by 2 % EtOH and treated with compound 1. Compound 1 significantly inhibited the EtOH induced nitric oxide production, apoptosis, and ROS generation in HepG2 cells. Encouraged by the in-vitro antioxidant and anti-inflammatory activities, compound 1 was then investigated for its protective effect in 2 % EtOH induced ALD zebrafish larva. Compound 1 protected the zebrafish larvae from liver injury by suppressing inflammatory (COX-2, TNF-α, and IL-1β) and lipogenic genes (C/EBP-α, SREBP1, and IL-1β) while upregulating the antioxidant gene. Our findings indicate that compound 1 synthesized from 6-gingerol ameliorated liver injury that likely, contributes to its potential antioxidant and anti-inflammatory properties.

Gallic acid inhibits neuroinflammation and reduces neonatal hypoxic-ischemic brain damages.

Neuroinflammation is a leading cause of secondary neuronal injury in neonatal hypoxic-ischemic encephalopathy (HIE). Regulation of neuroinflammation may be beneficial for treatment of HIE and its secondary complications. Gallic acid (GA) has been shown to have anti-inflammatory and antioxidant effects. In this report we found that oxygen-glucose deprivation and/reoxygenation (OGD/R)-induced cell death, and the generation of excessive reactive oxygen species (ROS) and inflammatory cytokines by microglia were inhibited by GA treatment. Furthermore, GA treatment reduced neuroinflammation and neuronal loss, and alleviated motor and cognitive impairments in rats with hypoxic-ischemic brain damage (HIBD). Together, our results reveal that GA is an effective regulator of neuroinflammation and has potential as a pharmaceutical intervention for HIE therapy.

Chronic stress but not acute stress decreases the seizure threshold in PTZ-induced seizure in mice: role of inflammatory response and oxidative stress.

Seizure is paroxysmal abnormal electrical discharges in the cerebral cortex. Inflammatory pathways and oxidative stress are involved in the pathophysiology of seizures. Stress can induce an oxidative stress state and increase the production of inflammatory mediators in the brain. We investigated the effects of acute and chronic stresses on the seizure threshold in pentylenetetrazol (PTZ)-induced seizures in mice, considering oxidative stress and inflammatory mediators in the prefrontal cortex. In this study, 30 male Naval Medical Research Institute (NMRI) mice were divided into 3 groups, including acute stress, chronic stress, and control groups. PTZ was used for the induction of seizures. The gene expression of inflammatory markers (IL-1β, TNF-α, NLRP3, and iNOS), malondialdehyde (MDA) level, nitrite level, and total antioxidant capacity (TAC) were assessed in the prefrontal cortex and serum. Our results showed that stress could increase the expression of inflammatory cytokines genes and oxidative stress in the prefrontal cortex of the brain and serum following PTZ-induced seizures, which is associated with increased seizure sensitivity and decreased the seizure threshold. The effects of chronic stress were much more significant than acute stress. We concluded that the effects of chronic stress on seizure sensitivity and enhancement of neuroinflammation and oxidative stress are much greater than acute stress.

Identification and Characterization of Cell Lines HepG2, Hep3B217 and SNU387 as Models for Porcine Epidemic Diarrhea Coronavirus Infection.

Porcine epidemic diarrhea virus (PEDV), a member of the genera alphacoronavirus, causes acute watery diarrhea and dehydration in suckling piglets and results in enormous economic losses in the swine industry worldwide. Identification and characterization of different cell lines are not only invaluable for PEDV entry and replication studies but also important for the development of various types of biological pharmaceuticals against PEDV. In this study, we present an approach to identify suitable permissive cell lines for PEDV research. Human cell lines were screened for a high correlation coefficient with the established PEDV infection model Huh7 based on RNA-seq data from the Cancer Cell Line Encyclopedia (CCLE). Experimentally testing permissiveness towards PEDV infection, three highly permissive human cell lines, HepG2, Hep3B217, and SNU387 were identified. The replication kinetics of PEDV in HepG2, Hep3B217, and SNU387 cells were similar to that in Vero and Huh7 cells. Additionally, the transcriptomes analysis showed robust induction of transcripts associated with the innate immune in response to PEDV infection in all three cell lines, including hundreds of inflammatory cytokine and interferon genes. Moreover, the expression of inflammatory cytokines and interferons were confirmed by qPCR assay. Our findings indicate that HepG2, Hep3B217, and SNU387 are suitable cell lines for PEDV replication and innate immune response studies.

