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Amporn Suphatrakul, Thaneeya Duangchinda, Natapong Jupatanakul, Kanjanawadee Prasittisa, Suppachoke Onnome, Jutharat Pengon, Bunpote Siridechadilok
Reporter virus is a versatile tool to visualize and to analyze virus infections. However, for flaviviruses, it is difficult to maintain the inserted reporter genes on the viral genome, limiting its use in several studies that require homogeneous virus particles and several rounds of virus replication. Here, we showed that flanking inserted GFP genes on both sides with ribosome-skipping 2A sequences improved the stability and the consistency of their fluorescent signals for dengue-virus-serotype 2 (DENV2) reporter viruses...
2018: PloS One
Keiichi Koizumi, Minako Hoshiai, Nobuyuki Katsumata, Takako Toda, Hiroaki Kise, Yohei Hasebe, Yosuke Kono, Yuto Sunaga, Masashi Yoshizawa, Atsushi Watanabe, Keiko Kagami, Masako Abe, Kanji Sugita
BACKGROUND: The effect of infliximab (IFX) on immune cells has not been fully reported in Kawasaki disease (KD). To investigate the mechanism of IFX in KD, we examined changes in the abundance of CD14+ CD16+ activated monocytes, regulatory T cells (Treg ), and T-helper cell type 17 (Th17) cells following treatment with IFX. METHODS: We collected peripheral blood from patients with intravenous immunoglobulin (IVIG)-resistant KD and analyzed the absolute counts of CD14+ CD16+ monocytes, Treg cells (CD4+ CD25+ FOXP3+ ) and Th17 cells (CD4+ IL-17A+ ) by flow cytometry...
March 15, 2018: Pediatrics International: Official Journal of the Japan Pediatric Society
Huan Yan, Hong Li, Linlin Zhu, Junjun Gao, Pengyun Li, Zhan Zhang
OBJECTIVE: Increased inflammation is considered as a risk factor and a promoter of preterm birth (PTB). Monocytes and neutrophils are the main sources of cytokines in the early inflammatory phase. So far, very few studies have indicated CD14/ TLR4 and TREM-1 on the monocytes and neutrophils as important targets in PTB. MATERIALS AND METHODS: TLR4 and TREM-1 on CD14+ maternal and cord blood monocytes and neutrophils were detected using flow cytometry in 48 normal term women, 48 PTB with chorioamnionitis (CCA) women and 40 PTB without CCA women...
March 13, 2018: Journal of Maternal-fetal & Neonatal Medicine
Mirhane Hassan, Hala M Raslan, Hesham Gamal Eldin, Eman Mahmoud, Hanaa Alm-Elhuda Abd Elwajed
INTRODUCTION: Type 1 Diabetes Mellitus (T1D) is an autoimmune disease that results from the destruction of insulin-producing beta cells of the pancreas by autoreactive T cells. Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells that can potently suppress T cell responses. AIM: To detect the presence of MDSCs in T1D and compare their percentage in T1D versus healthy individuals. METHOD: Thirty T1D patients were included in the study...
February 15, 2018: Open Access Macedonian Journal of Medical Sciences
Hsien-Tzung Liao, Tzu-Hao Li, Chun-Hsiung Chen, Hung-An Chen, Wei-Sheng Chen, Chien-Chih Lai, Chung-Tei Chou, Chang-Youh Tsai
BACKGROUND/PURPOSE: To investigate the Janus kinase-1 and 3 (JAK-1 and 3) expression in peripheral blood mononuclear cells (PBMCs) in ankylosing spondylitis (AS). METHODS: The levels of JAK-1 and JAK-3 mRNA in PBMCs, CD3+ T cells and CD14+ monocytes were measured by RT-PCR in 52 AS patients and 31 healthy controls (HCs). The demographic features, BASDAI, BASFI, HLA-B27, ESR, CRP and serum immunoglobulin A (IgA) level were recorded and correlated with the JAK-1 & JAK-3 transcripts in patients and HCs as appropriate...
