Pseudo-infectious reporter virus particles for measuring antibody-mediated neutralization and enhancement of dengue virus infection.
This chapter outlines methods for the production of dengue virus (DENV) reporter virus particles (RVPs) and their use in assays that measure antibody-mediated neutralization and enhancement of DENV infection. RVPs are pseudo-infectious virions produced by complementation of a self-replicating flavivirus replicon with the DENV structural genes in trans. RVPs harvested from transfected cells are capable of only a single round of infection and encapsidate replicon RNA that encodes a reporter gene used to enumerate infected cells. RVPs may be produced using the structural genes of different DENV serotypes, genotypes, and mutants by changing plasmids used for complementation. Further modifications are possible including generating RVPs with varying levels of uncleaved prM protein, which resemble either the immature or mature form of the virus. Neutralization potency is measured by incubating RVPs with serial dilutions of antibody, followed by infection of target cells that express DENV attachment factors. Enhancement of infection is measured similarly using Fc receptor-expressing cells capable of internalizing antibody-virus complexes.
Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Swati
S
Mukherjee
Theodore C
TC
Pierson
Kimberly A
KA
Dowd
D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D003715:Dengue / Q000276:immunology* / Q000821:virology*; D003716:Dengue Virus / Q000276:immunology*; D005816:Genetic Complementation Test; D006801:Humans; D009500:Neutralization Tests / Q000379:methods*; D012093:Replicon / Q000235:genetics; D019562:Viral Load; D014771:Virion / Q000276:immunology*
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral