Immune-mediated Inflammatory Response Research Articles

Calprotectin blockade inhibits long-term vascular pathology following peritoneal dialysis-associated bacterial infection.

Bacterial infections and the concurrent inflammation have been associated with increased long-term cardiovascular (CV) risk. In patients receiving peritoneal dialysis (PD), bacterial peritonitis is a common occurrence, and each episode further increases late CV mortality risk. However, the underlying mechanism(s) remains to be elucidated before safe and efficient anti-inflammatory interventions can be developed. Damage-Associated Molecular Patterns (DAMPs) have been shown to contribute to the acute inflammatory response to infections, but a potential role for DAMPs in mediating long-term vascular inflammation and CV risk following infection resolution in PD, has not been investigated. We found that bacterial peritonitis in mice that resolved within 24h led to CV disease-promoting systemic and vascular immune-mediated inflammatory responses that were maintained up to 28 days. These included higher blood proportions of inflammatory leukocytes displaying increased adhesion molecule expression, higher plasma cytokines levels, and increased aortic inflammatory and atherosclerosis-associated gene expression. These effects were also observed in infected nephropathic mice and amplified in mice routinely exposed to PD fluids. A peritonitis episode resulted in elevated plasma levels of the DAMP Calprotectin, both in PD patients and mice, here the increase was maintained up to 28 days. In vitro, the ability of culture supernatants from infected cells to promote key inflammatory and atherosclerosis-associated cellular responses, such as monocyte chemotaxis, and foam cell formation, was Calprotectin-dependent. In vivo, Calprotectin blockade robustly inhibited the short and long-term peripheral and vascular consequences of peritonitis, thereby demonstrating that targeting of the DAMP Calprotectin is a promising therapeutic strategy to reduce the long-lasting vascular inflammatory aftermath of an infection, notably PD-associated peritonitis, ultimately lowering CV risk.

Gene expression analysis in recurrent benign paroxysmal positional vertigo: a preliminary study.

This study aimed to determine the pathophysiology of recurrent benign paroxysmal positional vertigo (BPPV) in young patients using gene expression profiling combined with bioinformatics analysis. Total RNA was extracted from the whole blood of four young patients with recurrent BPPV and four controls. The differentially expressed genes (DEGs) between the groups were screened using a microarray analysis based on the cutoff criteria of |log2 fold change| > 1 and an adjusted p-value of < 0.05. Functional enrichment analysis of DEGs was performed using Gene Ontology analysis, and the protein-protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of the Interacting Genes database. A total of 39 DEGs were detected between the BPPV and control samples, comprising 33 upregulated DEGs and six downregulated DEGs in the BPPV group. Functional enrichment analysis indicated that the upregulated DEGs were significantly enriched in terms related to metabolic processes and the immune system. Two main pathways were extracted from the PPI network: one was associated with oxidative phosphorylation and stress and the other with the adaptive immune system and extracellular matrix degradation. The findings of our bioinformatics analysis indicated that oxidative stress or extracellular matrix degradation due to immune-mediated inflammatory responses may contribute to the development of recurrent BPPV in young patients.

Study of NLRP3 Single Nucleotide Polymorphisms in Juvenile Systemic Lupus Erythematosus (JSLE)

ABSTRACT Juvenile systemic lupus erythematosus (JSLE) is a multifaceted multifactorial disorder with an unclear etiopathogenesis. Environmental factors, genetic factors, and dysregulated and defective immune system responses are known to have a role in JSLE etiopathogenesis. NLRP3 inflammasome, as an important contributor to immune-mediated inflammatory responses, is assumed to be involved in JSLE etiopathogenesis. To determine whether the NLRP3 genetic variants are altered in patients with JSLE. Fifty-three patients diagnosed with JSLE and 56 healthy sex-matched controls were studied. NLRP3 (C/G rs10754558, C/T rs3806265, C/T rs4612666, A/C rs35829419) gene polymorphisms were evaluated using a TaqMan single-nucleotide polymorphism assay. C allele at position rs3806265 was detected in higher frequencies in patients than in the control group (37.74% vs 23.21%, P-value = .028). At the genotype level at the same position, CT has a significantly higher frequency in patients than the healthy subjects (75.47% vs 46.43%, P-value = .003). The NLRP3 rs3806265 CT genotype was detected at a higher frequency in patients with JSLE than in the healthy control group.

