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PI3K/Akt signaling pathway modulates influenza virus induced mouse alveolar macrophage polarization to M1/M2b.

Zhao X, Dai J, Xiao X, Wu L, Zeng J, Sheng J, Su J, Chen X, Wang G, Li K - PLoS ONE (2014)

Bottom Line: Protein expression assay showed similar results as the gene expression analysis for phenotype verification.Immunofluorescence showed that iNOS expression was up-regulated but not Arg1 expression.In conclusion, our results demonstrate the dynamic polarization of AM induced by influenza viruses, and suggested that PI3K/Akt signaling pathway modulates AM polarization to M1/M2b.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Shantou University Medical College, College, Shantou, Guangdong, China.

ABSTRACT
Macrophages polarized to M1 (pro-inflammation) or M2 (anti-inflammation) phenotypes in response to environmental signals. In this study, we examined the polarization of alveolar macrophage (AM), following induction by different influenza virus strains (ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2)). Macrophages from other tissues or cell line exert alternative responding pattern, and AM is necessary for investigating the respiratory system. AM polarized toward the M1 phenotype after 4 hours of infection by all three virus strains, and AM to presented M2b phenotype after 8 hours induction, and immunosuppressive phenotype after 24 hours of induction. Protein expression assay showed similar results as the gene expression analysis for phenotype verification. The ELISA assay showed that TNF-α secretion was up-regulated after 4 and 8 hours of infection by influenza viruses, and it returned to basal levels after 24 hours of infection. IL-10 expression was elevated after 8 and 24 hours of infection. Immunofluorescence showed that iNOS expression was up-regulated but not Arg1 expression. Influenza virus notably increased phospho-Akt but not phospho-Erk1/2 or phospho-p38, and the AM polarization pattern have been changed by LY294002 (PI3K inhibitor). In conclusion, our results demonstrate the dynamic polarization of AM induced by influenza viruses, and suggested that PI3K/Akt signaling pathway modulates AM polarization to M1/M2b.

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Related in: MedlinePlus

Elisa assay to detect the TNF-α and IL-10 expression.Cytokines in the AM culture supernatant were measured at the indicated times after influenza viruses infection at 2 MOI. (A) show the expression of TNF-α, (B) show the expression of IL-10. (C) indicate the trendline of the two cytokines expression. Experiments were performed in triplicate and data are expressed as the mean ± SEM, *p<0.05, **p<0.01. Three independent experiments have been processed.
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pone-0104506-g005: Elisa assay to detect the TNF-α and IL-10 expression.Cytokines in the AM culture supernatant were measured at the indicated times after influenza viruses infection at 2 MOI. (A) show the expression of TNF-α, (B) show the expression of IL-10. (C) indicate the trendline of the two cytokines expression. Experiments were performed in triplicate and data are expressed as the mean ± SEM, *p<0.05, **p<0.01. Three independent experiments have been processed.

Mentions: ELISA analysis showed that TNF-α and IL-10 have a similar change to gene expression. Figure 5 indicated that after 4 hours of induction, the virus infection caused dramatically elevated TNF-α expression compared with the control (P<0.01). The induction by ST169 (H1N1) is stronger, while ST602 (H3N2) and HKG9 (H9N2) have a weaker induction. After 8 hours of induction, the viruses induced an up-regulation in TNF-α expression compared with the control, and there was no difference between influenza virus strains. Similar to the results of the gene expression, TNF-α expression was suppressed after 24 hours of induction by the virus.


PI3K/Akt signaling pathway modulates influenza virus induced mouse alveolar macrophage polarization to M1/M2b.

Zhao X, Dai J, Xiao X, Wu L, Zeng J, Sheng J, Su J, Chen X, Wang G, Li K - PLoS ONE (2014)

Elisa assay to detect the TNF-α and IL-10 expression.Cytokines in the AM culture supernatant were measured at the indicated times after influenza viruses infection at 2 MOI. (A) show the expression of TNF-α, (B) show the expression of IL-10. (C) indicate the trendline of the two cytokines expression. Experiments were performed in triplicate and data are expressed as the mean ± SEM, *p<0.05, **p<0.01. Three independent experiments have been processed.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126709&req=5

pone-0104506-g005: Elisa assay to detect the TNF-α and IL-10 expression.Cytokines in the AM culture supernatant were measured at the indicated times after influenza viruses infection at 2 MOI. (A) show the expression of TNF-α, (B) show the expression of IL-10. (C) indicate the trendline of the two cytokines expression. Experiments were performed in triplicate and data are expressed as the mean ± SEM, *p<0.05, **p<0.01. Three independent experiments have been processed.
Mentions: ELISA analysis showed that TNF-α and IL-10 have a similar change to gene expression. Figure 5 indicated that after 4 hours of induction, the virus infection caused dramatically elevated TNF-α expression compared with the control (P<0.01). The induction by ST169 (H1N1) is stronger, while ST602 (H3N2) and HKG9 (H9N2) have a weaker induction. After 8 hours of induction, the viruses induced an up-regulation in TNF-α expression compared with the control, and there was no difference between influenza virus strains. Similar to the results of the gene expression, TNF-α expression was suppressed after 24 hours of induction by the virus.

Bottom Line: Protein expression assay showed similar results as the gene expression analysis for phenotype verification.Immunofluorescence showed that iNOS expression was up-regulated but not Arg1 expression.In conclusion, our results demonstrate the dynamic polarization of AM induced by influenza viruses, and suggested that PI3K/Akt signaling pathway modulates AM polarization to M1/M2b.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, Shantou University Medical College, College, Shantou, Guangdong, China.

ABSTRACT
Macrophages polarized to M1 (pro-inflammation) or M2 (anti-inflammation) phenotypes in response to environmental signals. In this study, we examined the polarization of alveolar macrophage (AM), following induction by different influenza virus strains (ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2)). Macrophages from other tissues or cell line exert alternative responding pattern, and AM is necessary for investigating the respiratory system. AM polarized toward the M1 phenotype after 4 hours of infection by all three virus strains, and AM to presented M2b phenotype after 8 hours induction, and immunosuppressive phenotype after 24 hours of induction. Protein expression assay showed similar results as the gene expression analysis for phenotype verification. The ELISA assay showed that TNF-α secretion was up-regulated after 4 and 8 hours of infection by influenza viruses, and it returned to basal levels after 24 hours of infection. IL-10 expression was elevated after 8 and 24 hours of infection. Immunofluorescence showed that iNOS expression was up-regulated but not Arg1 expression. Influenza virus notably increased phospho-Akt but not phospho-Erk1/2 or phospho-p38, and the AM polarization pattern have been changed by LY294002 (PI3K inhibitor). In conclusion, our results demonstrate the dynamic polarization of AM induced by influenza viruses, and suggested that PI3K/Akt signaling pathway modulates AM polarization to M1/M2b.

Show MeSH
Related in: MedlinePlus