Exploitation of herpesviral transactivation allows quantitative reporter gene-based assessment of virus entry and neutralization

Herpesviral entry is a highly elaborated process requiring many proteins to act in precise conjunction. Neutralizing antibodies interfere with this process to abrogate viral infection. Based on promoter transactivation of a reporter gene we established a novel method to quantify herpesvirus entry an...

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Bibliographic Details
Published in:PloS one 2011-01, Vol.6 (1), p.e14532-e14532
Main Authors: Reinhard, Henrike, Le, Vu Thuy Khanh, Ohlin, Mats, Hengel, Hartmut, Trilling, Mirko
Format: Article
Language:English
Subjects:
Adherent cells
Animals
Antibodies, Viral
Antigen presentation
Antigen-Antibody Reactions
Artificial chromosomes
Cloning
Congenital diseases
Cytomegalovirus
Dendritic cells
E coli
Escherichia coli
Exploitation
Gene expression
Genes
Genes, Reporter
Genomes
Glycoproteins
Health aspects
Herpes simplex
Herpes simplex virus 1
Herpes viruses
Herpesvirus 1, Human - genetics
Herpesvirus 1, Human - immunology
Herpesvirus 1, Human - physiology
Humans
IE1 protein
Immune serum
Immunoglobulins
Infection
Infections
Influenza
Luciferase
Mice
Monoclonal antibodies
Muromegalovirus
Neutralization
Neutralization Tests
Phosphorylation
Proteins
Reporter gene
Transcription factors
Transcriptional Activation
Virology
Virology/Diagnosis
Virology/Host Invasion and Cell Entry
Virus diseases
Virus Internalization
Viruses
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Description
Summary:Herpesviral entry is a highly elaborated process requiring many proteins to act in precise conjunction. Neutralizing antibodies interfere with this process to abrogate viral infection. Based on promoter transactivation of a reporter gene we established a novel method to quantify herpesvirus entry and neutralization by antibodies. Following infection with mouse and human cytomegalovirus and Herpes simplex virus 1 we observed promoter transactivation resulting in substantial luciferase expression (>1000-fold). No induction was elicited by UV-inactivated viruses. The response was MOI-dependent and immunoblots confirmed a correlation between luciferase induction and pp72-IE1 expression. Monoclonal antibodies, immune sera and purified immunoglobulin preparations decreased virus-dependent luciferase induction dose-dependently, qualifying this approach as surrogate virus neutralization test. Besides the reduced hands-on time, this assay allows analysis of herpesvirus entry in semi-permissive and non-adherent cells, which were previously non-assessable but play significant roles in herpesvirus pathology.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0014532