Abstract: Equine herpesvirus type 8 (EHV-8) is currently mainly isolated from donkeys. In order to study its biological characteristics and pathogenicity, the EHV-8 LCDC01 strain isolated in our laboratory was infected with Rabbit kidney cell (RK-13) cells and BALB/c mice, and the changes of viral proliferation, histopathology and proinflammatory factors after infection of this strain to RK-13 cells and mice were observed. The results showed that the strain can replicate in RK-13 and Baby Hamster Syrian Kidney (BHK-21) cells, with typical cytopathic effects on RK-13 and BHK-21 cells, such as shrinkage, fusion grape bunch-like appearance and shedding, with a viral titer of 103.75 TCID50/mL. The mice showed no clinical manifestations except weight loss after virus infection, and the quantitative polymerase chain reaction (qPCR) detected viral replication only in the lungs of the mice. The lung index of mice was highest at 1st day after infection, and there was no significant difference between the virus infection group and the control group at the 5th day. The secretion of proinflammatory cytokines IL-1β, IL-6, IL-8 and IFN-α in the serum of mice was not significantly different from the control, but the secretion in the lung was significantly increased. The above results indicated that the EHV-8 LCDC01 strain isolate was able to infect mice and mainly replicated in the lung of mice. This study provides data support for elucidating the pathogenicity of EHV-8.