Role Of MIF Research Articles

INVESTIGATION OF THE POSSIBLE ROLE OF MACROPHAGE MIGRATION INHIBITOR FACTOR (MIF) GENE -173G/C POLYMOPHISM IN ATHEROSCLEROSIS

Aim: Atherosclerosis is defined as an inflammatory disease that results in the formation of atherosclerotic plaques caused by the deposition of cholesterol in the arterial intima. As a result of damage to the intima layer of the vessel, foam cell formation following cholesterol accumulation and plaque development due to smooth muscle cell increase are observed. In different stages of atherosclerosis, migration of leukocytes to the vessel wall is provided by functional type chemokines; It was aimed to investigate the possible role of MIF -173G/C polymorphism in atherosclerosis disease depending on this function, since it has the same functional properties as chemokines.
 Material and Method: Thirty patients with 70% occlusion detected by angiography and 30 healthy individuals were included in the study. DNA isolation was performed from blood samples taken from individuals in EDTA tubes. Analysis of MIF -173G/C polymorphism was performed on Real Time PCR (LC480, Roche). Statistical analyzes were performed using the STATISTICA version 13.5.0.17 (TIBCO Software Inc. (2017)) program. 
 
 Results: In the MIF -173G/C polymorphism, the frequency of GG, GC and CC genotypes in the patient group was 55.26%, 41.18% and 66.66%, and 44.74%, 58.82% and 33.33% in the control group, respectively. Compared with the GG genotype, it was determined that those with the GC genotype had a 0.567-fold (p=0.3367) risk of developing the disease, and those with the CC genotype had a 1.6190-fold (p= 0.7038) risk of developing the disease.
 Conclusion: Further studies can be carried out by increasing the number of samples.

Salivary proteins as markers of radiation-related oral mucositis

Objectives The aim of this study was to characterize the salivary proteomic profile of patients treated for head and neck cancers (HNCs) and correlate it with the risk of developing severe radiation-related oral mucositis (OM). Study Design Forty-one patients with oral squamous cell carcinoma (OSCC) who underwent adjuvant radiotherapy (RT) or chemoradiotherapy (CRT) were included. All patients were submitted to dental conditioning protocols prior to RT. OM and OM-related pain were evaluated daily during RT and graded according to the Common Toxicity Criteria for Adverse Events (National Cancer Institute, version 4.0, 2010) and the visual analogue scale (VAS). For the molecular analysis, whole saliva was collected immediately before RT and subjected to proteomic analysis by means of liquid chromatography–mass spectrometry (LTQ Orbitrap Velos MS; Thermo Fisher Scientific, Bremen, Germany) and label-free protein quantification. The results obtained from the targeted proteomic analysis were compared with OM clinical outcomes. Statistical analysis was performed by using Wilcoxon's test. Results Of the study patients, 73% presented with stage III/IV OSCC; 58% were submitted to CRT protocols, with a mean RT dose of 66 Gy. Most of the patients (66%) had visible tumor areas at the time of saliva collection; 43.9% had grade 2 and 31.7% had grade 3 as the highest OM grade during RT, with a mean highest reported VAS score of 3.53. For the target proteomics analysis, a total of 65 proteins were observed to be mostly related to biologic processes, such as immune responses, peptidase inhibitor activity, and the inflammatory system. The macrophage migration inhibitory factor (MIF HUMAN) was statistically significant when correlated with OM grade. MIF was observed in patients with grades 1 to 4 OM and showed higher abundance for OM grades 3 and 4 compared with grades 1 and 2 (P = .04). Conclusions The correlation of MIF with the severity of OM is compatible with the role of MIF as a proinflammatory protein. This seems to be the first study to describe this association; therefore, future prospective studies would be ideal to observe pre- and post-RT alterations in salivary MIF levels to validate this result and better understand the role of MIF in the pathogenesis of OM.

