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 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.
Institute for Virology, Heinrich-Heine-University, Düsseldorf, Germany.
Henrike
H
Reinhard
Vu Thuy Khanh
VT
Le
Mats
M
Ohlin
Hartmut
H
Hengel
Mirko
M
Trilling
D000818:Animals; D000914:Antibodies, Viral; D000937:Antigen-Antibody Reactions; D003587:Cytomegalovirus; D017930:Genes, Reporter; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D006801:Humans; D051379:Mice; D018146:Muromegalovirus; D009500:Neutralization Tests; D015533:Transcriptional Activation; D053586:Virus Internalization
D000914:Antibodies, Viral
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