A rapid and affordable in vitro model to test SARS-CoV-2 susceptibility in animal species Coronavirus Disease COVID invitro SARSCoV2 biorxivpreprint USDA
By Dr. Chinta SidharthanApr 24 2023Reviewed by Benedette Cuffari, M.Sc. In a recent study posted to the bioRxiv* preprint server, researchers developed a novel in vitro model that expresses the angiotensin-converting enzyme 2 receptor from seven bar species in a non-permissive avian fibroblast cell line to study the susceptibility to and replication of the severe acute respiratory syndrome coronavirus 2 .
Background The etiological agent of the coronavirus disease 2019 , SARS-CoV-2, belongs to the Coronaviridae family and is an enveloped, single-stranded ribonucleic acid virus. Among the 1,400 species of bats found around the world, the susceptibility to SARS-CoV-2 has only been tested in four species. While testing the vulnerability of each of these bat species to SARS-CoV-2 might not be feasible, in vitro models that express ACE-2 receptors can be used to test the ability of the virus to replicate in specific bat species.
Fluorescence activation cell sorting was used to identify transgenic DF1 cells expressing human transmembrane serine protease 2 and bat ACE-2 receptor. Quantitative reverse transcription polymerase chain reaction was performed to determine the expression levels of bat ACE-2 in transgenic cells, while immunofluorescence and Western blot assays were used to measure the expression of TMPRSS2 and ACE-2.
Study findings The expression of ACE-2 receptors from seven bat species in DF1 cells caused the cells to be susceptible to three SARS-CoV-2 isolates. Notably, viral replication levels differed between bat species and the tested SARS-CoV-2 variants. Although little brown bats, Brazilian free-tailed bats, and Egyptian rousettes are believed to have more contact with humans, horseshoe bats are found in more rural areas with less contact with humans. The ACE-2 receptor from the Chinese rufous horseshoe bat, Pearson's horseshoe bat, and greater horseshoe bat allowed the entry and replication of all three SARS-CoV-2 variants.
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