Bovine Organospecific Microvascular Endothelial Cell Lines as New and Relevant In Vitro Models to Study Viral Infections

Abstract

Microvascular endothelial cells constitute potential targets for exogenous microorganisms, in particular for vector-borne pathogens. Their phenotypic and functional variations according to the organs they are coming from provide an explanation of the organ selectivity expressed in vivo by pathogens. In order to make available relevant tools for in vitro studies of infection mechanisms, our aim was to immortalize bovine organospecific endothelial cells but also to assess their permissivity to viral infection. Using transfection with SV40 large T antigen, six bovine microvascular endothelial cell lines from various organs and one macrovascular cell line from an umbilical cord were established. They display their own panel of endothelial progenitor/mature markers, as assessed by flow cytometry and RT-qPCR, as well as the typical angiogenesis capacity. Using both Bluetongue and foot-and-mouth disease viruses, we demonstrate that some cell lines are preferentially infected. In addition, they can be transfected and are able to express viral proteins such as BTV8-NS3. Such microvascular endothelial cell lines bring innovative tools for in vitro studies of infection by viruses or bacteria, allowing for the study of host-pathogen interaction mechanisms with the actual in vivo target cells. They are also suitable for applications linked to microvascularization, such as anti-angiogenic and anti-tumor research, growing fields in veterinary medicine.

Endothelial Cells as Tools to Model Tissue Microenvironment in Hypoxia-Dependent Pathologies

“…The feline skin endothelial cells (FSkMEC) derived from the microvasculature and the feline umbilical cord endothelial cells (FUmEC) derived from the macrovasculature were used to demonstrate the cellular process of specific EC skin bacterial invasion by Bartonella Hansellae in cats and the species specific transmission to human [ 105 ]. The similar cell line establishment strategies in bovine were performed to study, in proper models, the organ and specific molecular mechanisms of viral infections [ 106 ].…”

Host-Specific Interplay between Foot-and-Mouth Disease Virus 3D Polymerase and the Type-I Interferon Pathway

“…BUcEC, endothelial primary cells derived from bovine umbilical cord, were maintained in improved minimum essential media (OptiMEM) + GlutaMAX™ (Gibco) supplemented with 10% foetal calf serum (FCS, Eurobio), 100 mg/mL penicillin–streptomycin (Gibco) and 100 μg/mL streptomycin and 0.8 μg/mL fungizone (Gibco, ) at 37 °C and 5% CO 2 [ 34 ].…”

Bovine Airway Models: Approaches for Investigating Bovine Respiratory Disease

“…However, this study only focused on the carcinogenic effects and the life span of endothelial cells, rather than the effects of infection by a virus or bacterium . Bovine endothelial cell work has predominately been conducted for understanding diseased cardiac tissue or reproductive tracts. ,− For example, pulmonary vascular endothelial cells were used to investigate the effects of LPS on vascular thrombosis after Pasteurella haemolytica infection. , Another study showed that P. haemolytica infection on confluent monolayers of pulmonary artery endothelial cells with neutrophils had a protective effect, which was exhibited by lower levels of LPS . Additionally, adenomyosis, a condition that can lead to uterine dysfunction, was investigated in endothelial cells sourced from bovine uteri .…”