All-Trans Retinoic Acid Promotes M2 Macrophage Polarization in Vitro by Activating the p38MAPK/STAT6 Signaling Pathway

ABSTRACT Background M2-type macrophages are inflammation-suppressing cells that are differentiated after induction by cytokines such as IL-4 or IL-13, which play an important regulatory role in inflammation and influence the regression of inflammation-related diseases. All-trans retinoic acid (ATRA) has an important role in suppressing immune-mediated inflammatory responses but the effect and underlying mechanism of ATRA on the polarization of M2 macrophages remains unclear. Methods Macrophages were isolated from peritoneal wash fluid, and IL-4 (20 ng/mL) was used to construct a m2-type macrophage polarization model. The model was incubated with different concentrations of ATRA (15 µg/ml, 30 µg/ml, 45 µg/ml) for 24 h, and pretreated macrophages with p38MAPKα inhibitor SB202190 (20 μM). MTT, Trypan blue staining, Annexin V-PE/7-AAD staining, flow cytometry, real-time PCR and western blotting were used to investigate the effect and mechanism of ATRA on the polarization of M2 macrophages. Results Compared with the IL-4 group, the proportion of F4/80+CD206+ M2-type macrophages was significantly higher in the ATRA group (P < 0.01). mRNA and protein expression levels of Arg-1, IL-10 and TGF-β1 were as significantly higher (P < 0.01) in the ATRA group as phosphorylation levels of STAT6 and p38MAPK (P < 0.01). After pretreatment with the addition of the inhibitor SB202190, M2-type macrophages proportion and their associated factors expression were significantly (P < 0.01) reduced, as compared with those in the ATRA group, but they were comparable (P > 0.05) with the IL-4 group. Conclusion The combination of ATRA and IL-4 activated the p38MAPK/STAT6-signaling pathway to promote polarization of M2 macrophages.

Exploring the shared molecular mechanism of microvascular and macrovascular complications in diabetes: Seeking the hub of circulatory system injury.

Microvascular complications, such as diabetic retinopathy (DR) and diabetic nephropathy (DN), and macrovascular complications, referring to atherosclerosis (AS), are the main complications of diabetes. Blindness or fatal microvascular diseases are considered to be identified earlier than fatal macrovascular complications. Exploring the intrinsic relationship between microvascular and macrovascular complications and the hub of pathogenesis is of vital importance for prolonging the life span of patients with diabetes and improving the quality of life. The expression profiles of GSE28829, GSE30529, GSE146615 and GSE134998 were downloaded from the Gene Expression Omnibus database, which contained 29 atherosclerotic plaque samples, including 16 AS samples and 13 normal controls; 22 renal glomeruli and tubules samples from diabetes nephropathy including 12 DN samples and 10 normal controls; 73 lymphoblastoid cell line samples, including 52 DR samples and 21 normal controls. The microarray datasets were consolidated and DEGs were acquired and further analyzed by bioinformatics techniques including GSEA analysis, GO-KEGG functional clustering by R (version 4.0.5), PPI analysis by Cytoscape (version 3.8.2) and String database, miRNA analysis by Diana database, and hub genes analysis by Metascape database. The drug sensitivity of characteristic DEGs was analyzed. A total of 3709, 4185 and 8086 DEGs were recognized in AS, DN, DR, respectively, with 1820, 1666, 888 upregulated and 1889, 2519, 7198 downregulated. GO and KEGG pathway analyses of DEGs and GSEA analysis of common differential genes demonstrated that these significant sites focused primarily on inflammation-oxidative stress and immune regulation pathways. PPI networks show the connection and regulation on top-250 significant sites of AS, DN, DR. MiRNA analysis explored the non-coding RNA upstream regulation network and significant pathway in AS, DN, DR. The joint analysis of multiple diseases shows the common influenced pathways of AS, DN, DR and explored the interaction between top-1000 DEGs at the same time. In the microvascular and macrovascular complications of diabetes, immune-mediated inflammatory response, chronic inflammation caused by endothelial cell activation and oxidative stress are the three links linking atherosclerosis, diabetes retinopathy and diabetes nephropathy together. Our study has clarified the intrinsic relationship and common tissue damage mechanism of microcirculation and circulatory system complications in diabetes, and explored the mechanism center of these two vascular complications. It has far-reaching clinical and social value for reducing the incidence of fatal events and early controlling the progress of disabling and fatal circulatory complications in diabetes.

Curcumin Alleviates Hepatic Ischemia-Reperfusion Injury by Inhibiting Neutrophil Extracellular Traps Formation

Background Hepatic ischemia-reperfusion injury (IRI) is a common innate immune-mediated sterile inflammatory response in liver transplantation and liver tumor resection. Neutrophil extracellular traps (NETs) can aggravate liver injury and activates innate immune response in the process of liver IRI. However, Curcumin (Cur) can reverse this damage and reduce NETs formation. Nevertheless, the specific regulatory mechanism is still unclear in liver IRI. This study aimed to explore the potential mechanisms that how does Cur alleviate hepatic IRI by inhibits NETs production and develop novel treatment regimens. Methods We established a hepatic IRI model by subjecting C57BL/6J mice to 60 min of ischemia, followed by reperfusion for 2 h, 6 h, 12 h, and 24 h respectively. Subsequently, we were separated into 5 groups, namely the I/R group, Cur group, DNase-1 group, Cur + DNase1 group and sham operation group. Serum alanine aminotransferase (ALT) and aspartate transaminase (AST), Hematoxylin-eosin staining, immunofluorescence, and TUNEL analysis were applied to assess liver injury degree and NETs levels. Western blot assay was used to detect the protein levels of apoptosis-related proteins and MEK pathway proteins. Results Cur could alleviate hepatic IRI by inhibiting the generation of NETs via suppressing the MEK/ERK pathway. In addition, this study also revealed that DNase-1 is vital for alleviating hepatic IRI by reducing the generation of NETs. Conclusions Cur combined with DNase-1 was more effective than the two drugs administered alone in alleviating hepatic IRI by inhibiting the generation of NETs. These results also suggested that curcumin combined with DNase-1 was a potential therapeutic strategy to mitigate hepatic IRI.

A Graves&amp;apos; Disease Patient Diagnosed as Autoimmune Gastritis with Helicobacter pylori Infection

Autoimmune gastritis (AIG) is a type of atrophic gastritis characterized by destruction of parietal cells in the gastric fundus and body. These changes may be attributable to immune-mediated chronic inflammatory responses. AIG is characterized by extensive atrophy of the gastric body; therefore, endoscopic findings offer useful diagnostic clues. AIG is diagnosed based on serological and histopathological evaluation of endoscopic biopsy specimens; however, this condition may be accompanied by autoimmune diseases including autoimmune thyroid disease (ATD), and the opposite can be suspected. Diagnostic delays and misdiagnosis are common in patients with AIG owing to the nonspecific clinical presentation and accompanying autoimmune diseases. Additionally, confirmation of AIG based on serological atrophy or endoscopic findings is challenging in cases of active &lt;i&gt;Helicobacter pylori&lt;/i&gt; (&lt;i&gt;H. pylori&lt;/i&gt;) infection. We report a case of Graves’ disease (an ATD) in a patient diagnosed with AIG and concomitant &lt;i&gt;H. pylori&lt;/i&gt;-induced gastritis based on the rapid urease and serological test results and endoscopic biopsy findings.

Perinatal Cerebral Ischemic Lesion and SARS-CoV-2 Infection during Pregnancy: A Case Report and a Review of the Literature.

Perinatal stroke is related to possible differences in predisposing factors and outcomes between acutely and retrospectively diagnosed cases. In most cases, there are different risk factors and infections that could play an important role. Thus far, different clinical manifestations have been reported in children presenting with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), ranging from asymptomatic status to severe disease sustained by an immune-mediated inflammatory response. SARS-CoV-2 has been associated with severe neurological diseases including seizures and encephalitis in both adults and children. However, there are still few reports regarding the possible relation between SARS-CoV-2 infection of mothers during pregnancy and the neurologic outcome of the newborns. We described the case of a newborn diagnosed with a perinatal stroke, born at 35 weeks of gestation from a mother presenting with SARS- CoV-2 infection during the last months of pregnancy. We also added a brief review of the literature with similar cases. Close monitoring and early intervention in young children born to infected mothers would be highly recommended for the potential neurodevelopmental risk.