Prognostic role of macrophage migration inhibitory factor in patients with myocardial infarction with ST-segment elevation after percutaneous coronary intervention

The aim of the study was to determine the prediction role of MIF in development of STEMI complications within 6 months after the event.Materials and methods. The study included 45 people with confirmed myocardial infarction with ST segment elevation (STEMI) and successful restoration of blood flow - TIMI-III. The control group - 12 healthy volunteers. All patients underwent baseline investigation. The level of myocardial damage biomarkers MIF and troponin were determined before percutaneous coronary intervention (PCI), then within 24 hours. The pro-inflammatory CRP biomarker was determined before PCI and 5-7 days after the development of the coronary event. The patient follow-up period was 6 months, during which 8 patients reached the endpoint.Results and discussion. Statistically significant increase of MIF levels in STEMI patients was defined relative to the control group. During the observation period 19% of patients reached the endpoint. The MIFІ level significantly correlated with complications in the acute period of myocardial infarction, the class of acute heart failure according to Killip, with troponin levels, with active smoking, MIFII was correlated with stable angina pectoris before the index event, the size of the left atrium and the mass of the left ventricular myocardium. Assessment of blood glucose in with the MIFII was significant in predicting the development of an adverse outcome, in comparison with a separate biomarker definition, the diagnostic efficiency of the model was 88%.Conclusions. An early increase of MIF level was associated with an adverse course of the disease. Our prognostic model significantly improved the prediction of the risk of complications after STEMI

Using Intravital Microscopy to Study the Role of MIF in Leukocyte Trafficking In Vivo.

In vivo visualization of the microvasculature of the mouse cremaster muscle has been fruitful in the evaluation of the role of macrophage migration inhibitory factor in promotion of leukocyte trafficking. Here we explain how to undertake this preparation, including details on mouse anesthesia, securing intravenous access, and cremaster muscle exteriorization. We also provide information on the various microscopy modalities now available for imaging microvascular preparations of this nature.

Macrophage migration inhibitory factor interacting with Th17 cells may be involved in the pathogenesis of autoimmune damage in Hashimoto's thyroiditis.

Purpose. To explore the possible role of MIF and Th17 cells in the thyroid-specific autoimmune damage of Hashimoto's thyroiditis (HT). Material and Methods. We enrolled 40 HT patients and 30 healthy controls and divided HT patients into euthyroid subset (n = 22) and subclinical or overt hypothyroidism subset (n = 18). The percentages of Th17 cells and expressions of MIF, interleukin 17A (IL-17A) mRNA in PBMCs, as well as serum concentrations of MIF, and IL-17A, and thyroid functions, and thyroid-specific autoantibodies (TPOAb, TgAb) were detected by flow cytometry, real-time RT-PCR, ELISA, and ECLIA in all subjects. Results. MIF mRNA, IL-17A mRNA expressions and Th17 cells percentages, serum MIF, and IL-17A protein levels were all significantly higher in HT patients, even in euthyroid subgroup. Additionally, the differences became more obvious in dysfunction subgroup. Importantly, both MIF levels and Th17 cells percentage were positively correlated with serum TPOAb, TgAb, and thyrotropin (TSH) levels in HT patients. Conclusions. These data suggest that MIF and Th17 cells increased dynamically and positively correlated with the markers of thyroid autoimmune damage, which indicated that interaction between MIF and Th17 cells may participate in the pathogenesis and development of thyroid-specific autoimmunity in HT.

Macrophage migration inhibitory factor is required for recruitment of scar-associated macrophages during liver fibrosis.

