Entry of Scotophilus Bat Coronavirus-512 and Severe Acute Respiratory Syndrome Coronavirus in Human and Multiple Animal Cells

Abstract

Bats are natural reservoirs of severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV). Scotophilus bat CoV-512 demonstrates potential for cross-species transmission because its viral RNA and specific antibodies have been detected in three bat species of Taiwan. Understanding the cell tropism of Scotophilus bat CoV-512 is the first step for studying the mechanism of cross-species transmission. In this study, a lentivirus-based pseudovirus was produced using the spike (S) protein of Scotophilus bat CoV-512 or SARS-CoV as a surface protein to test the interaction between coronaviral S protein and its cell receptor on 11 different cells. Susceptible cells expressed red fluorescence protein (RFP) after the entry of RFP-bound green fluorescence protein (GFP)-fused S protein of Scotophilus bat CoV-512 (RFP-Sco-S-eGFP) or RFP-SARS-S pseudovirus, and firefly luciferase (FLuc) activity expressed by cells infected with FLuc-Sco-S-eGFP or FLuc-SARS-S pseudovirus was quantified. Scotophilus bat CoV-512 pseudovirus had significantly higher entry efficiencies in Madin Darby dog kidney epithelial cells (MDCK), black flying fox brain cells (Pabr), and rat small intestine epithelial cells (IEC-6). SARS-CoV pseudovirus had significantly higher entry efficiencies in human embryonic kidney epithelial cells (HEK-293T), pig kidney epithelial cells (PK15), and MDCK cells. These findings demonstrated that Scotophilus bat CoV-512 had a broad host range for cross-species transmission like SARS-CoV.Bats are the natural reservoirs of SARS-CoV, MERS-CoV, and HCoV-NL63. SARS-CoV and HCoV-NL63 gain cellular entry through angiotensin-converting enzyme 2 (ACE2) [9], whereas MERS-CoV utilizes dipeptidyl peptidase-4 (DPP4) as its entry receptor [10]. The entry receptor is considered a hallmark of coronaviral cross-species transmissibility. SARS-related CoV isolated from Rhinolophus bats can use the ACE2 of humans, civets, and Rhinolophus bats as its cell receptor to infect cells originated from human and many other animal species [11]. Both MERS-CoV and bat CoV-HKU4 can use bat and human DPP4 to infect cells originating from humans, camels, bats, and other animal species [12,13] and HCoV-NL63 can replicate in the lung cell line from tricolored bats [8].Little information is available regarding cross-species events for animal CoVs. A previous study detected Scotophilus bat CoV-512 in four different bat species along with Miniopterus bat CoV-1A and Rhinolophus SARS-related CoV in Taiwan through the reverse transcription polymerase chain reaction (RT-PCR) [14]. Antibodies specific to the nucleocapsid (N) protein fragments of Scotophilus bat CoV-512 were detected in serum collected from three bat species, namely Scotophilus kuhlii, Miniopterus fuliginosus, and Rhinolophus monoceros [15]. A close relationship and possible gene recombinants between Scotophilus bat CoV-512 and PEDV were observed through sequence alignments [14,16]. The results of the cell entry assay also indicated that PEDV can infe...

Full-Genome Sequences of Alphacoronaviruses and Astroviruses from Myotis and Pipistrelle Bats in Denmark

“…The six Pedacovirus-like BtCoV sequences described in this study are phylogenetically related to both PEDV and Scotophilus bat coronavirus 512. PEDV has been shown to bind to aminopeptidase-N (APN), but not necessarily as the only receptor, while Scotophilus bat coronavirus 512 is able to infect a wide range of cultured cell types without the use of APN [71]. To our knowledge, no cell receptors have been described for the Alphacoronavirus subgenera Nyctacovirus and Myotacovirus, but for other subgenera in this genus, APN, angiotensin-converting enzyme 2 (ACE2), N-glycolylneuraminic acid (Neu5Gc), and N-acetylneuraminic acid (Neu5Ac) are known receptors [72].…”

Antiviral performance of graphene-based materials with emphasis on COVID-19: A review

“…Here we briefly review these viruses. In terms of animal infectious coronaviruses, alphacoronaviruses include transmissible gastroenteritis virus [38] , porcine-respiratory coronavirus [39] and porcine-epidemic diarrhea virus [40] infecting pigs; feline-CoV affecting cats [41] ; bat coronaviruses such as Scotophilus bat CoV-512, Rhinolophus bat CoV-HKU2, Miniopterus bat CoV 1A/B and CoV-HKU8 infecting various species of bats [42] , [43] , [44] , [45] .…”

Replication of SARS-CoV-2 in cell lines used in public health surveillance programmes with special emphasis on biosafety

“…It has been shown that angiotensin-converting enzyme 2 (ACE2) receptors present on host cells are used by the SARS-CoV-2 for viral entry 25 . ACE2 receptors have been shown to be present on MDCK cells 23 26 . This probably indicates that post-adsorption mechanisms and/or cell machinery of the MDCK cell line do not allow replication of SARS-CoV-2.…”