Clostridioides difficile toxins enhanced the in vitro production of CXC chemokine ligand 2 and tumor necrosis factor-α via Toll-like receptors in macrophages

2021 ◽  
Vol 70 (4) ◽  
Author(s):  
Hiroe Konishi McKee ◽  
Chiaki Kajiwara ◽  
Tetsuo Yamaguchi ◽  
Yoshikazu Ishii ◽  
Norikazu Shimizu ◽  
...  
Keyword(s):  
Type Species ◽  
Toll Like Receptor ◽  
Hek 293 ◽  
Content Type ◽  
Clostridioides Difficile ◽  
Tnf Α ◽  
Chemokine Ligand ◽  
Factor Α ◽  
Chemokine Ligand 2 ◽  
Necrosis Factor

Introduction. Clostridioides difficile infection (CDI) causes toxin-mediated enteropathy, such as antibiotic-associated diarrhoea and pseudomembranous colitis. Rho-glucosylating toxin A (TcdA) and toxin B (TcdB) have been clearly implicated in pathogenesis, whereas the virulence of binary toxin (CDT) is still debated. Hypothesis statement. We hypothesized that CDT is involved in the host immune response and plays a pivotal role in establishing virulence by modulating pro-inflammatory cytokine production; this is achieved through the integral Toll-like receptor (TLR) signalling pathways. Aim. The aim of the present study was to determine whether and how CDT impacts macrophages compared to TcdA or TcdB by examining the induction of CXC chemokine ligand 2 (CXCL2) and tumour necrosis factor-α (TNF-α), both of which are crucial in mediating local and systematic inflammatory responses. Methodology. RAW264.7 cells or transfected human embryonic kidney (HEK) 293 T cells were incubated with TcdA, TcdB, or CDT. In some experiments, a neutralizing antibody against TLR2 or TLR4, or myeloid differentiation 88 inhibitory peptide were added. The amount of CXCL2 and TNF-α secreted was then measured. Results. In RAW264.7 macrophages, CXCL2 and TNF-α were produced via the Toll-like receptor 2 (TLR2) or Toll-like receptor 4 (TLR4) pathway in a TcdA, TcdB, or CDT dose-dependent manner. Interleukin-8 secretion was induced in TLR4/MD2/CD14-transfected, but not in TLR2-transfected, HEK 293 T cells following TcdB or CDT exposure. Conclusion. Our results showed that C. difficile toxins, including CDT, enhanced macrophage-mediated CXCL2 and TNF-α production via TLR2 and TLR4, indicating that CDT affects host immune responses.

scholarly journals Modification of Tumor Necrosis Factor-α and C-C Motif Chemokine Ligand 18 Secretion by Monocytes Derived from Patients with Diabetic Foot Syndrome

Biology ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 3
Author(s):  
Karine O. Galstyan ◽  
Ludmila V. Nedosugova ◽  
Narine S. Martirosian ◽  
Nikita G. Nikiforov ◽  
Natalia V. Elizova ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Diabetic Foot ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Tnf Α ◽  
Inflammatory Activation ◽  
Chemokine Ligand ◽  
Anti Inflammatory ◽  
Factor Α ◽  
Necrosis Factor

Background: This study involves the investigation of spontaneous and induced secretion of the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) and the anti-inflammatory chemokine C-C motif chemokine ligand 18 (CCL18) by monocytes isolated from blood of patients with long-term type 2 diabetes mellitus (T2DM), both with or without foot ulcers. Methods: A total of 121 patients with T2DM (79 without diabetic foot syndrome (DFS) and 42 patients with DFS) were included. Cluster of Differentiation 14 (CD14+) monocytes were isolated from patients’ blood and stimulated by interferon-γ (IFN-γ) and interleukin-4 (IL-4) for induction of pro- and anti-inflammatory monocyte activation, respectively. The concentrations of TNF-α and CCL18 in the culture medium were measured using ELISA on day 1 and day 6 after cell stimulation. Results: We found a correlation between glycated hemoglobin (HbA1c) and stimulated secretion levels of TNF-α (r = 0.726, p = 0.027) and CCL18 (r = –0.949, p = 0.051) in patients with DFS. There was an increase of pro- and anti-inflammatory activation of monocytes in all patients with different durations of DFS (p < 0.05). However, no stimulation of anti-inflammatory activation was detected in patients with DFS lasting more than 6 months (p = 0.033). Conclusions: Our study showed an increase in pro-inflammatory secretion and a decrease in anti-inflammatory secretion by monocytes isolated from blood of patients with T2DM depending on HbA1c levels and duration of the inflammatory process. These findings allow us to assume that monocytes isolated from T2DM patients are characterized by a biased ability to respond towards pro-inflammatory stimulation, contributing to the chronic wound process.

Mechanical stress induces tumor necrosis factor-α production through Ca2+ release-dependent TLR2 signaling

2008 ◽  
Vol 295 (2) ◽  
pp. C432-C439 ◽  
Author(s):  
Han Geun Kim ◽  
Joo Yun Kim ◽  
Min Geun Gim ◽  
Jung Min Lee ◽  
Dae Kyun Chung
Keyword(s):  
Tumor Necrosis Factor ◽  
Mechanical Stress ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Dominant Negative ◽  
Hek 293 ◽  
Tnf Α ◽  
Factor Α ◽  
Tlr2 Signaling ◽  
Necrosis Factor

We studied centrifugation-mediated mechanical stress-induced tumor necrosis factor-α (TNF-α) production in the monocyte-like cell line THP-1. The induction of TNF-α by mechanical stress was dependent on the centrifugation speed and produced the highest level of TNF-α after 1 h of stimulation. TNF-α production returned to normal levels after 24 h of stimulation. Mechanical stress also induced Toll-like receptor-2 (TLR2) mRNA in proportion to the expression of TNF-α. The inhibition of TLR2 signaling by dominant negative myeloid differentiation factor 88 (MyD88) blocked TNF-α expression response to mechanical stress. After transient overexpression of TLR2 in HEK-293 cells, mechanical stress induced TNF-α mRNA production. Interestingly, mechanical stress activated the c-Src-dependent TLR2 phosphorylation, which is necessary to induce Ca2+ fluxes. When THP-1 cells were pretreated with BAPTA-AM, thapsigargin, and NiCl2·6H2O, followed by mechanical stimulation, both TLR2 and TNF-α production were inhibited, indicating that centrifugation-mediated mechanical stress induces both TLR2 and TNF-α production through Ca2+ releases from intracellular Ca2+ stores following TLR2 phosphorylation. In addition, TNF-α treatment in THP-1 cells induced TLR2 production in response to mechanical stress, whereas the preincubation of anti-TNF-α antibody scarcely induced the mechanical stress-mediated production of TLR2, indicating that TNF-α produced by mechanically stimulated THP-1 cells affected TLR2 production. We concluded that TNF-α production induced by centrifugation-mediated mechanical stress is dependent on MyD88-dependent TLR2 signaling that is associated with Ca2+ release and that TNF-α production induced by mechanical stress affects TLR2 production.

Cyclosporine Ameliorates Silica-Induced Autoimmune Hepatitis in Rat Model by Altering the Expression of Toll-Like Receptor-4, Interleukin-2 and Tumor Necrosis Factor-α

2022 ◽  
Vol 22 ◽  
Author(s):  
Moustafa M. Mohamed ◽  
Ahmed M. M. Okasha ◽  
Amany H. Abdel Naiem ◽  
Reham F. Mohamed ◽  
Sayed F. Abdelwahab ◽  
...  
Keyword(s):  
Autoimmune Hepatitis ◽  
Interleukin 2 ◽  
Hepatic Function ◽  
Treated Group ◽  
Control Group ◽  
Toll Like Receptor ◽  
Hepatic Diseases ◽  
Tnf Α ◽  
Factor Α ◽  
Necrosis Factor

