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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
Sherwood, E. and Toliver-Kinsky, T. (2004) Mechanisms of the inflammatory response. Best Pract. Clin. Anaesthesiol. 18, 385–405.
Chandra, A., Enkhbaatar, P., Nakano, Y., Traber, L., and Traber, D. (2006) Sepsis: emerging role of nitric oxide and selectins. Clinics 61, 71–76.
Jaye, D. L. and Parkos, C. A. (2000) Neutrophil migration across intestinal epithelium. Ann. NY Acad. Sci. 915, 151–161.
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.
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.
Khair, O., Davies, R., and Devalia, J. (1996) Bacterial-induced release of inflammatory mediators by bronchial epithelial cells. Eur. Respir. J. 9, 1913–1922.
Mizgerd, J. (2002) Molecular mechanisms of neutrophil recruitment elicited by bacteria in the lungs. Semin. Immunol. 14, 123–132.
Patel, K., Cuvelier, S., and Wiehler, S. (2002) Selectins: critical mediators of leukocyte recruitment. Semin. Immunol. 14, 73–81.
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.
Luscinskas, F., Ma, S., Nusrat, A., Parkos, C., and Shaw, S. (2002) Leukocyte transendothelial migration: A junctional affair. Semin. Immunol. 14, 105–113.
Faurschou, M. and Borregaard, N. (2003) Neutrophil granules and secretory vesicles in inflammation. Microbe. Infect. 5, 1317–1327.
Roos, D., van Bruggen, R., and Meischl, C. (2003) Oxidative killing of microbes by neutrophils. Microbe. Infect. 5, 1307–1315.
Kobayashi, S., Voyich, J., and DeLeo, F. (2003) Regulation of the neutrophilmediated inflammatory response to infection. Microbe. Infect. 5, 1337–1344.
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.
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.
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.
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.
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.
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.
Boyden, S. (1962) The chemotactic effect of mixtures of antibody antigen on polymorphonuclear leucocytes. J. Exp. Med. 115, 453–466.
Ebrahimzadeh, P., Hogfors, C., and Braide, M. (2000) Neutrophil chemotaxis in moving gradients of fMLP. J. Leukoc. Biol. 67, 51–61.
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.
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.
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.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
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
Download citation
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