scholarly journals Cathepsin S provokes interleukin-6 (IL-6) trans-signaling through cleavage of the IL-6 receptor in vitro

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Charlotte M. Flynn ◽  
Yvonne Garbers ◽  
Stefan Düsterhöft ◽  
Rielana Wichert ◽  
Juliane Lokau ◽  
...  
Keyword(s):  
Interleukin 6 ◽  
Cleavage Site ◽  
Serum Levels ◽  
Biologically Active ◽  
Cathepsin S ◽  
Target Cells ◽  
Membrane Bound ◽  
Pleiotropic Functions

AbstractThe cytokine interleukin-6 (IL-6) fulfills its pleiotropic functions via different modes of signaling. Regenerative and anti-inflammatory activities are mediated via classic signaling, in which IL-6 binds to the membrane-bound IL-6 receptor (IL-6R). For IL-6 trans-signaling, which accounts for the pro-inflammatory properties of the cytokine, IL-6 activates its target cells via soluble forms of the IL-6R (sIL-6R). We have previously shown that the majority of sIL-6R in human serum originates from proteolytic cleavage and mapped the cleavage site of the IL-6R. The cleavage occurs between Pro-355 and Val-356, which is the same cleavage site that the metalloprotease ADAM17 uses in vitro. However, sIL-6R serum levels are unchanged in hypomorphic ADAM17ex/ex mice, making the involvement of ADAM17 questionable. In order to identify other proteases that could be relevant for sIL-6R generation in vivo, we perform a screening approach based on the known cleavage site. We identify several candidate proteases and characterize the cysteine protease cathepsin S (CTSS) in detail. We show that CTSS is able to cleave the IL-6R in vitro and that the released sIL-6R is biologically active and can induce IL-6 trans-signaling. However, CTSS does not use the Pro-355/Val-356 cleavage site, and sIL-6R serum levels are not altered in Ctss−/− mice. In conclusion, we identify a novel protease of the IL-6R that can induce IL-6 trans-signaling, but does not contribute to steady-state sIL-6R serum levels.

scholarly journals Elevation of interleukin-6 in response to a chronic inflammatory stimulus in mice: inhibition by indomethacin

Blood ◽  
1992 ◽  
Vol 80 (1) ◽  
pp. 194-202 ◽  
Author(s):  
E Shacter ◽  
GK Arzadon ◽  
J Williams
Keyword(s):  
Plasma Cell ◽  
Peritoneal Cavity ◽  
Interleukin 6 ◽  
Chronic Administration ◽  
Inflammatory Cells ◽  
Serum Levels ◽  
Chronic Inflammatory Response

Abstract Intraperitoneal (i.p.) injection of a mineral oil such as pristane induces a chronic inflammatory response in mice. This is characterized by a large influx of macrophages and other inflammatory cells into the peritoneal cavity for months after injection of the oil. By using the B9 cell bioassay, it was found that injection of pristane caused a marked and prolonged elevation of interleukin-6 (IL-6) levels in the peritoneal cavities of the mice. IL-6 was undetectable (less than 15 U/mL) in the peritoneal fluids of unprimed mice and during the first week after injecting pristane. From 4 to 20 weeks, the concentration of IL-6 increased to an apparent plateau with concentrations ranging from 200 to 2,000 U/mL. Increasing the dose of pristane did not substantially increase the peritoneal levels of IL-6 established at 20 weeks after pristane treatment. At later times (by day 250), the level decreased to 263 +/- 217 U/mL. However, mice that developed plasma cell tumors around day 300 showed high levels of IL-6 in the ascites fluid (650 to 2,400 U/mL). Serum levels of IL-6 were also elevated in pristane-primed mice but were substantially lower than those found in the peritoneal cavity. Chronic administration of the nonsteroidal anti- inflammatory drug indomethacin decreased the levels of IL-6 by 75% to 80%. Experiments performed in vitro showed that pristane-elicited macrophages secreted low levels of IL-6 constitutively and high levels of IL-6 in the presence of lipopolysaccharide. Both IL-6 and prostaglandin E2 production were inhibited by addition of indomethacin to macrophage cultures in vitro. Treatment of mice with pristane may provide a model system for studying the inflammatory pathways that control IL-6 levels in vivo. The relevance of these results to elucidation of the role of IL-6 in plasma cell tumorigenesis is discussed.

