scholarly journals The Increased RNase Activity of IRE1α in PBMCs from Patients with Rheumatoid Arthritis

2019 ◽  
Vol 9 (3) ◽  
pp. 505-509 ◽  
Author(s):  
Mahdieh Ahmadiany ◽  
Mahshid Alavi-Samani ◽  
Zahra Hashemi ◽  
Mohammad Amin Moosavi ◽  
Marveh Rahmati
Keyword(s):  
Rheumatoid Arthritis ◽  
Er Stress ◽  
Mononuclear Cells ◽  
Gradient Centrifugation ◽  
Rnase Activity ◽  
Healthy Controls ◽  
Peripheral Blood Mononuclear ◽  
Micro Rnas ◽  
Density Gradient Centrifugation Method ◽  
Gene Expression Levels

Purpose: Despite recent advances in the diagnosis and treatment of rheumatoid arthritis (RA), this inflammatory disease remains a challenge to patients and physicians. Recent evidence highlights the contribution of endoplasmic reticulum (ER) stress in the pathogenesis and treatment of RA. Herein, we study the expression of the ER stress sensor inositol-requiring enzyme 1α (IRE1α), as well as XBP1 splicing and the regulated IRE1-dependent decay (RIDD), in peripheral blood mononuclear cells (PBMCs) from patients with RA compared with healthy controls. Methods: The PBMCs from blood samples of RA patients and healthy volunteers were isolated by a density gradient centrifugation method using Ficoll. The gene expression levels of GRP78/ Bip, IRE1, XBP1s, micro-RNAs (miRNAs) were evaluated by real-time PCR. Results: The expression of GRP78, IRE1, and XBP1s were increased in PBMCs of RA patients compared with healthy controls. We further show that the RIDD targets (miRNA-17, -34a, -96, and -125b) were downregulated in RA samples. Conclusion: This study can expand our knowledge on the importance of RNase activity of IRE1α in RA and may offer new potentials for developing novel diagnostic and/or therapeutic biomarkers.

scholarly journals Using protein microarray reveals clinical correlation between self-perception of patient and the apoptosis-related proteins in rheumatoid arthritis

2020 ◽  
Author(s):  
Jian-ting Wen ◽  
Jian Liu ◽  
Hui Jiang ◽  
Lei Wan ◽  
Ling Xin ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Mononuclear Cells ◽  
Expression Profiles ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Self Perception ◽  
Roc Curve Analysis ◽  
Peripheral Blood Mononuclear ◽  
Related Proteins ◽  
Potential Biomarkers

Abstract Background: The most severe effects of rheumatoid arthritis (RA) are loss of physical function, which may have a significant impact on self-perception of patient (SPP). However, the inherent relationship between SPP and the key proteins is not clear. The aim of this study was to get an insight into SPP of RA in connection with the the apoptosis-related proteins. Methods: We set out to investigate changes of the apoptosis-related proteins expression in the peripheral blood mononuclear cells (PBMCs) of RA. Additionally, we aimed to correlate the apoptosis-related proteins expression profiles with SPP and clinical indexes. To this end, we employed antibody microarrays of the the apoptosis-related proteins in PBMCs from four RA patients and seven healthy controls. We used bioinformatics to screen several the apoptosis-related proteins. To validate key protein candidates, we performed Enzyme linked immunosorbent assay (ELISA) on 30 RA patients and 30 healthy controls. Results: We found the expression of ten the apoptosis-related proteins (caspase3, CD40, SMAC, HSP27, HTRA, IGFBP-1, IGFBP-6, sTNF-R1, sTNF-R2, TRAILR-3) were significantly altered in PBMCs of RA patients. Receiver operating characteristic (ROC) curve analysis suggested that these ten the apoptosis-related proteins are potential biomarkers of RA. Spearman Correlation analysis and Logistic-regression analysis revealed that the 10 selected the apoptosis-related proteins correlated with SPP and clinical indexes. Conclusion: Therefore, we highlight some the apoptosis-related proteins may serve as potential biomarkers in prediction of SPP for RA patients, although the underlying mechanisms need to be further explored.

scholarly journals miRNA-485-5p, an Inflammation-Related microRNA, May be a Diagnostic Biomarker of Rheumatoid Arthritis

2021 ◽  
Author(s):  
Dan Jiang ◽  
Ximing Zheng ◽  
Jian Chen ◽  
Aijun Zhang ◽  
Guangxian Xu
Keyword(s):  
Rheumatoid Arthritis ◽  
Inflammatory Cytokines ◽  
Mononuclear Cells ◽  
Roc Curves ◽  
Autoimmune Disorder ◽  
Healthy Controls ◽  
Diagnostic Biomarker ◽  
Peripheral Blood Mononuclear ◽  
Das 28 ◽  
The Impact

Abstract Background Rheumatoid arthritis (RA) is a chronic systematic autoimmune disorder that is characterized by symmetrical and inflammatory destruction of distal joints. Dysregulation of microRNAs(miRNAs) are frequently involved in inflammation, and can contribute to pathogenesis and progression of RA. This study aimed to investigate the expression of multiple inflammation-related miRNAs of RA patients and the latent mechanism, and identify novel diagnostic biomarkers. Methods Samples of 100 patients with RA and 72 healthy controls were included. The expression of predicted inflammation-related miRNAs, including miR-16, miR-17, miR-132, miR-140, miR-150, miR-181, miR-200-c, miR-203, miR-223 and miR-485-5p and RA associated genes, including IL-17, IL-18, DAS-28, MMP3, TLR-4, IRAK-4 in plasma and peripheral blood mononuclear cells (PBMCs) of RA patients compared with healthy controls (HC), were detected by qRT-PCR and ELISA. The interaction between miR-485-5p and TLR-4 or IRAK-4 was verified through dual luciferase report assays, western-blot and correlation analysis. The potential of miR-485-5p to be a biomarker for RA diagnostics was valued by ROC curves. Results Among the differentially expressed miRNAs, the expression of miR-485-5p exhibited significantly lower in plasma and PBMCs of the RA patients and was well relevant in the various body fluid samples. The expression of miR-485-5p was negatively correlated with the expression of DAS-28, IL-17, IL-18 and MMP-3, which are significant features of RA. Moreover, the ROC curve of plasma and PBMC miR-485-5p for RA revealed a high diagnostic accuracy. Furthermore, miR-485-5p could inhibit the expression of inflammatory cytokines in macrophages by targeting TLR4 and IRAK4, and the expression of miR-485-5p negatively correlated with the level of TLR4 and IRAK4 in the plasma of RA. Conclusion Collectively, our results indicated that down-expression of miR-485-5p was remarkably related to the deterioration of RA progression via the impact on inflammatory cytokines in macrophages, and may serve as a potential diagnostic biomarker and therapeutic target for RA.

scholarly journals Distinctive Expression of Bone Metabolism-related Genes between PBMCs from Condylar Hyperplasia, Rheumatoid Arthritis, and Ankylosing Spondylitis Patients

2020 ◽  
Author(s):  
Reza Amirzargar ◽  
Gholamreza Shirani ◽  
Shokoufeh Raisian ◽  
Maryam Davoudi ◽  
Saeed Aslani ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Ankylosing Spondylitis ◽  
Bone Metabolism ◽  
Bone Morphogenetic Proteins ◽  
Mononuclear Cells ◽  
Healthy Controls ◽  
Messenger Rnas ◽  
Healthy Individuals ◽  
Peripheral Blood Mononuclear ◽  
Condylar Hyperplasia

Bone morphogenetic proteins (BMPs) and wingless (Wnt) signaling molecules and their antagonists, such as sclerostin and noggin, have been identified to have different effects on bone metabolism. This research intended to evaluate the transcript levels of CTNNB1 (catenin beta 1protein), SOST (sclerostin protein), BMP4 (Bone Morphogenetic Protein 4 protein), and NOG (noggin protein) bone metabolism-related genes in peripheral blood mononuclear cells (PBMCs) from condylar hyperplasia (CH) patients in comparison to rheumatoid arthritis (RA), ankylosing spondylitis (AS), and healthy individuals. PBMCs were separated from blood samples of 10 patients with CH, AS, RA, and 10 healthy controls. SYBR Green real-time polymerase chain reaction (PCR) was used for quantitative analysis of CTNNB1, SOST, BMP4, and NOG messenger RNAs (mRNAs). The expression of CTNNB1 was significantly upregulated in CH and AS patients compared with healthy individuals and RA patients. The difference of SOST expression was not significant between all groups. The BMP4 expression was significantly downregulated in AS, CH, and RA patients compared with healthy controls. The NOG expression was downregulated in RA, AS, and CH groups, however, it was only significant in CH and RA patients compared with controls.CH and AS patients were distinguished from RA by the upregulatedCTNNB1 expression. These results demonstrated that CTNNB1, BMP4, and NOG, but not SOST, may contribute to the pathogenesis of CH, AS, and RA.

scholarly journals Fc Receptor-Like Gene Expression in Renal Transplantation Patients

