title: Use of vesicular stomatitis virus pseudotypes bearing hantaan or seoul virus envelope proteins in a rapid and safe neutralization test.
abstract: A vesicular stomatitis virus (VSV) pseudotype bearing hantavirus envelope glycoproteins was produced and used in a neutralization test as a substitute for native hantavirus. The recombinant VSV, in which the enveloped protein gene (G) was replaced by the green fluorescent protein gene and complemented with G protein expressed in trans (VSVDeltaG*G), was kindly provided by M. A. Whitt. 293T cells were transfected with plasmids for the expression of envelope glycoproteins (G1 and G2) of HTNV or SEOV and were then infected with VSVDeltaG*G. Pseudotype VSV with the Hantaan (VSVDeltaG*-HTN) or Seoul (VSVDeltaG*-SEO) envelope glycoproteins were harvested from the culture supernatant. The number of infectious units (IU) of the pseudotype VSVs ranged from 10(5) to 10(6)/ml. The infectivity of VSVDeltaG*-HTN and VSVDeltaG*-SEO was neutralized with monoclonal antibodies, immune rabbit sera, and sera from patients with hemorrhagic fever with renal syndrome, and the neutralizing titers were similar to those obtained with native hantaviruses. These results show that VSVDeltaG*-HTN and -SEO can be used as a rapid, specific, and safe neutralization test for detecting hantavirus-neutralizing antibodies as an effective substitute for the use of native hantaviruses. Furthermore, the IU of VSVDeltaG*-HTN and -SEO did not decrease by more than 10-fold when stored at 4 degrees C for up to 30 days. The stability of the pseudotype viruses allows distribution of the material to remote areas by using conventional cooling boxes for use as a diagnostic reagent.
authors.affiliation: Institute for Animal Experimentation, Hokkaido University Graduate School of Medicine, Sapporo 060-8638, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Ogino
authors.forename: Hideki
authors.initials: H
authors.lastname: Ebihara
authors.forename: Byoung-Hee
authors.initials: BH
authors.lastname: Lee
authors.forename: Koichi
authors.initials: K
authors.lastname: Araki
authors.forename: Ake
authors.initials: A
authors.lastname: Lundkvist
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawaoka
authors.forename: Kumiko
authors.initials: K
authors.lastname: Yoshimatsu
authors.forename: Jiro
authors.initials: J
authors.lastname: Arikawa
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D049452:Green Fluorescent Proteins; D018063:Hantaan virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006480:Hemorrhagic Fever with Renal Syndrome / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D008164:Luminescent Proteins / Q000235:genetics; D009500:Neutralization Tests / Q000379:methods*; D012328:RNA Viruses / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D029263:Seoul virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014162:Transfection; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007106:Immune Sera; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins