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Vagus nerve controls resolution and pro-resolving mediators of inflammation.

Mirakaj V, Dalli J, Granja T, Rosenberger P, Serhan CN - J. Exp. Med. (2014)

Bottom Line: In netrin-1(+/-) mice, resolvin D1 (RvD1) was less effective in reducing neutrophil influx promoting resolution of peritonitis compared with Ntn1(+/+).Human monocytes incubated with netrin-1 produced proresolving mediators, including resolvins and lipoxins.Netrin-1 and RvD1 displayed bidirectional activation in that they stimulated each other's expression and enhanced efferocytosis.

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Affiliation: Center for Experimental Therapeutics and Reperfusion Injury, Harvard Institutes of Medicine, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115 Clinic of Anesthesiology and Intensive Care Medicine, University Hospital Tübingen, Eberhard-Karls University, 72076 Tübingen, Germany.

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Netrin-1 reduces human PMN activation and increases Mϕ efferocytosis: Synergy with RvD1 pathway. (A) Isolated human PMN were incubated with either fMLP or netrin-1 for 30 min and the expression of CD11b (neo-epitope) and CD62L on PMN was measured by flow cytometry. (B) Isolated human PMN were placed on chemotaxis slides and exposed for 2 h to either fMLP or netrin-1. The cell migration was monitored for a 10-min period and analyzed using Image Pro Plus 7 software. (C) MΦ differentiated from peripheral blood monocytes were incubated with indicated concentrations of netrin-1. Carboxyfluorescein diacetate-labeled apoptotic PMN were added at 1:3 ratio (MΦ:PMN), and the rate of the efferocytosis was determined after 1 h by measurement of fluorescence. (D) MΦ differentiated from peripheral blood monocytes were incubated with netrin-1 in the absence or presence of RvD1. The degree of efferocytosis was performed using a fluorescent plate reader. (E) Peritoneal MΦ from WT or 12/15LO−/− mice were incubated with the indicated concentrations of netrin-1 (15 min, 37°C), the fluorescently labeled apoptotic PMN were added and the degree of efferocytosis was assessed after 1 h (37°C) using a fluorescent plate reader (see Materials and methods). Results are representative of five independent experiments (n = 5), mean ± SEM, in A–D. In E, results are representative of three independent experiments (n = 3 mice); *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig7: Netrin-1 reduces human PMN activation and increases Mϕ efferocytosis: Synergy with RvD1 pathway. (A) Isolated human PMN were incubated with either fMLP or netrin-1 for 30 min and the expression of CD11b (neo-epitope) and CD62L on PMN was measured by flow cytometry. (B) Isolated human PMN were placed on chemotaxis slides and exposed for 2 h to either fMLP or netrin-1. The cell migration was monitored for a 10-min period and analyzed using Image Pro Plus 7 software. (C) MΦ differentiated from peripheral blood monocytes were incubated with indicated concentrations of netrin-1. Carboxyfluorescein diacetate-labeled apoptotic PMN were added at 1:3 ratio (MΦ:PMN), and the rate of the efferocytosis was determined after 1 h by measurement of fluorescence. (D) MΦ differentiated from peripheral blood monocytes were incubated with netrin-1 in the absence or presence of RvD1. The degree of efferocytosis was performed using a fluorescent plate reader. (E) Peritoneal MΦ from WT or 12/15LO−/− mice were incubated with the indicated concentrations of netrin-1 (15 min, 37°C), the fluorescently labeled apoptotic PMN were added and the degree of efferocytosis was assessed after 1 h (37°C) using a fluorescent plate reader (see Materials and methods). Results are representative of five independent experiments (n = 5), mean ± SEM, in A–D. In E, results are representative of three independent experiments (n = 3 mice); *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: Assessment of the antiinflammatory and pro-resolving properties displayed by netrin-1 with human peripheral blood PMN revealed that netrin-1 reversed fMLP-induced CD62L shedding and CD11b neo-epitope exposure (Fig. 7 A). At concentrations as low as 1 nM, netrin-1 inhibited PMN migration (Fig. 7 B) and did not display chemotactic activities on its own in this concentration range (n = 4). RvD1 also stops PMN migration (Kasuga et al., 2008). Because resolution of inflammation is a dynamic process with one of the key steps being the clearance of apoptotic PMN by Mϕ (Serhan and Savill, 2005), we next assessed the ability of netrin-1 to regulate this pro-resolving mechanism. Netrin-1 stimulated the uptake of apoptotic PMN by human Mϕ in a dose-dependent manner (Fig. 7 C). These results were further corroborated in vivo, where netrin-1 enhanced Mϕ clearance of apoptotic PMN (unpublished data) in mice.