Abstract 15339: Er-Resident S1pr1 in Lung Endothelium Regulates Innate Immune Function and Mediates Inflammatory Lung Injury

Endothelial-cell surface localized sphingosine 1 phosphate receptor 1 (S1PR1) is known to promote anti-inflammatory and barrier enhancing niche upon ligating S1P. Recently we showed that S1P and TNFα, later being a well-known inflammatory agonist, phosphorylate S1PR1 at tyrosine 143 (Y 143 ) which functions as an endoplasmic reticulum (ER) import signal (Anwar et al, 2021). ER-retained S1PR1 instructs barrier disruptive signaling but the mechanism remains unclear. Here, we generated S1PR1 knock-in mice using CRISPR-Cas9 strategy to edit endogenous S1PR1 into Y 143 D-S1PR1 (phospho mimicking) or Y 143 F-S1PR1 (phosphodefective) to test the hypothesis that ER-localized S1PR1 subverts EC from anti-inflammatory to pro-inflammatory EC leading to vascular injury. Because EC constitutes about 50% of cells in the lungs, we assessed if knock-in of Y 143 D-S1PR1 impaired lung homeostasis. We show that editing of S1PR1 into Y 143 D- or Y 143 F-S1PR1 did not alter total S1PR1 expression. Interestingly, Y 143 D-S1PR1 knock-in mice showed marked vascular leak at homeostasis along with increased neutrophil influx and inflammatory cytokine generation including TNFα, IL1ß and MiP2 as compared to Y 143 F-S1PR1 or WT mice. We next challenge these mice with intratracheal LPS. LPS-induced non-resolvable vascular inflammatory injury in Y 143 D-S1PR1 mice. Surprisingly, Y 143 F-S1PR1 knock-in mice did not develop vascular inflammatory injury. Furthermore, NFκB activity, a predominant transcription factor inducing inflammatory EC phenotype, was increased in EC transducing Y 143 D-S1PR1 mutant as compared to WT. However, TNFα failed to induce NFkB activity in EC transducing Y 143 F-S1PR1 mutant. Together, these results show that ER-resident S1PR1 program endothelial niche into immune-active niche by activating NFkB pathway leading to irreparable lung injury. Further experiments are being done to assess epigenetic changes (ATACseq and ChIP-Seq) in EC to address the concept the ER-resident S1PR1 controls the fate of immune cells in the lungs. We believe that understanding how ER-resident S1PR1 programs EC into inflammatory phenotype would allow development of new targets for treating the inflammatory vascular diseases including lung injury, ARDS, and COVID-19

Transplantation of human adipose-derived stem cells overexpressing LIF/IFN-β promotes recovery in experimental autoimmune encephalomyelitis (EAE)

Multiple Sclerosis (MS) is the most common demyelinating disease with inflammatory demyelination in the central nerve system. Besides the defect in the myelin repair process, the balance change in inflammatory and anti- inflammatory cytokines is one of the most significant factors in MS pathogenesis. This study aimed at evaluating the effects of co-overexpressing beta interferon (IFN-β) and Leukemia inhibitory factor (LIF) in human adipose-derived stem cells (IFN-β/LIF-hADSCs) on the experimental autoimmune encephalomyelitis (EAE). 12 days after the induction of EAE on female mice C57Bl/6 with MOG35-55 and the emergence of primary clinical signs, the IFN-β/LIF-hADSCs were injected into the mice tail vein of the EAE mice. The mice were sacrificed after 32 days and the spinal cords of the experimental groups were dissected out for the histopathologic and real-time RT-PCR studies. Here, we showed that the clinical scores and infiltration of mononuclear cells of treated mice with IFN-β/LIF-hADSCs were decreased significantly. Demyelination and the number of Olig2+ and MBP+ cells were significantly increased in the test (IFN-β/LIF-hADSCs) group. The findings revealed that the pattern of inflammatory and anti- inflammatory cytokines gene expression in the IFN-β/LIF-hADSCs group was reversed compared to the control group. Overexpression of LIF as a neurotrophic and IFN-β as an anti-inflammatory cytokine in hADSCs increases the immunomodulatory effect of hADSCs reduces the extent of demyelination, improves the number of Olig2+ cells, and also increases the amount of MBP protein which can increase the production of myelin in EAE model. This, besides hADSCs capacity for proliferation and differentiation, might enhance the treatment efficacy and provide a promising candidate for stem cell-based gene therapy of MS therapy in the future.

