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Cited by 233Inflammasome and immune disordersAdenosine and Purinergic SignalingSepsis Diagnosis and Treatment

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P2X7 receptor induces mitochondrial failure in monocytes and compromises NLRP3 inflammasome activation during sepsis

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Martínez-García, Juan José, Martínez-Banaclocha, Helios, Angosto-Bazarra, Diego, de Torre-Minguela, Carlos, Baroja-Mazo, Alberto, Alarcón-Vila, Cristina, Martínez-Alarcón, Laura, Amores-Iniesta, Joaquín, Martín-Sánchez, Fátima, Ercole, Giovanni A., Martínez, Carlos M., González-Lisorge, Ada, Fernández-Pacheco, José, Martínez-Gil, Piedad, Adriouch, Sahil, Koch-Nolte, Friedrich, Luján, Juan, Acosta-Villegas, Francisco, Parrilla, Pascual, García-Palenciano, Carlos, Pelegrin, Pablo·2019·Nature Communications
DOI 10.1038/s41467-019-10626-xPMC PMC6586640
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Abstract

Sepsis is characterized by a systemic inflammatory response followed by immunosuppression of the host. Metabolic defects and mitochondrial failure are common in immunocompromised patients with sepsis. The NLRP3 inflammasome is important for establishing an inflammatory response after activation by the purinergic P2X7 receptor. Here, we study a cohort of individuals with intra-abdominal origin sepsis and show that patient monocytes have impaired NLRP3 activation by the P2X7 receptor. Furthermore, most sepsis-related deaths are among patients whose NLRP3 activation is profoundly altered. In monocytes from sepsis patients, the P2X7 receptor is associated with mitochondrial dysfunction. Furthermore, activation of the P2X7 receptor results in mitochondrial damage, which in turn inhibits NLRP3 activation by HIF-1α. We show that mortality increases in a mouse model of sepsis when the P2X7 receptor is activated in vivo. These data reveal a molecular mechanism initiated by the P2X7 receptor that contributes to NLRP3 impairment during infection.

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Figures

Fig. 1 Sepsis patients present with elevated inflammasome markers. a–c Plasma concentration of CRP, PCT a, IL-6, IL-8 b, IL-1β, IL-18, and HMGB1 c in healthy controls, abdominal surgery patients within the first 24 h after surgery and intra-abdominal origin septic patients within the first 24 h of admission to the surgical critical unit; dotted lines in a represent standard threshold for CRP and PCT in healthy population. d Percentage of monocytes from healthy controls and septic patient positives for active caspase-1 labeled with FLICA (left) or positives for intracellular ASC specks (right). e Fold increase of circulating ASC specks in the plasma of surgery controls and septic patients compared with healthy controls. Each dot represents an individual patient; average ± standard error is represented in all panels; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Mann–Whitney test for a; Kruskal–Wallis test for b–e
Fig. 1 Sepsis patients present with elevated inflammasome markers. a–c Plasma concentration of CRP, PCT a, IL-6, IL-8 b, IL-1β, IL-18, and HMGB1 c in healthy controls, abdominal surgery patients within the first 24 h after surgery and intra-abdominal origin septic patients within the first 24 h of admission to the surgical critical unit; dotted lines in a represent standard threshold for CRP and PCT in healthy population. d Percentage of monocytes from healthy controls and septic patient positives for active caspase-1 labeled with FLICA (left) or positives for intracellular ASC specks (right). e Fold increase of circulating ASC specks in the plasma of surgery controls and septic patients compared with healthy controls. Each dot represents an individual patient; average ± standard error is represented in all panels; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Mann–Whitney test for a; Kruskal–Wallis test for b–eCC BY · PubMed Central
