scholarly journals Meglumine acridone acetate, the ionic salt of CMA and N-methylglucamine, induces apoptosis in human PBMCs via the mitochondrial pathway

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Marina A. Plotnikova ◽  
Sergey A. Klotchenko ◽  
Artem A. Kiselev ◽  
Andrey N. Gorshkov ◽  
Anna-Polina S. Shurygina ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Viral Infections ◽  
Human Peripheral Blood ◽  
Antiviral Effect ◽  
Response To Treatment ◽  
Type I ◽  
Sequencing Analysis ◽  
Human Pbmcs ◽  
Peripheral Blood Mononuclear ◽  
Docking Simulations

AbstractMeglumine acridone acetate (MA) is used in Russia for the treatment of influenza and other acute respiratory viral infections. It was assumed, until recently, that its antiviral effect was associated with its potential ability to induce type I interferon. Advanced studies, however, have shown the failure of 10-carboxymethyl-9-acridanone (CMA) to activate human STING. As such, MA’s antiviral properties are still undergoing clarification. To gain insight into MA’s mechanisms of action, we carried out RNA-sequencing analysis of global transcriptomes in MA-treated (MA+) human peripheral blood mononuclear cells (PBMCs). In response to treatment, approximately 1,223 genes were found to be differentially expressed, among which 464 and 759 were identified as either up- or down-regulated, respectively. To clarify the cellular and molecular processes taking place in MA+ cells, we performed a functional analysis of those genes. We have shown that evident MA subcellular localizations are: at the nuclear envelope; inside the nucleus; and diffusely in perinuclear cytoplasm. Postulating that MA may be a nuclear receptor agonist, we carried out docking simulations with PPARα and RORα ligand binding domains including prediction and molecular dynamics-based analysis of potential MA binding poses. Finally, we confirmed that MA treatment enhanced nuclear apoptosis in human PBMCs. The research presented here, in our view, indicates that: (i) MA activity is mediated by nuclear receptors; (ii) MA is a possible PPARα and/or RORα agonist; (iii) MA has an immunosuppressive effect; and (iv) MA induces apoptosis through the mitochondrial signaling pathway.

scholarly journals The 2′-O-methylation status of a single guanosine controls transfer RNA–mediated Toll-like receptor 7 activation or inhibition

2012 ◽  
Vol 209 (2) ◽  
pp. 235-241 ◽  
Author(s):  
Stefanie Jöckel ◽  
Gernot Nees ◽  
Romy Sommer ◽  
Yang Zhao ◽  
Dmitry Cherkasov ◽  
...  
Keyword(s):  
Influenza A ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Thermus Thermophilus ◽  
Methylation Status ◽  
Transfer Rna ◽  
Type I ◽  
Peripheral Blood Mononuclear ◽  
Molecular Pattern

Foreign RNA serves as pathogen-associated molecular pattern (PAMP) and is a potent immune stimulator for innate immune receptors. However, the role of single bacterial RNA species in immune activation has not been characterized in detail. We analyzed the immunostimulatory potential of transfer RNA (tRNA) from different bacteria. Interestingly, bacterial tRNA induced type I interferon (IFN) and inflammatory cytokines in mouse dendritic cells (DCs) and human peripheral blood mononuclear cells (PBMCs). Cytokine production was TLR7 dependent because TLR7-deficient mouse DCs did not respond and TLR7 inhibitory oligonucleotides inhibited tRNA-mediated activation. However, not all bacterial tRNA induced IFN-α because tRNA from Escherichia coli Nissle 1917 and Thermus thermophilus were non-immunostimulatory. Of note, tRNA from an E. coli knockout strain for tRNA (Gm18)-2′-O-methyltransferase (trmH) regained immunostimulatory potential. Additionally, in vitro methylation of this immunostimulatory Gm18-negative tRNA with recombinant trmH from T. thermophilus abolished its IFN-α inducing potential. More importantly, Gm18-modified tRNA acted as TLR7 antagonist and blocked IFN-α induction of influenza A virus–infected PBMCs.

scholarly journals Differential In Vitro Cytokine Induction by the Species of Cryptococcus gattii Complex

2018 ◽  
Vol 86 (4) ◽  
Author(s):  
Patricia F. Herkert ◽  
Jessica C. dos Santos ◽  
Ferry Hagen ◽  
Fatima Ribeiro-Dias ◽  
Flávio Queiroz-Telles ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Sensu Stricto ◽  
Cryptococcus Gattii ◽  
The Other ◽  
Human Pbmcs ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Content Type ◽  
Tnf Α