Background: Autoimmune hepatitis (AIH) is an inflammatory liver disease which is characterized histologically by interface hepatitis, biochemically by elevated transaminase levels, and serologically by the presence of autoantibodies. Toll-like receptor (TLR)-4 is a TLR family member that, upon activation in hepatocytes, initiates a cascade of events. Interleukin-2 (IL-2) and tumour necrosis factor-α (TNF-α) are potent inflammatory cytokines secreted in AIH playing an important role in the early development of inflammation, and hepatocyte damage. Objective: This study examined cyclosporine role in AIH and tried to illustrate its actions on altered hepatic function in silica-induced AIH model. Methods: AIH was induced in Wester rats using Sodium Silicate. The rats were divided into four groups: control group, Silica-AIH group, cyclosporine-treated group, and prevention group. TLR-4, and IL-2 mRNA expression in liver tissues was tested by RT-PCR. Results: AIH was associated with up-regulation of liver enzymes, IL-2, and TLR-4 gene expression while cyclosporine significantly down-regulated the expression of both. The relative quantity of TLR-4 mRNA was 1±0, 13.57±1.91, 4±0.38, and 2±0 in the control, AIH, cyclosporine, and prevention groups, respectively (p<0.001). Also, the relative quantity of IL-2 mRNA was 1±0, 14.79±1.42, 7.07±0.96, and 3.4±0.55 in the same groups, respectively (p<0.001). Additionally, immuno-histochemical staining for TNF-α in liver sections was increased in the silica-AIH group but it was decreased in the cyclosporine-treated and prevention groups. Conclusion: This study advocates a therapeutic role of cyclosporine in treating immune-mediated hepatic diseases. Cyclosporine improves histological alterations in the liver and inhibits the production of proinflammatory cytokines.

scholarly journals Distinguishing rheumatoid arthritis from psoriatic arthritis

RMD Open ◽  
2018 ◽  
Vol 4 (2) ◽  
pp. e000656 ◽  
Author(s):  
Joseph F Merola ◽  
Luis R Espinoza ◽  
Roy Fleischmann
Keyword(s):  
Rheumatoid Arthritis ◽  
Psoriatic Arthritis ◽  
Bone Growth ◽  
Interferon Γ ◽  
Joint Involvement ◽  
Radiographic Findings ◽  
Tnf Α ◽  
Chemokine Ligand ◽  
Factor Α ◽  
Necrosis Factor

Rheumatoid arthritis (RA) and psoriatic arthritis (PsA) have key differences in clinical presentation, radiographic findings, comorbidities and pathogenesis to distinguish between these common forms of chronic inflammatory arthritis. Joint involvement is typically, but not always, asymmetric in PsA, while it is predominantly symmetric in RA. Bone erosions, without new bone growth, and cervical spine involvement are distinctive of RA, while axial spine involvement, psoriasis and nail dystrophy are distinctive of PsA. Patients with PsA typically have seronegative test findings for rheumatoid factor (RF) and cyclic citrullinated peptide (CCP) antibodies, while approximately 80% of patients with RA have positive findings for RF and CCP antibodies. Although there is overlap in the pathogenesis of PsA and RA, differences are also present that affect the efficacy of treatment. In PsA, levels of interleukin (IL)-1β, IL-6, IL-17, IL-22, IL-23, interferon-γ and tumour necrosis factor-α (TNF-α) are elevated, and in RA, levels of IL-1, IL-6, IL-22, IL-33, TNF-α, chemokine ligand 11 and chemokine C-X-C motif ligand 13 are elevated. Differences in the pathogenesis of RA and PsA translate into some variances in the specificity and efficacy of therapies.

scholarly journals Effects of a CCR2 antagonist on macrophages and Toll-like receptor 9 expression in a mouse model of diabetic nephropathy

2021 ◽  
Author(s):  
Seigo Ito ◽  
Hiroyuki Nakashima ◽  
Takuya Ishikiriyama ◽  
Masahiro Nakashima ◽  
Akira Yamagata ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Therapeutic Effects ◽  
Ros Production ◽  
Toll Like Receptor ◽  
Tlr9 Expression ◽  
Tnf Α ◽  
Factor Α ◽  
And Function ◽  
Ccr2 Antagonist ◽  
Necrosis Factor

The pathogenesis of diabetic nephropathy (DN) is related to macrophage (Mφ) recruitment to the kidneys, tumor necrosis factor-α (TNF-α) production, and oxidative stress. Toll-like receptor 9 (TLR9) activation is reportedly involved in systemic inflammation, and it exacerbates this condition in metabolic syndrome. Therefore, we hypothesized that TLR9 plays a role in the pathogenesis of DN. Two subsets of kidney macrophages in DN model (db/db) mice were analyzed using flow cytometry to evaluate their distribution and TLR9 expression and function. Mice were administered the CCR2 antagonist INCB3344 for 8 weeks; changes in macrophage distribution and function and its therapeutic effects on DN pathology were examined. Bone marrow-derived CD11bhigh (BM-) and tissue-resident CD11blow (Res-) Mφs were identified in the mouse kidneys. As DN progressed, the BM-Mφ number, TLR9 expression, and TNF-α production increased significantly. In Res-Mφs, reactive oxygen species (ROS) production and phagocytic activity were enhanced. INCB3344 decreased albuminuria, serum creatinine level, BM-Mφs abundance, TLR9 expression, and TNF-α production by BM-Mφs and ROS production by Res-Mφs. Both increased activation of BM-Mφs via TLR9 and TNF-α production and increased ROS production by Res-Mφs were involved in DN progression. Thus, inactivating macrophages and their TLR9 expressions by INCB3344 is a potential therapeutic strategy for diabetic nephropathy.