Analysis of local expression of inflammatory cytokines genes in patients with gastroesophageal reflux disease and its association with types and number of gastroesophageal refluxes

The aim of the study was to evaluate gene expression of proinflammatory cytokines in esophageal mucosa in patients with erosive esophagitis (EE) and non-erosive form (NERD) of gastroesophageal reflux disease (GERD) and perform correlation analysis between the genes expression and types and number of gastroesophageal refluxes. Methods. This was a single-center prospective study. Esophageal mucosa samples were taken from lower part of the esophagus during endoscopy. Expression of IL-1b, IL-10, IL-18, TNFA, TLR4, GATA3, CD68 and beta-2 macroglobulin genes was assessed with ImmunoQuantex assays. Multichannel intraluminal esophageal pH-impedance studies were performed with Ohmega recorder and 2pH-6impedance catheters. Results. Data of 60 patients (females - 34; mean age (M±SD) 54.6 years, 20 - with erosive esophagitis) were available for the analysis. In those with EE, there was higher expression of IL-18 (5.89±0.4 vs 5.28±1.1, p=0.04), GATA3 (2.92±0.86 vs 2.23±0.96, p=0.03) genes, and inflammatory index (92.12±32.41% vs 70.1±30.8%, p=0.02) compared to NERD. Direct correlation was found between acid exposure time and expression of IL-1b (Spearman rank R = 0.29), IL-18 (R=0.31), TNFA (R=0.35), GATA3 (R=0.34), TLR4 (R=0.29), CD68 (R=0.37) genes and inflammatory index (R=0.3). Indirect correlation was revealed between mean рН and expressions of IL-18 (R= -0.28), TNFA (R= -0.33), GATA3 (R= -0.28), TLR4 (R= -0.28), CD68 (R= -0.39) genes and inflammatory index (R= -0.17). No correlation was found between the genes’ expression and number of gastroesophageal refluxes. Conclusions: Expression of proinflammatory cytokines’ genes differ in patients with EE and NERD. There is a correlation between expression of studied genes and esophageal acidity.

The role of inflammation and innate immune cells in young myocardial infarction patients without traditional risk factors

