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Equid herpesvirus type 3 infection produces membrane-associated and secreted forms of glycoprotein G that are not required for efficient cell-to-cell spread of the virus in vitro

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Abstract

The ORF 70 gene of equid alphaherpesvirus type 3 (EHV-3) encodes glycoprotein G (gG), which is conserved in the majority of alphaherpesviruses. This glycoprotein is located in the viral envelope and has the characteristic of being secreted into the culture medium after proteolytic processing. It modulates the antiviral immune response of the host by interacting with chemokines. The aim of this study was to identify and characterize EHV-3 gG. By constructing viruses with HA-tagged gG, it was possible to detect gG in lysates of infected cells, their supernatants, and purified virions. A 100-, 60-, and 17-kDa form of the protein were detected in viral particles, while a 60-kDa form was identified in supernatants of infected cells. The role of EHV-3 gG in the viral infection cycle was assessed by the construction of a gG-minus EHV-3 mutant and its gG-positive revertant. When growth characteristics in an equine dermal fibroblast cell line were compared, the plaque size and the growth kinetics of the gG-minus mutant were similar to those of the revertant virus, suggesting that EHV-3 gG does not play a role in direct cell-to-cell transmission or virus proliferation of EHV-3 in tissue culture. The identification and characterization of EHV-3 gG described here provide a solid background for further studies to assess whether this glycoprotein has a function in modulating the host immune response.

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Data Availability

The data that support the findings of this study are available from the corresponding author, upon reasonable request.

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Acknowledgements

The authors thank the National Research Council (Conicet) and the University of Cuyo (UNCuyo) of Argentina for general support.

Funding

Financial support obtained from the Alberto J. Roemmers Foundation of Argentina made possible the completion of this work.

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Authors and Affiliations

  1. Instituto de Medicina y Biología Experimental de Cuyo IMBECU, CCT Mendoza, Área de Química Biológica, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, 5500, Mendoza, Argentina

    Antonella Losinno, Diego Sanchez & Armando Mario Damiani

  2. Instituto de Virología, CICVyA, INTA, Las Cabañas y Los Reseros s/n, 1712, Castelar, Argentina

    María Aldana Vissani

  3. Escuela de Veterinaria, Universidad del Salvador, Pilar, Buenos Aires, Argentina

    María Aldana Vissani

  4. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina

    Antonella Losinno, María Aldana Vissani, Diego Sanchez & Armando Mario Damiani

Authors
  1. Antonella Losinno
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  2. María Aldana Vissani
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  3. Diego Sanchez
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  4. Armando Mario Damiani
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, experiments, and analysis were performed by Antonella Losinno, Diego Sanchez, Maria Aldana Vissani, and Armando Mario Damiani. The first draft of the manuscript was written by Armando Mario Damiani, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Armando Mario Damiani.

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Handling Editor: Akbar Dastjerdi.

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Supplementary Information

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705_2023_5727_MOESM1_ESM.tif

Supplementary Fig. S1 Detection of EHV-3 gG products using a v_tagN-infected cell lysate (lane 1) and purified viral proteins from v_tagN (lanes 2 and 3) and v_tagC (lane 4). The upper panel shows detection of gG, and the lower panel shows β-actin detection in the v_tagN-infected cell lysate to rule out cell debris contamination in the v_tagN and v_tagC purified viral protein samples. (TIF 287 KB)

705_2023_5727_MOESM2_ESM.tif

Supplementary Fig. S2 Detection of 100- and 60-kDa forms of the EHV-3 gG protein in purified viral protein preparations (lanes 2, 3, and 4) and supernatants (lanes 5, 6, and 7) of infected cells. The upper panel shows detection of gG, and the lower panel shows β-actin detection in v_tagN-infected cell lysates (lane 1) to rule out cell debris contamination in v_tagN and v_tagC purified viral proteins and supernatant samples. (TIF 258 KB)

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Losinno, A., Vissani, M.A., Sanchez, D. et al. Equid herpesvirus type 3 infection produces membrane-associated and secreted forms of glycoprotein G that are not required for efficient cell-to-cell spread of the virus in vitro. Arch Virol 168, 122 (2023). https://doi.org/10.1007/s00705-023-05727-4

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