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Chemokines (CCL3, CCL4, and CCL5) Inhibit ATP-Induced Release of IL-1β by Monocytic Cells.
Amati, AL, Zakrzewicz, A, Siebers, R, Wilker, S, Heldmann, S, Zakrzewicz, D, Hecker, A, McIntosh, JM, Padberg, W, Grau, V
Mediators of inflammation. 2017;:1434872
Abstract
Chemokines and ATP are among the mediators of inflammatory sites that can enter the circulation via damaged blood vessels. The main function of chemokines is leukocyte mobilization, and ATP typically triggers inflammasome assembly. IL-1β, a potent inflammasome-dependent cytokine of innate immunity, is essential for pathogen defense. However, excessive IL-1β may cause life-threatening systemic inflammation. Here, we hypothesize that chemokines control ATP-dependent secretion of monocytic IL-1β. Lipopolysaccharide-primed human monocytic U937 cells were stimulated with the P2X7 agonist BzATP for 30 min to induce IL-1β release. CCL3, CCL4, and CCL5 dose dependently inhibited BzATP-stimulated release of IL-1β, whereas CXCL16 was ineffective. The effect of CCL3 was confirmed for primary mononuclear leukocytes. It was blunted after silencing CCR1 or calcium-independent phospholipase A2 (iPLA2) by siRNA and was sensitive to antagonists of nicotinic acetylcholine receptors containing subunits α7 and α9. U937 cells secreted small factors in response to CCL3 that mediated the inhibition of IL-1β release. We suggest that CCL chemokines inhibit ATP-induced release of IL-1β from U937 cells by a triple-membrane-passing mechanism involving CCR, iPLA2, release of small mediators, and nicotinic acetylcholine receptor subunits α7 and α9. We speculate that whenever chemokines and ATP enter the circulation concomitantly, systemic release of IL-1β is minimized.
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The Role of Extracellular Adenosine Triphosphate in Ischemic Organ Injury.
Zhao, H, Kilgas, S, Alam, A, Eguchi, S, Ma, D
Critical care medicine. 2016;(5):1000-12
Abstract
OBJECTIVES Ischemic tissue injury contributes to significant morbidity and mortality and is implicated in a range of pathologic conditions, including but not limited to myocardial infarction, ischemic stroke, and acute kidney injury. The associated reperfusion phase is responsible for the activation of the innate and adaptive immune system, further accentuating inflammation. Adenosine triphosphate molecule has been implicated in various ischemic conditions, including stroke and myocardial infarction. STUDY SELECTION Adenosine triphosphate is a well-defined intracellular energy transfer and is commonly referred to as the body's "energy currency." However, Laboratory studies have demonstrated that extracellular adenosine triphosphate has the ability to initiate inflammation and is therefore referred to as a damage-associated molecular pattern. Purinergic receptors-dependent signaling, proinflammatory cytokine release, increased Ca influx into cells, and subsequent apoptosis have been shown to form a common underlying extracellular adenosine triphosphate molecular mechanism in ischemic organ injury. CONCLUSIONS In this review, we aim to discuss the molecular mechanisms behind adenosine triphosphate-mediated ischemic tissue injury and evaluate the role of extracellular adenosine triphosphate in ischemic injury in specific organs, in order to provide a greater understanding of the pathophysiology of this complex process. We also appraise potential future therapeutic strategies to limit damage in various organs, including the heart, brain, kidneys, and lungs.
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An optimized protocol for adenosine triphosphate quantification in T lymphocytes of lymphopenic patients.