ABSTRACT Cryptococcal species vary in capsule and cell size, thermotolerance, geographic distribution, and affected populations. Cryptococcus gattii sensu stricto and C. deuterogattii affect mainly immunocompetent hosts; however, C. bacillisporus , C. decagattii , and C. tetragattii cause infections mainly in immunocompromised hosts. This study aimed to compare the capacities of different species of the C. gattii species complex to induce cytokines and antimicrobial molecules in human peripheral blood mononuclear cells (PBMCs). Cryptococcus bacillisporus and C. deuterogattii induced the lowest levels of tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and IL-6 among the five species of the C. gattii complex. Cryptococcus deuterogattii induced higher levels of IL-22 than those induced by C. tetragattii and the environmental species C. flavescens . In addition, C. bacillisporus and C. gattii sensu stricto proliferated inside human monocyte-derived macrophages after 24 h of infection. All Cryptococcus species were able to generate reactive oxygen species (ROS) in human PBMCs, with C. bacillisporus and C. deuterogattii being more efficient than the other species. In conclusion, C. bacillisporus and C. deuterogattii induce lower levels of the proinflammatory cytokines TNF-α, IL-1β, and IL-6 and higher ROS levels than those induced by the other species. Species of the Cryptococcus gattii complex have different abilities to induce cytokine and ROS production by human PBMCs.

scholarly journals Fulminant H1N1 and severe acute respiratory syndrome coronavirus-2 infections with a 4-year interval without an identifiable underlying cause: a case report

2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Terese L. Katzenstein ◽  
Sofie E. Jørgensen ◽  
Jann Mortensen ◽  
Marie Helleberg ◽  
Anna Kalhauge ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Mononuclear Cells ◽  
Viral Infections ◽  
Influenza Virus Infection ◽  
H1n1 Influenza ◽  
Asymptomatic Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Peripheral Blood Mononuclear ◽  
Genetic Changes

Abstract Background The clinical presentation of severe acute respiratory syndrome coronavirus-2 infection is highly variable from asymptomatic infection to fulminant disease. The reasons for the variation are only starting to unravel, with risk factors including age and certain comorbidities as well as genetic defects causing immunological perturbations in the interferon pathways. Case presentation We report the case of an otherwise healthy Caucasian man, who at ages 60 and 64 years suffered from severe H1N1 influenza virus infection and severe acute respiratory syndrome coronavirus-2 infections, respectively. In both cases, there were acute kidney impairment and the need for intensive care unit admission as well as mechanical ventilation. Fortunately, after both infections there was full clinical recovery. The severity of the infections indicates an underlying impairment in the ability to control these kinds of infections. Challenge of patient peripheral blood mononuclear cells showed impaired type I and III antiviral interferon responses and reduced interferon-stimulated gene expression. However, despite investigation of patient samples by whole exome sequencing and enzyme-linked immunosorbent assay, no known disease-causing genetic variants related to interferon pathways were found, nor were interferon autoantibodies demonstrated. Thus, any underlying immunological cause of this unusual susceptibility to severe viral infections remains unresolved. Conclusion The patient experienced very similar severe clinical pictures triggered by H1N1 and severe acute respiratory syndrome coronavirus-2 infections, indicating an underlying inability to contain these infections. We were unable to show that the patient had any of the currently known types of immune incompetence but identified genetic changes possibly contributing to the severe course of both infections. Further analyses to delineate contribution factors are needed.

Fibronectin modulates expression of interleukin-1 beta and its receptor antagonist in human mononuclear cells

1996 ◽  
Vol 271 (1) ◽  
pp. L61-L69 ◽  
Author(s):  
K. L. Graves ◽  
J. Roman
Keyword(s):  
Receptor Antagonist ◽  
Type I Collagen ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Interleukin 1 ◽  
Type I ◽  
Peripheral Blood Mononuclear ◽  
Human Mononuclear Cells ◽  
Naturally Occurring ◽  
Dose Dependent