scholarly journals The Delivery of Biologically Active Agents into the Nuclei of Target Cells for the Purposes of Translational Medicine

Acta Naturae ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 47-56
Author(s):  
A. S. Sobolev
Keyword(s):  
Targeted Delivery ◽  
Translational Medicine ◽  
Intracellular Transport ◽  
Biologically Active ◽  
Target Cells ◽  
Subcellular Compartment ◽  
Core Technology ◽  
Dividing Cells

Development of vehicles for the subcellular targeted delivery of biologically active agents is very promising for the purposes of translational medicine. This review summarizes the results obtained by researchers from the Laboratory of Molecular Genetics of Intracellular Transport, Institute of Gene Biology RAS, which allowed them to design the core technology: modular nanotransporters. This approach ensures high efficacy and cell specificity for different anti-cancer agents, as they are delivered into the most vulnerable subcellular compartment within the cells of interest and makes it possible for antibody mimetics to penetrate into a compartment of interest within the target cells (diving antibodies). Furthermore, polyplexes, complexes of polycationic block copolymers of DNA, have been developed and characterized. These complexes are efficient both in vitro and in vivo and demonstrate predominant transfection of actively dividing cells.

scholarly journals Granulocytic Myeloid-Derived Suppressor Cell Exosomal Prostaglandin E2 Ameliorates Collagen-Induced Arthritis by Enhancing IL-10+ B Cells

2020 ◽  
Vol 11 ◽  
Author(s):  
Xinyu Wu ◽  
Dongwei Zhu ◽  
Jie Tian ◽  
Xinyi Tang ◽  
Hongye Guo ◽  
...  
Keyword(s):  
B Cells ◽  
Prostaglandin E2 ◽  
Plasma Cells ◽  
Suppressor Cells ◽  
Suppressor Cell ◽  
Serum Levels ◽  
Biologically Active ◽  
Collagen Induced Arthritis

The results of recent studies have shown that granulocytic-myeloid derived suppressor cells (G-MDSCs) can secrete exosomes that transport various biologically active molecules with regulatory effects on immune cells. However, their roles in autoimmune diseases such as rheumatoid arthritis remain to be further elucidated. In the present study, we investigated the influence of exosomes from G-MDSCs on the humoral immune response in murine collagen-induced arthritis (CIA). G-MDSCs exosomes-treated mice showed lower arthritis index values and decreased inflammatory cell infiltration. Treatment with G-MDSCs exosomes promoted splenic B cells to secrete IL-10 both in vivo and in vitro. In addition, a decrease in the proportion of plasma cells and follicular helper T cells was observed in drainage lymph nodes from G-MDSCs exosomes-treated mice. Moreover, lower serum levels of IgG were detected in G-MDSCs exosomes-treated mice, indicating an alteration of the humoral environment. Mechanistic studies showed that exosomal prostaglandin E2 (PGE2) produced by G-MDSCs upregulated the phosphorylation levels of GSK-3β and CREB, which play a key role in the production of IL-10+ B cells. Taken together, our findings demonstrated that G-MDSC exosomal PGE2 attenuates CIA in mice by promoting the generation of IL-10+ Breg cells.

scholarly journals Inclusion of a Furin Cleavage Site Enhances Antitumor Efficacy against Colorectal Cancer Cells of Ribotoxin α-Sarcin- or RNase T1-Based Immunotoxins

Toxins ◽  
2019 ◽  
Vol 11 (10) ◽  
pp. 593 ◽  
Author(s):  
Javier Ruiz-de-la-Herrán ◽  
Jaime Tomé-Amat ◽  
Rodrigo Lázaro-Gorines ◽  
José G. Gavilanes ◽  
Javier Lacadena
Keyword(s):  
Cleavage Site ◽  
Tumor Antigens ◽  
Functional Characterization ◽  
Target Cells ◽  
Furin Cleavage ◽  
Ribonucleolytic Activity ◽  
Furin Cleavage Site ◽  
Potential Therapeutic Application