Fig. 2 Inflammasome activation is compromised in septic patients with high mortality. a ELISA for IL-1β in PBMC supernatants and percentage of monocytes with intracellular ASC specks from septic patients within the first 24 h of admission to the surgical critical unit and control groups after NLRP3 inflammasome activation by LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min) treatment. There is a group of septic patients that release low or no IL-1β (blue dots). b ELISA for IL-1β and HMGB1 in PBMC supernatants and percentage of monocytes with intracellular ASC specks from control groups and septic patients treated as in a, separating septic patients into two groups: NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). c Percentage of mortality in NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients with regard to the total mortality of septic patients. d Kaplan–Meier representation of NLRP3 non-immunocompromised (gray line) and immunocompromised (blue line) septic patients’ survival. e Concentration of C-reactive protein (CRP) in plasma of NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients at day 1; dotted lines represent threshold concentration of CRP for healthy population. f SOFA and APACHEII indexes in NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients at day 1. g ΔSOFA calculated as the variation in SOFA between day 5 and day 1, gray dotted line represents no change with regard to day 1. h Number of days admitted to the Surgical Critical Unit and number of days of mechanical ventilation for NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients. Each dot represents an individual patient; average ± standard error is represented in panels a, b, e–h; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test for a and b; Log-rank test for d; and Mann–Whitney test for e–h
Fig. 2 Inflammasome activation is compromised in septic patients with high mortality. a ELISA for IL-1β in PBMC supernatants and percentage of monocytes with intracellular ASC specks from septic patients within the first 24 h of admission to the surgical critical unit and control groups after NLRP3 inflammasome activation by LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min) treatment. There is a group of septic patients that release low or no IL-1β (blue dots). b ELISA for IL-1β and HMGB1 in PBMC supernatants and percentage of monocytes with intracellular ASC specks from control groups and septic patients treated as in a, separating septic patients into two groups: NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). c Percentage of mortality in NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients with regard to the total mortality of septic patients. d Kaplan–Meier representation of NLRP3 non-immunocompromised (gray line) and immunocompromised (blue line) septic patients’ survival. e Concentration of C-reactive protein (CRP) in plasma of NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients at day 1; dotted lines represent threshold concentration of CRP for healthy population. f SOFA and APACHEII indexes in NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients at day 1. g ΔSOFA calculated as the variation in SOFA between day 5 and day 1, gray dotted line represents no change with regard to day 1. h Number of days admitted to the Surgical Critical Unit and number of days of mechanical ventilation for NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar) septic patients. Each dot represents an individual patient; average ± standard error is represented in panels a, b, e–h; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test for a and b; Log-rank test for d; and Mann–Whitney test for e–hCC BY · PubMed Central
Fig. 3 NLRP3 immunoparalysis during sepsis is transitory. a Concentrations of C-reactive protein (CRP), procalcitonin (PCT), and IL-6 in plasma of septic patients at days 1, 3, and 5 during sepsis and at day 120 after sepsis recovery; dotted lines represent normal concentration of CRP and PCT. Levels of these markers in septic patients were compared with the abdominal surgery controls at 24 h after surgery. Control group and septic patients at day 1 correspond to patient data presented in Fig. 1a, b, and are shown here for comparison. b ELISA for IL-1β in PBMC supernatants (top) and percentage of monocytes with intracellular ASC specks (bottom) after NLRP3 inflammasome activation by LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min) treatment from control groups and NLRP3 non-immunocompromised (gray) or NLRP3 immunocompromised (blue) septic patients at day 1, 3, 5 during sepsis and at day 120 after sepsis recovery. Control groups and septic patients at day 1 correspond to patient data presented in Fig. 2a, b, and are shown here for comparison; each dot represents an individual patient; average ± standard error is represented in all panels; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test for a, b
