Home » Key Drug Discovery Articles » Human anti-varicella-zoster virus (VZV) recombinant monoclonal antibody produced after Zostavax immunization recognizes the gH/gL complex and neutralizes VZV infection

Human anti-varicella-zoster virus (VZV) recombinant monoclonal antibody produced after Zostavax immunization recognizes the gH/gL complex and neutralizes VZV infection

Significance Statement

Our paper features a method to analyze the immune response in humans following vaccination, which can easily be expanded to study any natural antibody response in humans. We identified a critical target for neutralizing antibodies on varicella zoster virus and highlight the need to include epitope conformation into antibody analysis. Our method also describes a reliable means to produce virtually unlimited amounts of specific and stable antibody to study virus pathogenesis and drug discovery.

 

Figure Legend

Human recombinant antibody (rec-RC IgG) recognizes a conform-depdentent antigen on the surface of varicella zoster virus (VZV) infected human lung fibroblast (a, red) and on the surface of human kidney (HEK 293) cells transfected with plasmid vectors expressing VZV glycoproteins gH, gL, gB and gE (b, green). Nuclear DNA is stained with DAPI (blue).

Human Anti Varicella-Zoster Virus (VZV) Recombinant Monoclonal Antibody Produced after Zostavax Immunization Recognizes the gH

Birlea M, Owens GP, Eshleman EM, Ritchie A, Traktinskiy I, Bos N, Seitz S, Azarkh Y, Mahalingam R, Gilden D, Cohrs RJ.

J Virol. 2013 Jan;87(1):415-21.

Department of Neurology, University of Colorado School of Medicine, Aurora, Colorado, USA.

 

Abstract

Varicella-zoster virus (VZV) is a ubiquitous, highly cell-associated, and exclusively human neurotropic alphaherpesvirus. Varicella-zoster virus infection is initiated by membrane fusion, an event dependent in part on Varicella-zoster virus glycoproteins gH and gL. Consistent with its location on the virus envelope, the gH/gL complexis a target of neutralizing antibodies produced after virus infection. One week after immunizing a 59-year-old VZV-seropositive man with Zostavax, we sorted his circulating blood plasma blasts and amplified expressed immunoglobulin variable domain sequences by single-cell PCR. Sequence analysis identified two plasma blast clones, one of which was used to construct a recombinant monoclonal antibody (rec-RC IgG). The rec-RC IgG colocalized with Varicella-zoster virus gE on the membranes of Varicella-zoster virus-infected cells and neutralized Varicella-zoster virus infection in tissue culture. Mass spectrometric analysis of proteins immunoprecipitated by rec-RC IgG identified both Varicella-zoster virus gH and gL. Transfection experiments showed that rec-RC IgG recognized a Varicella-zoster virus gH/gL protein complex but not individual  gH or gL proteins. Overall, our recombinant monoclonal anti-VZV antibody effectively neutralizes Varicella-zoster virus and recognizes a conformational epitope  within the Varicella-zoster virus gH/L protein complex. An unlimited supply of this antibody provides the opportunity to analyze membrane fusion events that follow virus attachment and to identify multiple epitopes on Varicella-zoster virus-specific proteins.

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