Circulating soluble RAGE and cell surface RAGE on peripheral blood mononuclear cells in healthy children

2018 ◽  
Vol 31 (6) ◽  
pp. 649-654 ◽  
Author(s):  
Alberto García-Salido ◽  
Gustavo Melen ◽  
Vanesa Gómez-Piña ◽  
Gonzalo Oñoro-Otero ◽  
Ana Serrano-González ◽  
...  
Keyword(s):  
Negative Correlation ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Adult Population ◽  
Critical Role ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Children ◽  
Peripheral Blood Mononuclear ◽  
Srage Level ◽  
Soluble Rage

Abstract Background: The receptor for advanced glycation end products (RAGE) has a critical role in the pathogenesis of inflammation. In healthy children, its basal expression on the peripheral blood mononuclear cell (PBMC) and the basal circulating soluble RAGE (sRAGE) levels are unknown. The aim of this study was to describe both. Methods: This is a monocentric, observational and descriptive study of samples obtained from healthy children. The RAGE expression on PBMC was analyzed using flow cytometry. The sRAGE values were determined with a specific sandwich enzyme-linked immunosorbent assay (ELISA) kit, later the relation between cellular RAGE and sRAGE was described. Results: Forty-three children were included. The median sRAGE level was 849.0±579.0 pg/mL. The RAGE mean fluorescence intensity (MFI) was 1382±506 in monocytes and 792±506 in lymphocytes. There were no differences between genders. A negative correlation was found between sRAGE and RAGE MFI in lymphocytes (r=−0.3; p=0.04). Conclusions: We describe for the first time the RAGE surface levels on PBMC in children. It showed a negative correlation with sRAGE. The sRAGE circulating level is lower than the sRAGE level described in adult population or non-healthy children. Our findings should be confirmed in order to apply them as reference values for future investigations.

Immune dysregulation in peripheral blood mononuclear cells from patients with liver cancer.

2018 ◽  
Vol 36 (5_suppl) ◽  
pp. 55-55
Author(s):  
Juhua Zhou
Keyword(s):  
Liver Cancer ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
Critical Role ◽  
Immune Dysregulation ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Underlying Mechanisms

55 Background: Immune regulation may play an important role in cancer development. The immune dysregulation and underlying mechanisms in patients with liver cancer have not fully understood. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from patients with liver cancer. Deep-sequencing, FACS analysis, ELISA and real-time PCR were used to analyze the immune dysregulation and underlying mechanisms in patients with liver cancer. Results: Our analysis discovered that the percentages of immune cell populations in PBMC from patients with liver cancer were significantly different from those in normal controls. Specifically, the percents of B cells and regulatory T cells were high in PBMC from patients with liver cancer. Expression of 161 genes such as EGF, IRF3, CD177, MAP2K2 and MMP9 was found to be significantly inhibited, but 66 genes including CD19, CD70, FOXP1 and IL-32 were significantly up-regulated in PBMC from patients with liver cancer. Further analysis showed that 190 long non-coding RNAs (lncRNAs) were significantly down-regulated; whereas150 lncRNAs were significantly up-regulated in PBMC from patients with liver cancer. Dysregulated lncRNAs were involved in the control of immune cell signaling, cell division and differentiation. Conclusions: The results suggest that the immune dysregulation may play a critical role in the pathogenesis of liver cancer. It also has a great implication in the development of immune therapeutic methods for patients with liver cancer.

scholarly journals Viability and Functionality of Cryopreserved Peripheral Blood Mononuclear Cells in Pediatric Dengue

2016 ◽  
Vol 23 (5) ◽  
pp. 417-426 ◽  
Author(s):  
Federico Perdomo-Celis ◽  
Doris M. Salgado ◽  
Diana M. Castañeda ◽  
Carlos F. Narváez
Keyword(s):  
Acute Phase ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Dengue Infection ◽  
Healthy Children ◽  
Peripheral Blood Mononuclear ◽  
Convalescent Phase ◽  
Blood Mononuclear Cells ◽  
Ifn Γ

