Retroviral Pseudotypes – From Scientific Tools to Clinical Utility

Abstract

Retroviral pseudotypes are important research and diagnostic tools for basic and clinical virology studies, facilitating the detailed investigation of individual genes, cellular receptors, antibody responses, serosurveillance and antiviral therapies. Importantly, pseudotypes enable the study of highly pathogenic viruses, without the need for high containment. Their use as gene therapy vectors is widely documented, but other uses, once less well known, are becoming more prominent. The substitution of envelope proteins expressed on the virion surface enables pseudotypes to be employed as surrogates for wildtype viruses in antibody neutralisation or antiviral screening assays and for the study of cell–virus receptor interactions. In addition, they are increasingly being utilised as vaccine immunogens, expressing the antigen either on the particle surface or as a transfer gene for cellular expression. These studies demonstrate the potential for using pseudotypes for both scientific and clinical applications.

Key Concepts

Students will learn:

  • What are retroviral pseudotypes.
  • How they are constructed and titrated.
  • The common elements that comprise a retroviral pseudotype.
  • How retroviral pseudotypes can be used to deliver genes.
  • How they can be used as tools for the study of viral attachment, entry and receptor identification.
  • About using pseudotypes as surrogate viruses in antibody neutralisation assays.
  • The use of pseudotypes to measure vaccine immunogenicity and screen for antiviral activity and resistance.
  • How pseudotypes can be used as experimental vaccines.

Keywords: lentiviral pseudotype; tropism; neutralisation assays; virus neutralisation; antibody neutralisation; virus envelope; serology; vaccines; antivirals; screening

Figure 1. Schematic of a triple‐plasmid co‐transfection system for generation of retrovirus pseudotypes, and use to measure antibody neutralisation. (a) Plasmid DNA expression vectors carrying the HIV gag‐pol ‘core’, envelope glycoprotein (env) and reporter genes (e.g. luciferase, GFP and β‐galactosidase) are transfected into producer cells (e.g. HEK293T). PRO indicates a (usually constitutive) gene promoter. LTR represents the retroviral long terminal repeats involved in vector integration into host cell DNA. (b) Pseudotype virus (PV) supernatants are harvested 48–72 h post‐transfection and relative PV titres are ascertained by measuring reporter expression in transduced target cells. (c) PVs can then be used in neutralisation assays (PNAs) to quantitate virus neutralising antibody (VNAb) responses in serum samples from individuals following natural infection or vaccination. The deletion and insertion of the packaging signal is indicated by the Ψ (psi) symbol. PRO, promoter; LTR, long terminal repeat.
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Further Reading

Centers for Disease Control and Prevention (2015) Diseases and Conditions. http://www.cdc.gov/DiseasesConditions/

Escors D, Breckpot K, Arce F, Kochan G and Stephenson H (2012) Lentiviral Vectors and Gene Therapy. Springer: Science.

Pöhlmann S and Simmonds G (2013) Viral Entry into Host Cells. Landes Bioscience (Austin, Texas) & Springer Science. New York: New York.

Temperton NJ and Wright E (2009) Retroviral Pseudotypes. Encyclopedia of Life Sciences. Chichester: John Wiley & Sons Ltd. DOI: 10.1002/9780470015902.a0021549.

Viral Pseudotype Unit (2015) http://www.viralpseudotypeunit.info

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Temperton, Nigel J, Wright, Edward, and Scott, Simon D(Jun 2015) Retroviral Pseudotypes – From Scientific Tools to Clinical Utility. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0021549.pub2]