2020 ◽  
Vol 9 ◽  
Author(s):  
Narges Jamshidian Tehrani ◽  
Zahra Amirghofran ◽  
Ali Reza Shamsaeefar ◽  
Aida Karachi ◽  
Mohammad Hossein Karimi
Keyword(s):  
Gene Expression ◽  
B Cells ◽  
Mononuclear Cells ◽  
Control Group ◽  
Healthy Controls ◽  
Peripheral Blood Mononuclear ◽  
Expression Levels ◽  
Transplant Patients ◽  
Blood Mononuclear Cells ◽  
Gene Expression Levels

Background: It has been well-documented that the Fc receptor-like (FCRL) molecule contributes to the pathogenesis of certain autoimmune disorders. FCRL molecules belong to the immunoglobulin superfamily produced by B cells. Also, these molecules induce activating or inhibitory signals of B cells.  According to this information and also considering the critical role of immune reactions in organ transplantation, the following experiment was performed to analyze the gene expression level of FCRLs in peripheral blood mononuclear cells of kidney transplant recipients. Materials and Methods: Blood samples were obtained from 32 renal transplant patients on days 1, 3, and 7 post-transplantations. Patients were divided into two groups according to the presence or absence of rejection. Also, 24 age-matched healthy subjects were enrolled as control group. After total RNA extraction from peripheral blood mononuclear cells (PBMC) and cDNA synthesis, the gene expression levels of FCRL1, FCRL2, and FCRL4 in each group were measured by real-time polymerase chain reaction. Results: Our results showed that FCRL1 expression levels in kidney transplant patients were significantly less than healthy controls. The overall FCRL2 expression level was not significantly different between them. However, at days 1 and 7, following transplantation in the non-rejected group FCRL2 level was significantly higher than the control group. Comparing the FCRL4 gene expression levels of both groups with healthy controls showed a significant decrease in the third and seventh days post-transplantation. Conclusion: It can be concluded that mononuclear cells, mainly B cells, have an essential role to play in kidney transplantation. [GMJ.2020;9:e1730]

scholarly journals Differentially Expressed Genes of Natural Killer Cells Can Distinguish Rheumatoid Arthritis Patients from Healthy Controls

Genes ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 492 ◽  
Author(s):  
Noha Mousaad Elemam ◽  
Mahmood Yaseen Hachim ◽  
Suad Hannawi ◽  
Azzam A. Maghazachi
Keyword(s):  
Rheumatoid Arthritis ◽  
Nk Cells ◽  
Natural Killer ◽  
Differentially Expressed Genes ◽  
Mononuclear Cells ◽  
Nk Cell ◽  
Protective Role ◽  
Differentially Expressed ◽  
Healthy Controls ◽  
Peripheral Blood Mononuclear

Rheumatoid arthritis (RA) is one of the most prevalent autoimmune diseases, while its molecular triggers are not fully understood. A few studies have shown that natural killer (NK) cells may play either a pathogenic or a protective role in RA. In this study, we sought to explore NK cell markers that could be plausibly used in evaluating the differences among healthy controls and RA patients. Publicly available transcriptome datasets from RA patients and healthy volunteers were analyzed, in order to identify differentially expressed genes (DEGs) between 1. different immune cells as compared to NK cells, and 2. NK cells of RA patients and healthy controls. The identified DEGs were validated using 16 healthy controls and 17 RA patients. Peripheral blood mononuclear cells (PBMCs) were separated by Ficoll density gradient method, while NK cells were isolated using RosetteSep technique. RNA was extracted and gene expression was assessed using RT-qPCR. All selected genes were differentially expressed in NK cells compared to PBMCs. CD56, CXCL16, PECAM-1, ITGB7, BTK, TLR10, and IL-1β were significantly upregulated, while CCL2, CCR4, RELA and IBTK were downregulated in the NK cells of RA patients when compared to healthy controls. Therefore, these NK specific genes might be used as promising biomarkers for RA diagnosis.

scholarly journals AB0107 INVESTIGATION OF THE EFFECTS OF KYNURENIC ACID ANALOGS IN RHEUMATOID ARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1352.1-1353
Author(s):  
B. Varga ◽  
A. Balog ◽  
F. Fülöp ◽  
L. Vécsei ◽  
Y. Mándi
Keyword(s):  
Rheumatoid Arthritis ◽  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Kynurenic Acid ◽  
Mononuclear Cells ◽  
Activity Score ◽  
Healthy Controls ◽  
Peripheral Blood Mononuclear ◽  
Tnf Α ◽  
Necrosis Factor