ABSTRACTCryopreserved peripheral blood mononuclear cells (PBMCs) are widely used in studies of dengue. In this disease, elevated frequency of apoptotic PBMCs has been described, and molecules such as soluble tumor necrosis factor (TNF)-related apoptosis-inducing ligands (sTRAIL) are involved. This effect of dengue may affect the efficiency of PBMC cryopreservation. Here, we evaluate the viability (trypan blue dye exclusion and amine-reactive dye staining) and functionality (frequency of gamma interferon [IFN-γ]-producing T cells after polyclonal stimulation) of fresh and cryopreserved PBMCs from children with dengue (in acute and convalescence phases), children with other febrile illnesses, and healthy children as controls. Plasma sTRAIL levels were also evaluated. The frequencies of nonviable PBMCs detected by the two viability assays were positively correlated (r= 0.74;P< 0.0001). Cryopreservation particularly affected the PBMCs of children with dengue, who had a higher frequency of nonviable cells than healthy children and children with other febrile illnesses (P≤ 0.02), and PBMC viability levels were restored in the convalescent phase. In the acute phase, an increased frequency of CD3+CD8+amine-positive cells was found before cryopreservation (P= 0.01). Except for B cells in the acute phase, cryopreservation usually did not affect the relative frequencies of viable PBMC subpopulations. Dengue infection reduced the frequency of IFN-γ-producing CD3+cells after stimulation compared with healthy controls and convalescent-phase patients (P≤ 0.003), and plasma sTRAIL correlated with this decreased frequency in dengue (rho = −0.56;P= 0.01). Natural dengue infection in children can affect the viability and functionality of cryopreserved PBMCs.

Cardiopulmonary Bypass Decreases G Protein–Coupled Receptor Kinase Activity and Expression in Human Peripheral Blood Mononuclear Cells

2003 ◽  
Vol 98 (2) ◽  
pp. 343-348 ◽  
Author(s):  
Scott A. Hagen ◽  
Amy L. Kondyra ◽  
Hilary P. Grocott ◽  
Habib El-Moalem ◽  
Daniel Bainbridge ◽  
...  
Keyword(s):  
Cardiopulmonary Bypass ◽  
G Protein ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
G Protein Coupled Receptor ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
G Protein Coupled

Background Cardiopulmonary bypass (CPB) has been implicated in the development of organ injury associated with cardiac surgery. At the molecular level, CPB is accompanied by a pronounced proinflammatory response including an increase in plasma interleukin (IL)-6. The IL-6 has been shown to be increased in rheumatoid arthritis, a chronic inflammatory disease, where it has been implicated in decreasing G protein-coupled receptor kinases (GRKs) in peripheral blood mononuclear cells. Since IL-6 is substantially increased after CPB, the study tested whether the increase of IL-6 during CPB leads to a decrease of GRKs in mononuclear cells. This is important because GRKs regulate the function of G protein-coupled receptors involved in inflammation. Methods Fifteen patients had blood withdrawn before CPB, 2 h after CPB, and on postoperative day one (POD1). Plasma IL-6 concentrations were determined by enzyme-linked immunosorbent assay. The GRK protein expression and activity were determined by Western blot and phosphorylation of rhodopsin using [gamma-(32)P] adenosine triphosphate, respectively. Results Plasma IL-6 increased over 20-fold after CPB and remained increased on POD1. Cytosolic GRK activity in mononuclear cells decreased by 39 +/- 29%; cytosolic GRK2 and membrane-bound GRK6 decreased by 90 +/- 15 and 65 +/- 43%, respectively. The GRK activity and expression of GRK2/GRK6 on POD1 returned to basal levels in many but not all patients. Conclusions The CPB causes a profound decrease in mononuclear cell GRKs, and the recovery of these kinases on POD1 is quite variable. The significance of the variable recovery of GRKs after CPB and their potential role as a marker of clinical outcome deserves further investigation.