March 9, 2018: Journal of the Formosan Medical Association, Taiwan Yi Zhi
Carlos Zamora, Elisabet Canto, Juan C Nieto, Ester Garcia-Planella, Jordi Gordillo, Mª Angels Ortiz, Xavier Suarez-Calvet, Lidia Perea, Germà Julia, Candido Juarez, Silvia Vidal
Background: Circulating monocytes from active ulcerative colitis (UC) patients produced high levels of tumor necrosis factor-alpha(TNFα) and interleukin(IL)-6 after Toll-like receptors (TLR) stimulation. Since platelets (PLT) can bind to leukocytes, thereby decreasing inflammatory cytokine production, UC patients may exhibit different levels of monocyte-platelet complexes depending on disease activity. Methods: We compared among healthy donors, active (onset flare and relapse), and inactive UC patients the presence of circulating monocyte-platelet complexes (CD14+PLT+) and membrane CD162 expression by flow cytometry...
February 23, 2018: Inflammatory Bowel Diseases
Masaru Nakagawa, Sigeyuki Uno, Noriyoshi Iriyama, Manabu Matsunawa, Makoto Makishima, Jin Takeuchi, Isao Tsuboi, Yoshihiro Hatta, Masami Takei
Benzo[a]pyrene (BaP) is an environmental pollutant found in cigarette smoke and is implicated as a causative agent of tobacco-related diseases, such as arteriosclerosis. In contrast, vitamin D signaling, which is principally mediated by conversion of vitamin D to the active form, 1α,25-dihydroxyvitamin D3 [1,25(OH)2 D3 ], decreases cardiovascular disease risk. However, combined treatment with BaP and 1,25(OH)2 D3 enhances BaP toxicity, including BaP-DNA adduct formation. We further investigated the cross-talk between BaP and 1,25(OH)2 D3 signaling pathways, and found that combined treatment with these compounds induces mRNA and protein expression of plasminogen activator inhibitor 1 (PAI-1) in monocyte/macrophage-derived THP-1 and U937 cells...
March 7, 2018: Toxicology and Applied Pharmacology
Saverio G Parisi, Samantha Andreis, Carlo Mengoli, Nicola Menegotto, Silvia Cavinato, Renzo Scaggiante, Massimo Andreoni, Giorgio Palù, Monica Basso, Anna Maria Cattelan
Soluble CD163, soluble CD14 and cellular HIV-1-DNA levels reflect two different aspects of HIV infection: immune activation and the reservoir of infected cells. The aim of this study was to describe their relationships in a cohort of HIV-HCV co-infected patients successfully treated for both HCV and HIV infections. Fifty-five patients were recruited and studied prior to the start of direct-acting antivirals (DAAs) (T0), at week 12 of DAA treatment (T1) and 24 weeks after T0 (T2). The subjects were classified as having undetectable plasma HIV viraemia (UV) or low-level viraemia (LLV) in the 18 months before T2...
March 9, 2018: Medical Microbiology and Immunology
Sally A F El-Sahrigy, Azza M O Abdel Rahman, Dalia Y Samaha, Nesrine A Mohamed, Sally M Saber, Hala A Talkhan, Ghada A Ismail, Essam M Ibraheem, Emad M Riad
INTRODUCTION: Tuberculosis (TB) remains a huge worldwide burden, despite extensive vaccination coverage with the Bacillus Calmette-Guérin (BCG), the only vaccine available against this disease, indicating that BCG-driven immunity is inadequate to protect the human population against TB. This underscore the critical necessitate to develop an improved TB vaccine, based on a better understanding of host-pathogen interactions and immune responses during mycobacterial infection. AIM OF THE WORK: To examine whether the exogenous addition of IFN-β could improve dendritic cell (DC) response to Mycobacterium bovis (M...
