Abstract
Haemophilus influenzae type b, a colonizer of the human respiratory tract, is capable of breaching the epithelial cellular barrier of this niche, as well as endothelial barriers of the vasculature to enter the blood system, prior to dissemination to other tissues including the brain (1). Besides typeable H. influenzae, acapsulate (nontypeable; NTHi) stains, especially those belonging to the clonal biogroup aegyptius, are also capable of causing disseminated infections (2). The specificity of the organism for human respiratory mucosa and the mechanisms that allow it to transmigrate across both epithelial and endothelial barriers are not fully understood. Bacteria may interact directly with host cells, or they may first interact with phagocytic cells and cross the barrier as passengers within phagocytes. In the former case, interactions with target cell receptors may lead to adhesion, followed by cell signaling events that allow either entry into the cells and transcytosis or opening of cell-cell junctions and paracytosis. Investigations at the cellular level that define the precise components involved in the interactions have been facilitated by the use of immortalized cell lines as in the case of epithelial cells or limited culture of primary human cells as in the case of endothelial cells. Although increasingly more endothelial cell lines are becoming available, human umbilical vein endothelial cells (HUVECs) remain a popular choice of primary endothelial cells and will be described here in detail.
References
Moxon E. R. and Ostrow P. T. (1977) Haemophilus influenzae meningitis in infant rats: role of bacteremia in pathogenesis of age-dependent inflammatory responses in cerebrospinal fluid. J. Infect. Dis. 135, 303–307.
Quinn F. D., Weyant R. S., Worley M. J., White E. H., Utt E. A., and Ades E. A. (1995) Human microvascular endothelial tissue culture cell model for studying pathogenesis of Brazilian purpuric fever. Infect. Immun. 63, 2317–2322.
Maruyama Y. (1963) The human endothelial cell in tissue culture. Z. Zellforsch. Mikrosk. Anat. 60, 69.
Fryer D. G., Birnbaum G., and Luttrell C. N. (1966) Human endothelium in cell culture. J. Atheroscler. Res. 6, 151.
Jaffe E. A., Nachman R. L., Becker, and Minick R. C. (1972) Culture of human endo thelial cells derived from human umbilical cord veins. Circulation 46II, 253.
Jaffe E. A., Hoyer L. W., and Nachman R. L. (1973) Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J. Clin. Invest. 52, 2757–2764.
Jaffe E. A, Nachman R. L., Becker C. G., and Minick C. R. (1973) Culture of human endothelial cells derived from umbilical veins. J. Clin. Invest. 52, 2745–2756.
Gimborne M. A., Cotran R. S., and Folkman J. (1974) Human vascular endothelial cells in culture. J. Cell Biol. 60, 673–684.
Bicknell R. (1996) Endothelial Cell Culture: Handbooks in Practical Animal Cell Biology, Cambridge University Press, pp. 1–5.
Booyse F. M., Sedlak B. J., and Rafelson M. E. (1975) Culture of arterial endothelial cells. Characterisation and growth of bovine aortic cells. Thromb. Diathes. Ahemorrh. 34, 825–839.
Weibel E. R. and Palade G. E. (1964) New cytoplasmic components in arterial endothelia. J. Cell Biol. 23, 101–112.
Scott P. A. E. and Bicknell R. (1993) The isolation and culture of microvascular endothelium. J. Cell Sci. 105, 269–273.
Drevets D. A. and Campbell P. A. (1991) Macrophage phagocytosis: use of fluorescence microscopy to distinguish between extracellular and intracellular bacteria. J. Immunol. Meth. 142, 31–38.
McNeil G., Virji M., and Moxon E. R. (1994) Interactions of Neisseria meningitides with human monocytes. Microbial pathogen. 16, 153–163.
Virji M., Kayhay H., Ferguson D. J. P., Alexandresc C., and Moxon E. R. (1991) Interactions of Haemophilus influenzae with cultured human endothelial cells. Microbial Pathogen. 10, 231–245.
Moxon E. R., et al. (1994) Adaptive evolution of highly mutable loci in pathogenic bacteria. Curr. Biol. 4, 24–33.
Prasadarao N. V., Lysenko E., Wass C. A., Kim K. S., and Weiser J. N. (1999) Opacity-associated protein A contributes to the binding of Haemophilus influenzae to change epithelial cells. Infect. Immun. 67, 4153–4160.
Virji M. and Serino L. (2001) Molecular recognition mechanisms of meningococci: identifying receptor-ligand pairs involved in microbe-host interactions, in Meningococcal Disease, Methods in Molecular Medicine, vol. 66 (Pollard A. J. and Maiden M. C. J., eds.), Humana Press, Totowa, NJ.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Virji, M., Hill, D.J. (2003). In Vitro Models of Infection II—Human Umbilical Vein Endothelial Cells (HUVECs) System. In: Herbert, M.A., Hood, D.W., Moxon, E.R. (eds) Haemophilus influenzae Protocols. Methods in Molecular Medicine™, vol 71. Humana Press. https://doi.org/10.1385/1-59259-321-6:297
Download citation
DOI: https://doi.org/10.1385/1-59259-321-6:297
Publisher Name: Humana Press
Print ISBN: 978-0-89603-928-5
Online ISBN: 978-1-59259-321-7
eBook Packages: Springer Protocols