Background:The investigation of anti-inflammatory and immunosuppressive functions of kynurenic acid (KYNA) is now in focus. Previously, we demonstrated the opposite effects of KYNA and different KYNA analogs on tumor necrosis factor-α (TNF-α) production and tumor necrosis factor-stimulated gene-6 (TSG-6) expression in U-937 monocytic cells. The potential effect of KYNA analogs on further immune mediators including alarmins (S100A12=EN-RAGE and S100A8/9=calprotectin), and on human neutrofil peptide 1-3(α-defensin) production has not been investigated.Objectives:Therefore, in the present study, we compared the effects of newly synthesized KYNA analog on the TNF-α, alarmins and α-defensin production, correlation with the effects on the TSG-6 expression in rheumatoid arthritis (RA).Methods:93 RA patients were involved and divided subgroups based on DAS28 activity score. Peripheral blood mononuclear cells (PBMC) was isolated from RA patients and healthy controls. As cytokine inducers heat inactivatedStaphylococcus aureus(SA1) were used. In parallel in vitro experiments, the SA1 induced PBMCs were pretreated with a newly synthesized KYNA analog (compound SZR-72 was synthesized by direct amidation of KYNA). The concentrations of the above mentioned inflammatory mediators in the supernatants were quantified by using ELISA kits and the TSG-6 expression was also determined by RTqPCR method.Results:The SA1 induced TNF-α, EN-RAGE, calprotectin and α-defensin production was significantly higher in RA patients’ group than in healthy controls. KYNA analog attenuated the SA1 induced TNF-α, EN-RAGE, calprotectin and α-defensin production, and increased TSG-6 production and TSG-6 mRNA expression in PBMC cells from RA patients. The SA1 induced TNF-α and TSG-6 production correlated with the DAS28 activity score. The TNF-α inhibitory effect of the KYNA analog correlated inversely with the TSG-6 stimulatory effect in all subgroups of RA patients based on DAS28 activity score.Conclusion:TSG-6 expression could participate in the suppression of inflammatory cytokines, such as TNF-α, EN-RAGE, calprotectin and α-defensin. We suppose that the elevation of the TSG-6 expression by KYNA and especially by new KYNA analogs might be one of the mechanisms that are responsible for their suppressive effect on TNF-α production as a feedback mechanism in RA. KYNA and KYNA analogs have an important role in influencing TSG-6 expression, and there is a possible benefit with potential therapeutic consequence of targeting TSG-6 expression by kynurenines in inflammatory conditions in RA.References:[1]Mándi Y, Endrész V, Mosolygó T, Burián K, Lantos I, Fülöp F, Szatmári I, Lőrinczi B, Balog A, Vécsei L, The Opposite Effects of Kynurenic Acid and Different Kynurenic Acid Analogs on Tumor Necrosis Factor-a (TNF-a) Production and Tumor Necrosis Factor-Stimulated Gene-6 (TSG-6) Expression. Frontiers in Immunology. doi: 10.3389/fimmu.2019.01406Acknowledgments:This work was supported by GINOP 2.3.2-2015-16-00034Disclosure of Interests:None declared

scholarly journals OP0033 REGULATORY T CELL CD39 EXPRESSION AS A PREDICTOR OF EARLY REMISSION-INDUCTION WITH METHOTREXATE IN NEW-ONSET RHEUMATOID ARTHRITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 18.2-18
Author(s):  
P. Brown ◽  
A. Anderson ◽  
B. Hargreaves ◽  
A. Morgan ◽  
J. D. Isaacs ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
T Cells ◽  
Cd4 T Cells ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Disease Modifying Therapy ◽  
Blood Mononuclear Cells ◽  
Treatment Naïve ◽  
Disease Modifying ◽  
Pre Treatment