Fig. 3 NLRP3 immunoparalysis during sepsis is transitory. a Concentrations of C-reactive protein (CRP), procalcitonin (PCT), and IL-6 in plasma of septic patients at days 1, 3, and 5 during sepsis and at day 120 after sepsis recovery; dotted lines represent normal concentration of CRP and PCT. Levels of these markers in septic patients were compared with the abdominal surgery controls at 24 h after surgery. Control group and septic patients at day 1 correspond to patient data presented in Fig. 1a, b, and are shown here for comparison. b ELISA for IL-1β in PBMC supernatants (top) and percentage of monocytes with intracellular ASC specks (bottom) after NLRP3 inflammasome activation by LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min) treatment from control groups and NLRP3 non-immunocompromised (gray) or NLRP3 immunocompromised (blue) septic patients at day 1, 3, 5 during sepsis and at day 120 after sepsis recovery. Control groups and septic patients at day 1 correspond to patient data presented in Fig. 2a, b, and are shown here for comparison; each dot represents an individual patient; average ± standard error is represented in all panels; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test for a, bCC BY · PubMed Central
Fig. 4 P2X7 receptor is transiently upregulated in monocytes during sepsis. a Representative histogram plot of surface P2X7 receptor staining in monocytes from healthy (white), septic patient (black) and non-stained monocytes (gray). b Quantification of P2X7 receptor mean fluorescence intensity (MFI, left) and percentage of positive monocytes for P2X7 receptor (right) in control and septic patients. c ELISA to quantify the concentration of soluble P2X7 receptor in plasma of control and septic patients. d Quantification of P2X7 receptor MFI at day 1 during sepsis and day 120 after recovery. e Quantification of P2X7 receptor MFI in monocytes from healthy donors treated with IFNγ, TNF-α, IL-6 (all at 20 ng/ml), or with increasing concentrations of LPS (10, 100, 1000 ng/ml) for 24 h. f Quantification of P2X7 receptor MFI in monocytes from healthy donors treated with LPS (1 μg/ml) for the indicated times. g Correlation between the concentration of IL-1β released from PBMCs treated with LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min), and the quantification of P2X7 receptor MFI in monocytes from the indicated control and septic individuals; IC: immunocompromised septic patients. Each dot represents an individual septic patient or healthy donor; average ± standard error is represented in panels b, c, e, f; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test was used in b; Mann–Whitney test for c, e, f; Pearson correlation was used in g
Fig. 4 P2X7 receptor is transiently upregulated in monocytes during sepsis. a Representative histogram plot of surface P2X7 receptor staining in monocytes from healthy (white), septic patient (black) and non-stained monocytes (gray). b Quantification of P2X7 receptor mean fluorescence intensity (MFI, left) and percentage of positive monocytes for P2X7 receptor (right) in control and septic patients. c ELISA to quantify the concentration of soluble P2X7 receptor in plasma of control and septic patients. d Quantification of P2X7 receptor MFI at day 1 during sepsis and day 120 after recovery. e Quantification of P2X7 receptor MFI in monocytes from healthy donors treated with IFNγ, TNF-α, IL-6 (all at 20 ng/ml), or with increasing concentrations of LPS (10, 100, 1000 ng/ml) for 24 h. f Quantification of P2X7 receptor MFI in monocytes from healthy donors treated with LPS (1 μg/ml) for the indicated times. g Correlation between the concentration of IL-1β released from PBMCs treated with LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min), and the quantification of P2X7 receptor MFI in monocytes from the indicated control and septic individuals; IC: immunocompromised septic patients. Each dot represents an individual septic patient or healthy donor; average ± standard error is represented in panels b, c, e, f; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test was used in b; Mann–Whitney test for c, e, f; Pearson correlation was used in gCC BY · PubMed Central
Fig. 5 P2X7 receptor induces mitochondrial membrane depolarization. a Mitochondrial membrane potential from blood monocytes stained with JC-10 in a control healthy donor (left), in a septic patient at day 1 (middle), and from healthy controls, abdominal surgery, and septic patients at day 1 or 120 (right); PE− cells represent the monocytes with mitochondrial depolarization. b Correlation between the percentage of monocytes with mitochondrial membrane depolarization and the quantification of P2X7 receptor MFI in monocytes from the septic patients; IC: immunocompromised septic patients. c Kinetics of mitochondrial depolarization after ATP (3 mM) stimulation of LPS-primed BMDMs incubated or not with the P2X7 receptor antagonist