Macrophage Migration Inhibitory Factor Research Articles

The Correlation Between the Level of Macrophages and Inflammatory Factors and the Clinical Prognosis of Acute Stroke

Atherosclerosis is the primary mechanism of acute ischemic stroke (AIS) due to its association with inflammation and its impact on the formation, development, and stability of arterial plaques. In addition, AIS is characterized by a higher risk of poor prognosis, although the exact mechanisms responsible for this remain unclear. Currently, there is a lack of laboratory indicators for early assessment of disease progression and prognosis in AIS. It is hypothesized that macrophages and inflammatory factors play a role in the subsequent deterioration of AIS, possibly through their impact on cellular processes. The purpose of this study was to investigate the correlation between macrophages and inflammatory cytokines levels and clinical outcomes in individuals with acute stroke. A retrospective analysis was conducted of clinical data obtained from 252 AIS patients who received treatment at our hospital. After a period of 6 months of treatment, the patients were categorized into groups with either a favorable or unfavorable prognosis, based on their scores on the modified Rankin scale (mRS). Subsequently, a comparative analysis was conducted on the clinical data, macrophage migration inhibitory factor (MIF), and inflammatory cytokines (hs-CRP, IL-6) between the two groups, followed by an assessment of the factors that significantly influenced poor prognosis in AIS using a logistic regression model. Finally, Spearman correlation analysis was employed to excavate the relationship between MIF, hs-CRP, IL-6, and mRS. Analysis revealed that among the 252 patients with AIS, 75 individuals exhibited a poor prognosis, resulting in an incidence rate of 29.76%. The analysis using a logistic regression model demonstrated that MIF, hs-CRP, and IL-6 independently contributed to an increased risk of poor prognosis in AIS (P < 0.05). When assessing the ability to predict poor prognosis in AIS, the AUC for MIF, hs-CRP, and IL-6 were found to be 0.964, 0.723, and 0.774, respectively. Moreover, the sensitivity and specificity values were obtained, with MIF exhibiting higher values of 0.867 and 0.944, respectively. hs-CRP had a sensitivity of 0.680 and a specificity of 0.701, while IL-6 demonstrated a specificity of 0.927 and a sensitivity of 0.413. The Spearman correlation analysis showed a positive correlation between MIF, hs-CRP, and IL-6 and mRS (r = 0.634, 0.546, 0.530, all P < 0.001). In conclusion, the findings suggest that MIF, hs-CRP, and IL-6 are associated with poor prognosis in AIS and have the potential to serve as biomarkers for early prediction of AIS prognosis.

Macrophage migration inhibitory factor may contribute to the occurrence of multiple primary lung adenocarcinomas.

This study aimed to identify the key genes involved in the development of multiple primary lung cancers. Differential expression analysis was performed, followed by comparing the infiltration levels of 22 immune cell types between multiple and single primary lung adenocarcinomas. Marker genes for epithelial cells with different proportions between the two types of lung adenocarcinomas were identified. The common genes between the marker genes and differentially expressed genes were identified. Finally, the effects of the key genes were tested on the in vitro proliferation, migration and morphology. The infiltration levels of helper follicular T cells, resting NK cells, activated NK cells, M2 macrophages and resting mast cells were higher in the patients with multiple than in those with single primary lung adenocarcinomas. A total of 1553 differentially expressed genes and 4414 marker genes of epithelial cells were identified. Logistic regression analysis was performed on the 164 resulting genes. The macrophage migration inhibitory factor expression was positively associated with the occurrence of multiple primary lung adenocarcinomas. Moreover, its signalling pathway was the key pathway among the epithelial cells and multiple and single primary lung adenocarcinoma cells, and it was upregulated in lung adenocarcinoma cells. It also increased the expression of lung cancer markers, including NES and CA125, induced morphological changes in alveolar epithelial type II cells, and promoted their proliferation, migration and invasion. Multiple and single primary lung adenocarcinomas have different tumour immune microenvironments, and migration inhibitory factor may be a key factor in the occurrence of multiple primary lung adenocarcinomas.

