, Volume 35, Issue 2, pp 409–419 | Cite as

Dynamic Mobility of Immunological Cells Expressing S100A8 and S100A9 in vivo: A Variety of Functional Roles of the two Proteins as Regulators in Acute Inflammatory Reaction

  • Akiko KoikeEmail author
  • Satoshi Arai
  • Sachiko Yamada
  • Akiko Nagae
  • Naoko Saita
  • Hiroshi Itoh
  • Shinji Uemoto
  • Masayuki Totani
  • Masaki Ikemoto


The immunological properties of rat S100A8 (r-S100A8) and S100A9 (r-S100A9) in immune cells are poorly understood. Enzyme-linked immunosorbent assay (ELISA) for r-S100A9 enabled us to discuss the differential functional roles of the two proteins, and their localization in the cells was observed microscopically. Recombinant human S100A8 (rh-S100A8) or S100A9 (rh-S100A9) were intravenously administrated into rats with LPS-induced liver damage. ELISA was used to measure the serum concentration of S100A9 in the rats. Western blotting and a preparative ELISA were used to prove specificity and avidity of monoclonal antibodies for r-S100A8 and r-S100A9. Immunohistochemical staining was carried out to visualize intracellular localization of the two proteins in the immune cells using the antibodies. When rh-S100A8 was intravenously injected in the rats (B group), the serum concentration of r-S100A9 apparently decreased as compared with that of the positive control rats (A group). The activities of AST, ALT, and LD in the rat sera (B group) also significantly went down in comparison with those of the rats (A group). Although both the S100A8 and S100A9 were abundantly expressed in activated immune cells, quite difference of not only their intracellular localization but also distribution of the cells expressing the two proteins was microscopically observed. In the rats (B group), less number of the immune cells or less amount of r-S100A8 and r-S100A9 in the cells than those of the rats (A group) was also seen. The r-S100A8 could serve as a regulator of acute inflammatory reaction in the rats with LPS-induced damage.


S100A8 S100A9 acute inflammation neutrophils macrophages lipopolysaccharide 


Rat S100A8


Rat S100A9


Recombinant human S100A8


Recombinant human S100A9







Horseradish peroxidase


Fluorescein 5-isothiocyanate


Polyacrylamide gel electrophoresis in the presence of SDS



We thank Dr Hiroshi Murayama (Yamasa Shoyu Co., Choshi, Chiba, Japan) for the gift of ELISA plates, and Dr Kyoichi Matsumoto (Mikuri Immunological Laboratory Co., Osaka, Japan) for his technical teaching in the preparation of monoclonal antibody. This work was supported by a Grant-in-Aid for Scientific Research (C: 20590567) from the Ministry of Education, Science, Sports and Culture of Japan (to M.I.)


  1. 1.
    Odink, K., N. Cerletti, J. Brüggen, R.G. Clerc, L. Tarcsay, G. Zwadlo, G. Gerhards, R. Schlegel, and C. Sorg. 1987. Two calcium-binding proteins in infiltrate macrophages of rheumatoid arthritis. Nature 330: 380–382.CrossRefGoogle Scholar
  2. 2.
    Rammes, A., J. Roth, M. Goebeler, M. Klempt, M. Hartmann, and C. Sorg. 1997. Myeloid-related protein (MRP) 8 and MRP14, calcium-binding proteins of the S100 family, are secreted by activated monocytes via novel, tubulin-dependent pathway. The Journal of Biological Chemistry 272: 9496–9502.PubMedCrossRefGoogle Scholar
  3. 3.
    Guignard, F., J. Mauel, and M. Markert. 1995. Identification and characterization of a novel human neutrophil protein related to the S100 family. The Biochemical Journal 309: 395–401.PubMedGoogle Scholar
  4. 4.
    Kerkhoff, C., M. Klempt, and C. Sorg. 1998. Novel insights into structure and function of MRP8 (S100A8) and MRP14 (S100A9). Biochimica et Biophysica Acta 1448: 200–211.PubMedCrossRefGoogle Scholar
  5. 5.
    Vogl, T., C. Pröpper, M. Hartmann, A. Strey, K. Strupat, C. van den Bos, C. Sorg, and J. Roth. 1999. S100A12 is expressed exclusively by granulocytes and acts independently from S100A8 and S100A9. The Journal of Biological Chemistry 274: 25291–25296.PubMedCrossRefGoogle Scholar
  6. 6.
    Nacken, W., C. Sopalla, C. Pröpper, C. Sorg, and C. Kerkhoff. 2000. Biochemical characterization of the murine S100A9 (MRP14) protein suggests that it is functionally equivalent to its human counterpart despite its low degree of sequence homology. European Journal of Biochemistry 267: 560–565.PubMedCrossRefGoogle Scholar
  7. 7.
    Nacken, W., J. Roth, C. Sorg, and C. Kerkhoff. 2003. S100A9/S100A8: myeloid representatives of the S100 protein family as prominent players in innate immunity. Microscopy Research and Technique 60: 569–580.PubMedCrossRefGoogle Scholar
  8. 8.
    Edgeworth, J., M. Gorman, R. Bennett, P. Freemont, and N. Hogg. 1991. Identification of p8, 14 as a highly abundant heterodimeric calcium binding protein complex of myeloid cells. The Journal of Biological Chemistry 266: 7706–7713.PubMedGoogle Scholar
  9. 9.
    Lim, S.Y., M.J. Raftery, J. Goyette, K. Hsu, and C.L. Geczy. 2009. Oxidative modifications of S100 proteins: functional regulation by redox. Journal of Leukocyte Biology 86: 577–587.PubMedCrossRefGoogle Scholar
  10. 10.
    Lim, S.Y., M.J. Raftery, H. Cai, K. Hsu, W.X. Yan, H.L. Hseih, R.N. Watts, D. Richardson, S. Thomas, M. Perry, and C.L. Geczy. 2008. S-Nitrosylated S100A8: Novel Anti-Inflammatory Properties. Journal of Immunology 181: 5627–5636.Google Scholar
  11. 11.
    Ikemoto, M., T. Tanaka, Y. Takai, H. Murayama, K. Tanaka, and M. Fujita. 2003. New ELISA system for myeloid-related protein complex (MRP8/14) and its clinical significance as a sensitive marker for inflammatory response associated with transplant rejection. Clinical Chemistry 49: 594–600.PubMedCrossRefGoogle Scholar
  12. 12.
    Ikemoto, M., H. Murayama, H. Itoh, M. Totani, and M. Fujita. 2007. Intrinsic function of S100A8/A9 complex as an anti-inflammatory protein in liver injury induced by lipopolysaccharide in rats. Clinica Chimica Acta 376: 197–204.CrossRefGoogle Scholar
  13. 13.
    Ryckman, C., K. Vandal, P. Rouleau, M. Talbot, and P.A. Tessier. 2003. Proinflammatory activities of S100: proteins S100A8, S100A9, and S100A8/A9 induce neutrophil chemotaxis and adhesion. Journal of Immunology 170: 3233–3242.Google Scholar
  14. 14.
    Ryckman, C., C. Gilbert, R. de Médicis, A. Lussier, K. Vandal, and P.A. Tessier. 2004. Monosodium urate monohydrate crystals induce the release of the proinflammatory protein S100A8/A9 from neutrophils. Journal of Leukocyte Biology 76: 433–440.PubMedCrossRefGoogle Scholar
  15. 15.
    Qing, G., L.C. Ma, A. Khorchid, G.V. Swapna, T.K. Mal, M. Takayama, B. Xia, S. Phadtare, H. ke, T. Acton, G.T. Montelione, M. Ikura, and M. Lnouyeet. 2004. Cold-shock induced high-yield protein production in Escherichia coli. Nature Biotechnology 22: 877–882.PubMedCrossRefGoogle Scholar
  16. 16.
    Namura, T., S. Arai, K. Okawa, A. Koike, S. Yamada, N. Saita, A. Nagae, H. Itoh, M. Totani, S. Uemoto, and M. Ikemoto. 2010. Identification of serum proteins that bind with S100A8, S100A9 and S100A8/A9: clinical significance of using proteins for monitoring the postoperative condition of liver recipients. Clinica Chimica Acta 411: 1766–1773.CrossRefGoogle Scholar
  17. 17.
    Imamichi, T., I. Uchida, S.M. Wahl, and N. MaCartney-Fransis. 1993. Expression and cloning of migration inhibitory factor-related protein (MRP)8 and MRP14 in arthritis-susceptible rats. Biochemical and Biophysical Research Communications 194: 819–825.PubMedCrossRefGoogle Scholar
  18. 18.
    Lowry, O.H., N.J. Rosebrough, A.L. Faee, and R.J. Randall. 1951. Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193: 265–275.PubMedGoogle Scholar
  19. 19.
    Towbin, H., T. Staehelin, and J. Gordon. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76: 4350–4354.PubMedCrossRefGoogle Scholar
  20. 20.
    Zanetti, M., L.V. d'Uscio, T.E. Peterson, Z.A. Katusic, and T. O'Brien. 2005. Analysis of superoxide anion production in tissue. Methods in Molecular Medicine 108: 65–72.PubMedGoogle Scholar
  21. 21.
    Itou, H., M. Yao, I. Fujita, N. Watanabe, M. Suzuki, J. Nishihira, and I. Tanaka. 2002. The crystal structure of human MRP14 (S100A9), a Ca(2+)-dependent regulator protein in inflammatory process. Journal of Molecular Biology 316: 265–276.PubMedCrossRefGoogle Scholar
  22. 22.
    Viemann, D., A. Strey, A. Janning, K. Jurk, K. Klimmek, T. Vogl, K. Hirono, F. Ichida, D. Foell, B. Kehrel, V. Gerke, C. Sorg, and J. Roth. 2005. Myeloid-related proteins 8 and 14 induce a specific inflammatory response in human microvascular endothelial cells. Blood 105: 2955–2962.PubMedCrossRefGoogle Scholar
  23. 23.
    Vandal, K., P. Rouleau, A. Boivin, C. Rickman, M. Talbot, and P.A. Tessier. 2003. Blockade of S100A8 and S100A9 suppresses neutrophil migration in response to lipopolysaccharide. Journal of Immunology 171: 2602–2609.Google Scholar
  24. 24.
    Vogl, T., K. Tenbrock, S. Ludwig, N. Leukert, C. Ehrhardt, M.A. van Zoelen, W. Nacken, D. Foell, T. van der Poll, C. Sorg, and J. Roth. 2007. Mrp8 and Mrp14 are endogenous activators of Toll-like receptor 4, promoting lethal, endotoxin-induced shock. Nature Medicine 13: 1042–1049.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Akiko Koike
    • 1
    Email author
  • Satoshi Arai
    • 2
  • Sachiko Yamada
    • 1
  • Akiko Nagae
    • 1
  • Naoko Saita
    • 1
  • Hiroshi Itoh
    • 1
  • Shinji Uemoto
    • 3
  • Masayuki Totani
    • 4
  • Masaki Ikemoto
    • 1
  1. 1.Human Health Sciences, Graduate School of MedicineKyoto UniversitySakyo-kuJapan
  2. 2.Immunological Laboratory, Diagnostic Division, Yamasa Shoyu Co., LtdChoshiJapan
  3. 3.Department of Transplantation UnitKyoto University HospitalKyotoJapan
  4. 4.Graduate School of Human Life ScienceShowa Women’s UniversityTokyoJapan

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