Abstract Background There is a group of patients that develops myocardial infarction (MI) in the absence of standard modifiable risk factors (SMuRFs) and at a young age. Identifying the mechanism underlying the development of atherosclerosis in these patients is paramount in order to develop adequate treatment strategies. Atherosclerosis is characterized as an inflammatory disease in which monocytes play a pivotal role. Circulating monocytes of atherosclerosis patients with SMuRFs have a pro-inflammatory phenotype. At present, the contribution of systemic inflammation, or innate immune cell activation in particular, in young SMuRFless MI patients is unknown. Purpose The aim of this study was to characterize the inflammatory profile of young SMuRFless MI patients, with an in-depth focus on monocyte phenotype, function and their epigenetic profiles. Methods We recruited 20 SMuRFless patients who suffered from a recent (1–4 year ago) MI and were <50 years old at the time. These patients were age- and sex matched to 18 healthy controls, who did not have atherosclerosis, as determined by coronary CT-scan. Blood samples were obtained after evening out any potential immunomodulating effect of statins and aspirin by interrupting statins for two weeks in patients and starting aspirin therapy for two weeks in controls. In both groups we studied circulating inflammatory markers with proximity extension assay, circulating monocyte (subset) phenotype with flow cytometry, and cytokine production of isolated peripheral blood mononuclear cells (PBMC) after stimulation with Toll-Like Receptor-agonists. Histone marks of chromatin accessibility were assessed on the promotor regions of several inflammatory cytokines in unstimulated isolated monocytes by chromatin immunoprecipitation (ChIP). Results PBMCs from patients showed elevated cytokine production after ex vivo stimulation compared to controls (P=0.046) (Figure 1). ChIP analysis of unstimulated monocytes revealed an overall enrichment of transcriptionally permissive tri-methylation of lysine 4 on histone H3 (H3K4me3) on the promoter regions of cytokines in patients compared to controls (Figure 2). Levels of H3K4me3 on IL-10 were significantly higher in patients (p=0.04). For TNF-α and IL-6 H3K4me3 levels did not reach statistical significance (p=0.07 and p=0.13 respectively). Despite the functional hyperresponsiveness, no differences were seen in monocyte phenotype on flow cytometry analysis (not shown). Additionally there were no differences in circulating high-sensitive C-reactive protein or other circulating inflammatory markers (not shown). Conclusions Circulating monocytes in SMuRFless MI patients are characterized by pro-inflammatory behavior, which is mediated by transcriptionally permissive epigenetic modifications at the promoters of key inflammatory cytokine genes. These findings are present in the absence of active systemic inflammation. Funding Acknowledgement Type of funding sources: None.

NOD1, NOD2, and NLRC5 Receptors in Antiviral and Antimycobacterial Immunity.

The innate immune system recognizes pathogen-associated molecular motifs through pattern recognition receptors (PRRs) that induce inflammasome assembly in macrophages and trigger signal transduction pathways, thereby leading to the transcription of inflammatory cytokine genes. Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) represent a family of cytosolic PRRs involved in the detection of intracellular pathogens such as mycobacteria or viruses. In this review, we discuss the role of NOD1, NOD2, and NLRC5 receptors in regulating antiviral and antimycobacterial immune responses by providing insight into molecular mechanisms as well as their potential health and disease implications.

Design, synthesis, and structure-activity relationship of novel RIPK2 inhibitors

The NOD1/2 (nucleotide-binding oligomerization domain-containing protein 1/2) signaling pathways are involved in innate immune control and host defense. NOD dysfunction can result in a variety of autoimmune disorders. NOD-induced generation of inflammatory cytokines is mediated by receptor-interacting protein kinase 2 (RIPK2), which has been considered as a promising therapeutic target. Herein, we disclose the design, synthesis, and SAR study of a series of RIPK2 inhibitors. The lead compound 17 displayed a high affinity for RIPK2 (Kd = 5.9 nM) and was capable of inhibiting RIPK2 kinase function in an ADP-Glo assay. In vitro DMPK studies showed that compound 17 had good metabolic stability and no CYP inhibition. Compound 17 effectively suppressed inflammatory cytokine production in both cells and animal model.

Clinical relevance and therapeutic potential of IL-38 in immune and non-immune-related disorders

Interleukin-38 (IL-38) is the most recent member of the IL-1 family that acts as a natural inflammatory inhibitor by binding to cognate receptors, particularly the IL-36 receptor. In vitro, animal and human studies on autoimmune, metabolic, cardiovascular and allergic diseases, as well sepsis and respiratory viral infections, have shown that IL-38 exerts an anti-inflammatory activity by modulating the generation and function of inflammatory cytokines (e.g. IL-6, IL-8, IL-17 and IL-36) and regulating dendritic cells, M2 macrophages and regulatory T cells (Tregs). Accordingly, IL-38 may possess therapeutic potential for these types of diseases. IL-38 down-regulates CCR3+ eosinophil cells, CRTH2+ Th2 cells, Th17 cells, and innate lymphoid type 2 cells (ILC2), but up-regulates Tregs, and this has influenced the design of immunotherapeutic strategies based on regulatory cells/cytokines for allergic asthma in future studies. In auto-inflammatory diseases, IL-38 alleviates skin inflammation by regulating γδ T cells and limiting the production of IL-17. Due to its ability to suppress IL-1β, IL-6 and IL-36, this cytokine could reduce COVID-19 severity, and might be employed as a therapeutic tool. IL-38 may also influence host immunity and/or the components of the cancer microenvironment, and has been shown to improve the outcome of colorectal cancer, and may participate in tumour progression in lung cancer possibly by modulating CD8 tumour infiltrating T cells and PD-L1 expression. In this review, we first briefly present the biological and immunological functions of IL-38, and then discuss the important roles of IL-38 in various types of diseases, and finally highlight its use in therapeutic strategies.

