A nano-luciferase expressing human coronavirus OC43 for countermeasure development

The genetic diversity of the coronavirus (CoV) family poses a significant challenge for drug discovery and development. Traditional antiviral drugs often target specific viral proteins from specific viruses which limits their use, especially against novel emerging viruses. Antivirals with broad-spectrum activity overcome this limitation by targeting highly conserved regions or catalytic domains within viral proteins that are essential for replication. For rapid identification of small molecules with broad antiviral activity, assays with viruses representing family-wide genetic diversity are needed. Viruses engineered to express a reporter gene (i.e. luminescence, fluorescence, etc.) can increase the efficiency, sensitivity or precision of drug screening over classical measures of replication like observation of cytopathic effect or measurement of infectious titers. We have previously developed reporter virus systems for multiple other endemic, pandemic, epidemic and enzootic CoV. Human CoV OC43 (HCoV-OC43) is a human endemic CoV that causes respiratory infection with age-related exacerbations of pathogenesis. Here, we describe the development of a novel recombinant HCoV-OC43 reporter virus that expresses nano-luciferase (HCoV-OC43 nLuc), and its potential application for screening of antivirals against CoV

Host range, transmissibility and antigenicity of a pangolin coronavirus

The pathogenic and cross-species transmission potential of SARS-CoV-2-related coronaviruses (CoVs) remain poorly characterized. Here we recovered a wild-type pangolin (Pg) CoV GD strain including derivatives encoding reporter genes using reverse genetics. In primary human cells, PgCoV replicated efficiently but with reduced fitness and showed less efficient transmission via airborne route compared with SARS-CoV-2 in hamsters. PgCoV was potently inhibited by US Food and Drug Administration approved drugs, and neutralized by COVID-19 patient sera and SARS-CoV-2 therapeutic antibodies in vitro. A pan-Sarbecovirus antibody and SARS-CoV-2 S2P recombinant protein vaccine protected BALB/c mice from PgCoV infection. In K18-hACE2 mice, PgCoV infection caused severe clinical disease, but mice were protected by a SARS-CoV-2 human antibody. Efficient PgCoV replication in primary human cells and hACE2 mice, coupled with a capacity for airborne spread, highlights an emergence potential. However, low competitive fitness, pre-immune humans and the benefit of COVID-19 countermeasures should impede its ability to spread globally in human populations

Engineered immunogens to elicit antibodies against conserved coronavirus epitopes

Immune responses to SARS-CoV-2 primarily target the receptor binding domain of the spike protein, which continually mutates to escape acquired immunity. Other regions in the spike S2 subunit, such as the stem helix and the segment encompassing residues 815-823 adjacent to the fusion peptide, are highly conserved across sarbecoviruses and are recognized by broadly reactive antibodies, providing hope that vaccines targeting these epitopes could offer protection against both current and emergent viruses. Here we employ computational modeling to design scaffolded immunogens that display the spike 815-823 peptide and the stem helix epitopes without the distracting and immunodominant receptor binding domain. These engineered proteins bind with high affinity and specificity to the mature and germline versions of previously identified broadly protective human antibodies. Epitope scaffolds interact with both sera and isolated monoclonal antibodies with broadly reactivity from individuals with pre-existing SARS-CoV-2 immunity. When used as immunogens, epitope scaffolds elicit sera with broad betacoronavirus reactivity and protect as “boosts” against live virus challenge in mice, illustrating their potential as components of a future pancoronavirus vaccine

Apatite and titanite from the Karrat Group, Greenland; implications for charting the thermal evolution of crust from the U-Pb geochronology of common Pb bearing phases

Titanite and apatite have Pb closure temperatures of ~700 °C and 450–550 °C, respectively, allowing different points on a cooling trajectory to be determined. However, both phases typically accommodate moderate to significant quantities of common Pb. Understanding the thermal diffusivity of a specific isotopic system in different minerals along with their apparent U-Pb age allows modelling of regional cooling trends. Such cooling trends may provide key evidence for correct interpretation of the measured geochronometer. Specifically, thermal history reconstruction may address questions related to the interpretation of an isotopic date as the time of crystallization versus cooling, or alternatively, as a resetting age. In this work, a case study from metavolcanic rocks of the Karrat Group, West Greenland, is used to inform the U-Pb geochronology of common Pb bearing phases, thermal modelling, and also the regional geology. Magmatic apatite yields a reset U-Pb age of 1826 ± 9 Ma, whereas titanite yields a mean U-Pb age of 1768 ± 8 Ma. The apatite age is interpreted as the time of total resetting during a > 485 °C event. In contrast, the titanite age is interpreted as the time of metamorphic crystallization, consistent with its REE chemistry. Thermal modelling indicates this metamorphic event did not exceed 452 °C. The resetting of the U-Pb system in magmatic apatite is interpreted as a response to the collision between the Rae Craton and the Superior Craton during the Trans-Hudson Orogeny. However, subsequent metamorphic titanite growth is interpreted as distal evidence of an event shared with the Nagssugtoqidian Orogen. The modelled thermal history implies over 100 million years of communal tectonic history between the Nagssugtoqidian and Rinkian Orogens. Of great significance is the fact that both apatite and titanite show distinctly different common Pb compositions. Apatite retains common Pb with a composition similar to ancient common Pb, whereas titanite retains common Pb with a lower 207 Pb/ 206 Pb ratio implying it was influenced also by Pb from recrystallized precursor U bearing minerals. The common Pb signature in minerals may assist in interpretation of the growth mechanism of the dated phase