Construction of “Coral” SERS sensor for ultrasensitive and rapid detection of harmful component macrophage migration inhibitory factor in Platelet-rich Plasma

Macrophage migration inhibitory factor (MIF) is a pro-inflammatory factor produced by residual red blood cell lysis, which can significantly influence the curative effect of Platelet-rich plasma (PRP) therapy used for osteoarthritis (OA) treatment. In this study, we proposed a novel approach for detecting the concentration of MIF in PRP using a dopamine-coated antibody-Au (core)-Ag (shell)-SERS sensor, which enables ultrasensitive and rapid detection of MIF. The best experimental conditions have a detection limit of only 90.05 pg/mL and a good linear relationship between 1-5000 ng/mL. In 40 PRP samples collected from actual clinical patients, we detected MIF concentrations ranging from 2.0-3.6 ng/mL. This indicated that the Coral SERS sensor not only allows for results highly consistent with the traditional ELISA method, but also costs less ($0.40-$0.70), needs shorter testing time (integration time is only 10s), and consumes less PRP that can greatly improve the sample quality and maximize the curative effect in clinical applications for OA treatment with PRP.

A Bidirectional Mendelian Randomization Analysis of Circulating Inflammatory Cytokines with Colon and Rectum Cancers

Chronic inflammation has been proposed to be associated with tumor development and progression. Dynamic changes in serum cytokines induced by feedback mechanisms from tumors make differentiation of cause and effect difficult. In this study, a bidirectional summary-level Mendelian randomization (MR) analysis was performed to elucidate the causal correlation between inflammatory regulators with colon (CC) and rectum (RC) carcinomas. Summary-level data on inflammation-related genetic variations were extracted from a genome-wide association meta-analysis. Corresponding data for CC and RC were obtained from the FinnGen (CC: 1396 cases vs. 174,006 controls; RC: 1,078 cases vs. 174,006 controls) and UK Biobank (CC: 2,226 cases vs. 358,968 controls; RC: 1,170 cases vs. 360,024 controls) consortiums. Inverse-variance weighted MR was used as the primary method, and sensitivity analysis was performed to assess the MR assumptions. After P value correction and meta-analysis, there was evidence of direct associations between macrophage migration inhibitory factor (MIF) and CC (95% confidence interval (CI):1.001-1.003; P = 8.77×10-4) and inverse correlations between Interleukin-17 (IL-17) and RC (95% CI: 0.997-0.999; P = 1.15×10-3). Conversely, interleukin-1 receptor antagonist (IL1ra) was downregulated in patients at a higher risk of RC (P = 0.006), and no reliable results revealed a causal association between other cytokines and CC and RC. Thus, this MR study suggests that downregulated serum MIF and elevated serum IL-17 concentrations may reduce the risk of CC and RC, respectively. Therefore, MIF and IL-17 may serve as potential therapeutic targets, and further exploration of their underlying mechanisms is warranted.

Deciphering Treg cell roles in esophageal squamous cell carcinoma: a comprehensive prognostic and immunotherapeutic analysis.

Background: Esophageal squamous cell carcinoma (ESCC) is a prevalent and aggressive form of cancer that poses significant challenges in terms of prognosis and treatment. Regulatory T cells (Treg cells) have gained attention due to their influential role in immune modulation within the tumor microenvironment (TME). Understanding the intricate interactions between Treg cells and the tumor microenvironment is essential for unraveling the mechanisms underlying ESCC progression and for developing effective prognostic models and immunotherapeutic strategies. Methods: A combination of single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq analysis was utilized to explore the role of Treg cells within the TME of ESCC. The accuracy and applicability of the prognostic model were assessed through multi-dimensional evaluations, encompassing an examination of the model's performance across various dimensions, such as the mutation landscape, clinical relevance, enrichment analysis, and potential implications for immunotherapy strategies. Results: The pivotal role of the macrophage migration inhibitory factor (MIF) signaling pathway within the ESCC TME was investigated, with a focus on its impact on Treg cells and other subpopulations. Through comprehensive integration of bulk sequencing data, a Treg-associated signature (TAS) was constructed, revealing that ESCC patients with elevated TAS (referred to as high-TAS individuals) experienced significantly improved prognoses. Heightened immune infiltration and increased expression of immune checkpoint markers were observed in high-TAS specimens. The model's validity was established through the IMvigor210 dataset, demonstrating its robustness in predicting prognosis and responsiveness to immunotherapy. Heightened therapeutic benefits were observed in immune-based interventions for high-TAS ESCC patients. Noteworthy differences in pathway enrichment patterns emerged between high and low-TAS cohorts, highlighting potential avenues for therapeutic exploration. Furthermore, the clinical relevance of key model genes was substantiated by analyzing clinical samples from ten paired tumor and adjacent tissues, revealing differential expression levels. Conclusion: The study established a TAS that enables accurate prediction of patient prognosis and responsiveness to immunotherapy. This achievement holds significant implications for the clinical management of ESCC, offering valuable insights for informed therapeutic interventions.

