West Nile virus-induced disruption of the blood–brain barrier in mice is characterized by the degradation of the junctional complex proteins and increase in multiple matrix metalloproteinases
West Nile virus (WNV) encephalitis is characterized by neuroinflammation, neuronal loss and blood–brain barrier (BBB) disruption. However, the mechanisms associated with the BBB disruption are unclear. Complex interactions between the tight junction proteins (TJP) and the adherens junction proteins (AJP) of the brain microvascular endothelial cells are responsible for maintaining the BBB integrity. Herein, we characterized the relationship between the BBB disruption and expression kinetics of key TJP, AJP and matrix metalloproteinases (MMPs) in the mice brain. A dramatic increase in the BBB permeability and extravasation of IgG was observed at later time points of the central nervous system (CNS) infection and did not precede virus–CNS entry. WNV-infected mice exhibited significant reduction in the protein levels of the TJP ZO-1, claudin-1, occludin and JAM-A, and AJP β-catenin and vascular endothelial cadherin, which correlated with increased levels of MMP-1, -3 and -9 and infiltrated leukocytes in the brain. Further, intracranial inoculation of WNV also demonstrated increased extravasation of IgG in the brain, suggesting the role of virus replication in the CNS in BBB disruption. These data suggest that altered expression of junction proteins is a pathological event associated with WNV infection and may explain the molecular basis of BBB disruption. We propose that WNV initially enters CNS without altering the BBB integrity and later virus replication in the brain initiates BBB disruption, allowing enhanced infiltration of immune cells and contribute to virus neuroinvasion via the ‘Trojan-horse’ route. These data further implicate roles of multiple MMPs in the BBB disruption and strategies to interrupt this process may influence the WNV disease outcome.
BovenL. A.,
MiddelJ.,
VerhoefJ.,
De GrootC. J.,
NottetH. S.2000; Monocyte infiltration is highly associated with loss of the tight junction protein zonula occludens in HIV-1-associated dementia. Neuropathol Appl Neurobiol 26:356–360 [View Article][PubMed]
CosbyS. L.,
BrankinB.1995; Measles virus infection of cerebral endothelial cells and effect on their adhesive properties. Vet Microbiol 44:135–139 [View Article][PubMed]
DallastaL. M.,
PisarovL. A.,
EsplenJ. E.,
WerleyJ. V.,
MosesA. V.,
NelsonJ. A.,
AchimC. L.1999; Blood-brain barrier tight junction disruption in human immunodeficiency virus-1 encephalitis. Am J Pathol 155:1915–1927 [View Article][PubMed]
DavisL. E.,
DeBiasiR.,
GoadeD. E.,
HaalandK. Y.,
HarringtonJ. A.,
HarnarJ. B.,
PergamS. A.,
KingM. K.,
DeMastersB. K.,
TylerK. L.2006; West Nile virus neuroinvasive disease. Ann Neurol 60:286–300 [View Article][PubMed]
GlassW. G.,
LimJ. K.,
CholeraR.,
PletnevA. G.,
GaoJ. L.,
MurphyP. M.2005; Chemokine receptor CCR5 promotes leukocyte trafficking to the brain and survival in West Nile virus infection. J Exp Med 202:1087–1098 [View Article][PubMed]
GralinskiL. E.,
AshleyS. L.,
DixonS. D.,
SpindlerK. R.2009; Mouse adenovirus type 1-induced breakdown of the blood-brain barrier. J Virol 83:9398–9410 [View Article][PubMed]
HayesE. B.,
GublerD. J.2006; West Nile virus: epidemiology and clinical features of an emerging epidemic in the United States. Annu Rev Med 57:181–194 [View Article][PubMed]
HuberJ. D.,
