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Model Systems to Investigate Neutrophil Adhesion and Chemotaxis

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Neutrophil Methods and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 412))

Abstract

Polymorphonuclear neutrophil (PMN) recruitment from the blood stream into surrounding tissues, followed by migration through the tissue with triggered release of oxidative enzymes or eventual clearance from the epithelial surface, involves a regulated series of events central to acute responses in host defense (1). Accumulations of large numbers of neutrophils within mucosal tissues are pathognomonic features of both acute and chronic inflammatory conditions including sepsis (2,3) and inflammatory bowel disease (4), but the precise signals governing neutrophil adhesion and transmigration remain to be fully characterized. Previous chapters examine methods employed for both neutrophil isolation and study of the mechanisms underlying regulation of PMN rolling behavior. Here, we describe in vitro experimental models for the examination of PMN adhesion to endothelial and epithelial monolayers as well as the characterization of signals influencing neutrophil migration, both along acellular matrices and across endothelial and epithelial monolayers, in the physiologically relevant directions. Studies employing these model systems allow further elucidation of the mechanisms governing PMN adhesion and transmigration.

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References

  1. Liu, Y., Shaw, S. K., Ma, S., Yang, L., Luscinskas, F. W., and Parkos, C. A. (2004) Regulation of leukocyte transmigration: cell surface interactions and signaling events. J. Immunol. 172, 7–13.

    PubMed  CAS  Google Scholar 

  2. Sherwood, E. and Toliver-Kinsky, T. (2004) Mechanisms of the inflammatory response. Best Pract. Clin. Anaesthesiol. 18, 385–405.

    Article  CAS  Google Scholar 

  3. Chandra, A., Enkhbaatar, P., Nakano, Y., Traber, L., and Traber, D. (2006) Sepsis: emerging role of nitric oxide and selectins. Clinics 61, 71–76.

    Article  PubMed  Google Scholar 

  4. Jaye, D. L. and Parkos, C. A. (2000) Neutrophil migration across intestinal epithelium. Ann. NY Acad. Sci. 915, 151–161.

    Article  PubMed  CAS  Google Scholar 

  5. Lewis, J., Lee, J., Underwood, J., Harris, A., and Lewis, C. (1999) Macrophage responses to hypoxia: relevance to disease mechanisms. J. Leukoc. Biol. 66, 889–900.

    PubMed  CAS  Google Scholar 

  6. Driscoll, K., Carter, J., Hassenbein, D., and Howard, B. (1997) Cytokines and particle-induced inflammatory cell recruitment. Environ. Health Perspect. 105(Suppl 5), 1159–1164.

    Article  PubMed  CAS  Google Scholar 

  7. Khair, O., Davies, R., and Devalia, J. (1996) Bacterial-induced release of inflammatory mediators by bronchial epithelial cells. Eur. Respir. J. 9, 1913–1922.

    Article  PubMed  CAS  Google Scholar 

  8. Mizgerd, J. (2002) Molecular mechanisms of neutrophil recruitment elicited by bacteria in the lungs. Semin. Immunol. 14, 123–132.

    Article  PubMed  CAS  Google Scholar 

  9. Patel, K., Cuvelier, S., and Wiehler, S. (2002) Selectins: critical mediators of leukocyte recruitment. Semin. Immunol. 14, 73–81.

    Article  PubMed  CAS  Google Scholar 

  10. Alon, R. and Feigelson, S. (2002) From rolling to arrest on blood vessels: leukocyte tap dancing on endothelial integrin ligands and chemokines at sub-second contacts. Semin. Immunol. 14, 93–104.

    Article  PubMed  CAS  Google Scholar 

  11. Luscinskas, F., Ma, S., Nusrat, A., Parkos, C., and Shaw, S. (2002) Leukocyte transendothelial migration: A junctional affair. Semin. Immunol. 14, 105–113.

    Article  PubMed  CAS  Google Scholar 

  12. Faurschou, M. and Borregaard, N. (2003) Neutrophil granules and secretory vesicles in inflammation. Microbe. Infect. 5, 1317–1327.

    Article  CAS  Google Scholar 

  13. Roos, D., van Bruggen, R., and Meischl, C. (2003) Oxidative killing of microbes by neutrophils. Microbe. Infect. 5, 1307–1315.