Host cell-type and pathogen-specific immunomodulatory functions of macrophage migration inhibitory factor (MIF) in infectious keratitis

Therapeutic management of inflammation in infectious keratitis (IK) requires new strategy and targets for selective immunomodulation. Targeting host cell-type specific inflammatory responses might be a viable strategy to curtail unnecessary inflammation and reduce tissue damage without affecting pathogen clearance. This study explores the possibility of pathogen and host cell-type dependent differences in the inflammatory pathways relevant in the pathogenesis of IK. Human corneal epithelial cell line (HCEC) and phorbol 12-myristate-13 acetate (PMA) differentiated THP-1 macrophage line were infected with either Aspergillus flavus conidia or Acanthamoeba castellanii trophozoites and the elicited inflammatory responses were studied in terms of gene expression and secretion of proinflammatory factors interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) and an upstream inflammatory regulator and mediator protein-the Macrophage Migration Inhibitory Factor (MIF). Given the pleotropic mode of MIF function in diverse cell types relevant in many human diseases, we tested if MIF driven responses to infection is different in HCECs and THP-1 macrophages by studying its expression, secretion and involvement in inflammation by siRNA mediated knockdown. We also examined IK patient tear samples for MIF levels. Infection with A. flavus or A. castellanii induced IL-8 and TNF-α responses in HCECs and THP-1 macrophages but to different levels. Our preliminary human data showed that the level of secreted MIF protein was elevated in IK patient tear, however, MIF secretion by the two cell types were strikingly different in-vitro, under both normal and infected conditions. We found that HCECs released MIF constitutively, which was significantly inhibited with infection, whereas THP-1 macrophages were stimulated to release MIF during infection. MIF gene expression remained largely unaffected by infection in both the cell lines. Although MIF in HCECs appeared to be intracellularly captured during infection, MIF knockdown in HCECs associated with a partial reduction of the IL-8 and TNF-α expression produced by either of the pathogens, suggesting a pro-inflammatory role for MIF in HCECs, independent of its canonical cytokine like function. In contrast, MIF knockdown in THP-1 macrophages accompanied a dramatic increase in IL-8 and TNF-α expression during A. castellanii infection, while the responses to A. flavus infection remained unchanged. These data imply a host cell-type and pathogen specific distinction in the MIF- related inflammatory signaling and MIF as a potential selective immunomodulatory target in infectious keratitis.

Assessing the reliability of serum macrophage migration inhibitory factor as a marker for diabetic nephropathy prediction in type 2 diabetes patients and the effect of ACE inhibitors on its level

Abstract Diabetic nephropathy (DN) is a prevalent chronic microvascular diabetic complication. As inflammation plays a vital role in the development and progress of DN the macrophages migration inhibitory factor (MIF), a proinflammatory multifunctional cytokine approved to play a critical function in inflammatory responses in various pathologic situations like DN. This study aimed To assess serum levels of MIF in a sample of Iraqi diabetic patients with nephropathy supporting its validity as a marker for predicting nephropathy in T2DM patients. In addition, to evaluate the nephroprotective effect of angiotensin-converting enzyme (ACE) inhibitors in terms of their influence on MIF levels. This is a case-control study involving ninety subjects that have been divided into three groups: twenty apparently healthy control group and seventy patients with type 2 diabetes mellitus divided into two equal groups according to the presence of diabetic nephropathy that has been further divided into two groups according to the use of ACE inhibitors or not. Serum MIF, urea, creatinine, RBS, HbA1c, BMI, eGFR, and urinary albumin to creatinine ratio have been measured for each subject. Serum MIF’s highest levels were observed in the diabetic nephropathy patients (24.9 ng/ml) followed by the diabetics (14. 1 ng/ml) with the lowest level observed in the control group (4.8 ng/ml). There was a significant relation between MIF levels and ACE inhibitors (p-value <0.05) with reduced MIF levels in ACE inhibitors users. The ROC curve showed that MIF has a good performance in disease prediction. These findings support the reliability of MIF as a biomarker for the prediction of diabetic nephropathy and the possible reducing effect of ACE inhibitors on MIF levels.

Highly Selective MIF Ketonase Inhibitor KRP-6 Diminishes M1 Macrophage Polarization and Metabolic Reprogramming.

