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Hydrogen Peroxide Induce Human Cytomegalovirus Replication through the Activation of p38-MAPK Signaling Pathway.

Xiao J, Deng J, Lv L, Kang Q, Ma P, Yan F, Song X, Gao B, Zhang Y, Xu J - Viruses (2015)

Bottom Line: In the present study, we found that H2O2 enhanced HCMV lytic replication through promoting major immediate early (MIE) promoter activity and immediate early (IE) gene transcription.The suppressive effect of NAC on CMV in an acute CMV-infected mouse model also showed a relationship between antioxidants and viral lytic replication.These results directly relate HCMV replication to H2O2, suggesting that treatment with antioxidants may be an attractive preventive and therapeutic strategy for HCMV.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing 100850, China. ammsxj@126.com.

ABSTRACT
Human cytomegalovirus (HCMV) is a major risk factor in transplantation and AIDS patients, which induces high morbidity and mortality. These patients infected with HCMV experience an imbalance of redox homeostasis that cause accumulation of reactive oxygen species (ROS) at the cellular level. H2O2, the most common reactive oxygen species, is the main byproduct of oxidative metabolism. However, the function of H2O2 on HCMV infection is not yet fully understood and the effect and mechanism of N-acetylcysteine (NAC) on H2O2-stimulated HCMV replication is unclear. We, therefore, examined the effect of NAC on H2O2-induced HCMV production in human foreskin fibroblast cells. In the present study, we found that H2O2 enhanced HCMV lytic replication through promoting major immediate early (MIE) promoter activity and immediate early (IE) gene transcription. Conversely, NAC inhibited H2O2-upregulated viral IE gene expression and viral replication. The suppressive effect of NAC on CMV in an acute CMV-infected mouse model also showed a relationship between antioxidants and viral lytic replication. Intriguingly, the enhancement of HCMV replication via supplementation with H2O2 was accompanied with the activation of the p38 mitogen-activated protein kinase pathway. Similar to NAC, the p38 inhibitor SB203580 inhibited H2O2-induced p38 phosphorylation and HCMV upregulation, while upregulation of inducible ROS was unaffected. These results directly relate HCMV replication to H2O2, suggesting that treatment with antioxidants may be an attractive preventive and therapeutic strategy for HCMV.

No MeSH data available.


Related in: MedlinePlus

ATA-induced intracellular H2O2, enhancing viral replication in HFF cells. Treatment of HFF cells with the catalase inhibitor 3-amino-1,2,4-triazole (ATA) (0, 1, 2, 4 mM) for 24 h reduced catalase activity and increased intracellular H2O2 level in a dose-dependent manner (A). Cells were cultured with ATA for 24 h. ATA-induced intracellular H2O2 increased MIE promoter activity (B) and HCMV IE1 transcription (C). Cells were infected with UV-HCMV (UV) or HCMV at an MOI of 0.5. An increase in pp72 (D) and pp65 (E) protein levels were detected by Western blotting under 0, 1, 2, 4 mM ATA treatments for 72 h, and β-actin was used to calibrate sample loading. (F) Relative virus titers were measured using TCID50 assay within five days. (G) Catalase, HCMV lytic protein pp72, pp65 and β-actin levels were determined by Western blotting after treatment with siRNA for five days. * p < 0.05; ** p < 0.01 or *** p < 0.001 for ATA-treated versus untreated cells.
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viruses-07-02748-f003: ATA-induced intracellular H2O2, enhancing viral replication in HFF cells. Treatment of HFF cells with the catalase inhibitor 3-amino-1,2,4-triazole (ATA) (0, 1, 2, 4 mM) for 24 h reduced catalase activity and increased intracellular H2O2 level in a dose-dependent manner (A). Cells were cultured with ATA for 24 h. ATA-induced intracellular H2O2 increased MIE promoter activity (B) and HCMV IE1 transcription (C). Cells were infected with UV-HCMV (UV) or HCMV at an MOI of 0.5. An increase in pp72 (D) and pp65 (E) protein levels were detected by Western blotting under 0, 1, 2, 4 mM ATA treatments for 72 h, and β-actin was used to calibrate sample loading. (F) Relative virus titers were measured using TCID50 assay within five days. (G) Catalase, HCMV lytic protein pp72, pp65 and β-actin levels were determined by Western blotting after treatment with siRNA for five days. * p < 0.05; ** p < 0.01 or *** p < 0.001 for ATA-treated versus untreated cells.

Mentions: Next, we verified whether an increase in intracellular H2O2 is sufficient to induce HCMV replication. Treatment of HFF cells with ATA, an inhibitor of the H2O2-scavenging enzyme catalase, reduced the activity of cellular catalase (Figure 3A left panel) and increased the intracellular H2O2 level (Figure 3A right panel). ATA increased the MIE promoter activities, the IE1 gene transcripts, the expression of HCMV pp72 and pp65, and production of infectious virions (Figure 3B–F).


