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Lymphotoxin β receptor signaling induces IL-8 production in human bronchial epithelial cells.

Mikami Y, Matsuzaki H, Horie M, Noguchi S, Jo T, Narumoto O, Kohyama T, Takizawa H, Nagase T, Yamauchi Y - PLoS ONE (2014)

Bottom Line: LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element.Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways.LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Asthma-related mortality has been decreasing due to inhaled corticosteroid use, but severe asthma remains a major clinical problem. One characteristic of severe asthma is resistance to steroid therapy, which is related to neutrophilic inflammation. Recently, the tumor necrosis factor superfamily member (TNFSF) 14/LIGHT has been recognized as a key mediator in severe asthmatic airway inflammation. However, the profiles and intracellular mechanisms of cytokine/chemokine production induced in cells by LIGHT are poorly understood. We aimed to elucidate the molecular mechanism of LIGHT-induced cytokine/chemokine production by bronchial epithelial cells. Human bronchial epithelial cells express lymphotoxin β receptor (LTβR), but not herpesvirus entry mediator, which are receptors for LIGHT. LIGHT induced various cytokines/chemokines, such as interleukin (IL)-6, oncostatin M, monocyte chemotactic protein-1, growth-regulated protein α and IL-8. Specific siRNA for LTβR attenuated IL-6 and IL-8 production by BEAS-2B and normal human bronchial epithelial cells. LIGHT activated intracellular signaling, such as mitogen-activated protein kinase and nuclear factor-κB (NF-κB) signaling. LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element. Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways. LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.

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LTβR signaling in bronchial epithelial cells induced by LIGHT.BEAS-2B cells were stimulated with 50 ng/ml LIGHT, and cell lysates were prepared 0, 5, 15, 20, 30 and 90 minutes later to evaluate phosphorylation of Erk, JNK, p38 and IκB. The MAPKs were phosphorylated for up to 15 minutes, and their signaling was activated. IκB was also phosphorylated at the same time and induced NF-κB release and translocation to nucleus. (A) BEAS-2B cells were pre-treated for 1 h with various inhibitors of phosphorylation of MAPKs. U0126 significantly inhibited IL-8 production by the cells. (B) BEAS-2B cells were pre-treated for 1 h with BAY11-7082. BAY11-7082 significantly inhibited IL-8 production. (C) We performed the same experiments using NHBE cells, which are primary normal human bronchial epithelial cells. LTβR siRNA, U0126 and BAY11-7082 significantly inhibited IL-8 production by NHBE cells in the same manner as seen for BEAS-2B cells.
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pone-0114791-g006: LTβR signaling in bronchial epithelial cells induced by LIGHT.BEAS-2B cells were stimulated with 50 ng/ml LIGHT, and cell lysates were prepared 0, 5, 15, 20, 30 and 90 minutes later to evaluate phosphorylation of Erk, JNK, p38 and IκB. The MAPKs were phosphorylated for up to 15 minutes, and their signaling was activated. IκB was also phosphorylated at the same time and induced NF-κB release and translocation to nucleus. (A) BEAS-2B cells were pre-treated for 1 h with various inhibitors of phosphorylation of MAPKs. U0126 significantly inhibited IL-8 production by the cells. (B) BEAS-2B cells were pre-treated for 1 h with BAY11-7082. BAY11-7082 significantly inhibited IL-8 production. (C) We performed the same experiments using NHBE cells, which are primary normal human bronchial epithelial cells. LTβR siRNA, U0126 and BAY11-7082 significantly inhibited IL-8 production by NHBE cells in the same manner as seen for BEAS-2B cells.

Mentions: In order to determine the mechanism of LIGHT-induced IL-8 production by bronchial epithelial cells, we evaluated MAPKs signaling and NF-κB signaling. As shown in Fig. 6A, LIGHT potently induced MAPKs signals, and also IκBα phosphorylation. Moreover, to examine whether induction of MAPKs and NF-kB signaling contributed to production of IL-8, we used specific inhibitors of signaling. U0126, but not SP600125 or SB203580, significantly inhibited IL-8 production (Fig. 6B). These results suggest that LIGHT can induce Erk1/2, p38 and JNK signaling, but the major contributor to IL-8 production is Erk1/2 signaling. BAY11-7082 also inhibited IL-8 production by the cells (Fig. 6C). We also examined the mechanism of IL-8 production by NHBE cells. As shown in Fig. 6D, U0126, BAY11-7082 and siRNA for LTβR each inhibited IL-8 production by NHBE cells, similar to BEAS-2B cells.


Lymphotoxin β receptor signaling induces IL-8 production in human bronchial epithelial cells.

