scholarly journals Platelet Toll-like Receptor 4-related Innate Immunity Potentially Participates in Transfusion Reactions Independent of ABO Compatibility: An ex Vivo Study

2021 ◽  
Author(s):  
Chien-Sung Tsai ◽  
Mei-Hua Hu ◽  
Yung-Chi Hsu ◽  
Go-Shine Huang
Keyword(s):  
Innate Immunity ◽  
Ex Vivo ◽  
Binding Protein ◽  
Blood Type ◽  
Saline Control ◽  
Dependent Manner ◽  
Tlr4 Expression ◽  
Tnf Α ◽  
Transfusion Reactions ◽  
Lps Binding

Abstract Purpose: The role of platelet TLR4 in transfusion reactions remains unclear. This study analyzed platelet TLR4, certain DAMPs, and the effect of ABO compatibility on TLR4 expression after a simulated transfusion ex vivo.Methods: Donor red blood cells were harvested from a blood bank. Recipient blood from patients undergoing cardiac surgery was processed to generate a washed platelet suspension. Donor blood was added to the washed platelets at 1%, 5%, or 10% (v/v). Blood mixing experiments were performed using four groups: 0.9% saline control group (n = 31); M, matched blood type mixing (n = 20); S, uncross-matched ABO type-specific mixing (n = 20); and I, ABO incompatible blood mixing (n = 20). Platelet TLR4 expression was determined after blood mixing. Levels of TLR4-binding DAMPs (HMGB1, S100A8, S100A9, and SAA) and that of LPS-binding protein and endpoint proteins (TNF-α, IL-1β, and IL-6) in the TLR4 signaling pathway were evaluated.Results: The 1%, 5%, and 10% blood mixtures significantly increased TLR4 expression in three groups (M, S, and I; all P < 0.001) in a concentration-dependent manner. TLR4 expression did not significantly differ among the three groups (P = 0.148). HMGB1, S100A8, and S100A9 showed elevated levels in response to blood mixing; SAA, LPS-binding protein, TNF-α, IL-1β, and IL-6 did not. Conclusion: Blood mixing may elicit innate immune responses by upregulating platelet TLR4 and DAMPs unassociated with ABO compatibility, suggesting that innate immunity through TLR4-mediated signaling may induce transfusion reactions. The trial was retrospectively registered at Chinese Clinical Trial Registry (ChiCTR2100045606) with date of registration on 19 April 2021.

scholarly journals Platelet Toll-Like Receptor 4–Related Innate Immunity Potentially Participates in Transfusion Reactions Independent of ABO Compatibility: An Ex Vivo Study

Biomedicines ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 29
Author(s):  
Chien-Sung Tsai ◽  
Mei-Hua Hu ◽  
Yung-Chi Hsu ◽  
Go-Shine Huang
Keyword(s):  
Innate Immunity ◽  
Ex Vivo ◽  
Binding Protein ◽  
Blood Type ◽  
Saline Control ◽  
Tlr4 Expression ◽  
Lipopolysaccharide Binding Protein ◽  
Group I ◽  
Transfusion Reactions ◽  
Lipopolysaccharide Binding

The role of platelet TLR4 in transfusion reactions remains unclear. This study analyzed platelet TLR4 and certain damage-associated molecular patterns (DAMPs) and evaluated how ABO compatibility affected TLR4 expression after a simulated ex vivo transfusion. A blood bank was the source of donor red blood cells. Blood from patients undergoing cardiac surgery was processed to generate a washed platelet suspension to which the donor blood was added in concentrations 1, 5, and 10% (v/v). Blood-mixing experiments were performed on four groups: a 0.9% saline control group (n = 31); a matched-blood-type mixing group (group M, n = 20); an uncross-matched ABO-specific mixing group (group S, n = 20); and an ABO-incompatible blood mixing group (group I, n = 20). TLR4 expression in the platelets was determined after blood mixing. We evaluated levels of TLR4-binding DAMPs (HMGB1, S100A8, S100A9, and SAA), lipopolysaccharide-binding protein, and endpoint proteins in the TLR4 signaling pathway. In the M, S, and I groups, 1, 5, and 10% blood mixtures significantly increased TLR4 expression (all p < 0.001) in a concentration-dependent manner. Groups M, S, and I were not discovered to have significantly differing TLR4 expression (p = 0.148). HMGB1, S100A8, and S100A9 levels were elevated in response to blood mixing, but SAA, lipopolysaccharide-binding protein, TNF-α, IL-1β, and IL-6 levels were not. Blood mixing may elicit innate immune responses by upregulating platelet TLR4 and DAMPs unassociated with ABO compatibility, suggesting that innate immunity through TLR4-mediated signaling may induce transfusion reactions.