A438079 (10 μM). d Percentage of human monocytes from healthy donors with mitochondrial membrane depolarization after ATP (3 mM, 30 min) stimulation and incubated or not with the P2X7 receptor antagonist AZ11645373 (10 μM). e Percentage of mouse BMDMs with mitochondrial membrane depolarization after ATP stimulation at the indicated concentrations for 30 min and incubated or not with anti-P2X7 nanobodies (13A7, blocking nanobody; 14D5, potentiating nanobody; each at 200 nM). f Mitochondrial ROS production from human monocytes isolated from healthy donors after ATP (3 mM, 30 min) stimulation. g Mitochondrial ROS from human monocytes isolated from surgery controls and septic patients (white bar); right bars separate the group of septic patients into NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). h Mitochondrial membrane depolarization from wild-type or knock-out BMDMs as indicated primed or not with LPS (4 h) and treated for 30 min with ATP (3 mM, green bars), ATP and A438079 (10 μM, white bars), or antimycin A (5 μM, gray bars). Each dot represents an individual patient in a, b, g, or an individual healthy donor in d, f, a single independent experiment in e, h, or average + SEM of two independent experiments in c; average ± standard error is represented in panels a (right), c–h; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Pearson correlation was used in b; Mann–Whitney test was used in f; and Kruskal–Wallis test was used in a, g, h
Fig. 5 P2X7 receptor induces mitochondrial membrane depolarization. a Mitochondrial membrane potential from blood monocytes stained with JC-10 in a control healthy donor (left), in a septic patient at day 1 (middle), and from healthy controls, abdominal surgery, and septic patients at day 1 or 120 (right); PE− cells represent the monocytes with mitochondrial depolarization. b Correlation between the percentage of monocytes with mitochondrial membrane depolarization and the quantification of P2X7 receptor MFI in monocytes from the septic patients; IC: immunocompromised septic patients. c Kinetics of mitochondrial depolarization after ATP (3 mM) stimulation of LPS-primed BMDMs incubated or not with the P2X7 receptor antagonist A438079 (10 μM). d Percentage of human monocytes from healthy donors with mitochondrial membrane depolarization after ATP (3 mM, 30 min) stimulation and incubated or not with the P2X7 receptor antagonist AZ11645373 (10 μM). e Percentage of mouse BMDMs with mitochondrial membrane depolarization after ATP stimulation at the indicated concentrations for 30 min and incubated or not with anti-P2X7 nanobodies (13A7, blocking nanobody; 14D5, potentiating nanobody; each at 200 nM). f Mitochondrial ROS production from human monocytes isolated from healthy donors after ATP (3 mM, 30 min) stimulation. g Mitochondrial ROS from human monocytes isolated from surgery controls and septic patients (white bar); right bars separate the group of septic patients into NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). h Mitochondrial membrane depolarization from wild-type or knock-out BMDMs as indicated primed or not with LPS (4 h) and treated for 30 min with ATP (3 mM, green bars), ATP and A438079 (10 μM, white bars), or antimycin A (5 μM, gray bars). Each dot represents an individual patient in a, b, g, or an individual healthy donor in d, f, a single independent experiment in e, h, or average + SEM of two independent experiments in c; average ± standard error is represented in panels a (right), c–h; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Pearson correlation was used in b; Mann–Whitney test was used in f; and Kruskal–Wallis test was used in a, g, hCC BY · PubMed Central
Fig. 6 P2X7 receptor stimulation impairs the NLRP3 inflammasome in monocytes. a, b Percentage of monocytes with ASC-specks (a) and release of IL-1β from PBMCs (b) isolated from healthy donor blood samples treated with ATP (3 mM, 30 min; ATP-pre), then washed and primed with or without LPS (1 μg/ml, 2 h) and then treated for 20 min with ATP (3 mM; ATP-post) or nigericin (10 μM) as indicated. c IL-1β release from wild type or P2rx7−/− BMDMs treated as in a, but with 4 h LPS priming and 30 min of ATP or nigericin. d IL-1β release from wild type or P2rx7−/− BMDMs treated for 30 min with antimycin A (5 μM) or FCCP (1 μM), and then washed and primed with LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min) as indicated. e Expression of Nrlp3 and Il1b genes analyzed by quantitative PCR from wild type or P2rx7−/− BMDMs treated with ATP (3 mM, 30 min; ATP-pre), then washed and primed with or without LPS (100 ng/ml, 4 h). f