Escherichia coli Maltose-Binding Protein Induces M1 Polarity of RAW264.7 Macrophage Cells via a TLR2- and TLR4-Dependent Manner
Abstract
:1. Introduction
2. Results
2.1. Maltose-Binding Protein (MBP) Enhances Nitric Oxide (NO) and Inflammatory Cytokine Secretion in RAW264.7 Cells
2.2. MBP Promotes Pinocytic Activities of RAW264.7 Cells with no Effect on Cell Viability
2.3. MBP Upregulates Expression of Markers for Macrophage Activation and M1 Polarization in RAW264.7 Cells
2.4. MBP Activates TLR2 and TLR4 Expressions on RAW264.7 Cells
2.5. MBP Activates NF-κB and p38 MAPK Signaling Pathways via TLR2 and TLR4 to Induce M1 Polarity
3. Discussion
4. Experimental Section
4.1. Reagents and Antibodies
4.2. Cell Culture
4.3. NO Production
4.4. Cytokine Assay
4.5. Pinocytosis Assay
4.6. Cell Viability Assay
4.7. Flow Cytometric Analysis
4.8. Blocking of TLR2/4
4.9. Western Blot Analysis
4.10. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Boos, W.; Shuman, H. Maltose/maltodextrin system of Escherichia coli: Transport, metabolism, and regulation. Microbiol. Mol. Biol. Rev. 1998, 62, 204–229. [Google Scholar] [PubMed]
- Riggs, P. Expression and purification of recombinant proteins by fusion to maltose-binding protein. Mol. Biotechnol. 2000, 15, 51–63. [Google Scholar] [CrossRef] [PubMed]
- Fox, J.D.; Kapust, R.B.; Waugh, D.S. Single amino acid substitutions on the surface of Escherichia coli maltose-binding protein can have a profound impact on the solubility of fusion proteins. Protein Sci. 2001, 10, 622–630. [Google Scholar] [CrossRef] [PubMed]
- Witteveldt, J.; Vlak, J.M.; van Hulten, M.C. Protection of Panaeus monodon against white spot syndrome virus using a WSSV subunit vaccine. Fish Shellfish Immunol. 2004, 16, 571–579. [Google Scholar] [CrossRef] [PubMed]
- Kang, Q.Z.; Duan, G.C.; Fan, Q.T.; Xi, Y.L. Fusion expression of Helicobacter pylori neutrophil-activating protein in E. coli. World J. Gastroenterol. 2005, 11, 454–456. [Google Scholar] [CrossRef]
- Du, Y.; Hashizume, T.; Kurita-Ochiai, T.; Yuzawa, S.; Abiko, Y.; Yamamoto, M. Nasal Immunization with a fusion protein consisting of the hemagglutinin A antigenic region and the maltose-binding protein elicits CD11c+ CD8+ dendritic cells for induced long-term protective immunity. Infect. Immun. 2011, 79, 895–904. [Google Scholar] [CrossRef] [PubMed]
- Fernandez, S.; Palmer, D.R.; Simmons, M.; Sun, P.; Bisbing, J.; McClain, S.; Mani, S.; Burgess, T.; Gunther, V.; Sun, W. Potential role for Toll-like receptor 4 in mediating Escherichia coli maltose-binding protein activation of dendritic cells. Infect. Immun. 2007, 75, 1359–1363. [Google Scholar] [CrossRef]
- Zhang, Q.Y.; Ni, W.H.; Zhao, X.X.; Wang, F.L.; Gao, Z.; Tai, G.X. Synergistic antitumor effects of Escherichia coli maltose binding protein and Bacillus Calmette-Guerin in a mouse lung carcinoma model. Immunol. Lett. 2011, 136, 108–113. [Google Scholar] [CrossRef] [PubMed]
- Ni, W.H.; Zhang, Q.Y.; Liu, G.M.; Wang, F.L.; Yuan, H.Y.; Zhang, X.; Xie, F.; Li, Q.S.; Tai, G.X. Escherichia coli maltose-binding protein activates mouse peritoneal macrophages and induces M1 polarization via TLR2/4 in vivo and in vitro. Int. Immunopharmacol. 2014, 21, 171–180. [Google Scholar] [CrossRef] [PubMed]
- Zhao, X.X.; Ni, W.H.; Zhang, Q.Y.; Wang, F.L.; Li, Y.Y.; Sun, X.X.; Liu, Z.H.; Tai, G.X. Maltose-binding protein isolated from Escherichia coli induces Toll-like receptor 2-mediated viability in U937 cells. Clin. Transl. Oncol. 2011, 13, 509–518. [Google Scholar] [CrossRef] [PubMed]
- Kim, J.H.; Kim, D.H.; Baek, S.H.; Lee, H.J.; Kim, M.R.; Kwon, H.J.; Lee, C.H. Rengyolone inhibits inducible nitric oxide synthase expression and nitric oxide production by down-regulation of NF-κB and p38 MAP kinase activity in LPS-stimulated RAW264.7 cells. Biochem. Pharmacol. 2006, 71, 1198–1205. [Google Scholar] [CrossRef]