scholarly journals Targeted Deletion of the Lipopolysaccharide (LPS)-binding Protein Gene Leads to Profound Suppression of LPS Responses Ex Vivo, whereas In Vivo Responses Remain Intact

1997 ◽  
Vol 186 (12) ◽  
pp. 2051-2056 ◽  
Author(s):  
Mark M. Wurfel ◽  
Brian G. Monks ◽  
Robin R. Ingalls ◽  
Russell L. Dedrick ◽  
Russell Delude ◽  
...  
Keyword(s):  
Lipid Transfer Protein ◽  
Ex Vivo ◽  
Binding Protein ◽  
Embryonic Stem ◽  
Cellular Responses ◽  
Tnf Α ◽  
Deficient Mice ◽  
Lps Binding

Gram-negative bacterial lipopolysaccharide (LPS) stimulates phagocytic leukocytes by interacting with the cell surface protein CD14. Cellular responses to LPS are markedly potentiated by the LPS-binding protein (LBP), a lipid-transfer protein that binds LPS aggregates and transfers LPS monomers to CD14. LBP also transfers LPS to lipoproteins, thereby promoting the neutralization of LPS. LBP present in normal plasma has been shown to enhance the LPS responsiveness of cells in vitro. The role of LBP in promoting LPS responsiveness in vivo was tested in LBP-deficient mice produced by gene targeting in embryonic stem cells. Whole blood from LBP-deficient animals was 1,000-fold less responsive to LPS as assessed by the release of tumor necrosis factor (TNF)-α. Blood from gene-targeted mice was devoid of immunoreactive LBP, essentially incapable of transferring LPS to CD14 in vitro, and failed to support cellular responses to LPS. These activities were restored by the addition of exogenous recombinant murine LBP to the plasma. Despite these striking in vitro findings, no significant differences in TNF-α levels were observed in plasma from wild-type and LBP-deficient mice injected with LPS. These data suggest the presence of an LBP-independent mechanism for responding to LPS. These LBP knockout mice may provide a tool for discovering the nature of the presumed second mechanism for transferring LPS to responsive cells.

Hemoglobin Enhances the Binding of Bacterial Endotoxin to Human Endothelial Cells

1996 ◽  
Vol 76 (02) ◽  
pp. 258-262 ◽  
Author(s):  
Robert I Roth
Keyword(s):  
Endothelial Cells ◽  
Binding Protein ◽  
Bacterial Endotoxin ◽  
Dependent Manner ◽  
Serum Free Medium ◽  
Free Medium ◽  
Human Endothelial Cells ◽  
Serum Factors ◽  
Serum Free ◽  
Lps Binding

SummaryHuman endothelial cells, when incubated with bacterial endotoxin (lipopolysaccharide, LPS), modify their surface in association with prominent production of procoagulant tissue factor (TF) activity. This deleterious biological effect of LPS has been shown previously to be enhanced approximately 10-fold by the presence of hemoglobin (Hb), a recently recognized LPS binding protein that causes disaggregation of LPS and increases the biological activity of LPS in a number of in vitro assays. The present study was performed to test the hypothesis that Hb enhances the LPS-induced procoagulant activity of human umbilical vein endothelial cells (HUVEC) by increasing LPS binding to the cells. The binding of 3H-LPS to HUVEC was determined in the absence or presence of Hb or two other known LPS-binding proteins, human serum albumin (HSA) and IgG. LPS binding was substantially increased in the presence of Hb, in a Hb concentration-dependent manner, but was not increased by HSA or IgG. Hb enhancement of LPS binding was observed in serum-free medium, indicating that there was no additional requirement for any of the serum factors known to participate in the interaction of LPS with cells (e.g., lipopolysaccharide (LPS)-binding protein (LBP) and soluble CD14 (sCD14)). Hb enhancement of LPS binding also was observed in the more physiologic condition of 100% plasma. LPS-induced TF activity was stimulated by Hb, but not by HSA or IgG. In serum-free medium, TF activity was not stimulated under any of the conditions tested. Ultrafiltration of LPS was dramatically increased after incubation with Hb but not with HSA or IgG, suggesting that LPS disaggregation by Hb was responsible for the enhanced binding of LPS to HUVEC and the subsequent stimulation of TF activity.