Recruitment of peripheral monocytes to the liver is a key contributor to the response to injury. MIF can act as a chemokine and cytokine, regulating innate immune responses in many tissues and cell types. We hypothesized that MIF contributes to the progression of CCl4-induced hepatic fibrosis by regulating recruitment of SAM. SAMs dynamically regulate HSC activation and ECM degradation. To gain insight into the role of MIF in progression of liver fibrosis, we investigated markers of fibrosis and immune responses after chronic CCl4 administration to female C57BL/6 and MIF(-/-) mice. Chronic CCl4 exposure increased activation of HSC in WT mice, indicated by increased expression of αSMA mRNA and protein, as well as mRNA for collagen 1α1; these responses were blunted in female MIF(-/-) mice. Despite lower activation of HSC in MIF(-/-) mice, accumulation of ECM was similar in WT and MIF(-/-)mice, suggesting a decreased rate of ECM degradation. Recruitment of SAMs was lower in MIF(-/-) mice compared with WT mice, both in their initial inflammatory phenotype, as well as in the later phase as proresolution macrophages. The decreased presence of resolution macrophages was associated with lower expression of MMP13 in MIF(-/-) mice. Taken together, these data indicate that MIF-dependent recruitment of SAMs contributes to degradation of ECM via MMP13, highlighting the importance of appropriate recruitment and phenotypic profile of macrophages in the resolution of fibrosis.

Coalescence of B cell receptor and invariant chain MHC II in a raft-like membrane domain

The BCR binds antigen for processing and subsequent presentation on MHC II molecules. Polyvalent antigen induces BCR clustering and targeting to endocytic processing compartments, which are also accessed by Ii-MHC II. Here, we report that clustered BCR is able to team up with Ii-MHC II already at the plasma membrane of mouse B-lymphocytes. Colocalization of BCR and Ii-MHC II on the cell surface required clustering of both types of molecules. The clustering of only one type did not trigger the recruitment of the other. Ii-bound MIF (a ligand of Ii) also colocalized with clustered BCR upon oligomerization of MIF on the surface of the B cell. Abundant surface molecules, such as B220 or TfnR, did not cocluster with the BCR. Some membrane raft-associated molecules, such as peptide-loaded MHC II, coclustered with the BCR, whereas others, such as GM1, did not. The formation of a BCR- and Ii-MHC II-containing membrane domain by antibody-mediated clustering was independent of F-actin and led to the coendocytosis of its constituents. With a rapid Brij 98 extraction method, it was possible to capture this membrane domain biochemically as a DRM. Ii and clustered BCR were present on the same DRM, as shown by immunoisolation. The coalescence of BCR and Ii-MHC II increased tyrosine phosphorylation, indicative of enhanced BCR signaling. Our work suggests a novel role for MIF and Ii-MHC II in BCR-mediated antigen processing.

May trophoblast participate in regulation of decidual cell survival? A putative role for MIF, the macrophage migratory inhibitory factor

May trophoblast participate in regulation of decidual cell survival? A putative role for MIF, the macrophage migratory inhibitory factor

MIF contributes to Type 1 diabetes favors insulitis by mediating macrophages and dendritic cells maturation (P4533)

Abstract Type 1 diabetes (T1D) is characterized by a cellular infiltrate in pancreatic islets where β cells are destroyed. The recognition of antigens and auto antigens takes place by macrophages (Mo) and dendritic cells (DCs). Previously we have showed that MIF favors the expression of co-stimulatory molecules on Mo and DCs on infection diseases. However, the role of MIF on Mo and CDs in T1D has not been explored. Here, we determined the expression of co-stimulatory molecules on Mo and CDs from WT or MIF-/- mice on experimental T1D induced by STZ. Cells extracted from pancreas and spleens were treated with antibodies anti- CD80, CD86, CD40, MHC-II and TLRs. Also we tested the number of CD4+CD25+FOXp3+ cells. The results show that MIF-/- mice not developed high glucose levels compared with WT mice. Pro-inflammatory cytokines in serum were diminished on MIF-/-STZ mice, while the anti-inflammatory cytokines were higher compared to WT STZ mice. Moreover, MIF-/-STZ mice had less CD80, CD86, MHCII, TLR-2 and TLR-4 in spleen and pancreatic islets Mo and CDs. In addition, CD4+CD25+ spleen cells had a high expression of FOXp3 transcription factor in MIF-/- healthy than MIF+/+ healthy mice. 8 weeks after of STZ induction the MIF-/-STZ mice present higher FOXp3 expression than WT STZ mice. Our results suggest that MIF favors the expression of co-stimulatory molecules in the Mo and CDs in T1D. Additionally we propose that MIF down-regulate the proliferation of regulatory T cells in T1D.

