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Gut phage protection from acute inflammatory response via macrophage (a) in vivo model of phage exposure followed by acute challenge with LPS lo (low) for measuring serum cytokines. (b) Serum pro-inflammatory cytokines levels are shown. Serum was collected after 3 h of LPS lo challenge for <t>analyzing</t> <t>IL-1β,</t> IL-12p40, IL-6 and <t>TNF-α</t> (n = 6 mice). Sterile PBS was used as a control. (c) monitoring IL-1β burst in PBS (control) or phage exposure mice followed by acute challenge with LPS lo. (d) and (e) quantification of IL-1β-driven luciferase activity (photons/sec, n = 3) of the whole body (d). Luciferase activity (e), color overlay represents photons/sec/cm2/steradian. (f) and (g) survival of WT mice pretreated with phages followed by secondary LPS hi (high) treatment (f) and survival curve (g). Data represent n = 9 mice from two independent experiments. (h) and (i) survival of Rag1-/- mice pretreated with phages followed by secondary LPS hi treatment (h) and survival curve (i). Data represent n = 8 mice from two independent experiments. (j) and (k) survival of macrophage-depleted mice pretreated with phages followed by secondary LPS hi treatment (J) and survival curve (k). Data represent n = 9 mice from two independent experiments. (l) Ex vivo peritoneal macrophage model of phage exposure followed by a secondary challenge with LPS. (m) peritoneal macrophage supernatants assayed for IL-1β, IL12p40, IL-6 and TNF-α (n = 5 mice). (b) and (m) one-way ANOVA with Tukey’s HSD test for statistical analysis. (d) Student’s’s t-test for statistical analysis. (g), (i) and (k) log-rank (mantel–cox) test for survival analysis. Data presented are group means ± SD. statistical significance was assessed by *p < 0.05,**p < 0.01, *** p <.001.
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Gut phage protection from acute inflammatory response via macrophage (a) in vivo model of phage exposure followed by acute challenge with LPS lo (low) for measuring serum cytokines. (b) Serum pro-inflammatory cytokines levels are shown. Serum was collected after 3 h of LPS lo challenge for <t>analyzing</t> <t>IL-1β,</t> IL-12p40, IL-6 and <t>TNF-α</t> (n = 6 mice). Sterile PBS was used as a control. (c) monitoring IL-1β burst in PBS (control) or phage exposure mice followed by acute challenge with LPS lo. (d) and (e) quantification of IL-1β-driven luciferase activity (photons/sec, n = 3) of the whole body (d). Luciferase activity (e), color overlay represents photons/sec/cm2/steradian. (f) and (g) survival of WT mice pretreated with phages followed by secondary LPS hi (high) treatment (f) and survival curve (g). Data represent n = 9 mice from two independent experiments. (h) and (i) survival of Rag1-/- mice pretreated with phages followed by secondary LPS hi treatment (h) and survival curve (i). Data represent n = 8 mice from two independent experiments. (j) and (k) survival of macrophage-depleted mice pretreated with phages followed by secondary LPS hi treatment (J) and survival curve (k). Data represent n = 9 mice from two independent experiments. (l) Ex vivo peritoneal macrophage model of phage exposure followed by a secondary challenge with LPS. (m) peritoneal macrophage supernatants assayed for IL-1β, IL12p40, IL-6 and TNF-α (n = 5 mice). (b) and (m) one-way ANOVA with Tukey’s HSD test for statistical analysis. (d) Student’s’s t-test for statistical analysis. (g), (i) and (k) log-rank (mantel–cox) test for survival analysis. Data presented are group means ± SD. statistical significance was assessed by *p < 0.05,**p < 0.01, *** p <.001.
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Gut phage protection from acute inflammatory response via macrophage (a) in vivo model of phage exposure followed by acute challenge with LPS lo (low) for measuring serum cytokines. (b) Serum pro-inflammatory cytokines levels are shown. Serum was collected after 3 h of LPS lo challenge for analyzing IL-1β, IL-12p40, IL-6 and TNF-α (n = 6 mice). Sterile PBS was used as a control. (c) monitoring IL-1β burst in PBS (control) or phage exposure mice followed by acute challenge with LPS lo. (d) and (e) quantification of IL-1β-driven luciferase activity (photons/sec, n = 3) of the whole body (d). Luciferase activity (e), color overlay represents photons/sec/cm2/steradian. (f) and (g) survival of WT mice pretreated with phages followed by secondary LPS hi (high) treatment (f) and survival curve (g). Data represent n = 9 mice from two independent experiments. (h) and (i) survival of Rag1-/- mice pretreated with phages followed by secondary LPS hi treatment (h) and survival curve (i). Data represent n = 8 mice from two independent experiments. (j) and (k) survival of macrophage-depleted mice pretreated with phages followed by secondary LPS hi treatment (J) and survival curve (k). Data represent n = 9 mice from two independent experiments. (l) Ex vivo peritoneal macrophage model of phage exposure followed by a secondary challenge with LPS. (m) peritoneal macrophage supernatants assayed for IL-1β, IL12p40, IL-6 and TNF-α (n = 5 mice). (b) and (m) one-way ANOVA with Tukey’s HSD test for statistical analysis. (d) Student’s’s t-test for statistical analysis. (g), (i) and (k) log-rank (mantel–cox) test for survival analysis. Data presented are group means ± SD. statistical significance was assessed by *p < 0.05,**p < 0.01, *** p <.001.