scholarly journals Platelet Toll-like receptor expression modulates lipopolysaccharide-induced thrombocytopenia and tumor necrosis factor-α production in vivo

Blood ◽  
2006 ◽  
Vol 107 (2) ◽  
pp. 637-641 ◽  
Author(s):  
Rukhsana Aslam ◽  
Edwin R. Speck ◽  
Michael Kim ◽  
Andrew R. Crow ◽  
K. W. Annie Bang ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Critical Role ◽  
Receptor Expression ◽  
Tlr4 Expression ◽  
Antiplatelet Antibody ◽  
Tnf Α ◽  
Intracellular Compartments ◽  
Thrombin Activation ◽  
Factor Α

AbstractToll-like receptors (TLRs) play a critical role in stimulating innate immunity by recognizing pathogen-associated molecular patterns (PAMPs) on invading microorganisms. Platelets also play a role in innate immunity, and we studied whether they express TLR. Results show that human and murine platelets variably expressed TLR2, TLR4, and TLR9 by flow cytometry and Western blotting. TLR4 expression was confirmed by demonstrating murine platelet binding to lipopolysaccharide (LPS). Thrombin activation of the platelets significantly enhanced the expression of TLR9, suggesting that at least some TLRs may derive from intracellular compartments. When LPS was administered to LPS-sensitive C3H/HeN and LPS-resistant C3H/HeJ mice, functional TLR4 expression in vivo was shown to be responsible for LPS-induced thrombocytopenia. However, when the C3H/HeN mice were first rendered thrombocytopenic by an antiplatelet antibody and then administered LPS, a significant reduction occurred in their ability to produce TNF-α. The decreased cytokine production in the thrombocytopenic mice was restored with platelet transfusion. These results suggest that platelets express various TLRs and that the functional significance of one of these, TLR4, appears to be a role in the modulation of LPS-induced thrombocytopenia and TNF-α production. This work implicates platelets as important mediators of innate immune responses against invading microorganisms.

Interleukin-18 binding protein therapy is protective in adriamycin nephropathy

2013 ◽  
Vol 304 (1) ◽  
pp. F68-F76 ◽  
Author(s):  
Kate R. Wyburn ◽  
Steven J. Chadban ◽  
Tony Kwan ◽  
Stephen I. Alexander ◽  
Huiling Wu
Keyword(s):  
Binding Protein ◽  
Kidney Injury ◽  
Saline Control ◽  
Interleukin 18 ◽  
Protein Therapy ◽  
Immune Mediators ◽  
Adriamycin Nephropathy ◽  
Tnf Α ◽  
Inflammatory Molecules ◽  
Oxide Synthase

Adriamycin nephropathy (AN) is an experimental model of focal segmental glomerulosclerosis (FSGS) in which macrophages are considered to play a pathogenic role. We hypothesize that interleukin-18 (IL-18), largely derived from macrophages, is a key contributor to kidney injury in AN and a potential therapeutic target. In this study, BALB/c mice received adriamycin (9.6 mg/kg) via tail vein injection and subsequently were treated with either neutralizing IL-18 binding protein (IL-18BP; 250 μg) or normal saline (control). At 5 wk, IL-18 was upregulated in AN, and IL-18BP therapy afforded significant protection against the development of AN, resulting in less proteinuria ( P < 0.01), kidney dysfunction ( P < 0.01), glomerulosclerosis ( P < 0.001), and interstitial accumulation of macrophages and T cells ( P < 0.001). Gene expression of IL-18 downstream inflammatory molecules, including inducible nitric oxide synthase ( P < 0.001), TNF-α ( P < 0.001), and IFN-γ ( P < 0.01); IL-17 ( P < 0.001) and the chemokines CCL2 ( P < 0.01) and CCL5 ( P < 0.005), was reduced. We demonstrate that IL-18 plays a significant role in the pathogenesis of AN. The protective effect of IL-18BP therapy illustrates the importance of immune mediators in chronic proteinuric kidney disease and highlights the potential of IL-18BP therapy.

scholarly journals Inflammation and LPS-Binding Protein Enable the Stimulatory Effect of Endotoxin on Prolactin Secretion in the Ovine Anterior Pituitary: Ex Vivo Study