Mo1760 High-Throughput 454 Pyrosequencing Analysis Reveals Dysbiosis of the Mucosa-Associated Microbiota in Dogs With Inflammatory Bowel Disease

G A A b st ra ct s enteropathy and ameliorates IBD but its role in T.gondii-induced IBD-like enteritis is less clear. MIF is a proinflammatory cytokine that plays a role in innate and adaptive immunity. Also, MIF is involved in IBD pathogenesis and in T.gondii and S.mansoni mediated immunopathology and disease severity. Our study examines the role of MIF in T.gondii-induced IBD-like enteritis and systemic inflammatory response in mice coinfected with S.mansoni. Methods: Wild type (wt) and MIF knock out (Mif ko) mice were infected by subcutaneous injection with 50 cercaries of BH strain of S.mansoni, and orally with 100 cysts of ME49 strain of T. gondii after 7-9 weeks post S.mansoni infection. Hemogram and alaninetransferase (ALT) levels were determined. Histopathology was analyzed and TNFα, IFNγ and IL-10 levels were measured by ELISA. Results: Histopathologycal analysis of wt mice monoinfected with T.gondii showed intense mononuclear cell infiltration and terminal ileum necrosis, which greatly reduced in the absence of MIF. When analyzing co-infection with S. mansoni, the intestinal mucosa showed a marked attenuation of the inflammation in wt, and an almost complete abrogation of the inflammatory response in Mif ko mice. However, despite profound reduction of inflammatory response in both coinfected groups, they presented increased mortality rates (n=6, Mif ko n=6, wt; p>0.05) though MIF-deficiency was protective in T.gondii single infection when compared to wt mice (n=5, Mif ko, n=5, wt, p<0.01) but not in S.mansoni monoinfection (n=11, Mif ko, n=11, wt, p<0.0001). Nonetheless, increased morbidity was observed in coinfected Mif ko mice which had augmented weight loss (18.6±2.1 x 23.5±3.5g; Mif ko x wt, p<0.05). Systemic response in mono and coinfected wt and Mif ko was determined. Histopathology showed severe liver mononuclear cell infiltration in wt coinfected when compared with monoinfected mice. In contrast, MIF deficiency resulted in amelioration of liver inflammation in mono and coinfected mice. Also, function was more preserved in Mif ko coinfected when compared with wt mice measured by ALT levels (113±13 UI/L x 68±6.3 UI/L; wt x Mif ko, p<0.01). IL-10 plasma levels were lower in coinfected Mif ko compared to wt mice (2.77± 0.67 x 0.56±0.46 ng/ml, wt x Mif ko, p<0.05). Conclusion: Our findings indicate that co-infection regulates MIF-dependent responses at intestinal mucosa but also profoundly affect systemic IL10-dependent and independent protective responses.

Mo1756 CD4+ T Cells Regulate the Multifactorial Gastrointestinal Inflammation Induced Following Colonization With Bacterial Provocateurs