Macrophage polarization is highly involved in autoimmunity. M1 polarized macrophages drive inflammation and undergo metabolic reprogramming, involving downregulation of mitochondrial energy production and acceleration of glycolysis. Macrophage migration inhibitory factor (MIF), an enigmatic tautomerase (ketonase and enolase), was discovered to regulate M1 polarization. Here, we reveal that KRP-6, a potent and highly selective MIF ketonase inhibitor, reduces MIF-induced human blood eosinophil and neutrophil migration similarly to ISO-1, the most investigated tautomerase inhibitor. We equally discovered that KRP-6 prevents M1 macrophage polarization and reduces ROS production in IFN-γ-treated cells. During metabolic reprogramming, KRP-6 improved mitochondrial bioenergetics by ameliorating basal respiration, ATP production, coupling efficiency and maximal respiration in LPS+IFN-γ-treated cells. KRP-6 also reduced glycolytic flux in M1 macrophages. Moreover, the selective MIF ketonase inhibitor attenuated LPS+IFN-γ-induced downregulation of PARP-1 and PARP-2 mRNA expression. We conclude that KRP-6 represents a promising novel therapeutic compound for autoimmune diseases, which strongly involves M1 macrophage polarization.

A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

BackgroundMantle cell lymphoma (MCL) is a rare B-cell non-Hodgkin lymphoma subtype which remains incurable despite multimodal approach including chemoimmunotherapy followed by stem cell transplant, targeted approaches such as the BTK inhibitor ibrutinib, and CD19 chimeric antigen receptor (CAR) T cells. CD74 is a nonpolymorphic type II integral membrane glycoprotein identified as an MHC class II chaperone and a receptor for macrophage migration inhibitory factor. Our group previously reported on CD74's abundant expression in MCL and its ability to increase via pharmacological inhibition of autophagosomal degradation. Milatuzumab, a fully humanized anti-CD74 monoclonal antibody, demonstrated significant activity in preclinical lymphoma models but failed to provide meaningful benefits in clinical trials mainly due to its short half-life. We hypothesized that targeting CD74 using a CAR-T cell would provide potent and durable anti-MCL activity.MethodsWe engineered a second generation anti-CD74 CAR with 4-1BB and CD3ζ signaling domains (74bbz). Through in silico and rational mutagenesis on the scFV domain, the 74bbz CAR was functionally optimized for superior antigen binding affinity, proliferative signaling, and cytotoxic activity against MCL cells in vitro and in vivo.ResultsFunctionally optimized 74bbz CAR-T cells (clone 42105) induced significant killing of MCL cell lines, and primary MCL patient samples including one relapse after commercial CD19 CAR-T cell therapy with direct correlation between antigen density and cytotoxicity. It significantly prolonged the survival of an animal model established in NOD-SCIDγc−/− (NSG) mice engrafted with a human MCL cell line Mino subcutaneously compared to controls. Finally, while CD74 is also expressed on normal immune cell subsets, treatment with 74bbz CAR-T cells resulted in minimal cytotoxicity against these cells both in vitro and in vivo.ConclusionsTargeting CD74 with 74bbz CAR-T cells represents a new cell therapy to provide a potent and durable and anti-MCL activity.

The impact of MIF deficiency on alterations of fecal microbiota in C57BL/6 mice induced by Trichinella spiralis infection.

