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Pseudo-infectious Reporter Virus Particles for Measuring Antibody-Mediated Neutralization and Enhancement of Dengue Virus Infection

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Dengue

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1138))

Abstract

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.

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Author information

Authors and Affiliations

  1. Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA

    Swati Mukherjee, Theodore C. Pierson & Kimberly A. Dowd

Authors
  1. Swati Mukherjee
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  2. Theodore C. Pierson
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  3. Kimberly A. Dowd
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Editors and Affiliations

  1. Georgetown University Medical Center Dept. of Microbiology and Immunology, Washington, District of Columbia, USA

    Radhakrishnan Padmanabhan

  2. Duke-NUS GMS Singapore Emerging Infectious Diseases, Singapore, Singapore

    Subhash G. Vasudevan

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Mukherjee, S., Pierson, T.C., Dowd, K.A. (2014). Pseudo-infectious Reporter Virus Particles for Measuring Antibody-Mediated Neutralization and Enhancement of Dengue Virus Infection. In: Padmanabhan, R., Vasudevan, S. (eds) Dengue. Methods in Molecular Biology, vol 1138. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0348-1_6

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  • DOI: https://doi.org/10.1007/978-1-4939-0348-1_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0347-4

  • Online ISBN: 978-1-4939-0348-1

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