Journal: Gut Microbes

Article Title: Gut bacteriophages induce specific-reprogramming of macrophages to generate a protective innate immunity response to lipopolysaccharide exposure

doi: 10.1080/19490976.2025.2524540

Figure Lengend Snippet: Gut phage protection from acute inflammatory response via macrophage (a) in vivo model of phage exposure followed by acute challenge with LPS lo (low) for measuring serum cytokines. (b) Serum pro-inflammatory cytokines levels are shown. Serum was collected after 3 h of LPS lo challenge for analyzing IL-1β, IL-12p40, IL-6 and TNF-α (n = 6 mice). Sterile PBS was used as a control. (c) monitoring IL-1β burst in PBS (control) or phage exposure mice followed by acute challenge with LPS lo. (d) and (e) quantification of IL-1β-driven luciferase activity (photons/sec, n = 3) of the whole body (d). Luciferase activity (e), color overlay represents photons/sec/cm2/steradian. (f) and (g) survival of WT mice pretreated with phages followed by secondary LPS hi (high) treatment (f) and survival curve (g). Data represent n = 9 mice from two independent experiments. (h) and (i) survival of Rag1-/- mice pretreated with phages followed by secondary LPS hi treatment (h) and survival curve (i). Data represent n = 8 mice from two independent experiments. (j) and (k) survival of macrophage-depleted mice pretreated with phages followed by secondary LPS hi treatment (J) and survival curve (k). Data represent n = 9 mice from two independent experiments. (l) Ex vivo peritoneal macrophage model of phage exposure followed by a secondary challenge with LPS. (m) peritoneal macrophage supernatants assayed for IL-1β, IL12p40, IL-6 and TNF-α (n = 5 mice). (b) and (m) one-way ANOVA with Tukey’s HSD test for statistical analysis. (d) Student’s’s t-test for statistical analysis. (g), (i) and (k) log-rank (mantel–cox) test for survival analysis. Data presented are group means ± SD. statistical significance was assessed by *p < 0.05,**p < 0.01, *** p <.001.

Article Snippet: After 3 h, mice were humanely euthanized, and blood was collected to assess serum IL-1β (Solarbio, SEKM-0002), IL-12P40 (Solarbio, SEKM-0012), IL-6 (TECAN, BE45061) and TNF (TECAN, BE45291).

Techniques: In Vivo, Sterility, Control, Luciferase, Activity Assay, Ex Vivo