title: High-throughput screening of viral entry inhibitors using pseudotyped virus.
abstract: Virus entry into a host cell is an attractive target for therapy because propagation of virus can be blocked at an early stage, minimizing chances for the virus to acquire drug resistance. Anti-infective drug discovery for BSL-4 viruses like Ebola or Lassa hemorrhagic fever virus presents challenges due to the requirement for a BSL-4 laboratory containment facility. Pseudotyped viruses provide a surrogate model in which the native envelope glycoprotein of a BSL-2 level virus (e.g., vesicular stomatitis virus) is replaced with envelope glycoprotein of a foreign BSL-4 virus (e.g., Ebola virus). Because the envelope glycoprotein determines interaction of virus with its cellular receptors, pseudotyped viruses can mimic the viral entry process of the original virus. Moreover, they are competent for only a single cycle of infection, and therefore can be used in BSL-2 facilities. Pseudotyped viruses have been used in high-throughput screening of entry inhibitors for a number of BSL-4 level viruses. This unit includes protocols for preparing pseudotyped viruses using lentiviral vectors and use of pseudotyped viruses for high-throughput screening of viral entry inhibitors.
authors.affiliation: Microbiotix, Worcester, Massachusetts, USA.
authors.forename: Arnab
authors.initials: A
authors.lastname: Basu
authors.forename: Debra M
authors.initials: DM
authors.lastname: Mills
authors.forename: Terry L
authors.initials: TL
authors.lastname: Bowlin
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D055808:Drug Discovery; D029043:Ebolavirus / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D016086:Lentivirus / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation; D053586:Virus Internalization / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D014759:Viral Envelope Proteins