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Sulfochitosan inhibits P-selectin-mediated HL-60 leukocyte adhesion under flow conditions

Abstract

Excessive trafficking of leukocytes can lead to serious tissue injury. Here, four regioselectively sulfated chitosans were assessed as inhibitors of HL-60 leukocyte binding to P-selectin, by investigating their effect on leukocyte adhesion to CHO cells expressing human P-selectin under static and flow conditions. The results show that the sulfochitosans exhibit inhibitory activity in this general order: heparin > N-sulfated/6-O-sulfated chitosan ≥ 3-O,6-O-sulfated chitosan > 6-O-sulfated chitosan N-sulfated chitosan. This suggests that the sulfation of the double site in chitosan is essential for efficient inhibition of P-selectin-mediated HL-60 leukocyte adhesion and that such sulfochitosans may have potential as therapeutic agents against inflammatory disease.

Abbreviations

AC1.2:

a non-blocking mAb against P-selectin

DMF:

dimethylformamide

FBS:

fetal bovine serum

HPLC:

high performance liquid chromatography

N-S-C:

N-sulfated chitosan

N,6-S-C:

N-sulfated/6-O-sulfated chitosan

P-Fc:

recombinant human P-selectin/Fc chimera protei

3,6-S-C:

3-O,6-Osulfated chitosan

6-S-C:

6-O-sulfated chitosan

9E1:

a leukocyte adhesion blocking IgG mAb against P-selectin

References

  1. Witko-Sarsat, V., Rieu, P., Descamps-Latscha, B., Lesavre, P. and Halbwachs-Mecarelli, L. Neutrophils: molecules, functions and pathophysiological aspects. Lab. Invest. 80 (2000) 617–653.

    PubMed  Article  CAS  Google Scholar 

  2. Albeda, S.M., Smith, C.W. and Ward, P.A. Adhesion molecules and inflammatory injury. FASEB J. 8 (1994) 504–512.

    Google Scholar 

  3. Lefer, A.M., Weyrich, A.S. and Buerke, M. Role of selectins, a new family of adhesion molecules, in ischaemia-reperfusion injury. Cardiovasc. Res. 28 (1994) 289–294.

    PubMed  Article  CAS  Google Scholar 

  4. Rossi, B. and Constantin, G. Anti-selectin therapy for the treatment of inflammatory diseases. Inflamm. Allergy Drug Targets 7 (2008) 85–93.

    PubMed  Article  CAS  Google Scholar 

  5. Etzioni, A. Adhesion molecules-Their role in health and disease. Pediatr. Res. 39 (1996) 191–198.

    PubMed  Article  CAS  Google Scholar 

  6. Springer, T.A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76 (1994) 301–314.

    PubMed  Article  CAS  Google Scholar 

  7. Sako, D., Chang, X.J., Barone, K.M., Vachino, G., White, H.M., Shaw, G., Veldman, G.M., Bean, K.M., Ahern, T.J., Furie, B., Cumming, D.A. and Larsen, G.R. Expression cloning of a functional glycoprotein ligand for P-selectin. Cell 75 (1993) 1179–1186.

    PubMed  Article  CAS  Google Scholar 

  8. McEver, R.P. and Cummings, R.D. Role of PSGL-1 binding to selectins in leukocyte recruitment. J. Clin. Invest. 100 (1997) S97–103.

    PubMed  Article  CAS  Google Scholar 

  9. Stoolman, L.M. Adhesion molecules controlling lymphocyte migration. Cell 56 (1989) 907–910.

    PubMed  Article  CAS  Google Scholar 

  10. Varki, A. Selectin ligands. Proc. Natl. Acad. Sci. USA 91 (1994) 7390–7397.

    PubMed  Article  CAS  Google Scholar 

  11. Nelson, R.M., Cecconi, O., Roberts, W.G., Aruffo, A., Linhardt, R.J. and Bevilacqua, M.P. Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation. Blood 82 (1993) 3253–3258.