    Article  CAS  Google Scholar 

  14. Kobayashi, S., Voyich, J., and DeLeo, F. (2003) Regulation of the neutrophilmediated inflammatory response to infection. Microbe. Infect. 5, 1337–1344.

    Article  CAS  Google Scholar 

  15. Zen, K., Babbin, B. A., Liu, Y., Whelan, J. B., Nusrat, A., and Parkos, C. A. (2004) JAM-C is a component of desmosomes and a ligand for CD11b/CD18-mediated neutrophil transepithelial migration. Mol. Biol. Cell. 15, 3926–3937.

    Article  PubMed  CAS  Google Scholar 

  16. Louis, N. A., Hamilton, K. E., Kong, T., and Colgan, S. P. (2005) HIF-dependent induction of apical CD55 coordinates epithelial clearance of neutrophils. FASEB J. 19, 950–959.

    Article  PubMed  CAS  Google Scholar 

  17. Liu, Y., Buhring, H. J., Zen, K., et al. (2002) Signal regulatory protein (SIRPalpha), a cellular ligand for CD47, regulates neutrophil transmigration. J. Biol. Chem. 277, 10,028–10,036.

    Article  PubMed  CAS  Google Scholar 

  18. Ward, P. A, Cochrane, C. G., and Müller-Eberhard, H. J. (1965) The role of serum complement in chemotaxis of leukocytes in vitro. J. Exp. Med. 122, 327–346.

    Article  PubMed  CAS  Google Scholar 

  19. Struyf, S., Gouwy, M., Dillen, C., Proost, P., Opdenakker, G., and Van Damme, J. (2005) Chemokines synergize in the recruitment of circulating neutrophils into inflamed tissue. Eur. J. Immunol. 35, 1583–1591.

    Article  PubMed  CAS  Google Scholar 

  20. Blancquaert, A. B., Colgan, S. P., and Bruyninckx, W. J. (1989) Influence of technical parameters on the in vitro motility of equine neutrophils in the presence of streptococcal supernatant. Vet. Immunol. Immunopathol. 23, 85–101.

    Article  PubMed  CAS  Google Scholar 

  21. Boyden, S. (1962) The chemotactic effect of mixtures of antibody antigen on polymorphonuclear leucocytes. J. Exp. Med. 115, 453–466.

    Article  PubMed  CAS  Google Scholar 

  22. Ebrahimzadeh, P., Hogfors, C., and Braide, M. (2000) Neutrophil chemotaxis in moving gradients of fMLP. J. Leukoc. Biol. 67, 51–61.

    Google Scholar 

  23. Parkos, C. A., Colgan, S. P., Liang, A., et al. (1996) CD 47 mediates post-adhesive events required for neutrophil migration across polarized intestinal epithelia. J. Cell Biol. 132, 437–450.

    Article  PubMed  CAS  Google Scholar 

  24. Lawrence, D. W., Bruyninckx, W. J., Louis, N. A., et al. (2003) Antiadhesive role of apical decay-accelerating factor (CD55) in human neutrophil transmigration across mucosal epithelia. J. Exp. Med. 198, 999–1010.

    Article  PubMed  CAS  Google Scholar 

  25. Nelson, R. D., Quie, P. G., and Simmons, R. L. (1975) Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leukocytes and monocytes. J. Immunol. 115, 1650–1656.

    PubMed  CAS  Google Scholar 

  26. Cereijido, M., Robbins, E., Dolan, W., Rotunno, C., and Sabatini, D. (1978) Polarized monolayers formed by epithelial cells on a permeable and translucent support. J. Cell Biol. 77, 853–880.

    Article  PubMed  CAS  Google Scholar 

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© 2007 Humana Press Inc., Totowa, NJ

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Louis, N.A., Campbell, E., Colgan, S.P. (2007). Model Systems to Investigate Neutrophil Adhesion and Chemotaxis. In: Quinn, M.T., DeLeo, F.R., Bokoch, G.M. (eds) Neutrophil Methods and Protocols. Methods in Molecular Biology™, vol 412. Humana Press. https://doi.org/10.1007/978-1-59745-467-4_17

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  • DOI: https://doi.org/10.1007/978-1-59745-467-4_17

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-788-4

  • Online ISBN: 978-1-59745-467-4

  • eBook Packages: Springer Protocols

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