Identification of factors that regulate production of proinflammatory cytokines may provide insight into mechanisms governing lung inflammation. One potential regulatory factor highly expressed in inflamed tissues is fibronectin (FN). To determine the potential effects of FN on interleukin (IL)-1 beta production, we exposed human peripheral blood mononuclear cells to soluble FN. This treatment resulted in the accumulation of IL-1 beta mRNA and enhancement of IL-1 beta protein synthesis and secretion. This effect was dose dependent and appeared to be mediated by the integrin alpha 5 beta 1. Treatment with FN also increased production of IL-1 receptor antagonist (IL-1ra), a naturally occurring inhibitor of IL-1 function. However, the stimulatory effect of FN on IL-1ra production was abolished by costimulation with type I collagen. We conclude that the increased deposition of FN in injured tissues may enhance the expression of IL-1 beta mRNA and augment the production and release of IL-1 beta protein by mononuclear cells. Differential expression of IL-1 beta and IL-1ra resulting in a high IL-1 beta-to-IL-1ra ratio in response to mixed matrices containing FN and type I collagen may be an important regulatory point in inflammation.

scholarly journals Alpha-Cyclodextrin Sulphate, an anti-HIV Agent, Retains its Antiviral Effect in the Presence of Hydrocortisol Phosphate

1993 ◽  
Vol 4 (1) ◽  
pp. 65-66 ◽  
Author(s):  
J. Pitha ◽  
R. Anand
Keyword(s):  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Antiviral Effect ◽  
Simultaneous Treatment ◽  
Peripheral Blood Mononuclear ◽  
Antiviral Compound ◽  
Cytopathic Effects ◽  
The Stability ◽  
Hiv 1

Cyclodextrin sulphates alpha (a) and beta (b) were found to be inhibitors of replication of human immunodeficiency virus (HIV-1) in phytohaemagglutinin (PHA)-stimulated human peripheral blood mononuclear cells (PBMC); the same cells were also stimulated to proliferate (Anand et al., 1990). B-Cyclodextrin sulphate was also found to stimulate another, more complex, cell proliferation-neovascularization process in rabbit cornea (Folkman et al., 1989). Since the process of neovascularization is generally associated with progression of tumour growth and other pathologies, it is obvious that its stimulation by an antiviral compound may be a serious side effect. Nevertheless endotoxin-induced neovascularization can be suppressed by simultaneous treatment with b-cyclodextrin sulphate and glucocorticoids (Folkman et al., 1989). The same co-treatment may possibly be used to improve antiviral action of cyclodextrin sulphates, but such a combination therapy may not be free of complications. Glucocorticoids also affect the stability of various cellular membranes. Treatment of cells in vitro by glucocorticoids is known to stabilize their lysozymes. Glucocorticoids affect both the proliferative capacity of the cells (Cristofalo, 1972) and cytopathic effects of rabies viral infection. Furthermore, it has been reported that the cytopathic effects of rabies and of yellow fever viruses were inhibited while those of the polio virus were only mildly affected (Hannoun et al., 1965). Consequently, in this work we evaluated effects of a glucocorticoid and of glucocorticoid-a-cyclodextrin sulphate combination on HIV-1 replication.

scholarly journals Neither Factor VIII Nor the Putative F8B Protein Is Detectable in Human Peripheral Blood Mononuclear Cells

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3507-3507
Author(s):  
Devi Gunasekera ◽  
Kenneth B. Lewis ◽  
Alexis A. Thompson ◽  
Ronald R, Louie ◽  
David W. Scott ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Hemophilia A ◽  
Human Peripheral Blood ◽  
Positive Control ◽  
Human Pbmcs ◽  
Peripheral Blood Mononuclear ◽  
Western Blots ◽  
Bhk Cells ◽  
Sds Page ◽  
Secondary Antibodies