scholarly journals Evaluation of the Effect of Epigallocatechin-3-gallate (EGCG) on HIF-1α Protein and RORC Gene Expression in Peripheral Blood Mononuclear Cells of Patients with Multiple Sclerosis

2021 ◽  
Author(s):  
Freshteh Alsahebfosoul ◽  
◽  
Boshra Afshar ◽  
Mazdak Ganjalikhani-Hakemi ◽  
Zahra Khalifezadeh Esfahani ◽  
...  
Keyword(s):  
Gene Expression ◽  
Multiple Sclerosis ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Th17 Cells ◽  
Mononuclear Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Anti Inflammatory

Background: Multiple sclerosis has been considered as chronic inflammation of the central nervous system (CNS) and autoimmune disease .MS is most widely considered to be mediated by activation of myelin-specific T CD4+ cells as well as TH1 and TH17 cells. TH17 cell has been involved in the pathogenesis of MS in various ways. HIF-1α and RORC are required for natural differentiation of TH17 and are essential transcription factors for the evolution of TH17 cells. Numerous studies indicate that epigallocatechin gallate (EGCG) has immunomodulatory and anti-inflammatory effects. Aims: This study investigated the effect of EGCG on normoxic HIF-1α and RORC2 expression in PBMCs of MS patients. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood of new cases MS patients. The cells cultured in the presence of a different concentration of EGCG (25, 50,100μM) for 18 and 48 hours. Afterward, HIF-1α and RORC2 level expressions were measured by enzyme-linked immunosorbent assay (ELISA) and Real-Time PCR, respectively. Result: The results showed that EGCG significantly decrease RORC2 gene expression. However, EGCG did not influence the level of HIF-1α. Our present data has led us to conclude that EGCG could be considered as an anti-inflammatory agent may serving as an achievable therapeutic agent for MS.

scholarly journals Simultaneous Detection of 15 Human Cytokines in a Single Sample of Stimulated Peripheral Blood Mononuclear Cells

2003 ◽  
Vol 10 (1) ◽  
pp. 133-139 ◽  
Author(s):  
Wilco de Jager ◽  
Henk te Velthuis ◽  
Berent J. Prakken ◽  
Wietse Kuis ◽  
Ger T. Rijkers
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Simultaneous Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Human Cytokines

ABSTRACT Cytokines secreted by cells of the immune system can alter the behavior and properties of immune or other cells. At a site of inflammation, sets of cytokines interact with immune cells, and their combined effect is often more important than the function of one isolated component. Conventional techniques, such as enzyme-linked immunosorbent assays, generally require large quantities of cells to characterize a complete cytokine profile of activated lymphocytes. The Bio-Plex system from Bio-Rad Laboratories combines the principle of a sandwich immunoassay with the Luminex fluorescent-bead-based technology. We developed a multiplex cytokine assay to detect different cytokines simultaneously in culture supernatant of human peripheral blood mononuclear cells stimulated with antigen and with mitogen. Fifteen human cytokines (interleukin 1α [IL-1α], IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-18, gamma interferon, and tumor necrosis factor alpha) were validated with a panel of healthy individuals, rheumatoid arthritis patients, and juvenile idiopathic arthritis patients. Comparing the multiplex assay with a regular enzyme-linked immunosorbent assay technique with this donor panel resulted in correlation coefficients for all cytokines ranging from 0.75 to 0.99. Intra-assay variance proved to be less then 10%, whereas interassay variability ranged between 10 and 22%. This multiplex system proved to be a powerful tool in the quantitation of cytokines. It will provide a more complete picture in differences between activated lymphocyte cytokine profiles from healthy individuals and those from patients with chronic inflammatory diseases.

scholarly journals Detection of Brucellae in peripheral blood mononuclear cells for monitoring therapeutic efficacy of brucellosis infection