Trichinellosis caused by Trichinella spiralis (T. spiralis) is a major food-borne parasitic zoonosis worldwide. Prevention of trichinellosis is an effective strategy to improve patient quality of life. Macrophage migration inhibitory factor (MIF) is closely related to the occurrence and development of several parasitic diseases. Studying the impact of MIF deficiency (Mif-/- ) on the alterations in host fecal microbiota due to T. spiralis infection may contribute to proposing a novel dual therapeutic approach for trichinellosis. To reveal the diversity and differences in fecal microbial composition, feces were collected from T. spiralis-uninfected and T. spiralis-infected wild-type (WT) and MIF knockout (KO) C57BL/6 mice at 0, 7, 14, and 35 days post-infection (dpi), and the samples were sent for 16S rRNA amplicon sequencing on the Illumina NovaSeq platform. Flow cytometry was used to determine the expression levels of IFN-γ and IL-4 in the CD4+ /CD8+ T-cell sets of mouse spleens. The results showed that operational taxonomic unit (OTU) clustering, relative abundance of microbial composition, alpha diversity, and beta diversity exhibited significant changes among the eight groups. The LEfSe analysis selected several potential biomarkers at the genus or species level, including Akkermansia muciniphila, Lactobacillus murinus, Coprococcus catus, Firmicutes bacterium M10_2, Parabacteroides sp. CT06, and Bacteroides between the KTs and WTs groups. The predicted bacterial functions of the fecal microbiota were mainly involved in metabolism, such as the metabolism of carbohydrates, amino acids, energy, cofactors, vitamins, nucleotides, glycans, and lipids. Flow cytometry revealed an increased CD3+ CD8- /CD3+ CD8+ T-cell ratio and increased IFN-γ and IL-4 levels in CD3+ CD8- T-cell sets from WT and MIF KO mice at 7 dpi. The results indicated that both MIF KO and infection time have a significant influence on the CD3+ CD8- IFN-γ+ and CD3+ CD8- IL-4+ response in mice after T. spiralis. In conclusion, this research showed alterations of the fecal microbiota and immune response in both WT and MIF KO mice before and after T. spiralis infection. These results revealed a potential role of MIF in regulating the pathogenesis of trichinellosis related to the intestinal microbiota. Importantly, the selected potential biomarkers combined with MIF will also offer a novel therapeutic approach to treat trichinellosis in the future.

Human macrophage migration inhibitory factor potentiates mesenchymal stromal cell efficacy in a clinically relevant model of allergic asthma

Current asthma therapies focus on reducing symptoms but fail to restore existing structural damage. Mesenchymal stromal cell (MSC) administration can ameliorate airway inflammation and reverse airway remodeling. However, differences in patient disease microenvironments seem to influence MSC therapeutic effects. A polymorphic CATT tetranucleotide repeat at position 794 of the human macrophage migration inhibitory factor (hMIF) gene has been associated with increased susceptibility to and severity of asthma. We investigated the efficacy of human MSCs in high- vs. low-hMIF environments and the impact of MIF pre-licensing of MSCs using humanized MIF mice in a clinically relevant house dust mite (HDM) model of allergic asthma. MSCs significantly attenuated airway inflammation and airway remodeling in high-MIF-expressing CATT7 mice but not in CATT5 or wild-type littermates. Differences in efficacy were correlated with increased MSC retention in the lungs of CATT7 mice. MIF licensing potentiated MSC anti-inflammatory effects at a previously ineffective dose. Mechanistically, MIF binding to CD74 expressed on MSCs leads to upregulation of cyclooxygenase 2 (COX-2) expression. Blockade of CD74 or COX-2 function in MSCs prior to administration attenuated the efficacy of MIF-licensed MSCs invivo. These findings suggest that MSC administration may be more efficacious in severe asthma patients with high MIF genotypes (CATT6/7/8).

Abstract PO-084: Deletion of macrophage migration inhibitory factor promotes antitumoral T cell infiltration and inhibits MDSC recruitment to the head and neck cancer microenvironment

Abstract Head and neck squamous cell carcinoma (HNSCC) is a significant public health concern worldwide. Immunomodulatory targets in the HNSCC tumor microenvironment are crucial to enhance the efficacy of HNSCC immunotherapy. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine that has been linked to worse prognosis in many cancers, but the mechanistic role of MIF in HNSCC remains unclear. Using a murine orthotopic oral cancer model in Mif+/+ or Mif− / − mice, we determined the function of host derived MIF in HNSCC tumor development, metastasis as well as localized and systemic tumor immune responses. We found that Mif− / − mice have decreased tumor growth and burden compared to their wild-type counterparts. Flow cytometric analysis of immune populations within the primary tumor site revealed increased T cell recruitment to the HNSCC tumor microenvironment, within the tumors of Mif− / − mice. MIF deletion also enhanced the effector function of anti-tumoral Th1 cells and decreased the accumulation of granulocytic MDSC in the tumor microenvironment. Furthermore, we show that isolated MDSC chemotactically respond to MIF in a dose dependent manner which is inhibited when MIF is depleted. Interestingly, MDSCs of tumor bearing Mif− / − mice expressed increased levels of PDL1 compared to Mif+/+ mice. However, Mif− / − and Mif+/+ MDSCs have comparable abilities to suppress T cell proliferation. Herein we describe a chemotactic and immunomodulatory role for MIF in the context of HNSCC. MIF targeted immunomodulation remains a potentially viable cancer therapeutic, however more remains to be understood about the cellular interaction of MIF within the tumor microenvironment before these targeted therapies can be effectively employed Citation Format: Steve Oghumu. Deletion of macrophage migration inhibitory factor promotes antitumoral T cell infiltration and inhibits MDSC recruitment to the head and neck cancer microenvironment [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-084.