Girardot, T, Mouillaux, J, Idealisoa, E, Poujol, F, Rouget, C, Rimmelé, T, Monneret, G, Textoris, J, Venet, F
Journal of immunological methods. 2016;:59-66
Abstract
In several clinical contexts, the measurement of ATP concentration in T lymphocytes has been proposed as a biomarker of immune status, predictive of secondary infections. However, the use of such biomarker in lymphopenic patients requires some adaptations in the ATP dosage protocol. We used blood from healthy volunteers to determine the optimal experimental settings. We investigated technical aspects such as the type of anticoagulant for blood sampling, the effect of freeze and thaw cycles, the reagent and sample mixing sequence, and the optimal dilution buffer. We also shortened the incubation time to 8h, and even showed that a 30min incubation may be sufficient. To evaluate the ATP rise upon lymphocyte activation, the optimal dose of stimulant was defined to be 4μg/mL of phytohaemagglutinin. Lastly, we determined that the number of T cells needed for this measurement was as low as 50,000, which is compatible with the existing lymphopenia in clinical settings. This optimized protocol appears ready to be assessed in lymphopenic patients to further investigate the interconnection between T lymphocyte metabolism and impaired phenotype and functions.
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Low adenosine triphosphate activity in CD4+ cells predicts infection in patients with lupus nephritis.
Liu, J, Pan, Y, Tang, LJ, Bao, JF, Hao, J, Yu, Q, Yuan, WJ, Jin, HM
Clinical and experimental rheumatology. 2014;(3):383-9
Abstract
OBJECTIVES The ImmuKnow (Cylex) assay has been reported to predict the risk of infection in some diseases; however, it is uncertain whether ImmuKnow can predict the risk of infection in lupus nephritis (LN) patients receiving immunosuppressive therapy. METHODS The ImmuKnow Immune Cell Function Assay (Cylex, Inc., Columbia, MD, USA) was applied to measure the activity of CD4+ T cells, as a marker of global immune-competence. The correlation between changes in T cell activation and the relative risk of over-immunosuppression as well as infection was studied. The amount of adenosine triphosphate (ATP) produced by CD4+ T cells in response to phytohemagglutinin (PHA) was measured for 74 LN patients without infection, 22 LN patients with severe infection (i.e. required hospitalisation), and 28 healthy controls. RESULTS No correlation was found between the ATP level and systemic lupus erythematosus (SLE) activity. The mean ATP level was significantly lower in LN patients with infection than that in healthy controls (p<0.01) and non-infected LN patients (p<0.01). The mean ATP level in non-infected LN patients was not significantly different compared to healthy controls. A cut-off ATP value of 300 ng/mL predicted infection in LN patients with a specificity of 77% and a sensitivity of 77%. Multi-variable partial correlation coefficient between the ATP assay and severe infection was r =-0.040, p<0.001; CRP was r=0.962, p<0.001. CONCLUSIONS The ImmuKnow assay may be effective in identifying an increased risk of infection in LN patients but is not correlated with SLE activity. Combined CRP value will increase the diagnostic rate of severe infection in SLE. Larger studies are required to establish clinical advantages of this assay in SLE treatment.
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Significant reduction of ATP production in PHA-activated CD4+ cells in 1-day-old blood from transplant patients.
Suviolahti, E, Petrosyan, A, Mirocha, J, Ge, S, Karasyov, A, Thomas, D, Galera, O, Lim, W, Jimenez, AM, Czer, LS, et al
Transplantation. 2012;(12):1243-9
Abstract
BACKGROUND Global immunosuppression can be measured by assessing adenosine triphospate (ATP) levels in mitogen-stimulated CD4+ T cells. METHODS We investigated the effect of storage time on ATP levels in 234 blood samples from 18 healthy individuals and 152 transplant patients. The difference between day 0 (<13 hours post-blood draw) and day 1 (24-37 hours) measurements was analyzed and compared with various factors; a subset of samples was also analyzed in 6-hour intervals. RESULTS The ATP levels were significantly lower on day 1 compared with that on day 0 in healthy individuals (279±159 vs 414±159 ng/mL, P<0.001) and patients (356±209 vs 455±221 ng/mL, P<0.0001). Of the 18 healthy individuals, 17 showed ATP reduction, whereas 192 (89%) of 216 patients did so on day 1 (24.8±24.1%). In the time course analysis, ATP levels decreased with the blood storage time in healthy and patient samples, and the reduction began as early as 7 hours post-blood draw. The reduction rate was significantly higher in patient samples with low day 0 ATP levels compared with samples with moderate or high levels (44.7±31.3% vs 23.2±23.6% or 18.7±15.7%; P<0.001). The reduction rate in patients treated with alemtuzumab induction was slightly higher than that in daclizumab-treated patients (28.8±24.6% vs 21.3±21.3%, P=0.09). CD4+ cell number did not change within 24 hours post-blood draw, but CD4 expression decreased 2.0±2.8% (P<0.05). CONCLUSIONS The ATP levels are significantly lower in 1-day-old blood compared with fresh blood, suggesting that fresh blood should be used for assessing the T cell immune function to obtain the most accurate results.
