Study of Andes virus entry and neutralization using a pseudovirion system.
Andes virus (ANDV), a member of the Hantavirus genus in the family Bunyaviridae, causes an acute disease characteristic of New-World hantaviruses called hantavirus pulmonary syndrome (HPS). HPS is a highly pathogenic disease with a case-fatality rate of 40%. ANDV is the only hantavirus reported to spread directly from human-to-human. The aim of the present study was to develop a quantitative and high-throughput pseudovirion assay to study ANDV infection and neutralization in biosafety level 2 facilities (BSL-2). This pseudovirion assay is based on incorporation of ANDV glycoproteins onto replication-defective vesicular stomatitis virus (VSV) cores in which the gene for the surface G protein has been replaced by that encoding Renilla luciferase. Infection by the pseudovirions can be quantified by luciferase activity of infected cell lysates. ANDV pseudovirions were neutralized by ANDV-specific antisera, and there was good concordance between specificity and neutralization titers of ANDV hamster sera as determined by our pseudovirion assay and a commonly used plaque reduction neutralization titer (PRNT) assay. In addition, the pseudovirions were used to evaluate the requirements for ANDV entry, like pH dependency and the role of beta3 integrin, the reported receptor for other pathogenic hantaviruses, on entry.
Ray
Neelanjana
N
Department of Microbiology, University of Pennsylvania, Philadelphia, PA 19104, USA.
Whidby
Jillian
J
Stewart
Shaun
S
Hooper
Jay W
JW
Bertolotti-Ciarlet
Andrea
A
D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D017930:Genes, Reporter; D005822:Genetic Vectors; D006476:Orthohantavirus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D049410:Luciferases, Renilla / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D009500:Neutralization Tests / Q000379:methods*; D011817:Rabbits; D013223:Statistics as Topic; D018116:Vesiculovirus / Q000235:genetics*; D010948:Viral Plaque Assay; D015678:Viral Structural Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*; D053586:Virus Internalization
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D015678:Viral Structural Proteins; D049410:Luciferases, Renilla