Mouse Anti-Virus PRV Hybridoma [4H0G4]

Gold nanoclusters (Au NCs) are widely used in biological imaging and antitumor treatment because of their excellent cell membrane permeability, good fluorescence properties, and high biocompatibility. However, their effects on viruses are still largely unknown. Here, pseudorabies virus (PRV) and porcine reproductive and respiratory syndrome virus (PRRSV) were used respectively as the models of DNA virus and RNA virus to investigate the influences of glutathione-stabilized fluorescent Au NCs on viruses by plaque assay, indirect immunofluorescence assay, quantitative realtime polymerase chain reaction assay, and Western blot assay. The experimental data indicated that Au NCs selectively inhibited proliferation and protein expression of PRRSV but not that of PRV. Mechanistically, Au NCs directly inactivated PRRSV and blocked viral adsorption but showed no effect on viral genome replication. These findings prompt the possibility of developing Au NCs as an effective specific antiviral nanomaterial against RNA virus infection in the future.

This protocol describes a footpad inoculation model used to study the initiation and development of neuroinflammatory responses during alphaherpesvirus infection in mice. As alphaherpesviruses are main invaders of the peripheral nervous system (PNS), this model is suitable to characterize the kinetics of viral replication, its spread from the PNS to CNS, and associated neuroinflammatory responses. The footpad inoculation model allows virus particles to spread from a primary infection site in the footpad epidermis to sensory and sympathetic nerve fibers that innervate the epidermis, sweat glands, and dermis. The infection spreads via the sciatic nerve to the dorsal root ganglia (DRG) and ultimately through the spinal cord to the brain. Here, a mouse footpad is inoculated with pseudorabies virus (PRV), an alphaherpesvirus closely related to herpes simplex virus (HSV) and varicella-zoster virus (VZV). This model demonstrates that PRV infection induces severe inflammation, characterized by neutrophil infiltration in the footpad and DRG. High concentrations of inflammatory cytokines are subsequently detected in homogenized tissues by ELISA. In addition, a strong correlation is observed between PRV gene and protein expression (via qPCR and IF staining) in DRG and the production of pro-inflammatory cytokines. Therefore, the footpad inoculation model provides a better understanding of the processes underlying alphaherpesvirus-induced neuropathies and may lead to the development of innovative therapeutic strategies. In addition, the model can guide research on peripheral neuropathies, such as multiple sclerosis and associated viralinduced damage to the PNS. Ultimately, it can serve as a cost-effective in vivo tool for drug development.