Immunotoxins are chimeric molecules that combine the specificity of an antibody to recognize and bind tumor antigens with the potency of the enzymatic activity of a toxin, thus, promoting the death of target cells. Among them, RNases-based immunotoxins have arisen as promising antitumor therapeutic agents. In this work, we describe the production and purification of two new immunoconjugates, based on RNase T1 and the fungal ribotoxin α-sarcin, with optimized properties for tumor treatment due to the inclusion of a furin cleavage site. Circular dichroism spectroscopy, ribonucleolytic activity studies, flow cytometry, fluorescence microscopy, and cell viability assays were carried out for structural and in vitro functional characterization. Our results confirm the enhanced antitumor efficiency showed by these furin-immunotoxin variants as a result of an improved release of their toxic domain to the cytosol, favoring the accessibility of both ribonucleases to their substrates. Overall, these results represent a step forward in the design of immunotoxins with optimized properties for potential therapeutic application in vivo.

scholarly journals G-CSF–stimulated Neutrophils Are a Prominent Source of Functional BLyS

2003 ◽  
Vol 197 (3) ◽  
pp. 297-302 ◽  
Author(s):  
Patrizia Scapini ◽  
Bernardetta Nardelli ◽  
Gianpaolo Nadali ◽  
Federica Calzetti ◽  
Giovanni Pizzolo ◽  
...  
Keyword(s):  
B Cell ◽  
B Lymphocyte ◽  
Serum Levels ◽  
Surface Expression ◽  
Biologically Active ◽  
Human Neutrophils ◽  
Activated Neutrophils ◽  
B Cell Homeostasis

B lymphocyte stimulator (BLyS) is a novel member of the TNF ligand superfamily that is important in B cell maturation and survival. We demonstrate that human neutrophils, after incubation with G-CSF or, less efficiently, IFNγ, express high levels of BLyS mRNA and release elevated amounts of biologically active BLyS. In contrast, surface expression of the membrane-bound BLyS was not detected in activated neutrophils. Indeed, in neutrophils, uniquely among other myeloid cells, soluble BLyS is processed intracellularly by a furin-type convertase. Worthy of note, the absolute capacity of G-CSF–stimulated neutrophils to release BLyS was similar to that of activated monocytes or dendritic cells, suggesting that neutrophils might represent an important source of BLyS. In this regard, we show that BLyS serum levels as well as neutrophil-associated BLyS are significantly enhanced after in vivo administration of G-CSF in patients. In addition, serum obtained from two of these patients induced a remarkable accumulation of neutrophil-associated BLyS in vitro. This effect was neutralized by anti–G-CSF antibodies, indicating that G-CSF, present in the serum, stimulated neutrophils to produce BLyS. Collectively, our findings suggest that neutrophils, through the production of BLyS, might play an unsuspected role in the regulation of B cell homeostasis.

scholarly journals Mesenchymal Stem Cell-Derived Extracellular Vesicles and Their Therapeutic Potential

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Ashley G. Zhao ◽  
Kiran Shah ◽  
Brett Cromer ◽  
Huseyin Sumer
Keyword(s):  
Tissue Engineering ◽  
Regenerative Medicine ◽  
Extracellular Vesicles ◽  
Therapeutic Potential ◽  
Target Cells ◽  
Therapeutic Effects ◽  
Multipotent Cells ◽  
Membrane Bound

Extracellular vesicles (EVs) are cell-derived membrane-bound nanoparticles, which act as shuttles, delivering a range of biomolecules to diverse target cells. They play an important role in maintenance of biophysiological homeostasis and cellular, physiological, and pathological processes. EVs have significant diagnostic and therapeutic potentials and have been studied both in vitro and in vivo in many fields. Mesenchymal stem cells (MSCs) are multipotent cells with many therapeutic applications and have also gained much attention as prolific producers of EVs. MSC-derived EVs are being explored as a therapeutic alternative to MSCs since they may have similar therapeutic effects but are cell-free. They have applications in regenerative medicine and tissue engineering and, most importantly, confer several advantages over cells such as lower immunogenicity, capacity to cross biological barriers, and less safety concerns. In this review, we introduce the biogenesis of EVs, including exosomes and microvesicles. We then turn more specifically to investigations of MSC-derived EVs. We highlight the great therapeutic potential of MSC-derived EVs and applications in regenerative medicine and tissue engineering.

