Characterization of herpes simplex virus type 1 recombinants that express and incorporate high levels of HCV E2-gC chimeric proteins

• Published in: Virus research 2007, Vol.123 (1), p.40-49
• Main Authors: Kouvatsis, V., Argnani, R., Tsitoura, E., Arsenakis, M., Georgopoulou, U., Mavromara, P., Manservigi, R.
• Format: Article
• Language: English
• Subjects: Animals; Antigens, CD - metabolism; Cell Line; Cercopithecus aethiops; gC:E2 chimeric proteins; HCD81; HCV; Heparan sulphate; Hepatitis C virus; Herpes simplex virus 1; Herpesvirus 1, Human - physiology; HSV-1; Humans; Protein Structure, Tertiary; Reassortant Viruses - physiology; Receptors, Virus - metabolism; Recombinant Proteins - biosynthesis; Tetraspanin 28; Vero Cells; Viral Envelope Proteins - biosynthesis; Viral Envelope Proteins - chemistry; Viral Envelope Proteins - genetics; Virus Replication
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/j.virusres.2006.07.017

We report the construction of two HSV-1 recombinants encoding chimeric forms of the E2 glycoprotein of HCV-1a composed of the ectodomain of E2 (aa384–611 or 384–711) fused to different parts of the transmembrane and cytoplasmic domain of the HSV-1 gC glycoprotein (gC). The parental HSV-1, known as KgBpK −gC −, is deleted for gC and the main heparan sulphate (HS) binding domain of gB, and it exhibits impaired binding (ca. 80%) to HS compared to the wild type virus KOS [Laquerre, S., Argnani, R., Anderson, D.B., Zucchini, S., Manservigi, R., Glorioso, J.C., 1998. Heparan sulphate proteoglycan binding by herpes simplex virus type 1 glycoproteins B and C, which differ in their contributions to virus attachment, penetration, and cell-to-cell spread. J. Virol. 72, 6119–6130]. We show that gC:E2 proteins are efficiently expressed and transported to the cell surface. We also demonstrate that HSV-1 can incorporate both gC:E2 chimeric proteins into particles and show that incorporation of both chimeric molecules in the viral envelope partially restored binding (ca. 20%) of the HSV-1 recombinants to heparan sulphate. Finally, we showed that the gC:E2ScaI chimeric glycoprotein was able to bind a recombinant form of hCD81 and virion-expressed gC:E2ScaI permitted the binding of the HSV-1 recombinant virus to the hCD81 molecule.