PRE- AND POSTOPERATIVE CYTOKINE RELEASE AFTER IN VITRO WHOLE BLOOD LIPOPOLYSACCHARIDE STIMULATION AND FREQUENT TOLL-LIKE RECEPTOR 4 POLYMORPHISMS

Abstract Background: In coronary in-stent restenosis (ISR), a substantial contribution of inflammation is assumed. We evaluated the association between polymorphisms in the Toll-like receptor 4 (TLR4) gene and cytokine response after lipopolysaccharide (LPS) challenge and the development of ISR. Methods: Patients were included after successful elective stent placement in a native coronary artery and were scheduled for follow-up angiography after 6 months. Quantitative coronary analysis was performed off-line. Patient whole blood was challenged with LPS for 24 h. Baseline and stimulated concentrations of interleukin (IL)-1β, IL-6, tumor necrosis factor-α, and IL-10 were assessed by ELISA. Two cosegregating single-nucleotide polymorphisms in the TLR4 gene (Asp299Gly and Thr399Ile) were analyzed by allele-specific PCR amplification of genomic DNA. Results: A total of 236 consecutive patients were included, and 40 (17%) developed ISR. Median baseline and stimulated cytokine concentrations did not differ between patients with and without ISR. In multivariate analysis, male sex, unstable angina, hypertension, and chronic total occlusion were predictors of ISR. TLR4 genotypes were not associated with baseline or stimulated cytokine concentrations or with angiographic variables at follow-up. Conclusions: In vitro cytokine response to LPS challenge is not increased in patients with ISR. Functionality of the TLR4 Asp299Gly polymorphism could not be demonstrated in this setting, and this polymorphism was not associated with angiographic outcome, calling into question its role in the progression of neointimal tissue growth.

Download Full-text

Abstract 197: Emerging Role of Toll-like Receptor 4 and Heat Shock Protein 60&70 in Ischaemia-Induced Skeletal Muscle Damage In Vitro

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.33.suppl_1.a197 ◽

2013 ◽

Vol 33 (suppl_1) ◽

Author(s):

Ali Navi ◽

Rebekah Yu ◽

Xu Shi-Wen ◽

Sidney Shaw ◽

Daryll Baker ◽

...

Keyword(s):

Skeletal Muscle ◽

Inflammatory Cytokine ◽

Caspase 3 ◽

Neutralizing Antibody ◽

Cytokine Release ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Skeletal Muscle Damage ◽

Tnf Α

OBJECTIVES The innate immune response contributes to the skeletal muscle damage in patients with critical limb ischaemia (CLI); however, the detailed signaling mechanisms are not fully understood. We hypothesized that simulated ischaemia induces inflammatory cytokine release from skeletal myotubes, via a mechanism that involves heat shock protein (HSP) 60&70, known endogenous ligands of Toll-like receptor 4 (TLR4), in vitro. METHODS Human gastrocnemius muscle biopsies were taken from patients with CLI undergoing major lower limb amputation and from patients with no peripheral arterial disease (PAD). Human myoblasts were isolated, cultured to myotubes and then pre-treated with TLR4 neutralizing antibody prior to exposure to simulated ischaemia. Fluorescent immunostaining was carried out to confirm cell differentiation; ELISA analysis were carried out to quantify IL6 and TNF-α release; and Western blot was used to assess expression of HSP60&70, TLR4 and cleaved caspase-3 as a marker of apoptosis. RESULTS Myotubes from patients with CLI expressed greater levels of cleaved caspase-3 and TLR4 as compared to those from patients with no PAD. When exposed to ischaemic conditions, increased IL6 and TNF-α release and upregulation of HSP60&70, cleaved caspase-3 and TLR4 were observed in myotubes from both groups of patients compared to culturing in normoxic conditions (P<0.05). Pre-treatment of myotubes from patients with CLI with TLR4 neutralizing antibody prior to simulated ischaemia was associated with reduced expression of HSP60&70, IL6, TNF-α and cleaved caspase-3 (P<0.05). CONCLUSIONS Increased cytokine release, apoptosis and expression of HSP60&70 and TLR4 occur in ischaemic skeletal muscle in vitro. TLR4 antagonism was associated with reduced apoptosis and inflammatory cytokine release and down-regulation of HSP60&70 expression. This suggests a potential pathway where TLR4 and its endogenous ligands contribute to a positive feedback loop to maintain a proinflammatory environment during ischaemia.

Download Full-text

Heterozygous Toll‐Like Receptor 4 Polymorphism Does Not Influence Lipopolysaccharide‐Induced Cytokine Release in Human Whole Blood

The Journal of Infectious Diseases ◽

10.1086/378095 ◽

2003 ◽

Vol 188 (6) ◽

pp. 938-943 ◽

Cited By ~ 57

Author(s):

Sonja von Aulock ◽

Nicolas W. J. Schröder ◽

Katja Gueinzius ◽

Stephanie Traub ◽

Sebastian Hoffmann ◽

...

Keyword(s):

Whole Blood ◽

Cytokine Release ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Human Whole Blood

Download Full-text

Serum Amyloid A1/Toll-Like Receptor-4 Axis, an Important Link between Inflammation and Outcome of TBI Patients

Biomedicines ◽

10.3390/biomedicines9060599 ◽

2021 ◽

Vol 9 (6) ◽

pp. 599

Author(s):

Víctor Farré-Alins ◽

Alejandra Palomino-Antolín ◽

Paloma Narros-Fernández ◽

Ana Belen Lopez-Rodriguez ◽

Céline Decouty-Perez ◽

...

