Abstract
Aim:
YCP, a novel (1,4)-α-D-glucan, was isolated from the mycelium of the marine filamentous fungus Phoma herbarum YS4108. In this work, we investigated a YCP-binding cellular receptor expressed by macrophages and the intracellular signal transduction pathways involved in YCP-induced macrophage activation.
Methods:
Fluorescence-labeled YCP (fl-YCP) was prepared using the CDAP-activation method. Fluorescence confocal laser microscopy and fluorescence-activated cell sorting (FACS) were used to analyze the effect of fl-YCP on macrophages. To characterize the properties of the YCP receptor, carbohydrates and antibodies were used to inhibit the binding of fl-YCP to macrophages. Moreover, we investigated the role of membrane receptors Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), Toll-like receptor 6 (TLR6) and complement receptor 3 (CR3). We also examined the role of the p38 kinase pathway in mediating nitric oxide (NO) production.
Results:
YCP had an in vitro stimulatory effect on the release of NO in macrophage, and fl-YCP can bind directly to receptors on the surface of macrophages in a time- and dose-dependent manner. Competition studies show that LPS, laminarin, anti-TLR4 antibody and anti-CD11b (CR3) antibody could inhibit fl-YCP binding to macrophages. Conversely, mannose, anti-TLR2 and anti-TLR6 antibody could not. Treatment of RAW264.7 cells with YCP resulted in significant activation of p38 in a time-dependent manner. The specific p38 inhibitor SB203580 abrogated YCP-induced NO generation. Treatment of RAW264.7 cells with anti-TLR4 antibody and anti-CR3 antibody significantly reduced YCP-induced NO production and p38 activation.
Conclusion:
We have demonstrated that YCP-induced NO production occurs through the TLR4 and CR3 membrane receptors in a p38 kinase-dependent manner in macrophages.
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References
Yang XB, Gao XD, Han F, Xu BS, Song YC, Tan RX . Purification, characterization and enzymatic degradation of YCP, a polysaccharide from marine filamentous fungus Phoma herbarum YS4108. Biochimie 2005; 87: 747–54.
Yao JF, Wu L, Wu XZ, Han F, Gao XD . Immuno-enhancing effect of YCP obtained from a marine filamentous fungus Keissleriella sp. YS 4108 on mice peritoneal macrophage. J China Pharm Univ 2004; 35: 573–5. Chinese.
Nair PKR, Melnick SJ, Ramachandran R, Escalon E, Ramachandran C . Mechanism of macrophage activation by (1,4)-α-D-glucan isolated from Tinospora cordifolia. Int Immunopharmacol 2006; 6: 1815–24.
Yun KJ, Kim JY, Kim JB, Lee KW, Jeong SY, Park HJ, et al. Inhibition of LPS-induced NO and PGE2 production by asiatic acid via NF-κB inactivation in RAW 264.7 macrophages: possible involvement of the IKK and MAPK pathways. Int Immunopharmacol 2008; 8: 431–41.
Yoon YD, Han SB, Kang JS, Lee CW, Park SK, Lee HS, et al. Toll-like receptor 4-dependent activation of macrophages by polysaccharide isolated from the radix of Platycodon grandiflorum. Int Immunopharmacol 2003; 3: 1873–82.
Jeon YJ, Han SB, Lee SH, Kim HC, Ahn KS, Kim HM . Activation of mitogen-activated protein kinase pathways by angelan in murine macrophages. Int Immunopharmacol 2001; 1: 237–45.
Yang XB, Gao XD, Han F, Tan RX . Sulfation of a polysaccharide produced by a marine filamentous fungus Phoma herbarum YS4108 alters its antioxidant properties in vitro. Biochim Biophys Acta 2005; 1725: 120–7.
Shafer DE, Toll B, Schuman RF, Nelson BL, Mond JJ, Lees A . Activation of soluble polysaccharides with 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) for use in protein-polysaccharide conjugate vaccines and immunological reagents. II. Selective crosslinking of proteins to CDAP-activated polysaccharides. Vaccine 2000; 18: 1273–81.
Bystrický S, Paulovicova E, Machova E . Candida albicans mannan-protein conjugate as vaccine candidate. Immunol Lett 2003; 85: 251–5.
Brown GD, Taylor PR, Reid DM, Willment JA, Williams DL, Pomares LM, et al. Dectin-1 is a major β-glucan receptor on macrophages. J Exp Med 2002; 196: 407–12.
