Studying latent Pseudorabies virus(PRV)infection has been limited by the lack of a suitable latent infection models.The aimed of this study is to establish an in vitro model for studying latent infection with PRV.Nerve cells were collected by isolating the dorsal root ganglion(DRG)of newborn mice,then indirect immunofluorescence(IFA)was used to identify that nerve cells in the DRG were successfully obtained.The nerve cells were infected by reporter recombinant virus(rPRV-EGFP),and Acyclovir(ACV)were used to inhibit viral replication to establish latent infection model.In addition,the conditions affecting latent infection,the multiplicity of infection(MOI)and the concentration of ACV were optimized.The results showed that with MOI 0.001 and 0.5mmol/L ACV,the PRV could stably establish latent infection on nerve cells.Then the latent infection was confirmed by genomic copies,virus titration and drug stimulation.The results showed that PRV could not replicate and no infectious virions were produced during latent infection in nerve cells.However,the PI3K inhibitor,LY294002,induced PRV reactivation and the production of large numbers of infectious virus particles.Taken together,this study established a PRV latent infection model in mouse primary nerve cells,which lays a foundation for further study on the latent infection of PRV.
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