title: West Nile virus infection modulates human brain microvascular endothelial cells tight junction proteins and cell adhesion molecules: Transmigration across the in vitro blood-brain barrier.
abstract: Neurological complications such as inflammation, failure of the blood-brain barrier (BBB), and neuronal death contribute to the mortality and morbidity associated with WNV-induced meningitis. Compromised BBB indicates the ability of the virus to gain entry into the CNS via the BBB, however, the underlying mechanisms, and the specific cell types associated with WNV-CNS trafficking are not well understood. Brain microvascular endothelial cells, the main component of the BBB, represent a barrier to virus dissemination into the CNS and could play key role in WNV spread via hematogenous route. To investigate WNV entry into the CNS, we infected primary human brain microvascular endothelial (HBMVE) cells with the neurovirulent strain of WNV (NY99) and examined WNV replication kinetics together with the changes in the expressions of key tight junction proteins (TJP) and cell adhesion molecules (CAM). WNV infection of HBMVE cells was productive as analyzed by plaque assay and qRT-PCR, and did not induce cytopathic effect. Increased mRNA and protein expressions of TJP (claudin-1) and CAM (vascular cell adhesion molecule and E-selectin) were observed at days 2 and 3 after infection, respectively, which coincided with the peak in WNV replication. Further, using an in vitro BBB model comprised of HBMVE cells, we demonstrate that cell-free WNV can cross the BBB, without compromising the BBB integrity. These data suggest that infection of HBMVE cells can facilitate entry of cell-free virus into the CNS without disturbing the BBB, and increased CAM may assist in the trafficking of WNV-infected immune cells into the CNS, via 'Trojan horse' mechanism, thereby contributing to WNV dissemination in the CNS and associated pathology.
authors.affiliation: Department of Tropical Medicine, Medical Microbiology and Pharmacology, Asia-Pacific Institute of Tropical Medicine and Infectious Diseases, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI 96813, USA.
authors.forename: Saguna
authors.initials: S
authors.lastname: Verma
authors.forename: Yeung
authors.initials: Y
authors.lastname: Lo
authors.forename: Moti
authors.initials: M
authors.lastname: Chapagain
authors.forename: Stephanie
authors.initials: S
authors.lastname: Lum
authors.forename: Mukesh
authors.initials: M
authors.lastname: Kumar
authors.forename: Ulziijargal
authors.initials: U
authors.lastname: Gurjav
authors.forename: Haiyan
authors.initials: H
authors.lastname: Luo
authors.forename: Austin
authors.initials: A
authors.lastname: Nakatsuka
authors.forename: Vivek R
authors.initials: VR
authors.lastname: Nerurkar
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000821:virology*; D001921:Brain / Q000098:blood supply / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D019108:Tight Junctions / Q000378:metabolism*; D014709:Vero Cells; D014901:West Nile Fever / Q000097:blood / Q000134:cerebrospinal fluid / Q000503:physiopathology*; D014902:West Nile virus / Q000276:immunology / Q000378:metabolism / Q000472:pathogenicity*
chemical_list: D015815:Cell Adhesion Molecules; D008565:Membrane Proteins