Parathyroid hormone induces hepatic production of bioactive interleukin-6 and its soluble receptor

2001 ◽  
Vol 280 (3) ◽  
pp. E405-E412 ◽  
Author(s):  
Mary Ann Mitnick ◽  
Andrew Grey ◽  
Urszula Masiukiewicz ◽  
Marcjanna Bartkiewicz ◽  
Laura Rios-Velez ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Interleukin 6 ◽  
Serum Levels ◽  
Soluble Receptor ◽  
Important Mediator ◽  
Dose Dependent ◽  
Circulating Levels

Interleukin-6 (IL-6) is an important mediator of parathyroid hormone (PTH)-induced bone resorption. Serum levels of IL-6 and its soluble receptor (IL-6sR) are regulated in part by PTH. The PTH/PTH-related protein type 1 receptor is highly expressed in the liver, and in the current study we investigated whether the liver produces IL-6 or IL-6sR in response to PTH. Perfusion of the isolated rat liver with PTH-(1-84) stimulated rapid, dose-dependent production of bioactive IL-6 and the IL-6sR. These effects were observed at near physiological concentrations of the hormone such that 1 pM PTH induced hepatic IL-6 production at a rate of ∼0.6 ng/min. In vitro, hepatocytes, hepatic endothelial cells, and Kupffer cells, but not hepatic stellate cells, were each found to produce both IL-6 and IL-6sR in response to higher (10 nM) concentrations of PTH. Our data suggest that hepatic-derived IL-6 and IL-6sR contribute to the increase in circulating levels of these cytokines induced by PTH in vivo and raise the possibility that PTH-induced, liver-derived IL-6 may exert endocrine effects on tissues such as bone.

scholarly journals Characterization, Stability, and In Vivo Efficacy Studies of Recombinant Human CNTF and Its Permeation into the Neural Retina in Ex Vivo Organotypic Retinal Explant Culture Models

Pharmaceutics ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 611 ◽  
Author(s):  
Jaakko Itkonen ◽  
Ada Annala ◽  
Shirin Tavakoli ◽  
Blanca Arango-Gonzalez ◽  
Marius Ueffing ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Disease Model ◽  
Neural Retina ◽  
Biologically Active ◽  
Target Cells ◽  
Therapeutic Effects ◽  
Duration Of Action ◽  
In Vivo Efficacy

Ciliary neurotrophic factor (CNTF) is one of the most studied neuroprotective agents with acknowledged potential in treating diseases of the posterior eye segment. Although its efficacy and mechanisms of action in the retina have been studied extensively, it is still not comprehensively understood which retinal cells mediate the therapeutic effects of CNTF. As with therapeutic proteins in general, it is poorly elucidated whether exogenous CNTF administered into the vitreous can enter and distribute into the retina and hence reach potentially responsive target cells. Here, we have characterized our purified recombinant human CNTF (rhCNTF), studied the protein’s in vitro bioactivity in a cell-based assay, and evaluated the thermodynamic and oligomeric status of the protein during storage. Biological activity of rhCNTF was further evaluated in vivo in an animal model of retinal degeneration. The retinal penetration and distribution of rhCNTF after 24 h was studied utilizing two ex vivo retina models. Based on our characterization findings, our rhCNTF is correctly folded and biologically active. Moreover, based on initial screening and subsequent follow-up, we identified two buffers in which rhCNTF retains its stability during storage. Whereas rhCNTF did not show photoreceptor preservative effect or improve the function of photoreceptors in vivo, this could possibly be due to the used disease model or the short duration of action with a single intravitreal injection of rhCNTF. On the other hand, the lack of in vivo efficacy was shown to not be due to distribution limitations; permeation into the retina was observed in both retinal explant models as in 24 h rhCNTF penetrated the inner limiting membrane, and being mostly observed in the ganglion cell layer, distributed to different layers of the neural retina. As rhCNTF can reach deeper retinal layers, in general, having direct effects on resident CNTF-responsive target cells is plausible.

