In Vitro and In Vivo Blood-Brain Barrier Models to Study West Nile Virus Pathogenesis

Methods Mol Biol. 2016:1435:103-13. doi: 10.1007/978-1-4939-3670-0_9.


The blood-brain barrier (BBB), a specialized interface between the peripheral blood circulation and the central nervous system, specifically regulates molecular and cellular flux between the two. It plays a critical role in the maintenance of brain hemostasis. The BBB restricts the entry of pathogens into the brain, and thus its permeability is a critical factor that determines their central effects. Once the permeability of BBB is compromised, it has serious implications in the etiology of many brain pathologies including West Nile virus (WNV) disease. In this chapter, we describe protocols for preparation, maintenance, infection and permeability measurement of monolayer and bilayer in vitro BBB models to study WNV pathogenesis. We also describe Evans blue dye assay, a well-established method to test vascular permeability in vivo after WNV infection.

Keywords: Blood–brain barrier; Transmigration; West Nile virus encephalitis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / cytology
  • Astrocytes / pathology
  • Astrocytes / virology
  • Blood-Brain Barrier / cytology*
  • Blood-Brain Barrier / virology*
  • Capillary Permeability
  • Cell Line
  • Disease Models, Animal
  • Humans
  • In Vitro Techniques
  • Mice, Inbred C57BL
  • Tight Junctions / metabolism
  • Tight Junctions / virology
  • West Nile Fever / blood
  • West Nile Fever / pathology*
  • West Nile virus / pathogenicity*