CHANGESINIMMUNOLOGY STATUS IN PREGNANT WOMEN WITH EARLY GESTOSES

One of the frequent complications of early terms of pregnancy is vomiting in pregnant women, which causes have not been studied sufficiently. The signs of early gestosis usually appear from the 4-9th week of pregnancy and decrease up to the 16th week. At the moment, there is no tendency for early gestoses to decrease their frequency. In the last years, huge significance for early gestosis lies in the disordered immune system in a pregnant woman. On the 4-5th week of pregnancy, the mother’s body gets antigens from the fetus that contributes to early gestosis.&#x0D; Aim of the research. Study features of immune-mediated inflammatory disorders during the early gestoses of different severity degrees.&#x0D; Materials and methods. We examined 120 females. The control group consisted of 30 women with the physiologic course of pregnancy. The main group had 90 pregnant females with present early gestoses of different severity degrees: the 1st one had 30 pregnant patients with present vomiting of a mild degree, the 2nd one – 30 females with early gestoses of moderate degree, the 3rd one – 30 women with early gestoses with the severe course. Inclusion criteria to the main group were considered the age of the patients from 18 to 40 years, present vomiting, present intrauterine singleton pregnancy, patient’s consent to the research, pregnancy term of 4-12 weeks. Exclusion criteria were gastrointestinal and hepatic diseases of different etiology in anamnesis, the patients younger than 18 years old and older than 40 years old, present oncologic diseases, ectopic pregnancy, patients with surgical intervention on the pelvic organs in anamnesis, pregnant women with IVF, present multiple pregnancy, present neuropsychiatric pathology, absent patient’s consent to the research, pregnancy term more than 12 weeks. In blood serum, the examinees had tested cytokine level, which regulated immune-mediated inflammatory response (IL-10, IL-6) on the STAT FAX 303/PLUS Microstrip Reader in the private certified laboratory PrimaMED (Ivano-Frankivsk).&#x0D; Results of the research. We tested levels of pro- and anti-inflammatory cytokines, vascular endothelial growth factors in blood serum in pregnant women with present early gestosis dependently on severity degree. Pregnant females with vomiting of several degrees have distorted humoral immune responses. This is seen provably by misbalanced pro- and anti-inflammatory cytokine levels in blood serum. The pro- and anti-inflammatory cytokine levels in pregnant patients with early gestoses depend on the severity of early gestoses. In the group of pregnant women with the mild course, we observed the lowest average values of IL-6 and the highest IL-10 level in blood serum. The females with moderate degree presented a probable decrease of IL-10 levels in 7.3 % and a significant increase of IL-6 levels in 5.5 % compared to the indices in females of the 1st group. The clearest misbalance in pro- and anti-inflammatory cytokines was registered in pregnant women of the 3rd group (severe degree). The women from this group presented a probable decrease of IL-10 level in 22.0% and a significant increase of IL-6 level in 14.8% compared to the indices in females from the 2nd group.&#x0D; Conclusions:&#x0D; &#x0D; The signs of early gestosis are associated with induction of inflammation.&#x0D; There are close direct correlations between severity degrees of early gestosis and IL-10 levels and inverse correlations with IL-6 levels. In females with signs of severe early gestosis in blood serum, we recorded a probable decrease of IL-10 levels and a probable increase of IL-6 levels compared to the values in females with mild and moderate early gestosis.&#x0D;

What Do We Know About Shoulder Injury Related to Vaccine Administration? An Updated Systematic Review.