2019 ◽  
Vol 8 (1) ◽  
Author(s):  
Heng Yang ◽  
Guoxia Zhang ◽  
Peifang Luo ◽  
Zuoping He ◽  
Feihuan Hu ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Significant Risk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Sensitive Assay ◽  
Peripheral Blood Mononuclear ◽  
Blood Samples ◽  
Blood Mononuclear Cells

Abstract Background Brucellosis is one of the most severe widespread zoonoses caused by the Gram-negative bacterium Brucella species. The diagnosis and clinical assessment of human brucellosis are very important for the management of patients, while there is a lack of effective methods to detect Brucellae. Classical culture of Brucella species is time consuming and often fails. A simple and sensitive assay is needed for diagnosis of Brucella infection and monitoring of treatment in man. Methods Blood samples and peripheral blood mononuclear cells (PBMCs) were collected from 154 patients hospitalized for brucellosis. Brucella antibodies were detected by Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (SAT) and enzyme-linked immunosorbent assay (ELISA). Intracellular Brucellae were detected by blood culture and immunofluorescence staining (IFS). Results Among 154 brucellosis patients, 59.7% (92/154) were antibody reactive by RBPT, 81.8% (126/154) by SAT and 95.5% (147/154) by ELISA, respectively. Only 3.2% (5/154) of patient blood samples resulted in positive Brucella culture, while 68.8% (106/154) carried IFS detectable Brucella antigens in PBMCs. Gender (P = 0.01) but not age (P > 0.05) was a significant risk factor. The frequency of intracellular Brucella antigens was similar between patients receiving different treatment regimens (P > 0.05). However, a significant decrease of intracellular Brucellae was observed only in patients with acute brucellosis after the third course of treatment (P < 0.05), suggesting that current regimens to treat chronic brucellosis were not effective. Conclusions IFS appears a sensitive assay for detection of Brucella antigens in PBMCs and could be used for diagnosis and therapeutic monitoring of brucellosis in clinical practice.

scholarly journals Coding and non-coding RNA interactions reveal immune-related pathways in peripheral blood mononuclear cells derived from patients with proliferative vitreoretinopathy

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yao Ni ◽  
Fangyuan Liu ◽  
Xiao Hu ◽  
Yingyan Qin ◽  
Zhaotian Zhang
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Proliferative Vitreoretinopathy ◽  
Mononuclear Cells ◽  
Critical Role ◽  
Differentially Expressed ◽  
Prognostic Indicators ◽  
Related Factors ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Abstract Background Peripheral immune response has been revealed to play a critical role in proliferative vitreoretinopathy (PVR). However, the reliable immune-related factors that are acting as prognostic indicators or therapeutic targets for PVR remain to explore further. Methods In the current study, we applied whole-transcriptome sequencing to profile peripheral blood mononuclear cells from PVR patients and also analyzed lncRNA-mRNA interactions in peripheral immune cells to explore the pathways that might mediate immunopathology and resultant retinal damage in PVR. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses and Ingenuity Pathway Analysis (IPA) were employed to classify the function of these differentially expressed genes. Results Compared to the controls, there were 319 genes upregulated, and 191 genes downregulated in PVR patients. GO, and KEGG enrichment analyses as well as IPA showed that these upregulated genes were significantly enriched in immune-related and infection-relate terms. Immune-related gene NFKBIA, CXCL2, and CXCL8 were detected as hub-genes in the co-expression network, while lncRNAs such as AC007032.1, AC037198.2, AL929472.2, and SLED1 were highly co-expressed with them. lncRNA-mRNA interactions analysis also showed that putative targeted genes of these differentially expressed lncRNAs were also significantly enriched in immune-related or infection-relate pathways. Conclusion Our study highlights the transformation of immune-related genes/pathways in PVR by comparing controls, and validates several critical genes and lncRNAs, which are serving as potential diagnostic markers for PVR patients.

scholarly journals Iron-Quercetin Complex Preconditioning Human Peripheral Blood Mononuclear Cells Accelerates Angiogenic and Wound Healing Efficacy

