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Role of JAK-STAT signaling in maturation of phagosomes containing Staphylococcus aureus.

Zhu F, Zhou Y, Jiang C, Zhang X - Sci Rep (2015)

Bottom Line: In this study, the S. aureus was engulfed by macrophages (RAW264.7 cells) but not digested by the cells, suggesting that the phagosomes did not maturate in macrophages.Further investigation revealed that peptidoglycan (PG) induced the phagosome maturation of macrophages, resulting in the eradication of S. aureus.Therefore, our study contributed evidence that revealed a novel aspect of PG-triggered JAK-STAT pathway in the phagosome maturation of macrophages.

View Article: PubMed Central - PubMed

Affiliation: Collaborative Innovation Center of Deep Sea Biology, Key Laboratory of Animal Virology of Ministry of Agriculture and College of Life Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Phagocytosis is a required mechanism for the defense against pathogens. Staphylococcus aureus, an important bacterial pathogen, can promptly escape from phagosomes and proliferate within the cytoplasm of host. However, the mechanism of phagocytosis against S. aureus has not been intensively investigated. In this study, the S. aureus was engulfed by macrophages (RAW264.7 cells) but not digested by the cells, suggesting that the phagosomes did not maturate in macrophages. Further investigation revealed that peptidoglycan (PG) induced the phagosome maturation of macrophages, resulting in the eradication of S. aureus. Genome-wide analysis and quantitative real-time PCR indicated that the JAK-STAT pathway was activated by PG during the phagosome maturation of macrophages against S. aureus. This finding presented that the PG-activated JAK-STAT pathway was required for phagosome maturation. Therefore, our study contributed evidence that revealed a novel aspect of PG-triggered JAK-STAT pathway in the phagosome maturation of macrophages.

No MeSH data available.


Related in: MedlinePlus

Analysis of signaling pathways required for the phagosome maturation.(A) The gene expression profiles were conducted with a DNA microarray using RAW264.7 cells treated with heat-inactivated S. aureus or heat-inactivated S. aureus and PG. Non-treated RAW264.7 cells were used as a control. Red indicated up-regulation of gene expression and green showed down-regulation of gene expression. (B) Venn diagram of differentially expressed genes. The differentially expressed genes were evaluated using DNA microarray data. The numbers represented the genes up-regulated or down-regulated (more than twofold) compared with the control. Arrows indicated up- or down-regulated genes. (C) Hierarchical cluster analysis of DNA microarray data. Clustering of 32 genes in JAK-STAT pathway in response to the challenge of heat-inactivated S. aureus or heat-inactivated S. aureus and PG. (D) The biological processes mediated by the differentially expressed genes. The 524 up-regulated genes in RAW264.7 cells treated with heat-inactivated S. aureus and PG were analyzed. The JAK-STAT pathway represented the key pathway. The red, blue and purple lines indicated the IntAct, MINT and DIP databases, respectively. The numbers indicated a protein’s interaction degree. (E) The expressions of genes from JAK-STAT pathway in RAW264.7 cells in response to PG challenge. At 1 h after challenge with heat-inactivated S. aureus, heat-inactivated S. aureus + PG and PG alone, the expression profiles of Jak2, Stat3, Stat5a, Cish, Csf1, IL-6, Socs3, Nfkb1, TNF-α and IFN-γ genes in RAW264.7 cells were characterized by quantitative real-time PCR. The GAPDH gene was used as a control. Statistically significant differences between treatments were indicated with asterisks (**P < 0.01). Control, RAW264.7 cells only.
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f2: Analysis of signaling pathways required for the phagosome maturation.(A) The gene expression profiles were conducted with a DNA microarray using RAW264.7 cells treated with heat-inactivated S. aureus or heat-inactivated S. aureus and PG. Non-treated RAW264.7 cells were used as a control. Red indicated up-regulation of gene expression and green showed down-regulation of gene expression. (B) Venn diagram of differentially expressed genes. The differentially expressed genes were evaluated using DNA microarray data. The numbers represented the genes up-regulated or down-regulated (more than twofold) compared with the control. Arrows indicated up- or down-regulated genes. (C) Hierarchical cluster analysis of DNA microarray data. Clustering of 32 genes in JAK-STAT pathway in response to the challenge of heat-inactivated S. aureus or heat-inactivated S. aureus and PG. (D) The biological processes mediated by the differentially expressed genes. The 524 up-regulated genes in RAW264.7 cells treated with heat-inactivated S. aureus and PG were analyzed. The JAK-STAT pathway represented the key pathway. The red, blue and purple lines indicated the IntAct, MINT and DIP databases, respectively. The numbers indicated a protein’s interaction degree. (E) The expressions of genes from JAK-STAT pathway in RAW264.7 cells in response to PG challenge. At 1 h after challenge with heat-inactivated S. aureus, heat-inactivated S. aureus + PG and PG alone, the expression profiles of Jak2, Stat3, Stat5a, Cish, Csf1, IL-6, Socs3, Nfkb1, TNF-α and IFN-γ genes in RAW264.7 cells were characterized by quantitative real-time PCR. The GAPDH gene was used as a control. Statistically significant differences between treatments were indicated with asterisks (**P < 0.01). Control, RAW264.7 cells only.

Mentions: To determine which genes and pathways were involved in phagosome maturation in RAW264.7 cells, a genome-wide analysis using oligonucleotide microarray was conducted after the cells were incubated with PG and inactivated S. aureus. The gene expression profiles of RAW264.7 cells treated with inactivated S. aureus and PG or inactivated S. aureus differed from untreated cells) (Fig. 2A). This finding suggested that PG could induce various receptors and pathways.