Increased Expression of Long Noncoding RNA LOC100506314 in T cells from Patients with Nonsegmental Vitiligo and Its Contribution to Vitiligo Pathogenesis.

This study aimed to identify the abnormal expression of long noncoding RNAs (lncRNAs) in T cells from patients with vitiligo and to investigate their functional roles in the immune system. Using microarray analysis, the expression levels of RNA transcripts in T cells from patients with vitiligo and controls were compared. We identified several genes and validated their expression levels in T cells from 41 vitiligo patients and 41 controls. The biological functions of the lncRNAs were studied in a transfection study using an RNA pull-down assay, followed by proteomic analysis and western blotting. The expression levels of 134 genes were significantly increased, and those of 142 genes were significantly decreased in T cells from vitiligo patients. After validation, six genes had increased expression, and three genes had decreased expression in T cells from patients with vitiligo. T-cell expression of LOC100506314 was increased in vitiligo, especially CD4+, but not CD8+ T cells. The expression levels of LOC100506314 in CD4+ T cells was positively and significantly associated with the severity of vitiligo. LOC100506314 was bound to the signal transducer and activator of transcription 3 (STAT3) and macrophage migration inhibitory factor (MIF). Enhanced expression of LOC100506314 inhibited the phosphorylation of STAT3, protein kinase B (AKT), and extracellular signal-regulated protein kinases (ERK), as well as the levels of nuclear protein of p65 and the expression of IL-6 and IL-17 in Jurkat cells and T cells from patients with vitiligo. In conclusion, this study showed that the expression of LOC100506314 was elevated in CD4+ T cells from patients with vitiligo and associated the severity of vitiligo. LOC100506314 interacted with STAT3 and MIF and inhibited IL-6 and IL-17 expression by suppressing the STAT3, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), AKT, and ERK pathways. Enhanced expression of LOC100506314 in T cells may be a potential treatment strategy for vitiligo.

ОЦЕНКА ЛОКАЛЬНОГО ПРОФИЛЯ ЦИТОКИНОВ, ЭКСПРЕССИРУЕМЫХ НАТИВНЫМИ КЛАПАНАМИ СЕРДЦА, ПОЛУЧЕННЫМИ ОТ ПАЦИЕНТОВ С ПРИОБРЕТЕННЫМИ ПОРОКАМИ СЕРДЦА

Doing. The pathogenesis of acquired heart defects is a complex process involving many different factors. One of the triggers for the development of dysfunctions of native heart valves can be an inflammatory process caused by various types of pathogens. The aim of the study was to evaluate the local profile of cytokines expressed by native heart valves obtained from patients with acquired heart defects during cardiac surgery. Materials and Methods: As a material for the study, biopsies of native mitral heart valves were obtained from 4 patients with infective endocarditis and from 10 patients with rheumatic heart disease. RNA was isolated using a commercial RNeasy® Plus Universal Mini Kit (Qiagen, Germany), protein extraction was performed using T-PER™ Tissue Protein Extraction Reagent lysis buffer (ThermoScientific, USA) with the addition of protease inhibitors Halt™ Protease Inhibitor Cocktail (ThermoScientific, USA) and 0.5M EDTA solution (ThermoScientific, USA). The level of cytokines expressed by native valves was determined by dot blotting using the Proteome Profiler™ Human Cytokine Array Kit (ARY005B) (R&D Systems, USA). Cytokine gene expression was assessed by qPCR using SYBR Green probes (JSC Evrogen, Russia) on a ViiA7 Real-Time PCR System amplifier (Applied Biosystems, USA). Statistical analysis of the results obtained was performed using the GraphPad Prism 8 program (GraphPad Software, USA). Results: The proteins MIF (macrophage migration inhibitory factor), PAI-1 (plasminogen activator inhibitor-1), ICAM-1 (intercellular adhesion molecule-1) and CXCL12 (stromal growth factor-1) were found to be registered in native valves, obtained from both patients with IE and patients with RHD. Semi-quantitative analysis showed that native valves obtained from patients with IE are characterized by a four-fold increase in the level of PAI-1 secretion and a two-fold decrease in the level of ICAM-1 and CXCL12 compared with the control. MIF was expressed at the same level in both studied groups. IL-1ra (interleukin 1 receptor antagonist), IL-6 (interleukin 6), IL-8 (interleukin 8), IL-16 (interleukin 16), CCL4 (chemokine ligand 4), CCL5 (chemokine ligand 5), and CXCL1 ( chemokine ligand 1) were found only in valves affected by IE. The expression of the MIF, IL6, and IL8 genes was increased in the experimental group relative to the control, and the expression of the PAI1, IL1RA, and CXCL1– genes was decreased in the experiment relative to the control. The gene expression of ICAM1, CXCL12, CCL4, CCL5 and IL16 was the same in both studied groups. Conclusions: The results of the study show that native heart valves affected by IE are characterized by a unique cytokine profile compared to valves obtained from patients with non-infectious endocardial pathology. Native heart valves affected by IE show nonspecific local inflammation, active neovascularization, and decreased resistance to pathogenic bacteremia.