scholarly journals Immunostimulatory Activity of Black Rice Bran in Cyclophosphamide-Induced Immunosuppressed Rats

2020 ◽  
Vol 15 (7) ◽  
pp. 1934578X2093491
Author(s):  
Young Mi Park ◽  
Hak Yong Lee ◽  
Dong Yeop Shin ◽  
Yang Hee Lee ◽  
Ye Jin Yang ◽  
...  
Keyword(s):  
Rice Bran ◽  
White Blood Cells ◽  
Serum Levels ◽  
Biologically Active ◽  
Black Rice ◽  
Immunostimulatory Activity ◽  
Thymic Tissue ◽  
Black Rice Bran

Black rice bran extract (BRBE), containing various biologically active compounds, such as anthocyanin, has antioxidant activity and numerous pharmacological effects. Here, we aimed to confirm the immunostimulatory effects of BRBE in cyclophosphamide (CP)-induced immunosuppressed cells. Our results confirmed that BRBE exerted an immunostimulatory effect. In vitro, BRBE treatment enhanced cell proliferation, activity of natural killer cells and cytotoxic T lymphocytes, and production of CP-repressed cytokines, such as tumor necrosis factor-α, interferon-γ, interleukin (IL)-2, and IL-12, and immunoglobulins G and A in isolated splenocytes. Additionally, in vivo, BRBE treatment increased the number of immune cells, such as white blood cells, lymphocyte counts, mid-range absolute counts, and neutrophils in CP-induced immunosuppressed rats. Furthermore, BRBE increased the serum levels of abovementioned inflammatory cytokines and immunoglobulins in CP-induced immunosuppressed rats. In addition, BRBE protected against CP-mediated spleen and thymic tissue damage. Our findings suggest that BRBE could be potentially used as a component of functional food for immunity enhancement.

scholarly journals gp130, The Cytokine Common Signal-Transducer of Interleukin-6 Cytokine Family, Is Downregulated in T Cells In Vivo by Interleukin-6

Blood ◽  
1998 ◽  
Vol 91 (9) ◽  
pp. 3308-3314 ◽  
Author(s):  
Xue-Jie Wang ◽  
Tetsuya Taga ◽  
Kanji Yoshida ◽  
Mikiyoshi Saito ◽  
Tadamitsu Kishimoto ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Interleukin 6 ◽  
Flow Cytometric Analysis ◽  
Serum Levels ◽  
T Cell Subsets ◽  
Common Signal ◽  
T Cell Subpopulations

gp130 is a common signal-transducing receptor component for the interleukin-6 (IL-6) family of cytokines. To investigate the expression of gp130 in T-cell subsets and its regulation, anti-murine gp130 monoclonal antibody (MoAb) was used for flow cytometric analysis. In normal mice, gp130 was differentially expressed in thymocyte and splenic T-cell subpopulations defined by CD4/CD8 expression. In aged MRL/lpr mice, although gp130 expression was detectable in splenic CD4+ or CD8+ T cells, gp130 expression was significantly downregulated. Because serum levels of IL-6 and soluble IL-6 receptor (sIL-6R) are elevated in these mice, we examined the possibility that the downregulation of gp130 expression on splenic T cells might be produced in response to continuous activation of gp130 by high levels of serum IL-6. In transgenic mice overexpressing IL-6, gp130 expression in the splenic T cells was significantly decreased. After stimulation with IL-6 in vitro, the level of gp130 on CD4+ or CD8+ splenic T cells from normal mice was significantly decreased. These results suggest that the expression of gp130 in splenic T cells could be downregulated by the IL-6 stimulation under physiological or pathological circumstances.

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