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Abstract
Echovirus-30 (E-30) is a non-polio enterovirus responsible for meningitis outbreaks in children worldwide. To gain access to the central nervous system (CNS), E-30 first has to cross the blood-brain barrier (BBB) or the blood-cerebrospinal fluid barrier (BCSFB). E-30 may use lipid rafts of the host cells to interact with and to invade the BCSFB. To study enteroviral infection of the BCSFB, an established in vitro model based on human immortalized brain choroid plexus papilloma (HIBCPP) cells has been used. Here, we investigated the impact of E-30 infection on the protein content of the lipid rafts at the BCSFB in vitro. Mass spectrometry analysis following E-30 infection versus uninfected conditions revealed differential abundancy in proteins implicated in cellular adhesion, cytoskeleton remodeling, and endocytosis/vesicle budding. Further, we evaluated the blocking of endocytosis via clathrin/dynamin blocking and its consequences for E-30 induced barrier disruption. Interestingly, blocking of endocytosis had no impact on the capacity of E-30 to induce loss of barrier properties in HIBCPP cells. Altogether, these data highlight the impact of E-30 on HIBCPP cells microdomain as an important factor for host cell alteration.
Highlights
Enterovirus infection outbreaks occur every year worldwide especially in the Asian-Pacific region
We observed a drastic decrease in the transepithelial electrical resistance (TEER) of human immortalized brain choroid plexus papilloma (HIBCPP) cells infected with E-30 at MOI 20 for 24 h compared to the uninfected control (** p < 0.001) (Supplemental Figure S1b)
These data show that E-30 infection with an MOI 20 for 24 h is well suited to perform a proteomic assay on the protein composition of the lipid rafts of HIBCPP cells, since these infection conditions have a high impact on the HIBCPP cell properties, lead to a high number of infected cells and a high amount of viral particles, as well as a low cytotoxicity in HIBCPP cells
Summary
Enterovirus infection outbreaks occur every year worldwide especially in the Asian-Pacific region. Amongst the most common epidemies, Echovirus-30 (E-30) infection in children rises every year in high and especially in low-income countries, which re-enforces the economic difficulties of the southern region [1]. The BCSFB is located at the choroid plexuses in the brain and is composed of epithelial cells tightly sealed together, providing a high transepithelial electrical resistance (TEER) and a low permeability for macromolecules [8,9]. The only functional human in vitro model of the BCSFB is the cell layer formed by HIBCPP (Human Immortalized Choroid Plexus Papilloma) cells [16]. Due to its strong barrier properties such as high TEER and low permeability, HIBCPP cells are a solid model to study viral infection and its impact on the BCSFB [17,18]
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