Development of a cell-based reporter assay for detection of Human alphaherpesviruses

• Published in: Molecular and cellular probes 2022-04, Vol.62, p.101806-101806, Article 101806
• Main Authors: Feltrin, Clarissa, Oliveira Simões, Cláudia Maria, Marques Sincero, Thaís Cristine
• Format: Article
• Language: English
• Subjects: Acyclovir - pharmacology; Acyclovir - therapeutic use; Animals; Antiviral screening; Cell-based reporter assay; Chlorocebus aethiops; GFP; Herpes Simplex - drug therapy; Herpesvirus 1, Human - genetics; Herpesvirus 2, Human - genetics; HSV-1; HSV-2; Human alphaherpesvirus; Humans; Vero Cells
• ISSN: 0890-8508; 1096-1194
• DOI: 10.1016/j.mcp.2022.101806

Immunosuppressed patients can suffer from Human alphaherpesvirus (HSV) infection with fast evolution, severe atypical symptomatology, and often-fatal outcome. Thus, the development and validation of new methods in vitro and in vivo to promote an early diagnosis and effective treatment of these patients are crucial. Therefore, this work aimed to develop a cell-based reporter assay for the detection of HSV through the transfection of Vero cells with the ICP10 promoter from HSV-2 linked to the pZsGreen1-1 plasmid. The assay was evaluated on Vero cells infected with HSV-1 or HSV-2 and followed by treating them with anti-HSV agents (acyclovir, gallic acid, convallatoxin, and Uncaria sp. extract) or with no anti-HSV activity agents (Passiflora edulis extract and cardenolide derivatives). The GFP expression was increased by both HSV cellular infection, which was detected by flow cytometry and fluorescence microscopy. F2R Zsgreen1-1 cells infection with 200 and 600 PFU/mL of HSV-2 increased the fluorescence intensity, when compared to the controls, by approximately 30% and 60%, respectively. Infection with 100 and 600 PFU/mL of HSV-1 also increased the fluorescence intensity by approximately 20% and 35%, when compared to the controls, respectively. The F2R ZsGreen1-1 system revealed to be an efficient assay, which can be used for clinical diagnosis, antiviral resistance evaluation, HSV cycle studies, and new antiviral drug research. [Display omitted] •A cell-based reporter assay based on GFP expression was developed.•GFP expression was induced by HSV-1 and HSV-2 infection.•GFP expression was proportional to viral MOI used.•Cell-based reporter assay can be used for antiviral screening.