G A A b st ra ct s enteropathy and ameliorates IBD but its role in T.gondii-induced IBD-like enteritis is less clear. MIF is a proinflammatory cytokine that plays a role in innate and adaptive immunity. Also, MIF is involved in IBD pathogenesis and in T.gondii and S.mansoni mediated immunopathology and disease severity. Our study examines the role of MIF in T.gondii-induced IBD-like enteritis and systemic inflammatory response in mice coinfected with S.mansoni. Methods: Wild type (wt) and MIF knock out (Mif ko) mice were infected by subcutaneous injection with 50 cercaries of BH strain of S.mansoni, and orally with 100 cysts of ME49 strain of T. gondii after 7-9 weeks post S.mansoni infection. Hemogram and alaninetransferase (ALT) levels were determined. Histopathology was analyzed and TNFα, IFNγ and IL-10 levels were measured by ELISA. Results: Histopathologycal analysis of wt mice monoinfected with T.gondii showed intense mononuclear cell infiltration and terminal ileum necrosis, which greatly reduced in the absence of MIF. When analyzing co-infection with S. mansoni, the intestinal mucosa showed a marked attenuation of the inflammation in wt, and an almost complete abrogation of the inflammatory response in Mif ko mice. However, despite profound reduction of inflammatory response in both coinfected groups, they presented increased mortality rates (n=6, Mif ko n=6, wt; p>0.05) though MIF-deficiency was protective in T.gondii single infection when compared to wt mice (n=5, Mif ko, n=5, wt, p<0.01) but not in S.mansoni monoinfection (n=11, Mif ko, n=11, wt, p<0.0001). Nonetheless, increased morbidity was observed in coinfected Mif ko mice which had augmented weight loss (18.6±2.1 x 23.5±3.5g; Mif ko x wt, p<0.05). Systemic response in mono and coinfected wt and Mif ko was determined. Histopathology showed severe liver mononuclear cell infiltration in wt coinfected when compared with monoinfected mice. In contrast, MIF deficiency resulted in amelioration of liver inflammation in mono and coinfected mice. Also, function was more preserved in Mif ko coinfected when compared with wt mice measured by ALT levels (113±13 UI/L x 68±6.3 UI/L; wt x Mif ko, p<0.01). IL-10 plasma levels were lower in coinfected Mif ko compared to wt mice (2.77± 0.67 x 0.56±0.46 ng/ml, wt x Mif ko, p<0.05). Conclusion: Our findings indicate that co-infection regulates MIF-dependent responses at intestinal mucosa but also profoundly affect systemic IL10-dependent and independent protective responses.

Macrophage migration inhibitory factor mediates the antidepressant actions of voluntary exercise

Voluntary exercise is known to have effects on various brain functions, including an antidepressant action. However, underlying mechanism of exercise effects on psychological disease is still ambiguous. We identified the candidate target genes for alleviating depression by the analysis of existing microarray data sets of mouse hippocampus mRNA from voluntary exercise and ECS (Electroconvulsive seizure) stimulated experiments. To validate the precise changes of those candidate genes, we conducted 28 day voluntary exercise and 10 day chronic ECS treatment with rats. Reverse transcription‐PCR and real‐time PCR analysis showed 5 genes; BDNF, COX‐2, SLC1A1, NPY and MIF were commonly induced by both ECS and voluntary exercise experiments. Furthermore, immunohistochemistry analysis firmly remarked that the amount of MIF, a proinflammatory cytokine, was also increased by voluntary exercise. We showed that recombinant MIF administration produced antidepressant‐like behavior in animal FST (Forced swim test). Moreover, MIF treatment for 1 hour increased the expression of TPH2, a rate‐limiting enzyme in synthesis of serotonin, and 12 hour treatment increased the amount of BDNF in Neuro‐2a neuroblastoma cell line. These results suggest the new role of MIF in relieving the symptom of depression by exercise, probably via enhancing neurotransmitter and neurotrophic factor in the brain.

Role of MIF and glutathione, in association with fetal ovine globin chain (Hbγ) and LPS, in induction of TNFα from cells of young and aged mice, and PBL from healthy human populations

Previous reports from our group have established that the fetal ovine gamma globin chain (Hbγ) and LPS can synergize in the induction of pro-inflammatory cytokines, especially TNFα, from mouse and human leukocytes. A fetal sheep liver extract (FSLE) which was observed to have marked immunoregulatory properties in vivo and in vitro had independently been observed to contain significant amounts of each of these molecules. However, the biological activity of this extract (hereafter FSLE) was not explained solely by its content of Hbγ and LPS, and independent analysis confirmed also the presence of migration inhibitory factor, MIF, and glutathione in FSLE. We have investigated whether MIF and the cellular anti-oxidant glutathione can further synergize with Hbγ and LPS in TNFα induction from human cells in vitro, and mouse cells activated in vivo/in vitro. Our data show that indeed there is evidence for such a synergy. Treatment or mouse cells with FSLE produced an enhanced TNFα production which could be inhibited independently both by anti-Hbγ and by anti-MIF, and optimally by a combination of these reagents.