- Chiou, W.F.; Don, M.J.; Liao, J.F.; Wei, B.L. Psoralidin inhibits LPS-induced iNOS expression via repressing Syk-mediated activation of PI3K-IKK-IκB signaling pathways. Eur. J. Pharmacol. 2011, 650, 102–109. [Google Scholar] [CrossRef] [PubMed]
- Wang, S.Y.; Tai, G.X.; Zhang, P.Y.; Mu, D.P.; Zhang, X.J.; Liu, Z.H. Inhibitory effect of activin A on activation of lipopolysaccharide-stimulated mouse macrophage RAW264.7 cells. Cytokine 2008, 42, 85–91. [Google Scholar]
- Ge, J.Y.; Wang, Y.N.; Feng, Y.; Liu, H.Y.; Cui, X.L.; Chen, F.F.; Tai, G.X.; Liu, Z.H. Direct effects of activin A on the activation of mouse macrophage RAW264.7 cells. Cell. Mol. Immunol. 2009, 6, 129–133. [Google Scholar] [CrossRef]
- Stout, R.D.; Jiang, C.; Matta, B.; Tietzel, I.; Watkins, S.K.; Suttles, J. Macrophages sequentially change their functional phenotype in response to changes in microenvironment influences. J. Immunol. 2005, 175, 342–349. [Google Scholar] [CrossRef] [PubMed]
- Akira, S.; Hoshino, K. Myeloid differentiation factor 88-dependent and -independent pathways in Toll-like receptor signaling. J. Infect. Dis. 2003, 187, S356–S363. [Google Scholar] [CrossRef] [PubMed]
- Bjorkbacka, H.; Fitzgerald, K.A.; Huet, F.; Li, X.; Gregory, J.A.; Lee, M.A.; Ordija, C.M.; Dowley, N.E.; Golenbock, D.T.; Freeman, M.W. The induction of macrophage gene expression by LPS predominantly utilizes Myd88-independent signaling cascades. Physiol. Genomics 2004, 19, 319–330. [Google Scholar] [CrossRef] [PubMed]
- Laird, M.; Rhee, S.H.; Perkins, D.J.; Medvedev, A.E.; Piao, W.; Fenton, M.J.; Vogel, S.N. TLR4/MyD88/PI3K interactions regulate TLR4 signaling. J. Leukoc. Biol. 2009, 85, 966–977. [Google Scholar] [CrossRef] [PubMed]
- Duff, G.W.; Atkins, E. The inhibitory effect of polymyxin B on endotoxin-induced endogenous pyrogen production. J. Immunol. Methods 1982, 52, 333–340. [Google Scholar] [CrossRef] [PubMed]
- Lien, E.; Means, T.K.; Heine, H.; Yoshimura, A.; Kusumoto, S.; Fukase, K.; Fenton, M.J.; Oikawa, M.; Qureshi, N.; Monks, B.; et al. Toll-like receptor 4 imparts ligand-specific recognition of bacterial lipopolysaccharide. J. Clin. Investig. 2000, 105, 497–504. [Google Scholar]
- Glushkova, O.V.; Parfenyuk, S.B.; Khrenov, M.O.; Novoselova, T.V.; Lunin, S.M.; Fesenko, E.E.; Novoselova, E.G. Inhibitors of TLR-4, NF-κB, and SAPK/JNK signaling reduce the toxic effect of lipopolysaccharide on RAW 264.7 cells. J. Immunotoxicol. 2013, 10, 133–140. [Google Scholar] [CrossRef]
- Yu, H.; Ha, T.; Liu, L.; Wang, X.; Gao, M.; Kelley, J.; Kao, R.; Williams, D.; Li, C. Scavenger receptor A (SR-A) is required for LPS-induced TLR4 mediated NF-κB activation in macrophages. Biochim. Biophys. Acta 2012, 1823, 1192–1198. [Google Scholar] [CrossRef] [PubMed]
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Wang, W.; Yuan, H.-Y.; Liu, G.-M.; Ni, W.-H.; Wang, F.; Tai, G.-X. Escherichia coli Maltose-Binding Protein Induces M1 Polarity of RAW264.7 Macrophage Cells via a TLR2- and TLR4-Dependent Manner. Int. J. Mol. Sci. 2015, 16, 9896-9909. https://doi.org/10.3390/ijms16059896
Wang W, Yuan H-Y, Liu G-M, Ni W-H, Wang F, Tai G-X. Escherichia coli Maltose-Binding Protein Induces M1 Polarity of RAW264.7 Macrophage Cells via a TLR2- and TLR4-Dependent Manner. International Journal of Molecular Sciences. 2015; 16(5):9896-9909. https://doi.org/10.3390/ijms16059896
Chicago/Turabian StyleWang, Wan, Hong-Yan Yuan, Guo-Mu Liu, Wei-Hua Ni, Fang Wang, and Gui-Xiang Tai. 2015. "Escherichia coli Maltose-Binding Protein Induces M1 Polarity of RAW264.7 Macrophage Cells via a TLR2- and TLR4-Dependent Manner" International Journal of Molecular Sciences 16, no. 5: 9896-9909. https://doi.org/10.3390/ijms16059896