The level of macrophage migration inhibitory factor is negatively correlated with the efficacy of PD-1 blockade immunotherapy combined with chemotherapy as a neoadjuvant therapy for esophageal squamous cell carcinoma

PurposeThis study aimed to screen biomarkers to predict the efficacy of programmed cell death 1 (PD-1) blockade immunotherapy combined with chemotherapy as neoadjuvant therapy for esophageal squamous cell carcinoma (ESCC). MethodsIn the first stage of the study, the baseline concentrations of 40 tumor-related chemokines in the serum samples of 50 patients were measured to screen for possible biomarkers. We investigated whether the baseline concentration of the selected chemokine was related to the therapeutic outcomes and tumor microenvironment states of patients treated with the therapy. In the second stage, the reliability of the selected biomarkers was retested in 34 patients. ResultsThe baseline concentration of macrophage migration inhibitory factor (MIF) was negatively correlated with disease-free survival (DFS) and overall survival (OS) in patients treated with the therapy. In addition, a low baseline expression level of MIF is related to a better tumor microenvironment for the treatment of ESCC. A secondary finding was that effective treatment decreased the serum concentration of MIF. ConclusionBaseline MIF levels were negatively correlated with neoadjuvant therapy efficacy. Thus, MIF may serve as a predictive biomarker for this therapy. The accuracy of the prediction could be improved if the serum concentration of MIF is measured again after the patient received several weeks of treatment.

Macrophage migration inhibitory factor exacerbates asthmatic airway remodeling via dynamin-related protein 1-mediated autophagy activation

BackgroundMacrophage migration inhibitory factor (MIF) and GTPase dynamin-related protein 1 (Drp1)-dependent aberrant mitochondrial fission are closely linked to the pathogenesis of asthma. However, it is unclear whether Drp1-mediated mitochondrial fission and its downstream targets mediate MIF-induced proliferation of airway smooth muscle cells (ASMCs) in vitro and airway remodeling in chronic asthma models. The present study aims to clarify these issues.MethodsIn this study, primary cultured ASMCs and ovalbumin (OVA)-induced asthmatic rats were applied. Cell proliferation was detected by CCK-8 and EdU assays. Western blotting was used to detect extracellular signal-regulated kinase (ERK) 1/2, Drp1, autophagy-related markers and E-cadherin protein phosphorylation and expression. Inflammatory cytokines production, airway reactivity test, histological staining and immunohistochemical staining were conducted to evaluate the development of asthma. Transmission electron microscopy was used to observe the mitochondrial ultrastructure.ResultsIn primary cultured ASMCs, MIF increased the phosphorylation level of Drp1 at the Ser616 site through activation of the ERK1/2 signaling pathway, which further activated autophagy and reduced E-cadherin expression, ultimately leading to ASMCs proliferation. In OVA-induced asthmatic rats, MIF inhibitor 4-iodo-6-phenylpyrimidine (4-IPP) treatment, suppression of mitochondrial fission by Mdivi-1 or inhibiting autophagy with chloroquine phosphate (CQ) all attenuated the development of airway remodeling.ConclusionsThe present study provides novel insights that MIF promotes airway remodeling in asthma by activating autophagy and degradation of E-cadherin via ERK/Drp1 signaling pathway, suggesting that targeting MIF/ERK/Drp1 might have potential therapeutic value for the prevention and treatment of asthma.

