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The role of toll-like receptor 9 in chronic stress-induced apoptosis in macrophage.

Xiang Y, Yan H, Zhou J, Zhang Q, Hanley G, Caudle Y, LeSage G, Zhang X, Yin D - PLoS ONE (2015)

Bottom Line: Toll-like receptors (TLRs) have been shown to play an essential role in modulating immune responses and cell survival.We have recently shown that TLR9 deficiency protects against lymphocyte apoptosis induced by chronic stress.TLR9 deficiency was also found to reverse elevation of plasma IL-1β, IL-10 and IL-17 levels and decrease of plasma IFN-γ level under the condition of chronic stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, College of Medicine, East Tennessee State University, Johnson City, Tennessee 37614, United States of America; Department of Pharmacology, Shandong University School of Medicine, Jinan, People's Republic of China.

ABSTRACT
Emerging evidence implied that chronic stress has been exerting detrimental impact on immune system functions in both humans and animals. Toll-like receptors (TLRs) have been shown to play an essential role in modulating immune responses and cell survival. We have recently shown that TLR9 deficiency protects against lymphocyte apoptosis induced by chronic stress. However, the exact role of TLR9 in stress-mediated change of macrophage function remains unclear. The results of the current study showed that when BALB/c mice were treated with restraint stress (12 h daily for 2 days), the number of macrophages recruited to the peritoneal cavity was obviously increased. Results also demonstrated that the sustained effects of stress elevated cytokine IL-1β, TNF-α and IL-10 production yet diminished IFN-γ production from macrophage, which led to apoptotic cell death. However, TLR9 deficiency prevented the chronic stress-mediated accumulation of macrophages. In addition, knocking out TLR9 significantly abolished the chronic stress-induced imbalance of cytokine levels and apoptosis in macrophage. TLR9 deficiency was also found to reverse elevation of plasma IL-1β, IL-10 and IL-17 levels and decrease of plasma IFN-γ level under the condition of chronic stress. These results indicated that TLR9-mediated macrophage responses were required for chronic stress-induced immunosuppression. Further exploration showed that TLR9 deficiency prevented the increment of p38 MAPK phosphorylation and reduction of Akt/Gsk-3β phosphorylation; TLR9 deficiency also attenuated the release of mitochondrial cytochrome c into cytoplasm, caused upregulation of Bcl-2/Bax protein ratio, downregulation of cleavage of caspase-3 and PARP, as well as decreased TUNEL-positive cells in macrophage of stressed mice. Collectively, our studies demonstrated that deficiency of TLR9 maintained macrophage function by modulating macrophage accumulation and attenuating macrophage apoptosis, thus preventing immunosuppression in restraint-stressed mice.

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A deficiency of TLR9 blocks chronic stress-induced accumulation of macrophages in peritoneal cavity TLR9 knockout mice or wild type BALB/c mice aged 6 to 8 weeks were subjected to a 12 h physical restraint daily.After 2 d stress, mice were sacrificed by cervical dislocation, and the peritoneal macrophages were harvested and the counts were performed. For TLR9 protein expression evaluating, the macrophages were harvested and cultured for 24 hours. The expression of TLR9 was analyzed by Western blot. Means and SEs were calculated from 7 mice per group. *p < 0.05, **p< 0.01 compared with indicated groups.
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pone.0123447.g001: A deficiency of TLR9 blocks chronic stress-induced accumulation of macrophages in peritoneal cavity TLR9 knockout mice or wild type BALB/c mice aged 6 to 8 weeks were subjected to a 12 h physical restraint daily.After 2 d stress, mice were sacrificed by cervical dislocation, and the peritoneal macrophages were harvested and the counts were performed. For TLR9 protein expression evaluating, the macrophages were harvested and cultured for 24 hours. The expression of TLR9 was analyzed by Western blot. Means and SEs were calculated from 7 mice per group. *p < 0.05, **p< 0.01 compared with indicated groups.

Mentions: Recent evidence showed that TLR9 is largely expressed on macrophages, however, the exact role of TLR9 in modulating macrophage function is not known yet [24]. Data suggests that chronic stress increases TLR9 expression in peritoneal macrophage (Fig 1A). We therefore asked whether TLR9 is involved in stress-mediated changes of macrophage function. Since macrophages in peritoneal cavity of chronic stress-induced mice, irrespective of their location, can significantly contribute to inflammation and immune response by producing cytokines and free oxygen radicals [25] [26], it is important to assess the total number of macrophages accumulated in peritoneal cavity. Therefore, we decided to examine the pattern of total macrophages increase after stress treatment in the peritoneal cavity of TLR9 knockout and wild type mice. We observed a robust accumulation of macrophages in peritoneal cavity 2 days after stress challenge, representing a > 2-fold increase over baseline number of cells; no significant change in number of peritoneal macrophages was observed after stress challenge compared with that in control group in TLR 9 knockout mice (Fig 1B). Therefore, TLR9 knockout mice lose their sensitivity to chronic stress-induced accumulation of macrophages, supporting a critical role of TLR9 in stress-induced immune response.


