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Literature detail

Entry, replication and innate immunity evasion of BANAL-236, a SARS-CoV-2-related bat virus, in Rhinolophus and human cells.

Ségolène Gracias1 Elodie Le Seac'h1 Samuel Donaire-Carpio1 Françoise Vuillier2 Léa Vendramini1,3 Adam Moundib1 Sarah Temmam3 Magdalena Rutkowska4,5 Flora Donati6 Anastasija Cupic4,5 Javier Juste7 Carles Martinez-Romero4 Nathalie Morel8 Olivier Schwartz9 Nevan J Krogan10,11,12 Lisa Miorin4,13 Marcel A Müller14,15 Caroline Demeret2 Sandie Munier16 Philippe Roingeard17 Jyoti Batra10,11,12 Adolfo Garcia-Sastre4,13,18,19 Vincent Caval1 Nolwenn Jouvenet1
Affiliations 19 institutions
  1. Institut Pasteur, Université Paris Cité, CNRS UMR 3569, Virus sensing and signaling Unit, Paris, France.
  2. Institut Pasteur, Université Paris Cité, Interactomics, RNA and Immunity Unit, Paris, France.
  3. Institut Pasteur, Université Paris Cité, Pathogen Discovery Laboratory, Paris, France.
  4. Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
  5. Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
  6. Institut Pasteur, National Reference Center for Respiratory Viruses, Paris, France.
  7. Estación biológica de doñana, Avda, Seville, and CIBER Epidemiology and Public Health, CIBERESP, Madrid, Spain.
  8. Département Médicaments et Technologies pour la Santé (DMTS), Service de Pharmacologie et Immunoanalyse, Université Paris Saclay, CEA, INRA, Gif-sur Yvette, France.
  9. Institut Pasteur, Université Paris Cité, Virus and Immunity Unit, Paris, France.
  10. Quantitative Biosciences Institute (QBI), University of California San Francisco, San Francisco, California United States of America.
  11. Gladstone Institute of Data Science and Biotechnology, J David Gladstone Institutes, San Francisco, California, United States of America.
  12. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, United States of America.
  13. Global Health Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
  14. Institute of Virology, Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
  15. German Center for Infection Research (DZIF), Partner Site Charité, Berlin, Germany.
  16. Institut Pasteur, Université Paris Cité, Lyssavirus Epidemiology and Neuropathology Unit, Paris, France.
  17. INSERM U1259 MAVIVH and INSERM US61, Université de Tours and CHU de Tours, Tours, France.
  18. The Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
  19. The Icahn Genomics Institute, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
PMID 42008572 2026 PLoS Pathog eng epublish
PubMed DOI Browse context

Article

Publication summary

Asian Rhinolophus bats are considered the natural reservoirs of an ancestral SARS-CoV-2. However, the biology of SARS-CoV-2-related viruses in bat cells is not well understood. Here, we investigated the replication of an isolate of BANAL-236, the only bat-derived SARS-CoV-2 relative isolated to date, in Rhinolophus ferrumequinum lungs (Rfe) cells. BANAL-236 did not replicate in wild-type Rhinolophus cell lines. Entry assays using pseudoviruses expressing the spike proteins (S) of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 (hACE2) and human TMPRSS2 (hTMPRSS2) in human and Rhinolophus cells. Through biochemical, virological, and electron microscopy analyses, we showed that BANAL-236 and SARS-CoV-2 completed their replication cycles in RFe cells engineered to express high levels of hACE2 and hTMPRSS2. Despite efficient viral replication in modified Rhinolophus and human cells, no induction of interferon (IFN)-stimulated genes was detected. Using a screening approach, we identified several BANAL-236 proteins that antagonize IFN production and signalling in human cells. Our findings thus show that BANAL-236 possesses critical features that enabled zoonotic spillover: hACE2 usage and potent evasion of human IFN responses. The Rhinolophus cellular model we established offers a platform for further investigating the interactions between bat sarbecoviruses and their reservoir hosts.