Kaplan–Meier representation of wild type (top) or P2rx7−/− (bottom) mice survival after sham operation (dotted line), or CLP operation (continuous line); some mice groups were i.p. injected with ATP (0.5 mg/g) 30 min before operation (dashed lines). Wild type CLP n = 10, CLP + ATP n = 8, sham n = 4, sham + ATP n = 4; P2rx7−/− CLP n = 4, CLP + ATP n = 6, sham n = 3. g IL-1β from peritoneal lavage (left) or bacterial load in blood (right) from wild type sham, CLP or CLP+ATP mice after 24 h. Each dot represents a single independent experiment (c, d), a sample from an individual healthy donor (a, b, e) or a single mouse (g); average ± standard error is represented in panels a–e, g; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ns, no significant difference (p > 0.05); Mann–Whitney test was used for a, b, d, e; Kruskal–Wallis test was used for c; Log-rank test for f
Fig. 6 P2X7 receptor stimulation impairs the NLRP3 inflammasome in monocytes. a, b Percentage of monocytes with ASC-specks (a) and release of IL-1β from PBMCs (b) isolated from healthy donor blood samples treated with ATP (3 mM, 30 min; ATP-pre), then washed and primed with or without LPS (1 μg/ml, 2 h) and then treated for 20 min with ATP (3 mM; ATP-post) or nigericin (10 μM) as indicated. c IL-1β release from wild type or P2rx7−/− BMDMs treated as in a, but with 4 h LPS priming and 30 min of ATP or nigericin. d IL-1β release from wild type or P2rx7−/− BMDMs treated for 30 min with antimycin A (5 μM) or FCCP (1 μM), and then washed and primed with LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min) as indicated. e Expression of Nrlp3 and Il1b genes analyzed by quantitative PCR from wild type or P2rx7−/− BMDMs treated with ATP (3 mM, 30 min; ATP-pre), then washed and primed with or without LPS (100 ng/ml, 4 h). f Kaplan–Meier representation of wild type (top) or P2rx7−/− (bottom) mice survival after sham operation (dotted line), or CLP operation (continuous line); some mice groups were i.p. injected with ATP (0.5 mg/g) 30 min before operation (dashed lines). Wild type CLP n = 10, CLP + ATP n = 8, sham n = 4, sham + ATP n = 4; P2rx7−/− CLP n = 4, CLP + ATP n = 6, sham n = 3. g IL-1β from peritoneal lavage (left) or bacterial load in blood (right) from wild type sham, CLP or CLP+ATP mice after 24 h. Each dot represents a single independent experiment (c, d), a sample from an individual healthy donor (a, b, e) or a single mouse (g); average ± standard error is represented in panels a–e, g; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ns, no significant difference (p > 0.05); Mann–Whitney test was used for a, b, d, e; Kruskal–Wallis test was used for c; Log-rank test for fCC BY · PubMed Central
Fig. 7 Mitochondrial dysfunction mediates P2X7 receptor-induced NLRP3 inflammasome impairment. a Mitochondrial membrane depolarization in BMDMs treated with ATP (3 mM, 30 min), then washed-out and incubated for the indicated times with or without LPS (1 μg/ml). b IL-1β release from wild-type BMDMs treated as in a, but after LPS priming cells were stimulated with nigericin (10 μM, 30 min). c Mitochondrial membrane depolarization in BMDMs treated with ATP (3 mM, 30 min) in the presence or absence pyrrolidine dithiocarbamate (PDTC, 40 μM). d IL-1β release from PBMC isolated from healthy donor blood samples treated with ATP (1 mM, 30 min; ATP-pre) with or without PDTC (10 μM), then washed and primed with or without LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min). e Representative histogram plot of HIF-1α staining (left) or quantification of mean intensity fluorescence increase (ΔMFI, right) in monocytes from healthy donor blood samples treated or not with ATP (1 mM, 30 min) in the presence or absence of AZ11645373 (10 μM) or PDTC (10 μM), and then washed and cultured for 2 h; non-stained monocytes (light gray, left). f IL-1β release from BMDM supernatants treated with ATP (3 mM, 30 min; ATP-pre) in the presence or absence of echinomycin (5 nM), then washed and primed with or without LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min). g Expression of HIF1A analyzed by qPCR from control surgery group and septic patients PBMCs; septic patients are separated into NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). h Correlation between HIF1A expression and IL-1β released from septic patients PBMCs treated with LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min); IC: immunocompromised septic patients. Each dot represents a single independent experiment or a sample from an individual healthy donor or septic patient; average ± standard error is represented in panels a–g; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test was used for b, c, f; Pearson correlation was used in h
