The Effect of Long-Term Betacoronavirus Infection on the Permeability of the Blood-Brain Barrier-In Vitro Model Studies.

The blood-brain barrier (BBB) is critical for central nervous system homeostasis, yet it is highly vulnerable to viral insults. While acute coronavirus infections are known to impair BBB integrity, the long-term impact of persistent infection on brain endothelial cells remains poorly understood. Using an in vitro BBB model, we examined the effects of a 12-week infection with the neurotropic murine coronavirus MHV-JHM. Structural and functional changes were assessed via fluorescein isothiocyanate (FITC)-dextran permeability assay, confocal imaging of mitochondria, actin cytoskeleton, reactive oxygen species (ROS), scanning electron microscopy (SEM) and RT-qPCR for viral RNA level. Long-term infection induced progressive mitochondrial fragmentation and sustained ROS overproduction. Permeability to 70 kDa dextran increased significantly at 48 h post-infection and exceeded control levels threefold by 168 h. SEM revealed gradual endothelial surface roughening, blebbing, and eventual monolayer collapse with extensive intercellular gaps by week 12. Our study demonstrates that long-term MHV-JHM infection profoundly alters brain endothelial cell structure and function, triggering a cascade of changes that culminate in the disintegration of the BBB model.