Lee KY, You HJ, Jeong HG, Kang JS, Kim HM, Rhee SD, et al. Polysaccharide isolated from Poria cocos sclerotium induces NF-κB/Rel activation and iNOS expression through the activation of p38 kinase in murine macrophages. Int Immunopharmacol 2004; 4: 1029–38.
Yoon YD, Kang JS, Han SB, Park SK, Lee HS, Kang JS, et al. Activation of mitogen-activated protein kinases and AP-1 by polysaccharide isolated from the radix of Platycodon grandiflorum in RAW 264.7 cells. Int Immunopharmacol 2004; 4: 1477–87.
Jeon YJ, Han SB, Lee SH, Kim HC, Ahn KS, Kim HM . Activation of mitogen-activated protein kinase pathways by angelan in murine macrophages. Int Immunopharmacol 2001; 1: 237–45.
Wang YD, Yin DK, Zhu J, Zhang H, Zhao H, Gao XD . Screening of the types of immunocytes bound to YCP obtained from marine filamentous Phoma herbarum YS4108 in vitro. J China Pharm Univ 2008; 39: 463–7. Chinese.
Sodhi A, Tarang S, Kesherwani V . Concanavalin A induced expression of Toll-like receptors in murine peritoneal macrophages in vitro. Int Immunopharmacol 2007; 7: 454–63.
Xu WY, Wang L, Wang HM, Wang YQ, Liang YF, Zhao TT, et al. TLR2 and TLR4 agonists synergistically up-regulate SR-A in RAW264.7 through p38. Mol Immunol 2007; 44: 2315–23.
Han SB, Yoon YD, Ahn HJ, Lee HS, Lee CW, Yoon WK, et al. Toll-like receptor-mediated activation of B cells and macrophages by polysaccharide isolated from cell culture of Acanthopanax senticosus. Int Immunopharmacol 2003; 3: 1301–12.
Feng J, Zhao LH, Yu QQ . Receptor-mediated stimulatory effect of oligochitosan in macrophages. Biochem Biophys Res Commun 2004; 317: 414–20.
Dobrovolskaia MA, Vogel SN . Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect 2002; 4: 903–14.
Kerrigan AM, Brown GD . C-type lectins and phagocytosis. Immunobiology 2009. doi: 10.1016/j.imbio.2008.11.003.
Kim JY, Yoon YD, Ahn JM, Kang JS, Park SK, Lee K, et al. Angelan isolated from Angelica gigas Nakai induces dendritic cell maturation through toll-like receptor 4. Int Immunopharmacol 2007; 7: 78–87.
Hawlisch H, Kohl J . Complement and Toll-like receptors: Key regulators of adaptive immune responses. Mol Immunol 2006; 43: 13–21.
Lee MH, Lee JM, Jun SH, Lee SH, Kim NW, Lee JH, et al. The anti-inflammatory effects of Pyrolae herba extract through the inhibition of the expression of inducible nitric oxide synthase (iNOS) and NO production. J Ethnopharmacol 2007; 112: 49–54.
Zhu XY, Liu YJ, Liu SJ, Diao F, Xu RB, Ni X . Lipopolysaccharide primes macrophages to increase nitric oxide production in response to Staphylococcus Aureus. Immunol Lett 2007; 112: 75–81.
Hosoi T, Wada S, Suzuki S, Okuma Y, Akira S, Matsuda T, et al. Bacterial endotoxin induces IL-20 expression in the glial cells. Mol Brain Res 2004; 130: 23–9.
Sato F, Imaizumi T, Sashinami H, Yoshida H, Kusumi T, Mori F, et al. Upregulation of vascular endothelial growth factor by heat-killed Listeria monocytogenes in macrophages. Biochem Biophys Res Commun 2007; 354: 608–12.
Kawahara KI, Biswas KK, Unoshimab M, Ito T, Kikuchi K, Morimoto Y, et al. CRP induces high-mobility group box-1 protein release through activation of p38 MAPK in macrophage RAW264.7 cells. Cardiovasc Pathol 2008; 17: 129–38.
Acknowledgements
This work was co-financed by the Ministry of Science and Technology (National '863' project 2006AA090302), the National Natural Science Foundation (No 30672479, 30873201), the Specialized Research Fund for the Doctoral Program of Higher Education (No 20060316001) and the National Found for Fostering Talents of Basic Science (No J0630858).
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Chen, S., Yin, Dk., Yao, Wb. et al. Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108. Acta Pharmacol Sin 30, 1008–1014 (2009). https://doi.org/10.1038/aps.2009.93
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DOI: https://doi.org/10.1038/aps.2009.93
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