Mikami Y, Matsuzaki H, Horie M, Noguchi S, Jo T, Narumoto O, Kohyama T, Takizawa H, Nagase T, Yamauchi Y - PLoS ONE (2014)

LTβR signaling in bronchial epithelial cells induced by LIGHT.BEAS-2B cells were stimulated with 50 ng/ml LIGHT, and cell lysates were prepared 0, 5, 15, 20, 30 and 90 minutes later to evaluate phosphorylation of Erk, JNK, p38 and IκB. The MAPKs were phosphorylated for up to 15 minutes, and their signaling was activated. IκB was also phosphorylated at the same time and induced NF-κB release and translocation to nucleus. (A) BEAS-2B cells were pre-treated for 1 h with various inhibitors of phosphorylation of MAPKs. U0126 significantly inhibited IL-8 production by the cells. (B) BEAS-2B cells were pre-treated for 1 h with BAY11-7082. BAY11-7082 significantly inhibited IL-8 production. (C) We performed the same experiments using NHBE cells, which are primary normal human bronchial epithelial cells. LTβR siRNA, U0126 and BAY11-7082 significantly inhibited IL-8 production by NHBE cells in the same manner as seen for BEAS-2B cells.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4263477&req=5

pone-0114791-g006: LTβR signaling in bronchial epithelial cells induced by LIGHT.BEAS-2B cells were stimulated with 50 ng/ml LIGHT, and cell lysates were prepared 0, 5, 15, 20, 30 and 90 minutes later to evaluate phosphorylation of Erk, JNK, p38 and IκB. The MAPKs were phosphorylated for up to 15 minutes, and their signaling was activated. IκB was also phosphorylated at the same time and induced NF-κB release and translocation to nucleus. (A) BEAS-2B cells were pre-treated for 1 h with various inhibitors of phosphorylation of MAPKs. U0126 significantly inhibited IL-8 production by the cells. (B) BEAS-2B cells were pre-treated for 1 h with BAY11-7082. BAY11-7082 significantly inhibited IL-8 production. (C) We performed the same experiments using NHBE cells, which are primary normal human bronchial epithelial cells. LTβR siRNA, U0126 and BAY11-7082 significantly inhibited IL-8 production by NHBE cells in the same manner as seen for BEAS-2B cells.
Mentions: In order to determine the mechanism of LIGHT-induced IL-8 production by bronchial epithelial cells, we evaluated MAPKs signaling and NF-κB signaling. As shown in Fig. 6A, LIGHT potently induced MAPKs signals, and also IκBα phosphorylation. Moreover, to examine whether induction of MAPKs and NF-kB signaling contributed to production of IL-8, we used specific inhibitors of signaling. U0126, but not SP600125 or SB203580, significantly inhibited IL-8 production (Fig. 6B). These results suggest that LIGHT can induce Erk1/2, p38 and JNK signaling, but the major contributor to IL-8 production is Erk1/2 signaling. BAY11-7082 also inhibited IL-8 production by the cells (Fig. 6C). We also examined the mechanism of IL-8 production by NHBE cells. As shown in Fig. 6D, U0126, BAY11-7082 and siRNA for LTβR each inhibited IL-8 production by NHBE cells, similar to BEAS-2B cells.

Bottom Line: LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element.Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways.LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.

ABSTRACT
Asthma-related mortality has been decreasing due to inhaled corticosteroid use, but severe asthma remains a major clinical problem. One characteristic of severe asthma is resistance to steroid therapy, which is related to neutrophilic inflammation. Recently, the tumor necrosis factor superfamily member (TNFSF) 14/LIGHT has been recognized as a key mediator in severe asthmatic airway inflammation. However, the profiles and intracellular mechanisms of cytokine/chemokine production induced in cells by LIGHT are poorly understood. We aimed to elucidate the molecular mechanism of LIGHT-induced cytokine/chemokine production by bronchial epithelial cells. Human bronchial epithelial cells express lymphotoxin β receptor (LTβR), but not herpesvirus entry mediator, which are receptors for LIGHT. LIGHT induced various cytokines/chemokines, such as interleukin (IL)-6, oncostatin M, monocyte chemotactic protein-1, growth-regulated protein α and IL-8. Specific siRNA for LTβR attenuated IL-6 and IL-8 production by BEAS-2B and normal human bronchial epithelial cells. LIGHT activated intracellular signaling, such as mitogen-activated protein kinase and nuclear factor-κB (NF-κB) signaling. LIGHT also induced luciferase activity of NF-κB response element, but not of activator protein-1 or serum response element. Specific inhibitors of phosphorylation of extracellular signal-regulated kinase (Erk) and that of inhibitor κB attenuated IL-8 production, suggesting that LIGHT-LTβR signaling induces IL-8 production via the Erk and NF-κB pathways. LIGHT, via LTβR signaling, may contribute to exacerbation of airway neutrophilic inflammation through cytokine and chemokine production by bronchial epithelial cells.

Show MeSH
Related in: MedlinePlus