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

Saguna Verma; Yeung Lo; Moti Chapagain; Stephanie Lum; Mukesh Kumar; Ulziijargal Gurjav; Haiyan Luo; Austin Nakatsuka; Vivek R. Nerurkar

doi:10.1016/j.virol.2008.11.047

Figure 3

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Immunoflourescence analysis of TJP in WNV-infected HBMVE cells

Confluent primary HBMVE cells were infected with WNV at MOI-5 and ZO-1 and claudin-1 expression, were determined using immunofluorescence. ZO-1 staining analyzed by epifluorescence microscopy at day 3 after infection was characterized by a staining pattern primarily confined to cell-cell borders and did not change as a result of WNV infection. Claudin-1 immunoreactivity analyzed by confocal microscopy was characterized by intracellular and junctional pattern in mock-infected HBMVE cells, while WNV-induced a significant increase in the overall immunoreactivity at days 3 and 4 after infection. The increase was evident in both, WNV-infected (inset, red) as well as neighboring un-infected HBMVE cells. The images shown are representative results of three independent experiments. Scale bar represents 10 μm at 63× magnification for ZO1 and 100 μm at 10× magnification for caludin-1.