An in vitro system to study trans-neuronal spread of pseudorabies virus infection

• Published in: Veterinary microbiology 2006-03, Vol.113 (3), p.193-197
• Main Authors: Ch’ng, T.H., Enquist, L.W.
• Format: Article
• Language: English
• Subjects: Animals; Aujeszky disease; Biological and medical sciences; cell culture; Cell Culture Techniques - methods; Cell Culture Techniques - veterinary; Cell–cell period; central nervous system; Compartmented chambers; cultured cells; disease course; epithelium; Fundamental and applied biological sciences. Psychology; Herpesvirus 1, Suid - metabolism; Herpesvirus 1, Suid - pathogenicity; Herpesvirus 1, Suid - physiology; in vitro studies; infection; latent period; microbial colonization; Microbiology; Microscopy, Confocal - veterinary; Miscellaneous; neurons; Neurons - ultrastructure; Neurons - virology; peripheral nervous system; Pseudorabies; Pseudorabies - pathology; Pseudorabies - virology; Pseudorabies virus; Rat PNS neurons; Rats; Suid herpesvirus 1; swine; viral fusion proteins; virion; Virion - ultrastructure; Virology; Virus Cultivation - methods; Virus Cultivation - veterinary; Virus Replication
• ISSN: 0378-1135; 1873-2542
• DOI: 10.1016/j.vetmic.2005.11.010

The neuronal spread of infection of alpha herpesviruses is controlled by unknown mechanisms. In the natural host, primary infection always leads to invasion of the peripheral nervous system, but rarely results in extensive invasion of the central nervous system. After reactivation of latent infection in the peripheral nervous system, virions are produced and shed from epithelial surfaces, but rarely invade the central nervous system. We have been studying two aspects of the general problem. First, using GFP and mRFP fusion proteins, we have used video confocal microscopy to assess mechanisms that influence spread of pseudorabies (PRV) virion components within axons. Second, and the subject of this report, is the development of a new in vitro cell culture system that enables the study of trans-neuronal spread of infection from neurons to non-neuronal cells similar to what happens after reactivation and spread to epithelial surfaces. We have developed a tissue culture system involving tri-chamber Teflon rings that enables facile analysis of trans-neuronal spread. The system duplicates all the known in vivo correlates of trans-neuronal spread and provides the opportunity to do both quantitative and qualitative assessment of spread of PRV infection from infected neurons to non-neuronal cells.