Background The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in late 2019 and spread globally, prompting an international effort to accelerate development of a vacci...

https://www.nejm.org/doi/pdf/10.1056/NEJMoa2022483

An mRNA Vaccine against SARS-CoV-2 — Preliminary Report

A novel severe acute respiratory syndrome (SARS)-like coronavirus (SARS-CoV-2) is causing the global coronavirus disease 2019 (COVID-19) pandemic. Understanding how SARS-CoV-2 enters human cells is a high priority for deciphering its mystery and curbing its spread. A virus surface spike protein mediates SARS-CoV-2 entry into cells. To fulfill its function, SARS-CoV-2 spike binds to its receptor human ACE2 (hACE2) through its receptor-binding domain (RBD) and is proteolytically activated by human proteases. Here we investigated receptor binding and protease activation of SARS-CoV-2 spike using biochemical and pseudovirus entry assays. Our findings have identified key cell entry mechanisms of SARS-CoV-2. First, SARS-CoV-2 RBD has higher hACE2 binding affinity than SARS-CoV RBD, supporting efficient cell entry. Second, paradoxically, the hACE2 binding affinity of the entire SARS-CoV-2 spike is comparable to or lower than that of SARS-CoV spike, suggesting that SARS-CoV-2 RBD, albeit more potent, is less exposed than SARS-CoV RBD. Third, unlike SARS-CoV, cell entry of SARS-CoV-2 is preactivated by proprotein convertase furin, reducing its dependence on target cell proteases for entry. The high hACE2 binding affinity of the RBD, furin preactivation of the spike, and hidden RBD in the spike potentially allow SARS-CoV-2 to maintain efficient cell entry while evading immune surveillance. These features may contribute to the wide spread of the virus. Successful intervention strategies must target both the potency of SARS-CoV-2 and its evasiveness.

https://www.pnas.org/content/pnas/117/21/11727.full.pdf

Cell entry mechanisms of SARS-CoV-2.

The coronavirus spike protein is a multifunctional molecular machine that mediates coronavirus entry into host cells. It first binds to a receptor on the host cell surface through its S1 subunit and then fuses viral and host membranes through its S2 subunit. Two domains in S1 from different coronaviruses recognize a variety of host receptors, leading to viral attachment. The spike protein exists in two structurally distinct conformations, prefusion and postfusion. The transition from prefusion to postfusion conformation of the spike protein must be triggered, leading to membrane fusion. This article reviews current knowledge about the structures and functions of coronavirus spike proteins, illustrating how the two S1 domains recognize different receptors and how the spike proteins are regulated to undergo conformational transitions. I further discuss the evolution of these two critical functions of coronavirus spike proteins, receptor recognition and membrane fusion, in the context of the corresponding fu...

Structure, Function, and Evolution of Coronavirus Spike Proteins.

A novel β-coronavirus (2019-nCoV) caused severe and even fetal pneumonia explored in a seafood market of Wuhan city, Hubei province, China, and rapidly spread to other provinces of China and other countries. The 2019-nCoV was different from SARS-CoV, but shared the same host receptor the human angiotensin-converting enzyme 2 (ACE2). The natural host of 2019-nCoV may be the bat Rhinolophus affinis as 2019-nCoV showed 96.2% of whole-genome identity to BatCoV RaTG13. The person-to-person transmission routes of 2019-nCoV included direct transmission, such as cough, sneeze, droplet inhalation transmission, and contact transmission, such as the contact with oral, nasal, and eye mucous membranes. 2019-nCoV can also be transmitted through the saliva, and the fetal–oral routes may also be a potential person-to-person transmission route. The participants in dental practice expose to tremendous risk of 2019-nCoV infection due to the face-to-face communication and the exposure to saliva, blood, and other body fluids, and the handling of sharp instruments. Dental professionals play great roles in preventing the transmission of 2019-nCoV. Here we recommend the infection control measures during dental practice to block the person-to-person transmission routes in dental clinics and hospitals.

/pdf/transmission-routes-of-2019-ncov-and-controls-in-dental-2v7tgt9gl9.pdf

Transmission routes of 2019-nCoV and controls in dental practice.

COVID-19 pathophysiology: A review.

Coronaviruses are enveloped positive-stranded RNA viruses that replicate in the cytoplasm. To deliver their nucleocapsid into the host cell, they rely on the fusion of their envelope with the host cell membrane. The spike glycoprotein (S) mediates virus entry and is a primary determinant of cell tropism and pathogenesis. It is classified as a class I fusion protein, and is responsible for binding to the receptor on the host cell as well as mediating the fusion of host and viral membranes—A process driven by major conformational changes of the S protein. This review discusses coronavirus entry mechanisms focusing on the different triggers used by coronaviruses to initiate the conformational change of the S protein: receptor binding, low pH exposure and proteolytic activation. We also highlight commonalities between coronavirus S proteins and other class I viral fusion proteins, as well as distinctive features that confer distinct tropism, pathogenicity and host interspecies transmission characteristics to coronaviruses.

/pdf/mechanisms-of-coronavirus-cell-entry-mediated-by-the-viral-3dun1p0bi2.pdf

Mechanisms of Coronavirus Cell Entry Mediated by the Viral Spike Protein

Feline coronaviruses (FCoV) exist as 2 biotypes: feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV). FECV causes subclinical infections; FIPV causes feline infectious peritonitis (FIP), a systemic and fatal disease. It is thought that mutations in FECV enable infection of macrophages, causing FIP. However, the molecular basis for this biotype switch is unknown. We examined a furin cleavage site in the region between receptor-binding (S1) and fusion (S2) domains of the spike of serotype 1 FCoV. FECV sequences were compared with FIPV sequences. All FECVs had a conserved furin cleavage motif. For FIPV, there was a correlation with the disease and >1 substitution in the S1/S2 motif. Fluorogenic peptide assays confirmed that the substitutions modulate furin cleavage. We document a functionally relevant S1/S2 mutation that arises when FIP develops in a cat. These insights into FIP pathogenesis may be useful in development of diagnostic, prevention, and treatment measures against coronaviruses.

/pdf/mutation-in-spike-protein-cleavage-site-and-pathogenesis-of-fuxau1umyw.pdf

Mutation in spike protein cleavage site and pathogenesis of feline coronavirus.

Canine enteric coronavirus (CCoV) is an alphacoronavirus infecting dogs that is closely related to enteric coronaviruses of cats and pigs. While CCoV has traditionally caused mild gastro-intestinal clinical signs, there are increasing reports of lethal CCoV infections in dogs, with evidence of both gastrointestinal and systemic viral dissemination. Consequently, CCoV is now considered to be an emerging infectious disease of dogs. In addition to the two known serotypes of CCoV, novel recombinant variants of CCoV have been found containing spike protein N-terminal domains (NTDs) that are closely related to those of feline and porcine strains. The increase in disease severity in dogs and the emergence of novel CCoVs can be attributed to the high level of recombination within the spike gene that can occur during infection by more than one CCoV type in the same host.

/pdf/canine-enteric-coronaviruses-emerging-viral-pathogens-with-2xu9vgyqn1.pdf

Canine Enteric Coronaviruses: Emerging Viral Pathogens with Distinct Recombinant Spike Proteins

Characterization of a recombinant canine coronavirus with a distinct receptor-binding (S1) domain.

Potential mosquito vectors of Dirofilaria immitis (Leidy) (Nematoda: Filarioidea), the causative agent of dog heartworm in the southeastern region of the United States, were collected with CDC light traps and gravid traps in seven counties in the state of Georgia, USA. The presence of D. immitis in these mosquitoes was detected by polymerase chain reaction using species-specific primers for the D. immitis surface or cuticular antigen, Overall, 1,574 mosquitoes of 13 species in seven genera were collected; 92% of the specimens were Aedes albopictus (Skuse), Aedes vexans (Meigen), or Anopheles punctipennis (Say). Ae. albopictus, An. punctipennis, and Anopheles crucians Wiedemann were positive for D. immitis DNA. Ae. albopictus had the highest maximum likelihood rate of infection (2.30%; 95% confidence interval [CI] = 1.15–4.00%) followed by An. crucians (1.38%: 95% CI = 0.04–6.93%), and An. punctipennis (0.85%: 95% CI 0.03–4.29%). The detection of D. immitis DNA in the heads and thoraxes of Ae. albopictus (0.40%; 95% CI = 0.12–2.02%) indicates that these mosquitoes can support the development of D. immitis to the infective stage 3 larvae.

/pdf/detection-of-dirofilaria-immitis-nematoda-filarioidea-by-3uzjwjj0ks.pdf

Detection of Dirofilaria immitis (Nematoda: Filarioidea) by polymerase chain reaction in Aedes albopictus, Anopheles punctipennis, and Anopheles crucians (Diptera: Culicidae) from Georgia, USA.

Beth N. Licitra

Papers

Mechanisms of Coronavirus Cell Entry Mediated by the Viral Spike Protein

Mutation in spike protein cleavage site and pathogenesis of feline coronavirus.

Canine Enteric Coronaviruses: Emerging Viral Pathogens with Distinct Recombinant Spike Proteins

Characterization of a recombinant canine coronavirus with a distinct receptor-binding (S1) domain.

Detection of Dirofilaria immitis (Nematoda: Filarioidea) by polymerase chain reaction in Aedes albopictus, Anopheles punctipennis, and Anopheles crucians (Diptera: Culicidae) from Georgia, USA.