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NOD2-mediated suppression of CD55 on neutrophils enhances C5a generation during polymicrobial sepsis.

Oh SJ, Kim JH, Chung DH - PLoS Pathog. (2013)

Bottom Line: Nucleotide-binding oligomerization domain (NOD) 2 is a cytosolic protein that plays a defensive role in bacterial infection by sensing peptidoglycans.Furthermore, we found that NOD2-mediated IL-10 production by neutrophils enhanced C5a generation by suppressing CD55 expression on neutrophils in IL-1β-dependent and/or IL-1β-independent manners, thereby aggravating CLP-induced sepsis.SB203580, a receptor-interacting protein 2 (RIP2) inhibitor downstream of NOD2, reduced C5a generation by enhancing CD55 expression on neutrophils, resulting in attenuation of polymicrobial sepsis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
Nucleotide-binding oligomerization domain (NOD) 2 is a cytosolic protein that plays a defensive role in bacterial infection by sensing peptidoglycans. C5a, which has harmful effects in sepsis, interacts with innate proteins. However, whether NOD2 regulates C5a generation during sepsis remains to be determined. To address this issue, cecal ligation & puncture (CLP)-induced sepsis was compared in wild type and Nod2-/- mice. Nod2-/- mice showed lower levels of C5a, IL-10, and IL-1β in serum and peritoneum, but higher survival rate during CLP-induced sepsis compared to wild type mice. Injection of recombinant C5a decreased survival rates of Nod2-/- mice rate during sepsis, whereas it did not alter those in wild type mice. These findings suggest a novel provocative role for NOD2 in sepsis, in contrast to its protective role during bacterial infection. Furthermore, we found that NOD2-mediated IL-10 production by neutrophils enhanced C5a generation by suppressing CD55 expression on neutrophils in IL-1β-dependent and/or IL-1β-independent manners, thereby aggravating CLP-induced sepsis. SB203580, a receptor-interacting protein 2 (RIP2) inhibitor downstream of NOD2, reduced C5a generation by enhancing CD55 expression on neutrophils, resulting in attenuation of polymicrobial sepsis. Therefore, we propose a novel NOD2-mediated complement cascade regulatory pathway in sepsis, which may be a useful therapeutic target.

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IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation during cecal ligation and puncture (CLP)-induced sepsis.(A) IL-1 and IL-10 receptor expression was estimated on total peritoneal cells in terms of mean fluorescence intensity (MFI) from WT (n = 3) and Nod2−/− (n = 3) mice 4 h and 24 h after CLP. (B) Serum and peritoneal C5a and C3a levels were measured 24 h after CLP in WT (n = 4) and Nod2−/− (n = 4) mice injected with recombinant IL-1β or IL-10 prior to CLP. (C) The survival percentages of WT and Nod2−/−mice injected with recombinant IL-1β or IL-10 were measured during CLP-induced sepsis (aP = 0.852 [not significant], bP = 0.002, cP = 0.092 [not significant],dP = 0.009, log-rank test; WT [n = 11], WT mice injected with recombinant IL-1β [n = 6] or IL-10 [n = 6], Nod2−/− [n = 10], and Nod2−/− mice injected with recombinant IL-1β [n = 8] or IL-10 [n = 8]). *P<0.05, **P<0.01, ***P<0.001 (one-way ANOVA [b]). Results shown are representative of three independent experiments except for (C) (mean and SEM).
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ppat-1003351-g003: IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation during cecal ligation and puncture (CLP)-induced sepsis.(A) IL-1 and IL-10 receptor expression was estimated on total peritoneal cells in terms of mean fluorescence intensity (MFI) from WT (n = 3) and Nod2−/− (n = 3) mice 4 h and 24 h after CLP. (B) Serum and peritoneal C5a and C3a levels were measured 24 h after CLP in WT (n = 4) and Nod2−/− (n = 4) mice injected with recombinant IL-1β or IL-10 prior to CLP. (C) The survival percentages of WT and Nod2−/−mice injected with recombinant IL-1β or IL-10 were measured during CLP-induced sepsis (aP = 0.852 [not significant], bP = 0.002, cP = 0.092 [not significant],dP = 0.009, log-rank test; WT [n = 11], WT mice injected with recombinant IL-1β [n = 6] or IL-10 [n = 6], Nod2−/− [n = 10], and Nod2−/− mice injected with recombinant IL-1β [n = 8] or IL-10 [n = 8]). *P<0.05, **P<0.01, ***P<0.001 (one-way ANOVA [b]). Results shown are representative of three independent experiments except for (C) (mean and SEM).

Mentions: To estimate cytokine-mediated effector functions of immune cells in sepsis, we measured the expression of IL-1β and IL-10 receptors on peritoneal cells. Both IL-1β and IL-10 receptors were similarly expressed on total peritoneal cells of WT and Nod2−/− mice with CLP (Fig. 3A). Next, to determine whether NOD2-mediated IL-1β and IL-10 production plays a critical role in C5a generation during sepsis, we administered rIL-1β or rIL-10 to WT or Nod2−/− mice 4 h or 12 h after CLP, respectively. Administration of rIL-1β or rIL-10 enhanced serum and peritoneal C5a, but not C3a, levels (Fig. 3B). Furthermore, rIL-1β or rIL-10 injection into Nod2−/− mice reduced survival rates during sepsis, whereas these recombinant cytokines did not affect the survival of WT mice (Fig. 3C). These findings indicate that NOD2-mediated IL-1β and IL-10 production by neutrophils contributes to the pathogenesis of sepsis by enhancing C5a generation.


NOD2-mediated suppression of CD55 on neutrophils enhances C5a generation during polymicrobial sepsis.