March 6, 2018: Journal of Immunological Methods
Xinhua Zhan, Boryana Stamova, Frank R Sharp
This review proposes that lipopolysaccharide (LPS, found in the wall of all Gram-negative bacteria) could play a role in causing sporadic Alzheimer's disease (AD). This is based in part upon recent studies showing that: Gram-negative E. coli bacteria can form extracellular amyloid; bacterial-encoded 16S rRNA is present in all human brains with over 70% being Gram-negative bacteria; ultrastructural analyses have shown microbes in erythrocytes of AD patients; blood LPS levels in AD patients are 3-fold the levels in control; LPS combined with focal cerebral ischemia and hypoxia produced amyloid-like plaques and myelin injury in adult rat cortex...
2018: Frontiers in Aging Neuroscience
Crystal C Walline, Janice S Blum, Tobyn Linton, Darrin Mangiacarne, Suthat Liangpunsakul
Excessive drinking can lead to the development of immune dysfunction. Our aim is to investigate the effect of alcohol on immune activation from circulating peripheral blood monocytes in excessive drinkers (EDs). Twenty-two EDs and healthy controls were enrolled. Time line follow-back was used to quantify the amount of alcohol consumed in the past 30 days before enrollment. Peripheral blood-derived CD14+ monocytes were isolated for gene expression analyses. Serum interleukin (IL)-6, IL-10 and lipopolysaccharides (LPS) were also measured...
March 8, 2018: Journal of Investigative Medicine: the Official Publication of the American Federation for Clinical Research
Luca Ferrari, Elena Canelli, Elena De Angelis, Alessia Catella, Giulia Ferrarini, Giulia Ogno, Luca Bonati, Roberto Nardini, Paolo Borghetti, Paolo Martelli
Highly pathogenic (HP) PRRSV isolates have been discovered within both PRRSV-1 and PRRSV-2 genotypes and investigated in recent years especially for their ability to cause extremely severe disease in conventional pig herds. The exacerbation of general and respiratory clinical signs has been attributed not only to an efficient replication (virulence) but also to the ability to dysregulate viral recognition and induce mechanisms of immune evasion or immune enhancement of humoral and cellular anti-viral responses differently from non-HP PRRSV isolates in terms of intensity and temporal onset...
March 2018: Veterinary Microbiology
Manli Zhang, Lili Ding, Xinrui Wang, Jie Hou, Man Li, Yanfang Jiang, Xiuting He, Meizi Cui, Feng Hu, Xiuna Zhang, Jingyuan Yang, Xiaohe Guo, Huiying Zhao, Pujun Gao
BACKGROUND: Despite the role of monocytes in the pathogenesis of severe acute pancreatitis (SAP), it remains unclear how different subtypes of monocytes regulate and contribute to this pathogenesis. METHODS: We examined the numbers of different subsets of monocytes by flow cytometry in 21 SAP, 15 mild acute pancreatitis (MAP) and 13 healthy controls (HC). The concentrations of plasma cytokines were assessed by cytometric bead array. Disease severity was evaluated based on the acute physiology and chronic health evaluation (APACHE) II score and plasma C-reactive proteins (CRP) levels...
March 5, 2018: International Immunopharmacology
K Tabeta, M Hosojima, M Nakajima, S Miyauchi, H Miyazawa, N Takahashi, Y Matsuda, N Sugita, Y Komatsu, K Sato, T Ishikawa, K Akiishi, K Yamazaki, K Kato, A Saito, H Yoshie
BACKGROUND AND OBJECTIVES: Previous reports suggest that several serum biomarkers play roles in the pathogenesis, inflammatory response, and oxidative stress in periodontitis caused by bacterial infections, linking chronic periodontitis to atherosclerotic vascular disease (ASVD). The aim of this preliminary study was to investigate, in a Japanese cross-sectional community survey, potential serum biomarkers of periodontitis that are associated with ASVD and chronic periodontitis. MATERIAL AND METHODS: The study cohort included a total of 108 male subjects who underwent annual health examinations...
March 8, 2018: Journal of Periodontal Research
Andrea N Loes, Jamie T Bridgham, Michael J Harms
Toll-like receptor 4 (TLR4) induces inflammation in response to both pathogen- and host-derived molecules. Lipopolysaccharide (LPS) recognition by TLR4 has been shown to occur across the amniotes, but endogenous signaling through TLR4 has not been validated outside of placental mammals. To determine whether endogenous danger signaling is also shared across amniotes, we studied the evolution of TLR4-activation by the calgranulin proteins (S100A8, S100A9, and S100A12), a clade of host molecules that potently activate TLR4 in placental mammals...