EgletonR. D.,
DavisT. P.2001; Molecular physiology and pathophysiology of tight junctions in the blood-brain barrier. Trends Neurosci 24:719–725 [View Article][PubMed]
KanlayaR.,
PattanakitsakulS. N.,
SinchaikulS.,
ChenS. T.,
ThongboonkerdV.2009; Alterations in actin cytoskeletal assembly and junctional protein complexes in human endothelial cells induced by dengue virus infection and mimicry of leukocyte transendothelial migration. J Proteome Res 8:2551–2562 [View Article][PubMed]
KleinR. S.,
LinE.,
ZhangB.,
LusterA. D.,
TollettJ.,
SamuelM. A.,
EngleM.,
DiamondM. S.2005; Neuronal CXCL10 directs CD8+ T-cell recruitment and control of West Nile virus encephalitis. J Virol 79:11457–11466 [View Article][PubMed]
LanciottiR. S.,
KerstA. J.,
NasciR. S.,
GodseyM. S.,
MitchellC. J.,
SavageH. M.,
KomarN.,
PanellaN. A.,
AllenB. C.other authors2000; Rapid detection of West Nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay. J Clin Microbiol 38:4066–4071[PubMed]
LimJ. K.,
ObaraC. J.,
RivollierA.,
PletnevA. G.,
KelsallB. L.,
MurphyP. M.2011; Chemokine receptor Ccr2 is critical for monocyte accumulation and survival in West Nile virus encephalitis. J Immunol 186:471–478 [View Article][PubMed]
LustigS.,
DanenbergH. D.,
KafriY.,
KobilerD.,
Ben-NathanD.1992; Viral neuroinvasion and encephalitis induced by lipopolysaccharide and its mediators. J Exp Med 176:707–712 [View Article][PubMed]
MishraM. K.,
DuttaK.,
SahebS. K.,
BasuA.2009; Understanding the molecular mechanism of blood-brain barrier damage in an experimental model of Japanese encephalitis: correlation with minocycline administration as a therapeutic agent. Neurochem Int 55:717–723 [View Article][PubMed]
MorreyJ. D.,
OlsenA. L.,
SiddharthanV.,
MotterN. E.,
WangH.,
TaroB. S.,
ChenD.,
RuffnerD.,
HallJ. O.2008; Increased blood-brain barrier permeability is not a primary determinant for lethality of West Nile virus infection in rodents. J Gen Virol 89:467–473 [View Article][PubMed]
Mun-BryceS.,
LukesA.,
WallaceJ.,
Lukes-MarxM.,
RosenbergG. A.2002; Stromelysin-1 and gelatinase A are upregulated before TNF-α in LPS-stimulated neuroinflammation. Brain Res 933:42–49 [View Article][PubMed]
SamuelM. A.,
DiamondM. S.2006; Pathogenesis of West Nile virus infection: a balance between virulence, innate and adaptive immunity, and viral evasion. J Virol 80:9349–9360 [View Article][PubMed]
SamuelM. A.,
WangH.,
SiddharthanV.,
MorreyJ. D.,
DiamondM. S.2007; Axonal transport mediates West Nile virus entry into the central nervous system and induces acute flaccid paralysis. Proc Natl Acad Sci U S A 104:17140–17145 [View Article][PubMed]
SchäferA.,
BrookeC. B.,
WhitmoreA. C.,
JohnstonR. E.2011; The role of the blood-brain barrier during Venezuelan equine encephalitis virus infection. J Virol 85:10682–10690 [View Article][PubMed]
WangP.,
DaiJ.,
BaiF.,
KongK. F.,
WongS. J.,
MontgomeryR. R.,
MadriJ. A.,
FikrigE.2008a; Matrix metalloproteinase 9 facilitates West Nile virus entry into the brain. J Virol 82:8978–8985 [View Article][PubMed]
WangS.,
WelteT.,
McGargillM.,
TownT.,
ThompsonJ.,
AndersonJ. F.,
FlavellR. A.,
FikrigE.,
HedrickS. M.,
WangT.2008b; Drak2 contributes to West Nile virus entry into the brain and lethal encephalitis. J Immunol 181:2084–2091[PubMed][CrossRef]
West Nile virus-induced disruption of the blood–brain barrier in mice is characterized by the degradation of the junctional complex proteins and increase in multiple matrix metalloproteinases