Differential Sensitivity of Bat Cells to Infection by Enveloped RNA Viruses: Coronaviruses, Paramyxoviruses, Filoviruses, and Influenza Viruses

Abstract

Bats (Chiroptera) host major human pathogenic viruses including corona-, paramyxo, rhabdo- and filoviruses. We analyzed six different cell lines from either Yinpterochiroptera (including African flying foxes and a rhinolophid bat) or Yangochiroptera (genera Carollia and Tadarida) for susceptibility to infection by different enveloped RNA viruses. None of the cells were sensitive to infection by transmissible gastroenteritis virus (TGEV), a porcine coronavirus, or to infection mediated by the Spike (S) protein of SARS-coronavirus (SARS-CoV) incorporated into pseudotypes based on vesicular stomatitis virus (VSV). The resistance to infection was overcome if cells were transfected to express the respective cellular receptor, porcine aminopeptidase N for TGEV or angiotensin-converting enzyme 2 for SARS-CoV. VSV pseudotypes containing the S proteins of two bat SARS-related CoV (Bg08 and Rp3) were unable to infect any of the six tested bat cell lines. By contrast, viral pseudotypes containing the surface protein GP of Marburg virus from the family Filoviridae infected all six cell lines though at different efficiency. Notably, all cells were sensitive to infection by two paramyxoviruses (Sendai virus and bovine respiratory syncytial virus) and three influenza viruses from different subtypes. These results indicate that bat cells are more resistant to infection by coronaviruses than to infection by paramyxoviruses, filoviruses and influenza viruses. Furthermore, these results show a receptor-dependent restriction of the infection of bat cells by CoV. The implications for the isolation of coronaviruses from bats are discussed.

Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses

“…5). None of the spike pseudotypes efficiently entered Rhinolophus cells, as was observed in previous studies using these cells 29,30 (Fig. 2c).…”

SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor

“…Analysis of the receptor binding motif (RBM), a portion of the receptor binding domain (RBD) that makes contact with ACE2 (Li et al, 2005a), revealed that most amino acid residues essential for ACE2 binding by SARS-S were conserved in SARS-2-S ( Figure 2B). In contrast, most of these residues were absent from S proteins of SARSr-CoV previously found not to use ACE2 for entry ( Figure 2B) (Ge et al, 2013;Hoffmann et al, 2013;Menachery et al, 2020). In agreement with these findings, directed expression of human and bat (Rhinolophus alcyone) ACE2 but not human DPP4, the entry receptor used by MERS-CoV (Raj et al, 2013), or human APN, the entry receptor used by HCoV-229E (Yeager et al, 1992), allowed SARS-2-S-and SARS-S-driven entry into otherwise non-susceptible BHK-21 cells ( Figure 3A).…”

“…Statistical significance was tested by two-way ANOVA with Dunnett posttest. dipeptidyl-peptidase 4 (DPP4) and human TMPRSS2 have been described elsewhere (Bertram et al, 2010;Brinkmann et al, 2017;Gierer et al, 2013;Hoffmann et al, 2013;Hofmann et al, 2005;Kleine-Weber et al, 2019). For generation of the expression plasmids for SARS-2-S with or without a C-terminal HA epitope tag we PCR-amplified the coding sequence of a synthetic, codon-optimized (for human cells) SARS-2-S DNA (GeneArt Gene Synthesis, ThermoFisher Scientific) based on the publicly available protein sequence in the National Center for Biotechnology Information database (NCBI Reference Sequence: YP_009724390.1) and cloned in into the pCG1 expression vector via BamHI and XbaI restriction sites.…”

Polymorphisms in dipeptidyl peptidase 4 reduce host cell entry of Middle East respiratory syndrome coronavirus

“…We further utilized pCAGGS-based expression vectors for vesicular stomatitis virus (VSV) glycoprotein (G), MERS-CoV S wildtype (WT) and MERS-CoV S (D510G) (the latter two either untagged or equipped with a C-terminal V5 epitope) that have been described elsewhere [35][36][37]. In addition, a previously described expression vector for angiotensin converting enzyme 2 was employed [38]. Similar to the strategy used for the generation of DPP4 mutants, we employed the overlap-extension PCR technique to introduce a single mutation into the MERS-CoV S open reading frame, thus generating untagged and V5-tagged MERS-CoV S (D539N).…”