title: Laboratory preparation of Varicella-Zoster Virus: concentration of virus-containing supernatant, use of a debris fraction and magnetofection for consistent cell-free VZV infections.
abstract: The research laboratory generation of free Varicella-Zoster Virus (VZV) from cultured yields results relatively low titers, with the result that most study of VZV infection utilizes cell-associated infection. However, important aspects of VZV-cell interaction, such as the entry mechanism and superinfection exclusion have not yet been studied in detail, in part due to the difficulty in obtaining a high titer cell free virus. Here, a method to generate relatively high-titer cell-free VZV, based on a combination of previously published techniques and subsequent concentration is described. VZV-infected cells are disrupted, sonicated and clarified by centrifugation. The cell-free virus in the supernatant is then concentrated to yield up to 10(5)PFU/ml. The cell debris pellet, which contains up to 10(6)PFU/ml can also be used for non cell-associated infection. Magnetic nanoparticles available commercially can be used to further enhance infection by cell-free-VZV. The tools described here hold promise for better understanding of important aspects of VZV-cell interactions such as entry and latency.
authors.affiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel. Electronic address: anna.sloutskin@gmail.com.
authors.forename: Anna
authors.initials: A
authors.lastname: Sloutskin
authors.affiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
authors.forename: Ronald S
authors.initials: RS
authors.lastname: Goldstein
mesh_terms: D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification*; D014771:Virion / Q000235:genetics / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
keywords: Cell-free virus; Magnetic infection; Varicella-Zoster Virus