Betacoronavirus Chiroptera Coronavirus Infections Immune Evasion Immunity, Innate SARS-CoV-2 Virus Internalization Virus Replication Angiotensin-Converting Enzyme 2 Animals Cell Line COVID-19 Humans Serine Endopeptidases Spike Glycoprotein, Coronavirus ACE2 protein, human TMPRSS2 protein, human

Structured evidence records

Evidence records

5 total
Host Range Experiment 2 records
Host Range Experiment Extraction confidence 0.88
Key finding

BANAL-236 failed to replicate in wild-type Rhinolophus cells but replicated in Rhinolophus ferrumequinum cells engineered to express human ACE2 and TMPRSS2, indicating human receptor-dependent host range expansion.

Virus
SARSr-CoV
Location
Not specified
Supporting text

We investigated the replication of an isolate of BANAL-236, the only bat-derived SARS-CoV-2 relative isolated to date, in Rhinolophus ferrumequinum lungs (Rfe) cells. BANAL-236 did not replicate in wild-type Rhinolophus cell lines. Entry assays using pseudoviruses expressing the spike proteins (S) of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 (hACE2) and human TMPRSS2 (hTMPRSS2) in human and Rhinolophus cells. BANAL-236 and SARS-CoV-2 completed their replication cycles in RFe cells engineered to express high levels of hACE2 and hTMPRSS2.

Method
replication assay; pseudovirus entry assay
Sample type
lung cells
Experimental system
in vitro cell culture
Host Range Experiment Extraction confidence 0.88
Key finding

BANAL-236 replicated in human cells expressing ACE2 and TMPRSS2, showing susceptibility of human cell systems to this bat sarbecovirus.

Virus
SARSr-CoV
Location
Not specified
Supporting text

Entry assays using pseudoviruses expressing the spike proteins (S) of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 (hACE2) and human TMPRSS2 (hTMPRSS2) in human and Rhinolophus cells. BANAL-236 completed its replication cycle in human cells expressing hACE2 and hTMPRSS2.

Method
replication assay; pseudovirus entry assay
Experimental system
in vitro cell culture
Molecular Adaptation 2 records
Molecular Adaptation Extraction confidence 0.90
Key finding

BANAL-236 spike protein mediates cell entry through human ACE2 and TMPRSS2, indicating receptor-binding adaptation that may facilitate human infection.

Virus
SARSr-CoV
Host
Not specified
Location
Not specified
Supporting text

Entry assays using pseudoviruses expressing the spike proteins of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 and human TMPRSS2 in human and Rhinolophus cells.

Genes or proteins
spike
Receptors
ACE2
Host factors
TMPRSS2
Mechanism types
receptor_binding; cell_entry
Molecular Adaptation Extraction confidence 0.90
Key finding

Multiple BANAL-236 proteins inhibit interferon production and signaling, showing molecular adaptation for innate immune evasion in human cells.

Virus
SARSr-CoV
Host
Not specified
Location
Not specified
Supporting text

Using a screening approach, we identified several BANAL-236 proteins that antagonize IFN production and signalling in human cells.

Mechanism types
immune_escape
Receptor Usage 1 records
Receptor Usage Extraction confidence 0.98
Key finding

BANAL-236 spike-mediated entry into human and Rhinolophus cells requires human ACE2 and TMPRSS2 expression.

Virus
SARSr-CoV
Location
Not specified
Supporting text

Entry assays using pseudoviruses expressing the spike proteins (S) of SARS-CoV-2, BANAL-236, and BANAL-52 revealed that efficient S-mediated entry depends on the expression of human ACE2 (hACE2) and human TMPRSS2 (hTMPRSS2) in human and Rhinolophus cells.

Method
pseudovirus assay; entry assay; biochemical analysis; electron microscopy
Receptors
human ACE2
Host factors
human TMPRSS2