A novel bioluminescent herpes simplex virus 1 for in vivo monitoring of herpes simplex encephalitis

Abstract

Herpes simplex virus 1 (HSV-1) is responsible for herpes simplex virus encephalitis (HSE), associated with a 70% mortality rate in the absence of treatment. Despite intravenous treatment with acyclovir, mortality remains significant, highlighting the need for new anti-herpetic agents. Herein, we describe a novel neurovirulent recombinant HSV-1 (rHSV-1), expressing the fluorescent tdTomato and Gaussia luciferase (Gluc) enzyme, generated by the Clustered regularly interspaced short palindromic repeats (CRISPR)—CRISPR-associated protein 9 (Cas9) (CRISPR-Cas9) system. The Gluc activity measured in the cell culture supernatant was correlated (P = 0.0001) with infectious particles, allowing in vitro monitoring of viral replication kinetics. A significant correlation was also found between brain viral titers and Gluc activity in plasma (R2 = 0.8510, P < 0.0001) collected from BALB/c mice infected intranasally with rHSV-1. Furthermore, evaluation of valacyclovir (VACV) treatment of HSE could also be performed by analyzing Gluc activity in mouse plasma samples. Finally, it was also possible to study rHSV-1 dissemination and additionally to estimate brain viral titers by in vivo imaging system (IVIS). The new rHSV-1 with reporter proteins is not only as a powerful tool for in vitro and in vivo antiviral screening, but can also be used for studying different aspects of HSE pathogenesis.

Single-cell transcriptomics of the ventral posterolateral nucleus-enriched thalamic regions from HSV-1-infected mice reveal a novel microglia/microglia-like transcriptional response

“…To determine whether HSV-1 invades the CNS following intranasal infection, we imaged C57BL/6N mice on day 6 after intranasal infection with 6 × 10 5 PFUs of rHSV-1 expressing Gaussia Luciferase using an intravital imaging system (IVIS) [ 29 ]. We first noted an ongoing HSV-1 infection in the nasal cavity.…”

“…Six-week-old male C57BL/6 N mice were purchased from Charles River, Canada. Mice were slightly anesthetized and infected with clinical HSV-1 strain H25 or recombinant HSV-1 (rHSV-1) in 20 µl of the minimum essential medium by intranasal inoculation [ 29 ]. Similar amounts of viral suspension were equitably administered to the right and the left mouse nostrils.…”

In vivo imaging reveals novel replication sites of a highly oncogenic avian herpesvirus in chickens

“…and Methicillin-resistant S. aureus (MRSA); and Plasmodium berghei (malaria), leishmania, and toxoplasma, respectively. Despite this broad body of evidence, bioluminescence imaging has mainly focused on the analysis of infections in mice, ferrets and fish [1][2][3][4][5][6], while no study to our knowledge has been reported in birds.…”

Bioluminescence Production by Turnip Yellows Virus Infectious Clones: A New Way to Monitor Plant Virus Infection

“…For this reason, bioluminescence is mainly used for in vitro measurements; macroscopic and microscopic observations are less common. Luciferase reporter viruses have been designed mostly for animal and human viruses, for example, hepatitis C virus, [6], herpes simplex virus 1 [7], HIV-1 [8], or dengue virus [9]. Examples of luciferase-tagged plant viruses are scarce.…”