Injection site vaccinology of a recombinant vaccinia-based vector reveals diverse innate immune signatures

Poxvirus systems have been extensively used as vaccine vectors. Herein a RNA-Seq analysis of intramuscular injection sites provided detailed insights into host innate immune responses, as well as expression of vector and recombinant immunogen genes, after vaccination with a new multiplication defective, vaccinia-based vector, Sementis Copenhagen Vector. Chikungunya and Zika virus immunogen mRNA and protein expression was associated with necrosing skeletal muscle cells surrounded by mixed cellular infiltrates. The multiple adjuvant signatures at 12 hours post-vaccination were dominated by TLR3, 4 and 9, STING, MAVS, PKR and the inflammasome. Th1 cytokine signatures were dominated by IFNγ, TNF and IL1β, and chemokine signatures by CCL5 and CXCL12. Multiple signatures associated with dendritic cell stimulation were evident. By day seven, vaccine transcripts were absent, and cell death, neutrophil, macrophage and inflammation annotations had abated. No compelling arthritis signatures were identified. Such injection site vaccinology approaches should inform refinements in poxvirus-based vector design.Jessamine E. Hazlewood, Troy Dumenil, Thuy T. Le, Andrii Slonchak, Stephen H. Kazakoff, Ann-Marie Patch ... et al

Reversible restructuring of supported Au nanoparticles during butadiene hydrogenation revealed by <em>operando</em> GISAXS/GIWAXS

Periodically arranged, monodisperse gold nanoparticles supported on flat silicon substrates were studied for the hydrogenation of 1,3-butadiene under operando conditions using Grazing Incidence Small- and Wide-Angle X-ray Scattering (GISAXS/GIWAXS). It was found that the composition and shape of the nanoparticles depends very much on the chemical environment; the particles are shown to be dynamic, undergoing reversible size and shape change particularly during catalytic reaction, highlighting a dynamism often not observed in traditional studies. Specifically, the size of the Au nanoparticles increases during butadiene hydrogenation and this is attributed to the partial removal of a Au2O3 at the metal–oxide interface and consequential shape change of the nanoparticle from a more hemispherical particle to a particle with a larger height to width ratio

Arthritogenic alphavirus vaccines: serogrouping versus cross-protection in mouse models

Chikungunya virus (CHIKV), Ross River virus (RRV), o’nyong nyong virus (ONNV), Mayaro virus (MAYV) and Getah virus (GETV) represent arthritogenic alphaviruses belonging to the Semliki Forest virus antigenic complex. Antibodies raised against one of these viruses can cross-react with other serogroup members, suggesting that, for instance, a CHIKV vaccine (deemed commercially viable) might provide cross-protection against antigenically related alphaviruses. Herein we use human alphavirus isolates (including a new human RRV isolate) and wild-type mice to explore whether infection with one virus leads to cross-protection against viremia after challenge with other members of the antigenic complex. Persistently infected Rag1(-/-) mice were also used to assess the cross-protective capacity of convalescent CHIKV serum. We also assessed the ability of a recombinant poxvirus-based CHIKV vaccine and a commercially available formalin-fixed, whole-virus GETV vaccine to induce cross-protective responses. Although cross-protection and/or cross-reactivity were clearly evident, they were not universal and were often suboptimal. Even for the more closely related viruses (e.g., CHIKV and ONNV, or RRV and GETV), vaccine-mediated neutralization and/or protection against the intended homologous target was significantly more effective than cross-neutralization and/or cross-protection against the heterologous virus. Effective vaccine-mediated cross-protection would thus likely require a higher dose and/or more vaccinations, which is likely to be unattractive to regulators and vaccine manufacturers.Wilson Nguyen, Eri Nakayama, Kexin Yan, Bing Tang, Thuy T. Le, Liang Liu, Tamara H. Cooper, John D. Hayball, Helen M. Faddy, David Warrilow, Richard J. N. Allcock, Jody Hobson-Peters, Roy A. Hall, Daniel J. Rawle, Viviana P. Lutzky, Paul Young, Nidia M. Oliveira, Gunter Hartel, Paul M. Howley, Natalie A. Prow, and Andreas Suhrbie

Early Science with the Large Millimeter Telescope: observations of dust continuum and CO emission lines of cluster-lensed submillimetre galaxies at z=2.0-4.7