Vagus nerve controls resolution and pro-resolving mediators of inflammation.

Mirakaj V, Dalli J, Granja T, Rosenberger P, Serhan CN - J. Exp. Med. (2014)

Netrin-1 reduces human PMN activation and increases Mϕ efferocytosis: Synergy with RvD1 pathway. (A) Isolated human PMN were incubated with either fMLP or netrin-1 for 30 min and the expression of CD11b (neo-epitope) and CD62L on PMN was measured by flow cytometry. (B) Isolated human PMN were placed on chemotaxis slides and exposed for 2 h to either fMLP or netrin-1. The cell migration was monitored for a 10-min period and analyzed using Image Pro Plus 7 software. (C) MΦ differentiated from peripheral blood monocytes were incubated with indicated concentrations of netrin-1. Carboxyfluorescein diacetate-labeled apoptotic PMN were added at 1:3 ratio (MΦ:PMN), and the rate of the efferocytosis was determined after 1 h by measurement of fluorescence. (D) MΦ differentiated from peripheral blood monocytes were incubated with netrin-1 in the absence or presence of RvD1. The degree of efferocytosis was performed using a fluorescent plate reader. (E) Peritoneal MΦ from WT or 12/15LO−/− mice were incubated with the indicated concentrations of netrin-1 (15 min, 37°C), the fluorescently labeled apoptotic PMN were added and the degree of efferocytosis was assessed after 1 h (37°C) using a fluorescent plate reader (see Materials and methods). Results are representative of five independent experiments (n = 5), mean ± SEM, in A–D. In E, results are representative of three independent experiments (n = 3 mice); *, P < 0.05; **, P < 0.01; ***, P < 0.001.
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fig7: Netrin-1 reduces human PMN activation and increases Mϕ efferocytosis: Synergy with RvD1 pathway. (A) Isolated human PMN were incubated with either fMLP or netrin-1 for 30 min and the expression of CD11b (neo-epitope) and CD62L on PMN was measured by flow cytometry. (B) Isolated human PMN were placed on chemotaxis slides and exposed for 2 h to either fMLP or netrin-1. The cell migration was monitored for a 10-min period and analyzed using Image Pro Plus 7 software. (C) MΦ differentiated from peripheral blood monocytes were incubated with indicated concentrations of netrin-1. Carboxyfluorescein diacetate-labeled apoptotic PMN were added at 1:3 ratio (MΦ:PMN), and the rate of the efferocytosis was determined after 1 h by measurement of fluorescence. (D) MΦ differentiated from peripheral blood monocytes were incubated with netrin-1 in the absence or presence of RvD1. The degree of efferocytosis was performed using a fluorescent plate reader. (E) Peritoneal MΦ from WT or 12/15LO−/− mice were incubated with the indicated concentrations of netrin-1 (15 min, 37°C), the fluorescently labeled apoptotic PMN were added and the degree of efferocytosis was assessed after 1 h (37°C) using a fluorescent plate reader (see Materials and methods). Results are representative of five independent experiments (n = 5), mean ± SEM, in A–D. In E, results are representative of three independent experiments (n = 3 mice); *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: Assessment of the antiinflammatory and pro-resolving properties displayed by netrin-1 with human peripheral blood PMN revealed that netrin-1 reversed fMLP-induced CD62L shedding and CD11b neo-epitope exposure (Fig. 7 A). At concentrations as low as 1 nM, netrin-1 inhibited PMN migration (Fig. 7 B) and did not display chemotactic activities on its own in this concentration range (n = 4). RvD1 also stops PMN migration (Kasuga et al., 2008). Because resolution of inflammation is a dynamic process with one of the key steps being the clearance of apoptotic PMN by Mϕ (Serhan and Savill, 2005), we next assessed the ability of netrin-1 to regulate this pro-resolving mechanism. Netrin-1 stimulated the uptake of apoptotic PMN by human Mϕ in a dose-dependent manner (Fig. 7 C). These results were further corroborated in vivo, where netrin-1 enhanced Mϕ clearance of apoptotic PMN (unpublished data) in mice.

Bottom Line: In netrin-1(+/-) mice, resolvin D1 (RvD1) was less effective in reducing neutrophil influx promoting resolution of peritonitis compared with Ntn1(+/+).Human monocytes incubated with netrin-1 produced proresolving mediators, including resolvins and lipoxins.Netrin-1 and RvD1 displayed bidirectional activation in that they stimulated each other's expression and enhanced efferocytosis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Experimental Therapeutics and Reperfusion Injury, Harvard Institutes of Medicine, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115 Clinic of Anesthesiology and Intensive Care Medicine, University Hospital Tübingen, Eberhard-Karls University, 72076 Tübingen, Germany.

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Related in: MedlinePlus