OmniVira

Literature detail

Mechanisms of zoonotic severe acute respiratory syndrome coronavirus host range expansion in human airway epithelium.

Timothy Sheahan1 Barry Rockx Eric Donaldson Amy Sims Raymond Pickles Davide Corti Ralph Baric
Affiliations 1 institutions
  1. Department of Epidemiology, 2107 McGavran-Greenberg, CB#7435, University of North Carolina, Chapel Hill, NC 27699-7435, USA.
PMID 18094188 2008 J Virol eng ppublish
PubMed DOI Browse context

Article

Publication summary

In 2003, severe acute respiratory syndrome coronavirus (SARS-CoV) emerged and caused over 8,000 human cases of infection and more than 700 deaths worldwide. Zoonotic SARS-CoV likely evolved to infect humans by a series of transmission events between humans and animals for sale in China. Using synthetic biology, we engineered the spike protein (S) from a civet strain, SZ16, into our epidemic strain infectious clone, creating the chimeric virus icSZ16-S, which was infectious but yielded progeny viruses incapable of propagating in vitro. After introducing a K479N mutation within the S receptor binding domain (RBD) of SZ16, the recombinant virus (icSZ16-S K479N) replicated in Vero cells but was severely debilitated in growth. The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells and on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2). The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2. Further, these viruses were neutralized by a human monoclonal antibody (MAb), S230.15, but the parent icSZ16-S K479N strain was eight times more resistant than the mutants. These data suggest that the human adaptation of zoonotic SARS-CoV strains may select for some variants that are highly susceptible to select MAbs that bind to RBDs. The epidemic, icSZ16-S K479N, and icSZ16-S K479N D22 viruses replicate similarly in the BALB/c mouse lung, highlighting the potential use of these zoonotic spike SARS-CoVs to assess vaccine or serotherapy efficacy in vivo.

Zoonoses Amino Acid Sequence Animals Base Sequence Chlorocebus aethiops DNA Primers Epithelial Cells Humans Immunohistochemistry Membrane Glycoproteins Mice Mice, Inbred BALB C Molecular Sequence Data Phylogeny Reverse Transcriptase Polymerase Chain Reaction Severe acute respiratory syndrome-related coronavirus Spike Glycoprotein, Coronavirus Trachea

Structured evidence records

Evidence records

5 total
Host Range Experiment 2 records
Host Range Experiment Extraction confidence 0.90
Key finding

Chimeric SARS-CoV spike constructs derived from civet and human strains showed limited replication initially but evolved mutations that enhanced growth on human airway epithelial cells and cells expressing human ACE2 receptor.

Virus
SARS-CoV
Location
Not specified
Supporting text

Using synthetic biology, we engineered the spike protein (S) from a civet strain, SZ16, into our epidemic strain infectious clone, creating the chimeric virus icSZ16-S, which was infectious but yielded progeny viruses incapable of propagating in vitro. After introducing a K479N mutation within the S receptor binding domain (RBD) of SZ16, the recombinant virus (icSZ16-S K479N) replicated in Vero cells but was severely debilitated in growth. The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells and on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2).

Method
synthetic biology; reverse genetics; in vitro evolution; replication assay
Sample type
airway epithelial cells
Experimental system
in vitro cell culture
Host Range Experiment Extraction confidence 0.90
Key finding

Engineered SARS-CoV spike variants exhibited similar replication levels in the lungs of BALB/c mice, demonstrating their utility for in vivo host range and therapeutic efficacy assessment.

Virus
SARS-CoV
Location
Not specified
Supporting text

The epidemic, icSZ16-S K479N, and icSZ16-S K479N D22 viruses replicate similarly in the BALB/c mouse lung, highlighting the potential use of these zoonotic spike SARS-CoVs to assess vaccine or serotherapy efficacy in vivo.

Method
experimental infection; replication assay
Sample type
lung
Experimental system
in vivo animal experiment
Genomic Evolution 1 records
Genomic Evolution Extraction confidence 0.80
Key finding

Experimental evolution of SARS-CoV spike identified adaptive RBD mutations Y442F and L472F that enhanced replication on human airway cells and modified hACE2 binding.

Virus
SARS-CoV
Location
Not specified
Supporting text

After introducing a K479N mutation within the S receptor binding domain (RBD) of SZ16, the recombinant virus (icSZ16-S K479N) replicated in Vero cells but was severely debilitated in growth. The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells ... The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2.

Genes or proteins
spike protein; receptor binding domain; ACE2
Analysis methods
in vitro evolution; sequence analysis; mutational analysis
Molecular Adaptation 1 records
Molecular Adaptation Extraction confidence 1.00
Key finding

Mutations Y442F and L472F in the SARS-CoV spike receptor binding domain enhanced replication on human airway epithelial cells by remodeling spike interactions with human ACE2, demonstrating host adaptation.

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

The in vitro evolution of icSZ16-S K479N on human airway epithelial (HAE) cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on HAE cells and on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2). The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2.

Genes or proteins
spike protein; receptor binding domain
Receptors
human angiotensin I converting enzyme 2 (hACE2)
Mutations
K479N; Y442F; L472F
Mechanism types
receptor_binding; replication_efficiency; host_range_expansion
Receptor Usage 1 records
Receptor Usage Extraction confidence 0.95
Key finding

Mutations Y442F and L472F in the spike receptor-binding domain of SARS-CoV variants altered interactions with human ACE2, promoting improved binding and replication in human airway epithelium and hACE2-expressing cells.

Virus
SARS-CoV
Location
Not specified
Supporting text

The in vitro evolution of icSZ16-S K479N on human airway epithelial cells produced two viruses (icSZ16-S K479N D8 and D22) with enhanced growth on delayed brain tumor cells expressing the SARS-CoV receptor, human angiotensin I converting enzyme 2 (hACE2). The icSZ16-S K479N D8 and D22 virus RBDs contained mutations in ACE2 contact residues, Y442F and L472F, that remodeled S interactions with hACE2.

Method
synthetic biology; in vitro evolution
Receptors
human angiotensin I converting enzyme 2 (hACE2)