2018: Frontiers in Immunology
J J Nieto-Fontarigo, F J Salgado, M E San-José, M J Cruz, A Casas-Fernández, M J Gómez-Conde, L Valdés-Cuadrado, M Á García-González, P Arias, M Nogueira, F J González-Barcala
LPS-ligation to CD14/TLR-4 on monocytes/macrophages triggers the production of IL-12-family cytokines. IL12/18 promote TH1 -differentiation, counteracting the TH2 -driven asthma. Therefore, CD14 modulation could alter the TH2 -differentiation and should be taken into account when studying asthma. To analyse the alteration in CD14 levels and its association with CD14 (-159 C/T) SNP (rs2569190) in Caucasian adults with stable allergic asthma, we performed a cross-sectional study (277 healthy subjects vs. 277 patients) where clinical parameters, CD14 values and the CD14 (-159 C/T) SNP were studied...
March 7, 2018: Scientific Reports
Ina Laura Pieper, Gemma Radley, Abigail Christen, Sabrina Ali, Owen Bodger, Catherine A Thornton
Ventricular assist devices (VADs) are a life-saving form of mechanical circulatory support in heart failure patients. However, VADs have not yet reached their full potential due to the associated side effects (thrombosis, bleeding, infection) related to the activation and damage of blood cells and proteins caused by mechanical stress and foreign materials. Studies of the effects of VADs on leukocytes are limited, yet leukocyte activation and damage including microparticle generation can influence both thrombosis and infection rates...
March 7, 2018: Artificial Organs
Nathalia Regina F Leal, Mariana Vieira Vigliano, Fabiana A Pinto, Thayane V de Sousa, Leosvaldo S M Velozo, Kátia C C Sabino, Maria da Graça Justo, Marsen Garcia Pinto Coelho
OBJECTIVES: To evaluate the anti-inflammatory potential of Pterodon polygalaeflorus hexane extract (HE) and its fractions on macrophage migration in vitro and in vivo. METHODS: Hexane extract from P. polygalaeflorus fruits was fractionated and yielded four fractions. RAW 264.7 cells were treated with samples to evaluate cell viability (MTT assay), cell migration (wound healing and transwell assays), CD14 expression (flow cytometry), iNOS and cytokine mRNA expression (RT-qPCR), NO (Griess reaction) and cytokine (ELISA) production...
March 7, 2018: Journal of Pharmacy and Pharmacology
Ana L Cardoso, Joana R Guedes
Analysis of miRNA expression in circulating immune cells, such as monocytes, using qRT-PCR arrays, allows the quantification of a wide range of miRNAs in easily accessible biosamples from Alzheimer's disease patients. This technique enables the identification of differentially expressed miRNAs and provides important clues for the discovery of new miRNA-based biomarkers. Here we describe how to isolate a specific lymphocyte population from human blood samples, CD14+ monocytes, and how to extract total RNA, containing short RNAs, from these cells, transcribe the RNA into cDNA and quantify a pre-set of specific miRNAs using customizable PCR plates of 96 or 384 wells...
2018: Methods in Molecular Biology
Won Seok Yang, Jin Ju Kim, Mee Jeong Lee, Eun Kyoung Lee, Su-Kil Park
BACKGROUND/AIMS: Lipopolysaccharide (LPS)-activated monocytes/macrophages develop endotoxin tolerance in part by reducing cell surface toll-like receptor 4 (TLR4) through cluster of differentiation 14 (CD14)-dependent endocytosis. In case of endothelial cells, CD14 is expressed in low copy numbers as compared with monocytes/macrophages. Thus, we explored how endothelial cells regulate TLR4 expression after LPS stimulation. METHODS: Cultured human aortic endothelial cells (HAECs) were treated with LPS...
February 28, 2018: Cellular Physiology and Biochemistry
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