We present Early Science observations with the Large Millimeter Telescope, AzTEC 1.1 mm continuum images and wide bandwidth spectra (73–111 GHz) acquired with the Redshift Search Receiver, towards four bright lensed submillimetre galaxies identified through the Herschel Lensing Survey-snapshot and the Submillimetre Common-User Bolometer Array-2 Cluster Snapshot Survey. This pilot project studies the star formation history and the physical properties of the molecular gas and dust content of the highest redshift galaxies identified through the benefits of gravitational magnification. We robustly detect dust continuum emission for the full sample and CO emission lines for three of the targets. We find that one source shows spectroscopic multiplicity and is a blend of three galaxies at different redshifts (z = 2.040, 3.252, and 4.680), reminiscent of previous high-resolution imaging follow-up of unlensed submillimetre galaxies, but with a completely different search method, that confirm recent theoretical predictions of physically unassociated blended galaxies. Identifying the detected lines as 12CO (Jup = 2–5) we derive spectroscopic redshifts, molecular gas masses, and dust masses from the continuum emission. The mean H2 gas mass of the full sample is (2.0 ± 0.2) × 1011 M⊙/μ, and the mean dust mass is (2.0 ± 0.2) × 109 M⊙/μ, where μ ≈ 2–5 is the expected lens amplification. Using these independent estimations we infer a gas-to-dust ratio of δGDR ≈ 55–75, in agreement with other measurements of submillimetre galaxies. Our magnified high-luminosity galaxies fall on the same locus as other high-redshift submillimetre galaxies, extending the L′CO–LFIR correlation observed for local luminous and ultraluminous infrared galaxies to higher far-infrared and CO luminosities

BET inhibition blocks inflammation-induced cardiac dysfunction and SARS-CoV-2 infection

Cardiac injury and dysfunction occur in COVID-19 patients and increase the risk of mortality. Causes are ill defined but could be through direct cardiac infection and/or inflammation-induced dysfunction. To identify mechanisms and cardio-protective drugs, we use a state-of-the-art pipeline combining human cardiac organoids with phosphoproteomics and single nuclei RNA sequencing. We identify an inflammatory "cytokine-storm", a cocktail of interferon gamma, interleukin 1β, and poly(I:C), induced diastolic dysfunction. Bromodomain-containing protein 4 is activated along with a viral response that is consistent in both human cardiac organoids (hCOs) and hearts of SARS-CoV-2-infected K18-hACE2 mice. Bromodomain and extraterminal family inhibitors (BETi) recover dysfunction in hCOs and completely prevent cardiac dysfunction and death in a mouse cytokine-storm model. Additionally, BETi decreases transcription of genes in the viral response, decreases ACE2 expression, and reduces SARS-CoV-2 infection of cardiomyocytes. Together, BETi, including the Food and Drug Administration (FDA) breakthrough designated drug, apabetalone, are promising candidates to prevent COVID-19 mediated cardiac damage.Richard J. Mills, Sean J. Humphrey, Patrick R.J. Fortuna, Mary Lor, Simon R. Foster, Gregory A. Quaife-Ryan, Rebecca L. Johnston, Troy Dumenil, Cameron Bishop, Rajeev Rudraraju, Daniel J. Rawle, Thuy Le, Wei Zhao, Leo Lee, Charley Mackenzie-Kludas, Neda R. Mehdiabadi, Christopher Halliday, Dean Gilham, Li Fu, Stephen J. Nicholls, Jan Johansson, Michael Sweeney, Norman C.W. Wong, Ewelina Kulikowski, Kamil A. Sokolowski, Brian W.C. Tse, Lynn Devilee, Holly K. Voges, Liam T. Reynolds, Sophie Krumeich, Ellen Mathieson, Dad Abu-Bonsrah, Kathy Karavendzas, Brendan Griffen, Drew Titmarsh, David A. Elliott, James McMahon, Andreas Suhrbier, Kanta Subbarao, Enzo R. Porrello, Mark J. Smyth, Christian R. Engwerda, Kelli P.A. MacDonald, Tobias Bald, David E. James, and James E. Hudso

Monoclonal Antibodies Specific for SARS-CoV-2 Spike Protein Suitable for Multiple Applications for Current Variants of Concern

The global coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spawned an ongoing demand for new research reagents and interventions. Herein we describe a panel of monoclonal antibodies raised against SARS-CoV-2. One antibody showed excellent utility for immunohistochemistry, clearly staining infected cells in formalin-fixed and paraffin embedded lungs and brains of mice infected with the original and the omicron variants of SARS-CoV-2. We demonstrate the reactivity to multiple variants of concern using ELISAs and describe the use of the antibodies in indirect immunofluorescence assays, Western blots, and rapid antigen tests. Finally, we illustrate the ability of two antibodies to reduce significantly viral tissue titers in K18-hACE2 transgenic mice infected with the original and an omicron isolate of SARS-CoV-2