Fig. 7 Mitochondrial dysfunction mediates P2X7 receptor-induced NLRP3 inflammasome impairment. a Mitochondrial membrane depolarization in BMDMs treated with ATP (3 mM, 30 min), then washed-out and incubated for the indicated times with or without LPS (1 μg/ml). b IL-1β release from wild-type BMDMs treated as in a, but after LPS priming cells were stimulated with nigericin (10 μM, 30 min). c Mitochondrial membrane depolarization in BMDMs treated with ATP (3 mM, 30 min) in the presence or absence pyrrolidine dithiocarbamate (PDTC, 40 μM). d IL-1β release from PBMC isolated from healthy donor blood samples treated with ATP (1 mM, 30 min; ATP-pre) with or without PDTC (10 μM), then washed and primed with or without LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min). e Representative histogram plot of HIF-1α staining (left) or quantification of mean intensity fluorescence increase (ΔMFI, right) in monocytes from healthy donor blood samples treated or not with ATP (1 mM, 30 min) in the presence or absence of AZ11645373 (10 μM) or PDTC (10 μM), and then washed and cultured for 2 h; non-stained monocytes (light gray, left). f IL-1β release from BMDM supernatants treated with ATP (3 mM, 30 min; ATP-pre) in the presence or absence of echinomycin (5 nM), then washed and primed with or without LPS (1 μg/ml, 4 h) and then stimulated with nigericin (10 μM, 30 min). g Expression of HIF1A analyzed by qPCR from control surgery group and septic patients PBMCs; septic patients are separated into NLRP3 non-immunocompromised (gray bar) and immunocompromised (blue bar). h Correlation between HIF1A expression and IL-1β released from septic patients PBMCs treated with LPS (1 μg/ml, 2 h) and ATP (3 mM, 30 min); IC: immunocompromised septic patients. Each dot represents a single independent experiment or a sample from an individual healthy donor or septic patient; average ± standard error is represented in panels a–g; exact n number for each panel is presented in Source Data file; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no significant difference (p > 0.05); Kruskal–Wallis test was used for b, c, f; Pearson correlation was used in hCC BY · PubMed Central

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Authors
Martínez-García, J. J., Martínez-Banaclocha, H., Angosto-Bazarra, D., de Torre-Minguela, C., Baroja-Mazo, A., Alarcón-Vila, C., Martínez-Alarcón, L., Amores-Iniesta, J., Martín-Sánchez, F., Ercole, G. A., Martínez, C. M., González-Lisorge, A., Fernández-Pacheco, J., Martínez-Gil, P., Adriouch, S., Koch-Nolte, F., Luján, J., Acosta-Villegas, F., Parrilla, P., García-Palenciano, C., & Pelegrin, P.
Affiliations
  • Instituto Murciano de Investigación Biosanitaria (ES)
  • Hospital Universitario Virgen de la Arrixaca (ES)
  • Inserm (FR)
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2019-06-20
Volume
10
Issue
1
Pages
2711-2711
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-019-10626-x
PMC
PMC6586640
OpenAlex
W2952125019
Cited by
233
References
63
Citations by year
2026: 16 · 2025: 16 · 2024: 44 · 2023: 48 · 2022: 39 · 2021: 34 · 2020: 33 · 2019: 3
FWCI
9.01
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Hispana (publishedVersion, cc-by)
  • HAL (submittedVersion, other-oa)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Institute for Research in Biomedicine
  • Ablynx
  • European Cooperation in Science and Technology (BM-1406)
  • Ministerio de Economía y Competitividad (FJCI-2014-22041, PI13/00174, CD13/00059)
  • Ministerio de Asuntos Económicos y Transformación Digital, Gobierno de España (SAF2017‐88276‐R)
  • European Regional Development Fund (PI13/00174)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
  • Health Sciences › Medicine › Critical Care and Intensive Care Medicine › Sepsis Diagnosis and Treatment
Keywords
P2X7 receptor, NLRP3 inflammasome, mitochondrial dysfunction, monocytes, sepsis, HIF-1α, purinergic signaling, immunosuppression, sepsis mortality, abdominal sepsis, mouse sepsis model
MeSH terms
NLR Family, Pyrin Domain-Containing 3 Protein, Adult, Aged, Aged, 80 and over, Animals, Disease Models, Animal, Female, Follow-Up Studies, Humans, Macrophages, Male, Middle Aged, Mitochondria, Monocytes, Up-Regulation, Sepsis, Mice, Hypoxia-Inducible Factor 1, alpha Subunit, Receptors, Purinergic P2X7, Inflammasomes, Mitochondrial Dynamics

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Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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