Synthesis and in vitro activity tests of N-benzoyl-N'-phenylthiourea derivatives as macrophage migration inhibitory factor

Context: The COVID-19 pandemic in 2020 resulted in widespread mortalities due to cytokine storms in the affected patients. Macrophage migration inhibitory factor (MIF) is one of the most interesting targets in developing anti-COVID-19 drugs. Some thiourea compounds have been identified as having potential as MIF inhibitors. Aims: To investigate MIF inhibitory activity of N-benzoyl-N'-phenylthiourea derivatives. Methods: The study consists of in-silico activity prediction of designed compounds using a molecular docking approach against MIF protein (PDB ID: 1LJT). Afterwards, the designed compounds were synthesized and tested in vitro using the tautomerase activity approach. Results: The molecular docking study showed that all designed compounds possess comparable docking scores to the native ligand of the protein. MIF Assay performed on compounds (1) and (2) indicated a decrease in tautomerase activity of the MIF target protein of only 10.1 and 6.2%, respectively, compared to the positive control. Conclusions: In silico results predicted better bioactivity against MIF protein, but the result does not translate to the in vitro assay, where two of the designed compounds possess only low inhibitory activity.

Macrophage migration inhibitory factor and pro-inflammatory cytokines have a main role in the infection of chorionic villi explants in vitro by Trypanosoma cruzi, the protozoan that cause congenital Chagas disease

Macrophage migration inhibitory factor and pro-inflammatory cytokines have a main role in the infection of chorionic villi explants in vitro by Trypanosoma cruzi, the protozoan that cause congenital Chagas disease

Exploring the anti-inflammatory activity of sulforaphane.

Dysregulation of innate immune responses can result in chronic inflammatory conditions. Glucocorticoids, the current frontline therapy, are effective immunosuppressive drugs but come with a trade-off of cumulative and serious side effects. Therefore, alternative drug options with improved safety profiles are urgently needed. Sulforaphane, a phytochemical derived from plants of the brassica family, is a potent inducer of phase II detoxification enzymes via nuclear factor-erythroid factor 2-related factor 2 (NRF2) signaling. Moreover, a growing body of evidence suggests additional diverse anti-inflammatory properties of sulforaphane through interactions with mediators of key signaling pathways and inflammatory cytokines. Multiple studies support a role for sulforaphane as a negative regulator of nuclear factor kappa-light chain enhancer of activated B cells (NF-κB) activation and subsequent cytokine release, inflammasome activation and direct regulation of the activity of macrophage migration inhibitory factor. Significantly, studies have also highlighted potential steroid-sparing activity for sulforaphane, suggesting that it may have potential as an adjunctive therapy for some inflammatory conditions. This review discusses published research on sulforaphane, including proposed mechanisms of action, and poses questions for future studies that might help progress our understanding of the potential clinical applications of this intriguing molecule.

Novel therapeutic for multiple sclerosis protects white matter function in EAE mouse model.

Multiple sclerosis (MS) is a chronic demyelinating disease with prominent axon dysfunction. Our previous studies in an MS mouse model, experimental autoimmune encephalomyelitis (EAE), demonstrated that major histocompatibility complex Class II constructs can reverse clinical signs of EAE. These constructs block binding and downstream signaling of macrophage migration inhibitory factors (MIF-1/2) through CD74, thereby inhibiting phosphorylation of extracellular signal-regulated kinase (ERK) activation and tissue inflammation and promoting remyelination. To directly assess the effects of a novel third generation construct, DRhQ, on axon integrity in EAE, we compared axon conduction properties using electrophysiology on corpus callosum slices and optic nerves. By using two distinct white matter (WM) tracts, we aimed to assess the impact of the EAE and the benefit of DRhQ on myelinated and unmyelinated axons as well as to test the clinical value of DRhQ on demyelinating lesions in CC and optic myelitis. Our study found that EAE altered axon excitability, delayed axon conduction and slowed spatiotemporal summation correlated with diffuse astrocyte and microglia activation. Because MS predisposes patients to stroke, we also investigated and showed that vulnerability to WM ischemia is increased in the EAE MS mouse model. Treatment with DRhQ after the onset of EAE drastically inhibited microglial and astrocyte activation, improved functional integrity of the myelinated axons and enhanced recovery after ischemia. These results demonstrate that DRhQ administered after the onset of EAE promotes WM integrity and function, and reduces subsequent vulnerability to ischemic injury, suggesting important therapeutic potential for treatment of progressive MS.