2021 ◽  
Author(s):  
Jiraporn Kantapan ◽  
Nampeung Anukul ◽  
Nipapan Leetrakool ◽  
Gwenaël Rolin ◽  
Jackie Vergote ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Network Formation ◽  
Mononuclear Cells ◽  
Bioactive Molecules ◽  
Enzyme Linked Immunosorbent Assay ◽  
Therapeutic Effects ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Abstract Background: Cells-based therapy is a highly promising treatment paradigm in ischemic disease due to its ability to repair tissues when implanted into a damaged site. These therapeutic effects have been involving a strong paracrine component resulting from the high levels of bioactive molecules they secrete in response to the local microenvironment. Therefore, the secreted therapeutic can be modulated by preconditioning the cells during in vitro culture. Herein, we investigated the potential use of magnetic resonance imaging (MRI) probes “Iron-Quercetin complex” or IronQ for preconditioning peripheral blood mononuclear cells (PBMCs) to expand proangiogenic cells and enhance their secreted therapeutic factors. Methods: PBMCs obtained from healthy donor blood were cultured in the presence of the Iron-Quercetin complex. Preconditioning-PBMCs differentiated cells were characterized by immunostaining. An enzyme-linked immunosorbent assay to describe the secreted cytokines. In vitro migration and tubular formation using human umbilical vein endothelial cells (HUVECs) were completed to investigate the proangiogenic efficacy.Results: IronQ significantly increased mononuclear progenitor cells' proliferation and differentiation into the spindle-shape-like cells, expressing both hematopoietic and stromal cell markers. The expansion increased the number of colony-forming units (CFU-Hill). The conditioned medium obtained from IronQ-treated PBMCs contained a high level of Interleukin (IL)-8, IL-10, urokinase-type-plasminogen-activator (uPA), matrix metalloproteinases-9 (MMP-9), and tumor necrosis factor-alpha (TNF-α), and augmented migration and capillary network formation of HUVEC and fibroblast cells in vitro.Conclusions: Our study demonstrated that the IronQ-precondition PBMCs protocol could enhance the angiogenic and reparative potential of non-mobilized PBMCs. This protocol can be used as an adjunctive strategy to improve cell therapy's efficacy of PBMCs for ischemic diseases and chronic wound.

scholarly journals Peripheral Blood NRF2 Expression as a Biomarker in Human Health and Disease

Antioxidants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 28
Author(s):  
Lee E. Neilson ◽  
Joseph F. Quinn ◽  
Nora E. Gray
Keyword(s):  
Clinical Trials ◽  
Peripheral Blood ◽  
Target Genes ◽  
Mononuclear Cells ◽  
Critical Role ◽  
Related Factor ◽  
Peripheral Blood Mononuclear ◽  
Duration Of Action ◽  
Nrf2 Activation ◽  
Health And Disease

Nuclear factor erythroid 2-related factor 2 (NRF2), a transcription factor which plays a critical role in maintenance of cellular redox, has been identified as a therapeutic target in a number of human diseases. Several reports have demonstrated beneficial effects of NRF2 manipulation in animal models of disease, and one NRF2-activating drug, dimethyl fumarate, is already approved for the treatment of multiple sclerosis. However, drug discovery is slowed due to a dearth of biomarkers which can inform target engagement and magnitude and duration of action. Peripheral blood mononuclear cells (PBMCs) are an accessible, minimally-invasive source of biomarkers which can be readily assayed and objectively monitored as a surrogate endpoint of NRF2 activation in clinical trials. We undertook a review of the literature on PBMC NRF2 measurements in human studies to explore its role as a suitable biomarker in various contexts of health and disease. It is clear that NRF2 and its target genes can be readily assayed from PBMCs in multiple disease contexts and may track with disease progression. Further work needs to be undertaken to evaluate its stability but should be considered as an exploratory marker in clinical trials targeting NRF2 activation.

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