Shoulder injury related to vaccine administration (SIRVA) is postulated to be an immune-mediated inflammatory response to a vaccine antigen injected into or near the subacromial bursae or synovium, leading to shoulder pain and dysfunction. The number of studies on this topic is rapidly increasing. Recent comparative studies have reported conflicting conclusions, which suggests that a systematic review of the best-available evidence may be helpful. In this systematic review, we asked: What are the (1) clinical characteristics, (2) diagnoses, and (3) management approaches and outcomes reported in association with SIRVA? A search was performed on October 4, 2021, of the PubMed and Medline databases for studies related to SIRVA. Inclusion criteria were English-language comparative studies, case series, and case reports that involved shoulder pain occurring after vaccination. Studies of exclusively neurologic conditions after vaccination were excluded. Forty-two studies met the eligibility criteria, including three retrospective comparative studies (72 patients and 105 controls), five database case series (2273 patients), and 34 case reports (49 patients). Study quality was assessed for the database case series and retrospective comparative studies using the Methodological Index for Non-randomized Studies tool. Among patients in the case reports, the median age was 51 years (range 15-90 years), and 73% (36 of 49) were women. BMI was reported for 24% of patients (12 of 49) in case reports, with a median of 23.5 kg/m2 (range 21-37.2 kg/m2). The most common symptoms were shoulder pain and reduced ROM. The most common diagnoses were shoulder bursitis, adhesive capsulitis, and rotator cuff tears. The most frequent management modalities included physical or occupational therapy, NSAIDs, and steroid injections, followed by surgery, which was generally used for patients whose symptoms persisted despite nonsurgical management. Full resolution of symptoms was reported in 2.9% to 56% of patients. The association between inflammatory conditions of the shoulder (such as bursitis) and vaccination appears to be exceedingly rare, occurring after approximately 1:130,000 vaccination events according to the best-available comparative study. Currently, there is no confirmatory experimental evidence supporting the theory of an immune-mediated inflammatory response to vaccine antigens. Although the clinical evidence is limited, similar to any bursitis, typical treatments appear effective, and surgery should rarely be performed. Additional research is needed to determine the best injection technique or evaluate alternate injection sites such as the anterolateral thigh that do not involve positioning a needle close to the shoulder.

Immune-Mediated Inflammatory Responses of Alveolar Epithelial Cells: Implications for COVID-19 Lung Pathology.

Background. Clinical and experimental evidence point to a dysregulated immune response caused by SARS-CoV-2 as the primary mechanism of lung disease in COVID-19. However, the pathogenic mechanisms underlying COVID-19-associated ARDS (Acute Respiratory Distress Syndrome) remain incompletely understood. This study aims to explore the inflammatory responses of alveolar epithelial cells to either the spike S1 protein or to a mixture of cytokines secreted by S1-activated macrophages. Methods and Results. The exposure of alveolar A549 cells to supernatants from spike-activated macrophages caused a further release of inflammatory mediators, with IL-8 reaching massive concentrations. The investigation of the molecular pathways indicated that NF-kB is involved in the transcription of IP-10 and RANTES, while STATs drive the expression of all the cytokines/chemokines tested, with the exception of IL-8 which is regulated by AP-1. Cytokines/chemokines produced by spike-activated macrophages are also likely responsible for the observed dysfunction of barrier integrity in Human Alveolar Epithelial Lentivirus-immortalized cells (hAELVi), as demonstrated by an increased permeability of the monolayers to mannitol, a marked decrease of TEER and a disorganization of claudin-7 distribution. Conclusion. Upon exposure to supernatants from S1-activated macrophages, A549 cells act both as targets and sources of cytokines/chemokines, suggesting that alveolar epithelium along with activated macrophages may orchestrate lung inflammation and contribute to alveolar injury, a hallmark of ARDS.

Topical Cyclosporine-A for Management of Epiphora in Eyes with Acquired Punctal Stenosis

Background: Cyclosporine A (CsA) is an inhibitor of calcineurin that prevents T-cell production of inflammatory cytokines and disrupts the immune-mediated inflammatory response that plays a crucial mechanism in punctal stenosis. Purpose: To evaluate the clinical outcomes and tolerances of CsA in treating epiphora in eyes with punctal stenosis. Study Design: Prospective study. Methods: The study included patients who presented with symptomatic epiphora associated with lower punctal stenosis during the period between July 2019 and December 2020. Patients were treated with topical 0.05% CsA on twice daily dose with topical preservative-free artificial tears Q.I. D. Patients were followed up monthly for at least 3 months by Munk epiphora grading, Fluorescein dye disappearance test (FDT) and evaluation of the patient satisfaction. Results: A total of 26 patients with 47 eyes were included in the study with a mean age of 55.1 ± 10.24 years. All the eyes had grade 0 lower punctal stenosis with grade 4 on Munk grading and grade 3 in FDT. There was a statistically significant difference in the Munk grading and FDT along the follow-up period compared to the baseline values (p rd month. None of the patients encountered any complications during treatment. Conclusion: CsA can control the inflammation of the conjunctival sac and restore the integrity of the ocular surface with subsequent symptomatic relief of the epiphora in eyes with punctal stenosis.