Keyword(s):

Injury Severity ◽

White Blood Cells ◽

Serum Amyloid ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Important Link ◽

Tlr4 Mrna ◽

Serum Amyloid A1

Traumatic brain injury (TBI) is one of the leading causes of mortality and disability worldwide without any validated biomarker or set of biomarkers to help the diagnosis and evaluation of the evolution/prognosis of TBI patients. To achieve this aim, a deeper knowledge of the biochemical and pathophysiological processes triggered after the trauma is essential. Here, we identified the serum amyloid A1 protein-Toll-like receptor 4 (SAA1-TLR4) axis as an important link between inflammation and the outcome of TBI patients. Using serum and mRNA from white blood cells (WBC) of TBI patients, we found a positive correlation between serum SAA1 levels and injury severity, as well as with the 6-month outcome of TBI patients. SAA1 levels also correlate with the presence of TLR4 mRNA in WBC. In vitro, we found that SAA1 contributes to inflammation via TLR4 activation that releases inflammatory cytokines, which in turn increases SAA1 levels, establishing a positive proinflammatory loop. In vivo, post-TBI treatment with the TLR4-antagonist TAK242 reduces SAA1 levels, improves neurobehavioral outcome, and prevents blood–brain barrier disruption. Our data support further evaluation of (i) post-TBI treatment in the presence of TLR4 inhibition for limiting TBI-induced damage and (ii) SAA1-TLR4 as a biomarker of injury progression in TBI patients.

Download Full-text

Detrimental Role of Heat Shock Protein 60&70 and Toll-like Receptor 4 in Skeletal Muscle Ischaemia In Vitro

Journal of Vascular Surgery ◽

10.1016/j.jvs.2013.02.134 ◽

2013 ◽

Vol 57 (5) ◽

pp. 77S

Author(s):

Ali Navi ◽

Rebekah Yu ◽

Xu Shi-Wen ◽

Sidney Shaw ◽

George Hamilton ◽

...

Keyword(s):

Skeletal Muscle ◽

Heat Shock ◽

Heat Shock Protein ◽

Heat Shock Protein 60 ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Muscle Ischaemia

Download Full-text

Ginsenoside Rb1 exerts anti-inflammatory effects in vitro and in vivo by modulating toll-like receptor 4 dimerization and NF-kB/MAPKs signaling pathways

Phytomedicine ◽

10.1016/j.phymed.2020.153197 ◽

2020 ◽

Vol 69 ◽

pp. 153197 ◽

Cited By ~ 5

Author(s):

Hongwei Gao ◽

Naixin Kang ◽

Chao Hu ◽

Ziyu Zhang ◽

Qiongming Xu ◽

...

Keyword(s):

Signaling Pathways ◽

Ginsenoside Rb1 ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Anti Inflammatory

Download Full-text

Micrococcus luteus Teichuronic Acids Activate Human and Murine Monocytic Cells in a CD14- and Toll-Like Receptor 4-Dependent Manner

Infection and Immunity ◽

10.1128/iai.69.4.2025-2030.2001 ◽

2001 ◽

Vol 69 (4) ◽

pp. 2025-2030 ◽

Cited By ~ 32

Author(s):

Shuhua Yang ◽

Shunji Sugawara ◽

Toshihiko Monodane ◽

Masahiro Nishijima ◽

Yoshiyuki Adachi ◽

...

Keyword(s):

Lipid A ◽

Micrococcus Luteus ◽

Dependent Manner ◽

Toll Like Receptor ◽

Toll Like Receptor 4 ◽

Necrosis Factor Alpha ◽

Monocytic Cells ◽

Tnf Α

ABSTRACT Teichuronic acid (TUA), a component of the cell walls of the gram-positive organism Micrococcus luteus (formerlyMicrococcus lysodeikticus), induced inflammatory cytokines in C3H/HeN mice but not in lipopolysaccharide (LPS)-resistant C3H/HeJ mice that have a defect in the Toll-like receptor 4 (TLR4) gene, both in vivo and in vitro, similarly to LPS (T. Monodane, Y. Kawabata, S. Yang, S. Hase, and H. Takada, J. Med. Microbiol. 50:4–12, 2001). In this study, we found that purified TUA (p-TUA) induced tumor necrosis factor alpha (TNF-α) in murine monocytic J774.1 cells but not in mutant LR-9 cells expressing membrane CD14 at a lower level than the parent J774.1 cells. The TNF-α-inducing activity of p-TUA in J774.1 cells was completely inhibited by anti-mouse CD14 monoclonal antibody (MAb). p-TUA also induced interleukin-8 (IL-8) in human monocytic THP-1 cells differentiated to macrophage-like cells expressing CD14. Anti-human CD14 MAb, anti-human TLR4 MAb, and synthetic lipid A precursor IVA, an LPS antagonist, almost completely inhibited the IL-8-inducing ability of p-TUA, as well as LPS, in the differentiated THP-1 cells. Reduced p-TUA did not exhibit any activities in J774.1 or THP-1 cells. These findings strongly suggested that M. luteus TUA activates murine and human monocytic cells in a CD14- and TLR4-dependent manner, similar to LPS.

Download Full-text