Abstract Factor VIII (FVIII) is encoded by 26 exons comprising the F8 gene. The F8 mRNA transcript encodes the full-length FVIII protein, which consists of 2332 amino acids. A CpG island within intron 22 of F8 includes an initiation site for a second, shorter transcript termed F8B. F8B contains a short exon spliced in-frame to F8 exons 23-26, and this mRNA transcript is expressed in multiple human tissues. The putative protein encoded by this gene corresponds to 8 amino acids encoded by the exon within F8 intron 22, followed by the FVIII C2 domain sequence. Earlier studies have unambiguously demonstrated FVIII expression in human endothelial cells, including liver and lung. Animal model studies have indicated FVIII is expressed, possibly exclusively, in endothelial cells. Other recent reports have presented evidence for FVIII and F8B protein expression in human peripheral blood mononuclear cells (PBMCs). In this study, we utilized several methods to test the hypothesis that FVIII and/or the putative F8B protein is expressed in human PBMCs. Frozen PBMCs from healthy human subjects were thawed and then washed 5-10X to ensure removal of plasma-derived FVIII. Samples containing 50 million PBMCs were lysed in buffer containing protease inhibitors, centrifuged, and the supernatants analyzed by immunoprecipitation (IP) followed by SDS-PAGE and Western blots. The IP experiments utilized both polyclonal anti-FVIII antibodies and cocktails of monoclonal antibodies (mAbs) specific for different FVIII domains, and Western blots used several combinations of primary and secondary antibodies. Negative controls consisted of BHK cell lysates, and positive controls were purified FVIII and lysates of BHK cells transduced with a plasmid encoding FVIII. Western blots carried out with sensitive chemiluminescence detection identified the expected bands in the positive control lanes. Bands at expected positions for FVIII or F8B proteins (~240kDa, 75 kDa, 25-30 kDa) visualized by Coomassie-blue staining of lysates analyzed by SDS-PAGE were cut from the gels, subjected to trypsin digestion and then analyzed by mass spectrometry (LC-MS). FVIII-derived peptides were detected only in the positive control samples. Intracellular staining (ICS) of permeabilized PBMCs was carried out using mAbs specific for the FVIII A1, A2, C1, and C2 domains and a mAb specific for the FVIII light chain. The mAbs were covalently labeled with Alexa-Fluor 488 to avoid possible nonspecific binding of labeled secondary antibodies. PBMCs from (1) normal control subjects; (2) a severe hemophilia A subject with an intron-22 inversion mutation and (3) a severe hemophilia A subject with a deletion of F8 exons 3-6 were analyzed. All 5 anti-FVIII mAbs detected FVIII protein in FVIII-expressing BHK cells, while none detected FVIII in BHK cells or in any of the human PBMC samples. Our results are consistent with several recent studies reporting proteomics analysis of human PBMCs, which have identified no FVIII peptides or proteins. We conclude that if FVIII or F8B proteins are expressed in human PBMCs, their concentration is below the detection limits of these assays. Disclosures No relevant conflicts of interest to declare.

NCS 613 exhibits anti-inflammatory effects on PBMCs from lupus patients by inhibiting p38 MAPK and NF-κB signalling pathways while reducing proinflammatory cytokine production

2013 ◽  
Vol 91 (5) ◽  
pp. 353-361 ◽  
Author(s):  
Issaka Yougbaré ◽  
Gilles Boire ◽  
Michelle Roy ◽  
Claire Lugnier ◽  
Éric Rouseau
Keyword(s):  
P38 Mapk ◽  
Lupus Erythematosus ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Pde4 Inhibitor ◽  
Human Pbmcs ◽  
Peripheral Blood Mononuclear ◽  
Healthy Donors ◽  
Tnf Α ◽  
Anti Inflammatory

Systemic lupus erythematosus (SLE) is a polymorphic and multigenic autoimmune disease that evolves into progressive and chronic inflammation of multiple joints and organs. Phosphorylation and activation of p38 MAPK, along with the resulting overproduction of interleukin (IL)-1β, IL-6, and tumour necrosis factor (TNF)-α is a hallmark of inflammatory disorders. Here, we investigated the anti-inflammatory pathway modulated by NCS 613, a specific PDE4 inhibitor, on human peripheral blood mononuclear cells (PBMCs) from 5 healthy donors and 12 SLE patients. PDE4 subtypes, p38 MAPK, and IκBα protein levels were analyzed by Western blot, while NF-κB and PDE4B immunostaining was assessed in control and lipopolysaccharide (LPS) -pretreated PBMCs. Proinflammatory cytokines were quantified by ELISA, while IL-1β mRNA was resolved by RT–qPCR. NCS 613 treatment decreased PDE4B and upregulated PDE4C in human PBMCs from healthy donors and SLE patients. LPS stimulation increased p38 MAPK phosphorylation and NF-κB translocation to the nucleus, which was abolished by NCS 613 treatment. Concomitantly, NCS 613 restored IκBα detection levels in human PBMCs from both healthy donors and SLE patients. This compound also abolished LPS-induced inflammation in PBMCs by reducing IL-6, IL-8, and TNF-α cytokines. NCS 613 is a small molecule displaying anti-inflammatory properties that may provide an alternative or complementary strategy for SLE management.

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