Background:The long term outcomes for patients with rheumatoid arthritis (RA) depend on early and effective disease control. Methotrexate remains the key first line disease modifying therapy for the majority of patients, with 40% achieving an ACR50 on monotherapy(1). There are at present no effective biomarkers to predict treatment response, preventing effective personalisation of therapy. A putative mechanism of action of methotrexate, the potentiation of anti-inflammatory adenosine signalling, may inform biomarker discovery. By antagonism of the ATIC enzyme in the purine synthesis pathway, methotrexate has been proposed to increase the release of adenosine moieties from cells, which exert an anti-inflammatory effect through interaction with ADORA2 receptors(2). Lower expression of CD39 (a cell surface 5-’ectonucleotidase required for the first step in the conversion of ATP to adenosine) on circulating regulatory T-Lymphocytes (Tregs) was previously identified in patients already established on methotrexate who were not responding (DAS28 >4.0 vs <3.0)(3). We therefore hypothesised that pre-treatment CD39 expression on these cells may have clinical utility as a predictor of early methotrexate efficacy.Objectives:To characterise CD39 expression in peripheral blood mononuclear cells in RA patients naïve to disease modifying therapy commencing methotrexate, and relate this expression to 4 variable DAS28CRP remission (<2.6) at 6 months.Methods:68 treatment naïve early RA patients starting methotrexate were recruited from the Newcastle Early Arthritis Clinic and followed up for 6 months. Serial blood samples were taken before and during methotrexate therapy with peripheral blood mononuclear cells isolated by density centrifugation. Expression of CD39 by major immune subsets (CD4+ and CD8+ T-cells, B-lymphocytes, natural killer cells and monocytes) was determined by flow cytometry. The statistical analysis used was binomial logistic regression with baseline DAS28CRP used as a covariate due to the significant association of baseline disease activity with treatment response.Results:Higher pre-treatment CD39 expression was observed in circulating CD4+ T-cells of patients who subsequently achieved clinical remission at 6 months versus those who did not (median fluorescence 4854.0 vs 3324.2; p = 0.0108; Figure 1-A). This CD39 expression pattern was primarily accounted for by the CD4+CD25 high sub-population (median fluorescence 9804.7 vs 6455.5; p = 0.0065; Figure 1-B). These CD25 high cells were observed to have higher FoxP3 and lower CD127 expression than their CD39 negative counterparts, indicating a Treg phenotype. No significant associations were observed with any other circulating subset. A ROC curve demonstrates the discriminative utility of differential CD39 expression in the CD4+CD25 high population for the prediction of DAS28CRP remission in this cohort, showing greater specificity than sensitivity for remission prediction(AUC: 0.725; 95% CI: 0.53 - 0.92; Figure 1-C). Longitudinally, no significant induction or suppression of the CD39 marker was observed amongst patients who did or did not achieve remission over the 6 months follow-up period.Figure 1.Six month DAS28CRP remission versus pre-treatment median fluorescence of CD39 expression on CD4+ T-cells (A); CD25 High expressing CD4+ T-cells (B); and ROC curve of predictive utility of pre-treatment CD39 expression on CD25 High CD4+ T-cells (C).Conclusion:These findings support the potential role of CD39 in the mechanism of methotrexate response. Expression of CD39 on circulating Tregs in treatment-naïve RA patients may have particular value in identifying early RA patients likely to respond to methotrexate, and hence add value to evolving multi-parameter discriminatory algorithms.References:[1]Hazlewood GS, et al. BMJ. 2016 21;353:i1777[2]Brown PM, et al. Nat Rev Rheumatol. 2016;12(12):731-742[3]Peres RS, et al. Proc Natl Acad Sci U S A. 2015;112(8):2509-2514Disclosure of Interests:None declared

scholarly journals Impact of SARS-CoV-2 infection on the recovery of peripheral blood mononuclear cells by density gradient

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Maria D. I. Manunta ◽  
Giuseppe Lamorte ◽  
Francesca Ferrari ◽  
Elena Trombetta ◽  
Mario Tirone ◽  
...  
Keyword(s):  
Density Gradient ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Early Phase ◽  
Mononuclear Cells ◽  
Early Stage ◽  
Gradient Centrifugation ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Circulating Lymphocytes

AbstractSARS-CoV-2 virus infection is responsible for coronavirus disease (COVID-19), which is characterised by a hyperinflammatory response that plays a major role in determining the respiratory and immune-mediated complications of this condition. While isolating peripheral blood mononuclear cells (PBMCs) from whole blood of COVID-19 patients by density gradient centrifugation, we noticed some changes in the floating properties and in the sedimentation of the cells on density medium. Investigating this further, we found that in early phase COVID-19 patients, characterised by reduced circulating lymphocytes and monocytes, the PBMC fraction contained surprisingly high levels of neutrophils. Furthermore, the neutrophil population exhibited alterations in the cell size and in the internal complexity, consistent with the presence of low density neutrophils (LDNs) and immature forms, which may explain the shift seen in the floating abilities and that may be predictive of the severity of the disease. The percentage of this subset of neutrophils found in the PBMC band was rather spread (35.4 ± 27.2%, with a median 28.8% and IQR 11.6–56.1, Welch’s t-test early phase COVID-19 versus blood donor healthy controls P < 0.0001). Results confirm the presence of an increased number of LDNs in patients with early stage COVID-19, which correlates with disease severity and may be recovered by centrifugation on a density gradient together with PBMCs.

Sign in / Sign up

Export Citation Format

Share Document