Hydrogen Peroxide Induce Human Cytomegalovirus Replication through the Activation of p38-MAPK Signaling Pathway.

Xiao J, Deng J, Lv L, Kang Q, Ma P, Yan F, Song X, Gao B, Zhang Y, Xu J - Viruses (2015)

ATA-induced intracellular H2O2, enhancing viral replication in HFF cells. Treatment of HFF cells with the catalase inhibitor 3-amino-1,2,4-triazole (ATA) (0, 1, 2, 4 mM) for 24 h reduced catalase activity and increased intracellular H2O2 level in a dose-dependent manner (A). Cells were cultured with ATA for 24 h. ATA-induced intracellular H2O2 increased MIE promoter activity (B) and HCMV IE1 transcription (C). Cells were infected with UV-HCMV (UV) or HCMV at an MOI of 0.5. An increase in pp72 (D) and pp65 (E) protein levels were detected by Western blotting under 0, 1, 2, 4 mM ATA treatments for 72 h, and β-actin was used to calibrate sample loading. (F) Relative virus titers were measured using TCID50 assay within five days. (G) Catalase, HCMV lytic protein pp72, pp65 and β-actin levels were determined by Western blotting after treatment with siRNA for five days. * p < 0.05; ** p < 0.01 or *** p < 0.001 for ATA-treated versus untreated cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488715&req=5

viruses-07-02748-f003: ATA-induced intracellular H2O2, enhancing viral replication in HFF cells. Treatment of HFF cells with the catalase inhibitor 3-amino-1,2,4-triazole (ATA) (0, 1, 2, 4 mM) for 24 h reduced catalase activity and increased intracellular H2O2 level in a dose-dependent manner (A). Cells were cultured with ATA for 24 h. ATA-induced intracellular H2O2 increased MIE promoter activity (B) and HCMV IE1 transcription (C). Cells were infected with UV-HCMV (UV) or HCMV at an MOI of 0.5. An increase in pp72 (D) and pp65 (E) protein levels were detected by Western blotting under 0, 1, 2, 4 mM ATA treatments for 72 h, and β-actin was used to calibrate sample loading. (F) Relative virus titers were measured using TCID50 assay within five days. (G) Catalase, HCMV lytic protein pp72, pp65 and β-actin levels were determined by Western blotting after treatment with siRNA for five days. * p < 0.05; ** p < 0.01 or *** p < 0.001 for ATA-treated versus untreated cells.
Mentions: Next, we verified whether an increase in intracellular H2O2 is sufficient to induce HCMV replication. Treatment of HFF cells with ATA, an inhibitor of the H2O2-scavenging enzyme catalase, reduced the activity of cellular catalase (Figure 3A left panel) and increased the intracellular H2O2 level (Figure 3A right panel). ATA increased the MIE promoter activities, the IE1 gene transcripts, the expression of HCMV pp72 and pp65, and production of infectious virions (Figure 3B–F).

Bottom Line: In the present study, we found that H2O2 enhanced HCMV lytic replication through promoting major immediate early (MIE) promoter activity and immediate early (IE) gene transcription.The suppressive effect of NAC on CMV in an acute CMV-infected mouse model also showed a relationship between antioxidants and viral lytic replication.These results directly relate HCMV replication to H2O2, suggesting that treatment with antioxidants may be an attractive preventive and therapeutic strategy for HCMV.

View Article: PubMed Central - PubMed

Affiliation: Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing 100850, China. ammsxj@126.com.

ABSTRACT
Human cytomegalovirus (HCMV) is a major risk factor in transplantation and AIDS patients, which induces high morbidity and mortality. These patients infected with HCMV experience an imbalance of redox homeostasis that cause accumulation of reactive oxygen species (ROS) at the cellular level. H2O2, the most common reactive oxygen species, is the main byproduct of oxidative metabolism. However, the function of H2O2 on HCMV infection is not yet fully understood and the effect and mechanism of N-acetylcysteine (NAC) on H2O2-stimulated HCMV replication is unclear. We, therefore, examined the effect of NAC on H2O2-induced HCMV production in human foreskin fibroblast cells. In the present study, we found that H2O2 enhanced HCMV lytic replication through promoting major immediate early (MIE) promoter activity and immediate early (IE) gene transcription. Conversely, NAC inhibited H2O2-upregulated viral IE gene expression and viral replication. The suppressive effect of NAC on CMV in an acute CMV-infected mouse model also showed a relationship between antioxidants and viral lytic replication. Intriguingly, the enhancement of HCMV replication via supplementation with H2O2 was accompanied with the activation of the p38 mitogen-activated protein kinase pathway. Similar to NAC, the p38 inhibitor SB203580 inhibited H2O2-induced p38 phosphorylation and HCMV upregulation, while upregulation of inducible ROS was unaffected. These results directly relate HCMV replication to H2O2, suggesting that treatment with antioxidants may be an attractive preventive and therapeutic strategy for HCMV.

No MeSH data available.


Related in: MedlinePlus