Immunotherapy-Induced Acute Gastritis: First Case Report of Nivolumab Re-Challenge after Severe Immune-Mediated Inflammatory Response

Background: The use of Checkpoint Inhibitors (CPIs) exposes patients to CPI immune-related adverse events (irAEs) that affect a variety of body organs and systems. IrAEs of the upper gastrointestinal tract and gastritis are rare manifestations of CPI-related IrAEs, with only a few cases reported in the literature. Currently, there are no guidelines for the evaluation, treatment and re-challenge after CPI-mediated gastritis.

Incidence of Skeletal Muscle Injury with SARS-CoV-2 Infection in Tertiary Care Hospitals in Qatar

Background: SARS-CoV-2 can contribute to a number of neurological manifestations. 19.3% of patients with severe COVID-19 had evidence of marked skeletal muscle injury and 13.7% of patients with COVID-19 have high CK levels. Methodology: We retrospectively analyzed 413 COVID-19 patients. We reviewed electronic medical records and collected data. Skeletal muscle injury was defined as skeletal muscle pain and elevated serum creatine kinase level. The primary objective to determine the incidence of skeletal muscle injury in COVID-19 patients. Results: The most common risk factors were hypertension diabetes and obesity and neurological symptoms at onset of illness were myalgia. Multi-organ systems involvement including 47.8% had skeletal muscle injury, 15.7% had rhabdomyolysis, 36.7% had acute kidney injury, 27.5% had acute liver injury, 23.9% had acute myocardial injury. Discussion: In addition to respiratory system involvement, other extrapulmonary multiorgan systems involvement including skeletal muscle injury, acute kidney injury, acute liver injury, acute myocardial injury, rhabdomyolysis, acute heart failure, acute pancreatitis and post ICU critical illness neuromyopathy. SARSCoV- 2 may direct invade skeletal muscle, hematogenous dissemination, immunemediated skeletal muscles injury and inflammatory response with cytokine storm may lead to pathological changes in skeletal muscle tissue including muscle fiber proteolysis and fibrosis. Conclusion: Skeletal muscles injury with COVID-19 patient’s manifest as myalgia, generalized fatigue, myositis, rhabdomyolysis, dermatomyositis-like interferonopathy 1, immune-mediated necrotizing myopathy and respiratory failure. Further studies are requiring with focus on long term outcomes of skeletal muscles injury and benefits of using post COVID rehabilitation programs.

Leg paralysis after AstraZeneca COVID-19 vaccination diagnosed as neuralgic amyotrophy of the lumbosacral plexus: a case report

The ongoing global administration of vaccines for coronavirus disease 2019 (COVID-19) means that increasing numbers of patients are likely to present with post-vaccination complications. We describe the first reported case of neuralgic amyotrophy (NA) involving the lumbosacral plexus occurring after AstraZeneca COVID-19 vaccination. The patient presented with acute-onset leg paralysis following administration of the vaccine. Based on the clinical, electrodiagnostic, and radiologic findings, the patient was diagnosed with post-vaccination NA. We speculate that the COVID-19 vaccine elicited an immune-mediated inflammatory response to the injected antigen due to inflammatory immunity in a patient with predisposed susceptibility to NA.

Chia oil prevents chemical and immune-mediated inflammatory responses in mice: Evidence for the underlying mechanisms