Role of JAK-STAT signaling in maturation of phagosomes containing Staphylococcus aureus.

Zhu F, Zhou Y, Jiang C, Zhang X - Sci Rep (2015)

Analysis of signaling pathways required for the phagosome maturation.(A) The gene expression profiles were conducted with a DNA microarray using RAW264.7 cells treated with heat-inactivated S. aureus or heat-inactivated S. aureus and PG. Non-treated RAW264.7 cells were used as a control. Red indicated up-regulation of gene expression and green showed down-regulation of gene expression. (B) Venn diagram of differentially expressed genes. The differentially expressed genes were evaluated using DNA microarray data. The numbers represented the genes up-regulated or down-regulated (more than twofold) compared with the control. Arrows indicated up- or down-regulated genes. (C) Hierarchical cluster analysis of DNA microarray data. Clustering of 32 genes in JAK-STAT pathway in response to the challenge of heat-inactivated S. aureus or heat-inactivated S. aureus and PG. (D) The biological processes mediated by the differentially expressed genes. The 524 up-regulated genes in RAW264.7 cells treated with heat-inactivated S. aureus and PG were analyzed. The JAK-STAT pathway represented the key pathway. The red, blue and purple lines indicated the IntAct, MINT and DIP databases, respectively. The numbers indicated a protein’s interaction degree. (E) The expressions of genes from JAK-STAT pathway in RAW264.7 cells in response to PG challenge. At 1 h after challenge with heat-inactivated S. aureus, heat-inactivated S. aureus + PG and PG alone, the expression profiles of Jak2, Stat3, Stat5a, Cish, Csf1, IL-6, Socs3, Nfkb1, TNF-α and IFN-γ genes in RAW264.7 cells were characterized by quantitative real-time PCR. The GAPDH gene was used as a control. Statistically significant differences between treatments were indicated with asterisks (**P < 0.01). Control, RAW264.7 cells only.
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f2: Analysis of signaling pathways required for the phagosome maturation.(A) The gene expression profiles were conducted with a DNA microarray using RAW264.7 cells treated with heat-inactivated S. aureus or heat-inactivated S. aureus and PG. Non-treated RAW264.7 cells were used as a control. Red indicated up-regulation of gene expression and green showed down-regulation of gene expression. (B) Venn diagram of differentially expressed genes. The differentially expressed genes were evaluated using DNA microarray data. The numbers represented the genes up-regulated or down-regulated (more than twofold) compared with the control. Arrows indicated up- or down-regulated genes. (C) Hierarchical cluster analysis of DNA microarray data. Clustering of 32 genes in JAK-STAT pathway in response to the challenge of heat-inactivated S. aureus or heat-inactivated S. aureus and PG. (D) The biological processes mediated by the differentially expressed genes. The 524 up-regulated genes in RAW264.7 cells treated with heat-inactivated S. aureus and PG were analyzed. The JAK-STAT pathway represented the key pathway. The red, blue and purple lines indicated the IntAct, MINT and DIP databases, respectively. The numbers indicated a protein’s interaction degree. (E) The expressions of genes from JAK-STAT pathway in RAW264.7 cells in response to PG challenge. At 1 h after challenge with heat-inactivated S. aureus, heat-inactivated S. aureus + PG and PG alone, the expression profiles of Jak2, Stat3, Stat5a, Cish, Csf1, IL-6, Socs3, Nfkb1, TNF-α and IFN-γ genes in RAW264.7 cells were characterized by quantitative real-time PCR. The GAPDH gene was used as a control. Statistically significant differences between treatments were indicated with asterisks (**P < 0.01). Control, RAW264.7 cells only.
Mentions: To determine which genes and pathways were involved in phagosome maturation in RAW264.7 cells, a genome-wide analysis using oligonucleotide microarray was conducted after the cells were incubated with PG and inactivated S. aureus. The gene expression profiles of RAW264.7 cells treated with inactivated S. aureus and PG or inactivated S. aureus differed from untreated cells) (Fig. 2A). This finding suggested that PG could induce various receptors and pathways.

Bottom Line: In this study, the S. aureus was engulfed by macrophages (RAW264.7 cells) but not digested by the cells, suggesting that the phagosomes did not maturate in macrophages.Further investigation revealed that peptidoglycan (PG) induced the phagosome maturation of macrophages, resulting in the eradication of S. aureus.Therefore, our study contributed evidence that revealed a novel aspect of PG-triggered JAK-STAT pathway in the phagosome maturation of macrophages.

View Article: PubMed Central - PubMed

Affiliation: Collaborative Innovation Center of Deep Sea Biology, Key Laboratory of Animal Virology of Ministry of Agriculture and College of Life Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Phagocytosis is a required mechanism for the defense against pathogens. Staphylococcus aureus, an important bacterial pathogen, can promptly escape from phagosomes and proliferate within the cytoplasm of host. However, the mechanism of phagocytosis against S. aureus has not been intensively investigated. In this study, the S. aureus was engulfed by macrophages (RAW264.7 cells) but not digested by the cells, suggesting that the phagosomes did not maturate in macrophages. Further investigation revealed that peptidoglycan (PG) induced the phagosome maturation of macrophages, resulting in the eradication of S. aureus. Genome-wide analysis and quantitative real-time PCR indicated that the JAK-STAT pathway was activated by PG during the phagosome maturation of macrophages against S. aureus. This finding presented that the PG-activated JAK-STAT pathway was required for phagosome maturation. Therefore, our study contributed evidence that revealed a novel aspect of PG-triggered JAK-STAT pathway in the phagosome maturation of macrophages.

No MeSH data available.


Related in: MedlinePlus