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.
Highlights
Microvascular endothelial cells (ECs) form a continuous monolayer lining the inner face of vascular walls throughout the body and play crucial roles in the regulation of vascular functions [1,2]
Immortalization, and characterization of six microvascular and one macrovascular bovine endothelial cell lines, that were immortalized in the same way as previous human, murine, and feline ECs, their successful infection by two already mentioned viruses, BTV and FMDV, and their ability to produce viral proteins upon DNA transfection
Our work shows that the six microvascular endothelial cell lines tested allow BTV replication, even if the cells derived from mesenteric lymph nodes and umbilical cord seem the most susceptible to infection
Summary
Microvascular endothelial cells (ECs) form a continuous monolayer lining the inner face of vascular walls throughout the body and play crucial roles in the regulation of vascular functions (homeostasis, transport of nutrients or hormones, leukocyte adhesion and trafficking across ECs and angiogenesis) [1,2]. ECs are not always the main target cells of pathogens but might represent the site of initial replication after vector-borne inoculation, and before infection of blood cells. They could constitute a reservoir for the pathogen during persistent infection by playing the role of niche cells, protecting the pathogen from the immune system [6]
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