scholarly journals Toll-like receptor-4, but not toll-like receptor-2 mediates secretion of tumour necrosis factor α and interleukin-8 in lipopolysaccharide-stimulated mouse mammary epithelial cells

2013 ◽  
Vol 57 (3) ◽  
pp. 393-397 ◽  
Author(s):  
Chang-Liang He ◽  
Qiong Yi ◽  
Yuan-Fang Li ◽  
Hang Yang ◽  
Lu Wang
Keyword(s):  
Epithelial Cells ◽  
Tumour Necrosis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Tumour Necrosis Factor Α ◽  
Toll Like Receptor ◽  
Tnf Α ◽  
Toll Like Receptor 2 ◽  
Factor Α ◽  
Necrosis Factor

Abstract Mammary epithelial cells (MECs) from Kunming mice were isolated and stimulated in vitro with 10 μg/mL of Escherichia coli lipopolysaccharide (LPS). The release of tumour necrosis factor α (TNF-α) and interleukin-8 (IL-8) into culture supernatants was measured by ELISA. Furthermore, blocking experiments with Toll-like receptor 2 (TLR2) and TLR4 antibodies were performed to verify whether cytokine secretion depended on LPS-induced activation of TLR2 or TLR4. The results revealed that LPS-stimulated mouse MECs significantly secreted TNF-α and IL-8. Blocking of the TLR4 pathway inhibited the secretion of TNF-α and IL-8, while inhibition of LPS-induced TNF-α and IL-8 production was not observed when TLR2 was blocked. Thus, TLR4 can mediate the LPS-induced expression of cytokines such as TNF-α and IL-8 in mouse MECs.

scholarly journals Subinhibitory Doses of Aminoglycoside Antibiotics Induce Changes in the Phenotype of Mycobacterium abscessus

2015 ◽  
Vol 59 (10) ◽  
pp. 6161-6169 ◽  
Author(s):  
Sheng-Hui Tsai ◽  
Hsin-Chih Lai ◽  
Shiau-Ting Hu
Keyword(s):  
Mycobacterium Abscessus ◽  
Aminoglycoside Antibiotics ◽  
Toll Like Receptor ◽  
Necrosis Factor Alpha ◽  
Content Type ◽  
Tnf Α ◽  
Bacterial Morphology ◽  
Toll Like Receptor 2 ◽  
Factor Alpha ◽  
Necrosis Factor

ABSTRACTSubinhibitory doses of antibiotics have been shown to cause changes in bacterial morphology, adherence ability, and resistance to antibiotics. In this study, the effects of subinhibitory doses of aminoglycoside antibiotics onMycobacterium abscessuswere investigated. The treatment ofM. abscessuscells with subinhibitory doses of amikacin was found to change their colony from a smooth to a rough morphotype and increase their ability to adhere to a polyvinylchloride plate, aggregate in culture, and resist phagocytosis and killing by macrophages.M. abscessuscells treated with a subinhibitory dose of amikacin also became more potent in Toll-like receptor 2 (TLR-2) stimulation, leading to increased tumor necrosis factor alpha (TNF-α) production by macrophages. The MAB_3508c gene was shown to play a role in mediating these phenotypic changes, as its expression inM. abscessuscells was increased when they were treated with a subinhibitory dose of amikacin. In addition, overexpression of MAB_3508c inM. abscessuscells caused changes similar to those induced by subinhibitory doses of amikacin, including a switch from smooth to rough colony morphology, increased ability to aggregate in liquid culture, decreased motility, and increased resistance to killing by macrophages. These findings suggest the importance of using sufficient doses of antibiotics for the treatment ofM. abscessusinfections.

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