Oh SJ, Kim JH, Chung DH - PLoS Pathog. (2013)

IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation during cecal ligation and puncture (CLP)-induced sepsis.(A) IL-1 and IL-10 receptor expression was estimated on total peritoneal cells in terms of mean fluorescence intensity (MFI) from WT (n = 3) and Nod2−/− (n = 3) mice 4 h and 24 h after CLP. (B) Serum and peritoneal C5a and C3a levels were measured 24 h after CLP in WT (n = 4) and Nod2−/− (n = 4) mice injected with recombinant IL-1β or IL-10 prior to CLP. (C) The survival percentages of WT and Nod2−/−mice injected with recombinant IL-1β or IL-10 were measured during CLP-induced sepsis (aP = 0.852 [not significant], bP = 0.002, cP = 0.092 [not significant],dP = 0.009, log-rank test; WT [n = 11], WT mice injected with recombinant IL-1β [n = 6] or IL-10 [n = 6], Nod2−/− [n = 10], and Nod2−/− mice injected with recombinant IL-1β [n = 8] or IL-10 [n = 8]). *P<0.05, **P<0.01, ***P<0.001 (one-way ANOVA [b]). Results shown are representative of three independent experiments except for (C) (mean and SEM).
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ppat-1003351-g003: IL-1β-dependent IL-10 production mediated by nucleotide-binding oligomerization domain (NOD) 2 enhances C5a generation during cecal ligation and puncture (CLP)-induced sepsis.(A) IL-1 and IL-10 receptor expression was estimated on total peritoneal cells in terms of mean fluorescence intensity (MFI) from WT (n = 3) and Nod2−/− (n = 3) mice 4 h and 24 h after CLP. (B) Serum and peritoneal C5a and C3a levels were measured 24 h after CLP in WT (n = 4) and Nod2−/− (n = 4) mice injected with recombinant IL-1β or IL-10 prior to CLP. (C) The survival percentages of WT and Nod2−/−mice injected with recombinant IL-1β or IL-10 were measured during CLP-induced sepsis (aP = 0.852 [not significant], bP = 0.002, cP = 0.092 [not significant],dP = 0.009, log-rank test; WT [n = 11], WT mice injected with recombinant IL-1β [n = 6] or IL-10 [n = 6], Nod2−/− [n = 10], and Nod2−/− mice injected with recombinant IL-1β [n = 8] or IL-10 [n = 8]). *P<0.05, **P<0.01, ***P<0.001 (one-way ANOVA [b]). Results shown are representative of three independent experiments except for (C) (mean and SEM).
Mentions: To estimate cytokine-mediated effector functions of immune cells in sepsis, we measured the expression of IL-1β and IL-10 receptors on peritoneal cells. Both IL-1β and IL-10 receptors were similarly expressed on total peritoneal cells of WT and Nod2−/− mice with CLP (Fig. 3A). Next, to determine whether NOD2-mediated IL-1β and IL-10 production plays a critical role in C5a generation during sepsis, we administered rIL-1β or rIL-10 to WT or Nod2−/− mice 4 h or 12 h after CLP, respectively. Administration of rIL-1β or rIL-10 enhanced serum and peritoneal C5a, but not C3a, levels (Fig. 3B). Furthermore, rIL-1β or rIL-10 injection into Nod2−/− mice reduced survival rates during sepsis, whereas these recombinant cytokines did not affect the survival of WT mice (Fig. 3C). These findings indicate that NOD2-mediated IL-1β and IL-10 production by neutrophils contributes to the pathogenesis of sepsis by enhancing C5a generation.

Bottom Line: Nucleotide-binding oligomerization domain (NOD) 2 is a cytosolic protein that plays a defensive role in bacterial infection by sensing peptidoglycans.Furthermore, we found that NOD2-mediated IL-10 production by neutrophils enhanced C5a generation by suppressing CD55 expression on neutrophils in IL-1β-dependent and/or IL-1β-independent manners, thereby aggravating CLP-induced sepsis.SB203580, a receptor-interacting protein 2 (RIP2) inhibitor downstream of NOD2, reduced C5a generation by enhancing CD55 expression on neutrophils, resulting in attenuation of polymicrobial sepsis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Seoul National University College of Medicine, Seoul, Korea.

ABSTRACT
Nucleotide-binding oligomerization domain (NOD) 2 is a cytosolic protein that plays a defensive role in bacterial infection by sensing peptidoglycans. C5a, which has harmful effects in sepsis, interacts with innate proteins. However, whether NOD2 regulates C5a generation during sepsis remains to be determined. To address this issue, cecal ligation & puncture (CLP)-induced sepsis was compared in wild type and Nod2-/- mice. Nod2-/- mice showed lower levels of C5a, IL-10, and IL-1β in serum and peritoneum, but higher survival rate during CLP-induced sepsis compared to wild type mice. Injection of recombinant C5a decreased survival rates of Nod2-/- mice rate during sepsis, whereas it did not alter those in wild type mice. These findings suggest a novel provocative role for NOD2 in sepsis, in contrast to its protective role during bacterial infection. Furthermore, we found that NOD2-mediated IL-10 production by neutrophils enhanced C5a generation by suppressing CD55 expression on neutrophils in IL-1β-dependent and/or IL-1β-independent manners, thereby aggravating CLP-induced sepsis. SB203580, a receptor-interacting protein 2 (RIP2) inhibitor downstream of NOD2, reduced C5a generation by enhancing CD55 expression on neutrophils, resulting in attenuation of polymicrobial sepsis. Therefore, we propose a novel NOD2-mediated complement cascade regulatory pathway in sepsis, which may be a useful therapeutic target.

Show MeSH
Related in: MedlinePlus