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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

AM was infected by influenza viruses at 2 MOI, and immunostained for iNOS and Arg1 at the indicated times.The nucleus was stained with Hoechst (blue). Exposure to negative control (virus culture medium) resulted in an iNOSlowArg1low phenotype at the three time pointes. Viruses infected AM induced iNOS (green) but not Arg1 (red) expression after 4 hours and 8 hours induction, after 24 hours induction, virus induced AM an iNOSlowArg1low phenotype similar to negative control.
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pone-0104506-g006: AM was infected by influenza viruses at 2 MOI, and immunostained for iNOS and Arg1 at the indicated times.The nucleus was stained with Hoechst (blue). Exposure to negative control (virus culture medium) resulted in an iNOSlowArg1low phenotype at the three time pointes. Viruses infected AM induced iNOS (green) but not Arg1 (red) expression after 4 hours and 8 hours induction, after 24 hours induction, virus induced AM an iNOSlowArg1low phenotype similar to negative control.

Mentions: Immunofluorescence detection showed that after both 4 and 8 hours of virus challenge, the expression of iNOS was elevated; iNOS expression returned to control levels by 24 hours after induction (Figure 6). Arg1 expression was not notably changed after virus infection. The positive control (Figure S2 in File S1) was processed following Shikah Arora's method [31]. Protein level of markers in influenza virus induced AM polarization consistent with gene expression results. TNF-αhigh iNOShigh IL-10low Arglow showed compatible with a M1 phenotype; whereas; TNF-αhigh iNOShigh IL-10high Arglow showed compatible with a M2b phenotype; and all four proteins expression showed down-regulation after 24 hours induction.


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)

AM was infected by influenza viruses at 2 MOI, and immunostained for iNOS and Arg1 at the indicated times.The nucleus was stained with Hoechst (blue). Exposure to negative control (virus culture medium) resulted in an iNOSlowArg1low phenotype at the three time pointes. Viruses infected AM induced iNOS (green) but not Arg1 (red) expression after 4 hours and 8 hours induction, after 24 hours induction, virus induced AM an iNOSlowArg1low phenotype similar to negative control.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104506-g006: AM was infected by influenza viruses at 2 MOI, and immunostained for iNOS and Arg1 at the indicated times.The nucleus was stained with Hoechst (blue). Exposure to negative control (virus culture medium) resulted in an iNOSlowArg1low phenotype at the three time pointes. Viruses infected AM induced iNOS (green) but not Arg1 (red) expression after 4 hours and 8 hours induction, after 24 hours induction, virus induced AM an iNOSlowArg1low phenotype similar to negative control.
Mentions: Immunofluorescence detection showed that after both 4 and 8 hours of virus challenge, the expression of iNOS was elevated; iNOS expression returned to control levels by 24 hours after induction (Figure 6). Arg1 expression was not notably changed after virus infection. The positive control (Figure S2 in File S1) was processed following Shikah Arora's method [31]. Protein level of markers in influenza virus induced AM polarization consistent with gene expression results. TNF-αhigh iNOShigh IL-10low Arglow showed compatible with a M1 phenotype; whereas; TNF-αhigh iNOShigh IL-10high Arglow showed compatible with a M2b phenotype; and all four proteins expression showed down-regulation after 24 hours induction.

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