Location: Arthropod-borne Animal Diseases Research
Project Number: 3020-32000-020-006-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Apr 3, 2023
End Date: Apr 2, 2025
Objective:
Although bluetongue virus and epizootic hemorrhagic disease virus are closely related orbiviruses and cause similar disease in a variety of domestic and wild animals, they have different host ranges, cell/tissue tropism, rates of replication, and levels of viremia. These characteristics ultimately determine whether a given species will act as a viral reservoir for midge transmission. Specific interactions between the viral capsid proteins and as yet undetermined cell surface receptor ligands confer susceptibility of a species. The aim of this study is to identify cellular receptors of both mammalian and insect host cells for bluetongue outer capsid proteins, providing insight into the molecular mechanisms of host range specificity, tissue tropism, and viremia which are critical for the transmission cycle between midges and livestock.
Approach:
Plasma membrane preparations from ovine, bovine, and midge cells will be tested by far-western blotting for the ability to bind cloned, expressed viral capsid proteins. Mass spectroscopy will be used to identify the receptor ligands. Receptor binding activity will be confirmed with 1) loss of function studies by targeting the receptors in the wild type cell lines with clustered regularly interspaced short palindromic repeats (CRISPR) to create genetic knock-down lines and testing for decreased virus binding activity; and 2) gain of function studies by transducing a refractory cell line with receptor ligands and testing for the ability to bind virus.