A randomized, controlled trial on the effectiveness of a proprietary marine lipo-peptide formula vs omega-3 on cytokines profile, anxiety, and pain symptoms in patients with fibromyalgia

Objective: The aim of the present study in an RCT manner (physicians and patients) a novel lipo-peptide marine compound, LD-1227, on physical-, emotional- and functional-related symptomatic complaints in fibromyalgia patients as well as inflammatory cytokines profile and gene expression while using omega-3 as a control group.Methods: The following questionnaire-based or clinical evaluation-based parameters were evaluated: widespread pain index [WPI] patient global impression of change, total tender points [TTP], fibromyalgia impact questionnaire, Beck depression inventory, fatigue severity ratings, cognitive symptom severity, symptom severity score [SSS] and weekly pain intensity ratings. Additional biochemical and gene expression analysis of cytokines (IL6, TNF-α, IL-1β, MCP-1, IL-8, IL-13, IL-1α, and GM-CSF) was performed as well. Data were analyzed with either a paired t-test or the Wilcoxon signed rank test depending on the parametric or non-parametric distribution.Results: Comparing the data from before and after treatment for Group B indicated a statistically significant reduction (p=0.05) in TTP, WPI score, and SSS score. These data suggest a positive effect of a 3-month treatment with the LD-1227 but not omega-3 treatment on Fibromyalgia pain and related anxiety/depressive symptoms. Inspections of HRV and Cytokines found a statistically significant improvement after LD1227 treatment. Unlike the group supplemented with omega-3, the treatment with LD-1227 brought about a decrease in WPI and weekly pain intensity symptoms for the majority of participants. The pre-and post-treatment data for Group B indicated a statistically significant reduction (p=0.05) in TPC, WPI, and SSS scores. No adverse events were reported.Conclusion: These results provide the first indications that the LD-1227 treatment has a statistically significant effect on the recognized fibromyalgia diagnosis metrics of WPI, TTP, and SSS as well on inflammatory markers and parasympathetic balance.Keywords: fibromyalgia, marine compound, lipopeptide, cytokines, anxiety, pain, LD-1227.

Anti-inflammatory effects of neutral lipids, glycolipids, phospholipids from Halocynthia aurantium tunic by suppressing the activation of NF-κB and MAPKs in LPS-stimulated RAW264.7 macrophages.

Halocynthia aurantium is a marine organism that has been considered a promising source for bio-functional materials. Total lipids were extracted from H. aurantium tunic, and then they were separated into neutral lipids, glycolipids, and phospholipids. In the present study, fatty acid profiles of three lipids and their anti-inflammatory effects in RAW264.7 cells were investigated. Among the lipid classes, phospholipids showed the diversity of fatty acid constituents, compared with the glycolipids and neutral lipids. Three lipids contain different contents of fatty acids depending on the kinds of lipids. The most contents were saturated fatty acids (SFAs, 53-69% of the fatty acids) and monounsaturated fatty acids (MUFAs, 15-17% of fatty acids) and polyunsaturated fatty acids (PUFAs, 14-32% of fatty acids) are followed. H. aurantium lipids not only dose-dependently inhibited nitric oxide production but also reduced the expression of inflammatory cytokine genes such as TNF-α, IL-1β, and IL-6 in LPS-stimulated macrophages. It was also demonstrated that the expression of COX-2 was dose-dependently suppressed. Moreover, H. aurantium lipids decreased phosphorylation of NF-κB p-65, p38, ERK1/2, and JNK, suggesting that three lipids from H. aurantium tunic provide anti-inflammatory effects through NF-κB and MAPK signaling. These results indicate that H. aurantium is a potential source for anti-inflammation.