2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Dorota Tomaszewska-Zaremba ◽  
Karolina Haziak ◽  
Monika Tomczyk ◽  
Andrzej P. Herman
Keyword(s):  
Anterior Pituitary ◽  
Ex Vivo ◽  
Binding Protein ◽  
Hormone Secretion ◽  
Prolactin Secretion ◽  
Cell Surface Receptor ◽  
Bacterial Endotoxin ◽  
Pituitary Cells ◽  
Immune Stress ◽  
Lps Binding

Prolactin is a hormone that plays an important role in the regulation of many physiological processes including lactation, reproduction, fat metabolism, and immune response. The secretion of prolactin could be disturbed by an immune stress commonly accompanying infection. This study was designed to determine the influence of bacterial endotoxin—lipopolysaccharide (LPS)—on prolactin gene (PRL) expression and prolactin release from the ovine anterior pituitary (AP) explants collected from saline- and LPS-treated ewes in the follicular phase. The expressions of toll-like receptor 4 (TLR4) and proinflammatory cytokines interleukin- (IL-) 1β, IL-6, and tumor necrosis factor- (TNF-) α genes were also assayed. The results of the study showed that LPS stimulates prolactin secretion and IL-6 gene expression in the AP explants, but its action on lactotrophs depends on the immunological status of animal. It was demonstrated that an important role in enhancing the effect of LPS on the pituitary in the saline-treated ewes is played by LPS-binding protein (LBP)- “adapter molecule” for LPS binding to the cell surface receptor CD14 and then to TLR4. Also, it was found that bacterial endotoxin acting on the anterior pituitary cells may enhance prolactin secretion, and this effect of LPS could be mediated by IL-6 which is known as prolactin-releasing factor. Identification of the neuroendocrine and immune interactions in the regulation of prolactin secretion could be helpful in developing newer and more effective treatments for dysfunctions connected with disorders in this hormone secretion.

Interleukin-11 attenuates pulmonary inflammation and vasomotor dysfunction in endotoxin-induced lung injury

1999 ◽  
Vol 277 (5) ◽  
pp. L861-L867 ◽  
Author(s):  
Brett C. Sheridan ◽  
Charles A. Dinarello ◽  
Daniel R. Meldrum ◽  
David A. Fullerton ◽  
Craig H. Selzman ◽  
...  
Keyword(s):  
Lung Injury ◽  
Lung Tumor ◽  
Binding Protein ◽  
Saline Control ◽  
Myeloperoxidase Activity ◽  
Neutrophil Sequestration ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Relaxation Responses ◽  
Vasomotor Dysfunction

Interleukin (IL)-11, like other members of the gp130 receptor class, possesses anti-inflammatory properties. We hypothesized that IL-11 pretreatment would attenuate endotoxin [lipopolysaccharide (LPS)]-induced lung inflammation and diminish injury to endothelium-dependent and -independent mechanisms of pulmonary vasorelaxation that require cGMP in Sprague-Dawley rats. LPS (20 mg/kg ip) increased lung tumor necrosis factor (TNF)-α compared with the saline control (0.7 ± 0.15 ng/g lung wet wt for control vs. 3.5 ± 0.09 ng/g lung wet wt for LPS; P < 0.05). IL-11 (200 mg/kg ip) injected 10 min before LPS administration attenuated the LPS-induced lung TNF-α levels (1.6 ± 0.91 ng/g lung wet wt; P < 0.05 vs. LPS). IL-11 also diminished LPS-induced lung neutrophil sequestration as assessed by myeloperoxidase units (2.1 ± 0.25 U/g lung wet wt for saline and 15.6 ± 2.02 U/g lung wet wt for LPS vs. 7.07 ± 1.65 U/g lung wet wt for LPS plus IL-11; P < 0.05). Similarly, TNF-α binding protein (175 mg/kg) attenuated LPS-induced myeloperoxidase activity (6.04 ± 0.14 U/g lung wet wt; P < 0.05). Both IL-11 and TNF-α binding protein similarly attenuated LPS-induced endothelium-dependent vasomotor dysfunction with improved relaxation responses to 10−7and 10−6M acetylcholine and A-23187 in phenylephrine-preconstricted isolated pulmonary artery rings ( P < 0.05 vs. LPS). Endothelium-independent relaxation responses to sodium nitroprusside were also improved after LPS at 10−6M ( P < 0.05 vs. LPS). Moreover, IL-11 decreased endotoxin-induced mortality in CF1 mice from 90 to 50% ( P ≤ 0.05 vs. LPS). Therefore, IL-11 prevents LPS-induced lung TNF-α production, neutrophil sequestration, and pulmonary vasomotor dysfunction. We conclude that IL-11 possesses anti-inflammatory activity that protects against LPS-induced lung injury and lethality.