The role of toll-like receptor 9 in chronic stress-induced apoptosis in macrophage.

Xiang Y, Yan H, Zhou J, Zhang Q, Hanley G, Caudle Y, LeSage G, Zhang X, Yin D - PLoS ONE (2015)

A deficiency of TLR9 blocks chronic stress-induced accumulation of macrophages in peritoneal cavity TLR9 knockout mice or wild type BALB/c mice aged 6 to 8 weeks were subjected to a 12 h physical restraint daily.After 2 d stress, mice were sacrificed by cervical dislocation, and the peritoneal macrophages were harvested and the counts were performed. For TLR9 protein expression evaluating, the macrophages were harvested and cultured for 24 hours. The expression of TLR9 was analyzed by Western blot. Means and SEs were calculated from 7 mice per group. *p < 0.05, **p< 0.01 compared with indicated groups.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401452&req=5

pone.0123447.g001: A deficiency of TLR9 blocks chronic stress-induced accumulation of macrophages in peritoneal cavity TLR9 knockout mice or wild type BALB/c mice aged 6 to 8 weeks were subjected to a 12 h physical restraint daily.After 2 d stress, mice were sacrificed by cervical dislocation, and the peritoneal macrophages were harvested and the counts were performed. For TLR9 protein expression evaluating, the macrophages were harvested and cultured for 24 hours. The expression of TLR9 was analyzed by Western blot. Means and SEs were calculated from 7 mice per group. *p < 0.05, **p< 0.01 compared with indicated groups.
Mentions: Recent evidence showed that TLR9 is largely expressed on macrophages, however, the exact role of TLR9 in modulating macrophage function is not known yet [24]. Data suggests that chronic stress increases TLR9 expression in peritoneal macrophage (Fig 1A). We therefore asked whether TLR9 is involved in stress-mediated changes of macrophage function. Since macrophages in peritoneal cavity of chronic stress-induced mice, irrespective of their location, can significantly contribute to inflammation and immune response by producing cytokines and free oxygen radicals [25] [26], it is important to assess the total number of macrophages accumulated in peritoneal cavity. Therefore, we decided to examine the pattern of total macrophages increase after stress treatment in the peritoneal cavity of TLR9 knockout and wild type mice. We observed a robust accumulation of macrophages in peritoneal cavity 2 days after stress challenge, representing a > 2-fold increase over baseline number of cells; no significant change in number of peritoneal macrophages was observed after stress challenge compared with that in control group in TLR 9 knockout mice (Fig 1B). Therefore, TLR9 knockout mice lose their sensitivity to chronic stress-induced accumulation of macrophages, supporting a critical role of TLR9 in stress-induced immune response.

Bottom Line: Toll-like receptors (TLRs) have been shown to play an essential role in modulating immune responses and cell survival.We have recently shown that TLR9 deficiency protects against lymphocyte apoptosis induced by chronic stress.TLR9 deficiency was also found to reverse elevation of plasma IL-1β, IL-10 and IL-17 levels and decrease of plasma IFN-γ level under the condition of chronic stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, College of Medicine, East Tennessee State University, Johnson City, Tennessee 37614, United States of America; Department of Pharmacology, Shandong University School of Medicine, Jinan, People's Republic of China.

ABSTRACT
Emerging evidence implied that chronic stress has been exerting detrimental impact on immune system functions in both humans and animals. Toll-like receptors (TLRs) have been shown to play an essential role in modulating immune responses and cell survival. We have recently shown that TLR9 deficiency protects against lymphocyte apoptosis induced by chronic stress. However, the exact role of TLR9 in stress-mediated change of macrophage function remains unclear. The results of the current study showed that when BALB/c mice were treated with restraint stress (12 h daily for 2 days), the number of macrophages recruited to the peritoneal cavity was obviously increased. Results also demonstrated that the sustained effects of stress elevated cytokine IL-1β, TNF-α and IL-10 production yet diminished IFN-γ production from macrophage, which led to apoptotic cell death. However, TLR9 deficiency prevented the chronic stress-mediated accumulation of macrophages. In addition, knocking out TLR9 significantly abolished the chronic stress-induced imbalance of cytokine levels and apoptosis in macrophage. TLR9 deficiency was also found to reverse elevation of plasma IL-1β, IL-10 and IL-17 levels and decrease of plasma IFN-γ level under the condition of chronic stress. These results indicated that TLR9-mediated macrophage responses were required for chronic stress-induced immunosuppression. Further exploration showed that TLR9 deficiency prevented the increment of p38 MAPK phosphorylation and reduction of Akt/Gsk-3β phosphorylation; TLR9 deficiency also attenuated the release of mitochondrial cytochrome c into cytoplasm, caused upregulation of Bcl-2/Bax protein ratio, downregulation of cleavage of caspase-3 and PARP, as well as decreased TUNEL-positive cells in macrophage of stressed mice. Collectively, our studies demonstrated that deficiency of TLR9 maintained macrophage function by modulating macrophage accumulation and attenuating macrophage apoptosis, thus preventing immunosuppression in restraint-stressed mice.

Show MeSH
Related in: MedlinePlus