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Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model
Published on: August 29, 2012
Feline immunodeficiency virus can productively infect cultured endothelial cells from cat brain microvessels
A M Steffan1, M E Lafon, J L Gendrault
1Intitut National de la Santé et de la Recherche Médicale U74, Institut de Virologie de la Faculté de Médecine, Strasbourg, France.
Abstract:
Feline immunodeficiency virus (FIV) provokes a disease in cats characterized by histopathological lesions similar to those observed in AIDS patients. In order to determine whether endothelial cells from brain microvessels are involved in the central nervous system disease to the same extent as macrophages and microglia, cells were isolated from healthy cat brains, cultured and infected in vitro with the FIV Villefranche IFFA 1/88 strain. The isolated cells displayed typical endothelial cell ultrastructural features and were characterized further by von Willebrand factor-labelling and the binding of specific lectins such as Ulex europaeus lectin on their membrane. They were also able to take up acetylated low density lipoproteins. Two weeks after infection, significant amounts of FIV p24 antigen were detected by indirect immunofluorescence in syncytia and single cells. Concomitantly, the same antigen could be detected in the culture medium of the infected cells by an ELISA technique. Numerous viral particles as well as different steps in the process of viral budding were observed under transmission electron microscopy. The synthesis of FIV p24 antigens still occurred in cells in which replication was blocked in the G2 phase with taxol. Our results suggest the possibility of a productive infection of brain microvascular endothelial cells by FIV in vivo, which could lead to important perturbations in the functions of the blood-brain barrier.
Insights
Feline immunodeficiency virus (FIV) infects brain microvascular endothelial cells in cats. This productive infection may disrupt the blood-brain barrier, similar to feline AIDS.
Area of Science:
- Neuroscience
- Virology
- Immunology
Background:
- Feline immunodeficiency virus (FIV) causes feline AIDS-like lesions.
- The role of brain microvascular endothelial cells in FIV-induced central nervous system disease is unclear.
Purpose of the Study:
- To investigate the susceptibility of feline brain microvascular endothelial cells to FIV infection in vitro.
- To determine if FIV can productively infect these cells and impact blood-brain barrier function.
Main Methods:
- Isolation and in vitro culture of feline brain microvascular endothelial cells.
- Infection with FIV (Villefranche IFFA 1/88 strain).
- Characterization of infected cells using ultrastructural analysis, immunofluorescence (FIV p24 antigen), ELISA, and transmission electron microscopy.
Main Results:
- Isolated cells exhibited endothelial cell features and specific markers.
- FIV p24 antigen was detected in infected cells and culture medium.
- Viral particles and budding were observed via electron microscopy.
- FIV antigen synthesis continued even when viral replication was inhibited.
Conclusions:
- Feline brain microvascular endothelial cells are susceptible to productive FIV infection in vitro.
- FIV infection of these cells may lead to blood-brain barrier dysfunction in cats.