IMPROVED SURVIVAL AND INCREASED HEPATIC TNF-α GENE EXPRESSION IN LPS BINDING PROTEIN (LBP) KNOCKOUT MICE GIVEN SYSTEMIC BUT NOT LOCALIZED GENE THERAPY IN A PNEUMONIA MODEL.

Shock ◽  
2002 ◽  
Vol 17 (Supplement) ◽  
pp. 30
Author(s):  
M H Fan ◽  
L Steinstraesser ◽  
L U Lahoda ◽  
R D Klein ◽  
A C Merry ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Therapy ◽  
Knockout Mice ◽  
Binding Protein ◽  
Tnf Α ◽  
Lps Binding

scholarly journals Binding of Plasminogen to Streptococcus suis Protein Endopeptidase O Facilitates Evasion of Innate Immunity in Streptococcus suis

2021 ◽  
Vol 12 ◽  
Author(s):  
Yang Zhou ◽  
Kang Yan ◽  
Chengfeng Sun ◽  
Feng Liu ◽  
Wei Peng ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Immune Evasion ◽  
Binding Protein ◽  
Bacterial Species ◽  
Gain Control ◽  
Streptococcus Suis ◽  
Dependent Manner ◽  
Deficient Mutant ◽  
Wildtype Strain ◽  
Type Plasminogen Activator

The Gram-positive bacterial species Streptococcus suis is an important porcine and human pathogen that causes severe life-threatening diseases associated with high mortality rates. However, the mechanisms by which S. suis evades host innate immunity remain elusive, so identifying novel virulence factors involved in immune evasion is crucial to gain control over this threatening pathogen. Our previous work has shown that S. suis protein endopeptidase O (SsPepO) is a novel fibronectin-binding protein. Here, we identified that recombinant SsPepO binds human plasminogen in a dose-dependent manner. Moreover, the binding of SsPepO and plasminogen, upon the activation of urokinase-type plasminogen activator, generated plasmin, which could cleave complement C3b, thus playing an important role in complement control. Additionally, a SspepO-deficient mutant showed impaired adherence to plasminogen as well as impaired adherence to and invasion of rat brain microvascular endothelial cells compared with the wildtype strain. We further found that the SspepO-deficient mutant was efficiently killed by human serum and blood. We also confirmed that the SspepO-deficient mutant had a lower mortality rate than the wildtype strain in a mouse model. In conclusion, these results indicate that SsPepO is a novel plasminogen-binding protein that contributes to S. suis immune evasion.

scholarly journals Novel LysM motifs for antigen display on lactobacilli for mucosal immunization

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Fernanda Raya-Tonetti ◽  
Melisa Müller ◽  
Jacinto Sacur ◽  
Haruki Kitazawa ◽  
Julio Villena ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Dependent Manner ◽  
The Novel ◽  
Destination Vector ◽  
Serum Igg ◽  
Tnf Α ◽  
Fluorescence Protein ◽  
Serum Igg Levels ◽  
Dose Dependent ◽  
Harsh Conditions

AbstractWe characterized two LysM domains of Limosilactobacillus fermentum, belonging to proteins Acglu (GenBank: KPH22907.1) and Pgb (GenBank: KPH22047.1) and bacterium like particles (BLP) derived from the immunomodulatory strain Lacticaseibacillus rhamnosus IBL027 (BLPs027) as an antigen display platform. The fluorescence protein Venus fused to the novel LysM domains could bind to the peptidoglycan shell of lactobacilli and resisted harsh conditions such as high NaCl and urea concentrations. Acglu with five LysM domains was a better anchor than Pgb baring only one domain. Six-week-old BALB/c mice were nasally immunized with the complex Venus-Acglu-BLPs027 at days 0, 14 and 28. The levels of specific serum IgG, IgG1 and IgG2a and the levels of total immunoglobulins (IgT) and IgA in broncho-alveolar lavage (BAL) were evaluated ten days after the last boosting. Venus-Acglu-BLPs027, nasally administered, significantly increased specific BAL IgT and IgA, and serum IgG levels. In addition, spleen cells of mice immunized with Venus-Acglu-BLPs027 secreted TNF-α, IFN-γ and IL-4 when stimulated ex vivo in a dose-dependent manner. We constructed a Gateway compatible destination vector to easily fuse the selected LysM domain to proteins of interest for antigen display to develop mucosal subunit vaccines.

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