Chia (Salvia hispanica L.) is an herbaceous plant used as omega-3 polyunsaturated fatty acid (ω-3 PUFA) source that presents a range of beneficial effects on human health. Herein, it was used a chia oil containing over than 62% of α-linolenic acid (ALA), a compound widely related to anti-inflammatory actions. Chia oil effect was tested using paw edema and mechanical hyperalgesia induced by carrageenan, and ear edema induced by croton oil, histamine, and capsaicin. Croton oil was used in both preventive and therapeutic treatment schedules of chia oil while histamine and capsaicin were used only in preventive treatment schedule. Chia oil mechanism of action was investigated using nociception and paw edema response induced by intraplantar injection of acidified saline (ASIC activator), PGE2 (prostaglandin pathway), cinnamaldehyde (TRPA1 activator), bradykinin (BK pathway), menthol (TRPM8 activator), and capsaicin (TRPV1 activator). Further, RT-PCR for inflammatory mediators (TRPA1, NF-κB, PPAR-γ, COX-2, IL-6, TNF, FPR2, FAAH, MAGL, and IL-12A) induced by carrageenan, NLRP3 inflammasome activation, and the cell viability were then accessed. Later, chia oil actions were evaluated in the experimental autoimmune encephalomyelitis (EAE), a multiple sclerosis (MS) model. Chia oil showed anti-edematogenic and anti-hyperalgesic effects when administered 1 h before pro-inflammatory stimulus – particularly carrageenan and croton oil. Moreover, chia oil upregulated the mRNA levels of COX-2 and formyl peptide receptor 2 (FPR2) while reduced IL-6 expression in the spinal cord of mice submitted to i.pl. injection of carrageenan. Interestingly, chia oil mediates antinociceptive effects in mice decreasing the nociceptive response induced by acidified saline, PGE2, and cinnamaldehyde, but not by bradykinin, menthol, and capsaicin. On the EAE model, chia oil preventively administered attenuated EAE-induced motor deficits and mechanical hyperalgesia in mice, suggesting a valuable effect of chia oil supplementation in regulating inflammatory responses and some immune functions during immune-mediated inflammatory disorders (IMID). Nonetheless, additional reports will need to assess the effect of chia oil in well-controlled clinical trials performed in MS patients.

Antimicrobial REG3A and Network Inflammatory Proteins Reflect Stroke Severity and Functional Impairment During Ischemic Stroke

Introduction Regenerating Family Member 3 Alpha (REG3A) is an antimicrobial protein secreted by the intestine and pancreas and is involved in immune-mediated inflammatory responses. In healthy individuals, REG3A is minimally expressed. However, acute-phase inflammatory mediators such as IL6 upregulate REG3A during inflammation, trauma, and cell dysregulation. The University of Kentucky Blood and Clot Thrombectomy Registry and Collaboration (BACTRAC) protocol utilizes thrombectomy to isolate intracranial arterial blood (i.e. distal to thrombus) and systemic arterial blood (i.e. carotid) to better understand the pathophysiology of stroke. Initial proteomic analyses of 184 plasma proteins sampled during thrombectomy found that systemic arterial REG3A was increased during large vessel occlusion. The function of REG3A in neuroinflammation sequela of ischemic stroke has yet to be elucidated. Here, we examined the association of plasma REG3A levels with other signaling proteins in systemic plasma during stroke, and its correlation with clinical outcomes of stroke. Methods Systemic arterial plasma samples from 35 thrombectomy subjects underwent Proximity Extension Assay via Olink Proteomics. REG3A levels and inflammatory markers were examined using bivariate regressions. Biserial correlation examined the relationship between the National Institutes of Health Stroke Scale (NIHSS) upon hospital admission and REG3A. Multilinear regression determined predictability of infarct volume and edema volume by REG3A. Results: Systemic arterial levels of IL6 and REG3A were positively correlated (R=0.371, P=0.031). Higher levels of REG3A correlated with increased infarct and edema volume (R=0.416, P=0.014; R=0.418, P=0.014). Multilinear regression predicting infarct (R2=0.985,P<0.001) and edema volume (R2=0.998, P<0.001) each yielded a model with similar shared proteins, REG3A, CXCL11, TGFβ, IL17A, and NCAM1. NIHSS score at admission positively correlated with REG3A( R= 0.373, P=0.032). Conclusions Within a network of proteins, REG3A was predictive of infarct severity and increased neurologic deficits in patients with ELVO. Results of IL6 and REG3A uphold similar associations reported within the literature. Models predicting stroke severity yielded potential protein partners with REG3A, such as IL17, CXCL11, and TGFβ, which interact with REG3A in other inflammatory processes. REG3A function in stroke has yet to be understood, however this study reports novel findings demonstrating the relationship between REG3A and outcomes of stroke. Examining these associations will be critical in defining REG3A function during neuroinflammation and as a potential biomarker of large vessel occlusive stroke predicting clinical outcomes.

Tissue Engineering for Mimics and Modulations of Immune Functions.