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Stimulation of P2 receptors causes release of IL-1beta-loaded microvesicles from human dendritic cells.
Pizzirani, C, Ferrari, D, Chiozzi, P, Adinolfi, E, Sandonà, D, Savaglio, E, Di Virgilio, F
Blood. 2007;(9):3856-64
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Abstract
Dendritic cells (DCs) are professional antigen-presenting cells that initiate the immune response by activating T lymphocytes. DCs express plasma membrane receptors for extracellular nucleotides named P2 receptors (P2Rs). Stimulation of P2Rs in these cells is known to cause chemotaxis, cytokine release, and cell death and to modulate LPS-dependent differentiation. Here we show that stimulation of the P2X(7) receptor subtype (P2X(7)R) causes fast microvesicle shedding from DC plasma membrane. Vesicle release occurs from both immature and mature DCs; however, only vesicles from mature DCs, due to their previous exposure to LPS, contain IL-1beta. Microvesicles, whether from immature or mature DCs, also contain caspase-1 and -3 and cathepsin D. They also express the P2X(7)R in addition to other P2Rs and known markers of immune cells such as major histocompatibility complex II (MHC II) and CD39. Activation of the P2X(7)R by extracellular ATP causes IL-1beta release from the vesicle lumen. Previous studies demonstrated that high extracellular K(+) inhibits IL-1beta processing and release; here we show that high ionic strength reduces microvesicle shedding when compared with a low ionic strength medium but strongly increases microvesicle IL-1beta loading.
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Adenosine 5'-triphosphate and adenosine as endogenous signaling molecules in immunity and inflammation.
Bours, MJ, Swennen, EL, Di Virgilio, F, Cronstein, BN, Dagnelie, PC
Pharmacology & therapeutics. 2006;(2):358-404
Abstract
Human health is under constant threat of a wide variety of dangers, both self and nonself. The immune system is occupied with protecting the host against such dangers in order to preserve human health. For that purpose, the immune system is equipped with a diverse array of both cellular and non-cellular effectors that are in continuous communication with each other. The naturally occurring nucleotide adenosine 5'-triphosphate (ATP) and its metabolite adenosine (Ado) probably constitute an intrinsic part of this extensive immunological network through purinergic signaling by their cognate receptors, which are widely expressed throughout the body. This review provides a thorough overview of the effects of ATP and Ado on major immune cell types. The overwhelming evidence indicates that ATP and Ado are important endogenous signaling molecules in immunity and inflammation. Although the role of ATP and Ado during the course of inflammatory and immune responses in vivo appears to be extremely complex, we propose that their immunological role is both interdependent and multifaceted, meaning that the nature of their effects may shift from immunostimulatory to immunoregulatory or vice versa depending on extracellular concentrations as well as on expression patterns of purinergic receptors and ecto-enzymes. Purinergic signaling thus contributes to the fine-tuning of inflammatory and immune responses in such a way that the danger to the host is eliminated efficiently with minimal damage to healthy tissues.