Virus entry and replication in the brain precedes blood-brain barrier disruption during intranasal alphavirus infection

- 2P microscopic demonstration of VEEV TC83 CNS entry into wild-type and IFIT1-/- mice via axons of the olfactory route.
- Viral dissemination within the CNS by 2 dpi was associated BBB disruption only in the olfactory bulb, regardless of viral loads.
- BBB permeability increased subsequent to interaction of CX3CR1+ and CCR2+ inflammatory monocytes within venules at 6 dpi.
- TC83 gains CNS access via anterograde axonal migration without directly altering BBB function.
- Mononuclear and endothelial cell interactions may underlie BBB disruption during alphavirus encephalitis.

Viral infections of the central nervous system (CNS) are often associated with blood-brain barrier (BBB) disruption, yet the impact of virus replication and immune cell recruitment on BBB integrity are incompletely understood. Using two-photon microscopy, we demonstrate that Venezuelan equine encephalitis virus (VEEV) strain TC83-GFP, a GFP expressing, attenuated strain with a G3A mutation within the 5′ UTR that is associated with increased sensitivity to type I interferons (IFNs), does not directly impact BBB permeability. Following intranasal infection of both wild-type and IFN-induced protein with tetratricopeptide repeats 1 (IFIT1)-deficient mice, which fail to block TC83-specific RNA translation, virus spreads to the olfactory bulb and cortex via migration along axonal tracts of neurons originating from the olfactory neuroepithelium. Global dissemination of virus in the CNS by 2 days post-infection (dpi) was associated with increased BBB permeability in the olfactory bulb, but not in the cortex or hindbrain, where permeability only increased after the recruitment of CX3CR1+ and CCR2+ mononuclear cells on 6 dpi, which corresponded with tight junction loss and claudin 5 redistribution. Importantly, despite higher levels of viral replication, similar results were obtained in IFIT1-deficient mice. These findings indicate that TC83 gains CNS access via anterograde axonal migration without directly altering BBB function and that mononuclear and endothelial cell interactions may underlie BBB disruption during alphavirus encephalitis.

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