Limits...
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.

Show MeSH

Related in: MedlinePlus

Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 4 hours.Results are expressed as a ratio to mock-inoculated cells after 4 hours induction. ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2) promote M1 polarization of AM. mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by qRT-PCR. All genes were normalized to GAPDH expression. 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
getmorefigures.php?uid=PMC4126709&req=5

pone-0104506-g002: Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 4 hours.Results are expressed as a ratio to mock-inoculated cells after 4 hours induction. ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2) promote M1 polarization of AM. mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by qRT-PCR. All genes were normalized to GAPDH expression. 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: Macrophages undergoing M1 or M2 polarization have characteristic profiles of marker expression. qRT-PCR test showed that after 4 hours of ST169 (H1N1) infection, compared with the 4 hours control, AM expressed dramatically higher levels of M1 markers, including TNF-α, MCP-1, iNOS, IL6 and IL-12. IL-10 (an M2 marker) was also up-regulated by 2.56-fold compared with the control. We infer that ST169 (H1N1) infection a M1 phenotype in the AM (Figure 2). ST602 (H3N2) induced a similar phonotype to ST169 (H1N1) in AM but with a slightly higher IL-10 expression (4.35-fold increased). Conversely, HKG9 (H9N2) induced dramatically higher levels of MCP-1 and IL-6, but lower levels of IL-10, which is compatible with a M1 phenotype. In addition, TLRs expression showed ST169 (H1N1) induced higher levels of TLR5 and TLR2 expression in AM, ST602 (H3N2) has the similar result, whereas HKG9 (H9N2) caused TLR5 expression up-regulation (Figure 2). When comparing the three influenza virus strains, we infer that the influenza virus induced AM to polarize to the M1 phenotype in the early stages (4 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)

Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 4 hours.Results are expressed as a ratio to mock-inoculated cells after 4 hours induction. ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2) promote M1 polarization of AM. mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by qRT-PCR. All genes were normalized to GAPDH expression. 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-g002: Gene expression of AM infected by influenza viruses at 2 MOI in vitro after 4 hours.Results are expressed as a ratio to mock-inoculated cells after 4 hours induction. ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2) promote M1 polarization of AM. mRNA levels of M1, M2 and Toll like receptors genes of AM was analyzed by qRT-PCR. All genes were normalized to GAPDH expression. 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: Macrophages undergoing M1 or M2 polarization have characteristic profiles of marker expression. qRT-PCR test showed that after 4 hours of ST169 (H1N1) infection, compared with the 4 hours control, AM expressed dramatically higher levels of M1 markers, including TNF-α, MCP-1, iNOS, IL6 and IL-12. IL-10 (an M2 marker) was also up-regulated by 2.56-fold compared with the control. We infer that ST169 (H1N1) infection a M1 phenotype in the AM (Figure 2). ST602 (H3N2) induced a similar phonotype to ST169 (H1N1) in AM but with a slightly higher IL-10 expression (4.35-fold increased). Conversely, HKG9 (H9N2) induced dramatically higher levels of MCP-1 and IL-6, but lower levels of IL-10, which is compatible with a M1 phenotype. In addition, TLRs expression showed ST169 (H1N1) induced higher levels of TLR5 and TLR2 expression in AM, ST602 (H3N2) has the similar result, whereas HKG9 (H9N2) caused TLR5 expression up-regulation (Figure 2). When comparing the three influenza virus strains, we infer that the influenza virus induced AM to polarize to the M1 phenotype in the early stages (4 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