Replication and physical parameters important for preparing purified Junin virus.
Junin virus (JV) is an Arenavirus and the causative agent of Argentine hemorrhagic fever (AHF), an often fatal human disease. The attenuated strain XJ-clone 3 (XJC13) of JV, after being tested in humans, has been considered a promising vaccine. We found that synthesis of JV XJC13 reaches a peak 2 days after infection and the kinetics of synthesis are little affected by the multiplicity of infection (MOI) in a range from 0.125 to 1.00. Virus synthesis is sensitive to actinomycin D, indicating that cellular biosynthesis is required for viral replication. Combined precipitation and ultracentrifugation of supernatant from virus-infected cells yielded large amounts of concentrated and purified virion that banded in sucrose as a single peak with average density 1.177 +/- 0.015 g/ml. Purified virions have an average diameter of 203 +/- 23 nm by electron microscopy and an average sedimentation coefficient of 454 +/- 27 S. The results from the present study should assist in the preparation of large amounts of attenuated Junin virus which are required for vaccination against and diagnosis of Argentine hemorrhagic fever.
Bushar
G
G
Molecular Biology Branch, Food and Drug Administration, Rockville, MD 20852.
Sagripanti
J L
JL
D000818:Animals; D006477:Arenaviruses, New World / Q000302:isolation & purification* / Q000502:physiology / Q000648:ultrastructure; D003609:Dactinomycin / Q000494:pharmacology; D007700:Kinetics; D008854:Microscopy, Electron; D014461:Ultracentrifugation; D014709:Vero Cells; D014776:Virus Cultivation; D014779:Virus Replication / Q000187:drug effects
D003609:Dactinomycin