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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

Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 8 hours.Results are expressed as a ratio to mock-inoculated cells after 8 hours induction. Influenza viruses promote M2b polarization of AM, mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by quantitative PCR. Set as 1 and indicated by the horizontal X axis, four duplication per gene was detected. ↑ =  mild upregulated (P<0.05), ↓ = mild downregulated (P<0.05); ↑↑ = dramatically upregulated (p<0.01), ↓↓ = dramatically downregulated (P<0.01), the significant fold change were numbered. Three independent experiments have been processed.
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pone-0104506-g003: Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 8 hours.Results are expressed as a ratio to mock-inoculated cells after 8 hours induction. Influenza viruses promote M2b polarization of AM, mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by quantitative PCR. Set as 1 and indicated by the horizontal X axis, four duplication per gene was detected. ↑ =  mild upregulated (P<0.05), ↓ = mild downregulated (P<0.05); ↑↑ = dramatically upregulated (p<0.01), ↓↓ = dramatically downregulated (P<0.01), the significant fold change were numbered. Three independent experiments have been processed.

Mentions: Figure 3 showed that, compared with the 8 hours control, three influenza virus strains induced AM to polarize toward the typical M2b phenotype. In this stage, all influenza viruses induced dramatically higher levels of TNF-α, STAT1 and iNOS, lower levels of IL-12 (M1 markers), and dramatically higher levels of the M2 marker IL-10. Our results showed that AM phenotypes were dynamically transferred from M1 to M2b after 8 hours induction. In addition, all three influenza virus strains induced TLR2 expression have also higher levels compared with the control.


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)

Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 8 hours.Results are expressed as a ratio to mock-inoculated cells after 8 hours induction. Influenza viruses promote M2b polarization of AM, mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by quantitative PCR. Set as 1 and indicated by the horizontal X axis, four duplication per gene was detected. ↑ =  mild upregulated (P<0.05), ↓ = mild downregulated (P<0.05); ↑↑ = dramatically upregulated (p<0.01), ↓↓ = dramatically downregulated (P<0.01), the significant fold change were numbered. Three independent experiments have been processed.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104506-g003: Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 8 hours.Results are expressed as a ratio to mock-inoculated cells after 8 hours induction. Influenza viruses promote M2b polarization of AM, mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by quantitative PCR. Set as 1 and indicated by the horizontal X axis, four duplication per gene was detected. ↑ =  mild upregulated (P<0.05), ↓ = mild downregulated (P<0.05); ↑↑ = dramatically upregulated (p<0.01), ↓↓ = dramatically downregulated (P<0.01), the significant fold change were numbered. Three independent experiments have been processed.
Mentions: Figure 3 showed that, compared with the 8 hours control, three influenza virus strains induced AM to polarize toward the typical M2b phenotype. In this stage, all influenza viruses induced dramatically higher levels of TNF-α, STAT1 and iNOS, lower levels of IL-12 (M1 markers), and dramatically higher levels of the M2 marker IL-10. Our results showed that AM phenotypes were dynamically transferred from M1 to M2b after 8 hours induction. In addition, all three influenza virus strains induced TLR2 expression have also higher levels compared with the control.

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