In the field of regenerative medicine, advances in tissue engineering have surpassed the reconstruction of individual tissues or organs and begun to work towards engineering systemic factors such as immune objects and functions. The immune system plays a crucial role in protecting and regulating systemic functions in the human body. Engineered immune tissues and organs have shown potential in recovering dysfunctions and aplasia of the immune system and the evasion from immune-mediated inflammatory responses and rejection elicited by engineered implants from allogeneic or xenogeneic sources are also being pursued to facilitate clinical transplantation of tissue engineered grafts. Here, current progress in tissue engineering to mimic or modulate immune functions is reviewed and elaborated from two perspectives: 1) engineering of immune tissues and organs per se and 2) immune evasion of host immunoinflammatory rejection by tissue-engineered implants.

Abstract P798: Antimicrobial Protein Reg3a and Network Inflammatory Proteins Are Predictive of Infarct Volume and Functional Impairment in Ischemic Stroke

Introduction: Regenerating Family Member 3 Alpha (REG3A) is an antimicrobial protein secreted by the intestine and pancreas. REG3A is involved in immune-mediated inflammatory responses and helps maintain homeostasis of the gut microbiome. REG3A is elevated in patients with ischemic stroke, but its role within the gut-brain axis during stroke remains unclear. Our aim was to examine the association of plasma REG3A levels with other signaling proteins in systemic plasma during stroke, and its correlation with clinical outcomes of ischemic stroke. Methods: Intracranial and systemic plasma samples from n=25 thrombectomy subjects underwent Proximity Extension Assay via Olink Proteomics. REG3A levels and inflammatory markers were examined using bivariate regressions. Stepwise regression determined the predictability of infarct volume by REG3A. Two-tailed t-tests were used to examine the relationship between the National Institutes of Health Stroke Scale (NIHSS) and levels of REG3A. Results: Increased expression of systemic REG3A correlated with increases in infarct volume (p=.009, R=.514). Stepwise regression predicting infarct volume yielded a model including REG3A and systemic proteins such as CCL19, IL15, and IL1α (P&lt;.001, R=.980). NIHSS scores corresponding to moderate-severe/severe strokes had increased intracranial REG3A (p&lt;.05) and increased ratios of intracranial:systemic REG3A expression (p=.05). Conclusions: Within a network of proteins, REG3A is predictive of increased infarct volume and decreased function. Studies suggest that IL17 and IL6 induce REG3A expression. Subsequent REG3A can interact with NFkB, which transcriptionally regulates CCL19, IL1α, and IL15. In the intestine, REG3A regulates microbial synthesis of fatty acids and inflammatory processes. Fatty acids suppress cytokines such as IL6 and IL1α and are reportedly neuroprotective in stroke. Increase of systemic REG3A may reflect cellular responses to gut dysbiosis and inflammation triggered from ischemic stroke. Future studies will examine prognostic and therapeutic potential of the REG3A pathway in large vessel occlusive stroke.

Prevalence and Characteristics of Hypoxic Hepatitis in COVID-19 Patients in the Intensive Care Unit: A First Retrospective Study.

Purpose: Coronavirus disease 2019 (COVID-19) has been associated with acute liver injury in reports worldwide. But no studies to date have described hypoxic hepatitis (HH) in patients with COVID-19. We aim to identify the prevalence of and possible mechanisms of HH in COVID-19 patients in the Intensive Care Unit (ICU).Methods: This retrospective study was conducted on 51 patients with confirmed SARS-CoV-2 infection in the ICU at Zhongnan Hospital of Wuhan University from December 21, 2019, to March 11, 2020. Information on clinical features of enrolled patients was collected for analysis.Results: HH was observed in 5.88% of the ICU patients with SARS-CoV-2 infection. All HH patients were progressing to respiratory failure and peak alanine aminotransferase (ALT) values were 1665, 1414, and 1140 U/L during hospitalization, respectively. All patients with HH died as a result of the deterioration of multiple organ failure (MOF). The dynamic changes of ALT, aspartate transaminase (AST), and total bilirubin (TBIL) levels were more dramatic in HH groups. Levels of TBIL, C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6(IL-6) showed statistically significant elevation in HH cases compared with that in non-HH cases (P < 0.001). Besides, the median survival time of the HH group was significantly shorter than the non-HH group (P < 0.05).Conclusions: In ICU, HH was not a rare condition in patients with severe COVID-19 and has a high mortality. The main causes of HH are respiratory and cardiac failure and may be associated with the immune-mediated inflammatory response. Clinicians should search for any underlying hemodynamic or respiratory instability even in patients with normal ALT levels on admission.