    PubMed  CAS  Google Scholar 

  12. Wei, M., Tai, G., Gao, Y., Li, N., Huang, B., Zhou, Y., Hao, S. and Zeng, X. Modified heparin inhibits P-selectin-mediated cell adhesion of human colon carcinoma cells to immobilized platelets under dynamic flow conditions. J. Biol. Chem. 279 (2004) 29202–29210.

    PubMed  Article  CAS  Google Scholar 

  13. Wang, L., Brown, J.R., Varki, A. and Esko, J.D. Heparin’s antiinflammatory effects require glucosamine 6-O-sulfation and are mediated by blockade of L- and P-selectins. J. Clin. Invest. 110 (2002) 127–136.

    PubMed  CAS  Google Scholar 

  14. Wang, R., Huang, J., Wei, M. and Zeng, X. The synergy of 6-O-sulfation and N- or 3-O-sulfation of chitosan is required for efficient inhibition of Pselectin-mediated human melanoma A375 cell adhesion. Biosci. Biotechnol. Biochem. 74 (2010) 1697–1700.

    PubMed  Article  CAS  Google Scholar 

  15. Holme, K.R. and Perlin, A.S. Chitosan N-sulfate. A water-soluble polyelectrolyte. Carbohydr. Res. 302 (1997) 7–12.

    PubMed  Article  CAS  Google Scholar 

  16. Focher, B., Massoli, A., Torri, G., Gervasini, A. and Morazzoni, F. High molecular weight chitosan 6-O-sulfate. Synthesis, ESR and NMR characterization. Macromol. Chem. Phys. 187 (1986) 2609–2620.

    Article  CAS  Google Scholar 

  17. Baumann, H. and Faust, V. Concepts for improved regioselective placement of O-sulfo, N-sulfo, N-acetyl, and N-carboxymethyl groups in chitosan derivatives. Carbohydr. Res. 331 (2001) 43–57.

    PubMed  Article  CAS  Google Scholar 

  18. Gao, Y., Li, N., Fei, R., Chen, Z., Zheng, S. and Zeng, X. P-Selectinmediated acute inflammation can be blocked by chemically modified heparin, RO-heparin. Mol. Cells 19 (2005) 350–355.

    PubMed  CAS  Google Scholar 

  19. Shodai, T., Suzuki, J., Kudo, S., Itoh, S., Terada, M., Fujita, S., Shimazu, H. and Tsuji, T. Inhibition of P-selectin-mediated cell adhesion by a sulfated derivative of sialic acid. Biochem. Biophys. Res. Commun. 312 (2003) 787–793.

    PubMed  Article  CAS  Google Scholar 

  20. Harlan, J.M. and Winn, R.K. Leukocyte-endothelial interactions: Clinical trials of anti-adhesion therapy. Crit. Care Med. 30 (2002) S214–219.

    PubMed  Article  CAS  Google Scholar 

  21. Fritzsche, J., Alban, S., Ludwig, R.J., Rubant, S., Boehncke, W.H., Schumacher, G. and Bendas, G. The influence of various structural parameters of semisynthetic sulfated polysaccharides on the P-selectin inhibitory capacity. Biochem. Pharmacol. 72 (2006) 474–485.

    PubMed  Article  CAS  Google Scholar 

  22. Kim, M.S., Sung, M.J., Seo, S.B., Yoo, S.J., Lim, W.K. and Kim, H.M. Water-soluble chitosan inhibits the production of pro-inflammatory cytokine in human astrocytoma cells activated by amyloid beta peptide and interleukin-1beta. Neurosci. Lett. 321 (2002) 105–109.

    PubMed  Article  CAS  Google Scholar 

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Correspondence to Xianlu Zeng or Min Wei.

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Huang, J., Wang, R., Liu, X. et al. Sulfochitosan inhibits P-selectin-mediated HL-60 leukocyte adhesion under flow conditions. Cell Mol Biol Lett 18, 200–208 (2013). https://doi.org/10.2478/s11658-013-0084-1

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  • DOI: https://doi.org/10.2478/s11658-013-0084-1

Key words

  • Chitosan
  • Sulfated chitosan
  • P-selectin
  • Inhibition
  • Cell adhesion
  • Flow condition
  • Heparin