Immunological study of COVID-19 vaccine candidate based on recombinant spike trimer protein from different SARS-CoV-2 variants of concern

The emergency of new SARS-CoV-2 variants that feature increased immune escape marks an urgent demand for better vaccines that will provide broader immunogenicity. Here, we evaluated the immunogenic capacity of vaccine candidates based on the recombinant trimeric spike protein (S) of different SARS-CoV-2 variants of concern (VOC), including the ancestral Wuhan, Beta and Delta viruses. In particular, we assessed formulations containing either single or combined S protein variants. Our study shows that the formulation containing the single S protein from the ancestral Wuhan virus at a concentration of 2µg (SW2-Vac 2µg) displayed in the mouse model the highest IgG antibody levels against all the three (Wuhan, Beta, and Delta) SARS-CoV-2 S protein variants tested. In addition, this formulation induced significantly higher neutralizing antibody titers against the three viral variants when compared with authorized Gam-COVID-Vac-rAd26/rAd5 (Sputnik V) or ChAdOx1 (AstraZeneca) vaccines. SW2-Vac 2µg was also able to induce IFN-gamma and IL-17, memory CD4 populations and follicular T cells. Used as a booster dose for schedules performed with different authorized vaccines, SW2-Vac 2µg vaccine candidate also induced higher levels of total IgG and IgG isotypes against S protein from different SARS-CoV-2 variants in comparison with those observed with homologous 3-dose schedule of Sputnik V or AstraZeneca. Moreover, SW2-Vac 2µg booster induced broadly strong neutralizing antibody levels against the three tested SARS-CoV-2 variants. SW2-Vac 2µg booster also induced CD4+ central memory, CD4+ effector and CD8+ populations. Overall, the results demonstrate that SW2-Vac 2 µg is a promising formulation for the development of a next generation COVID-19 vaccine.Fil: Rudi, Erika. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Martin Aispuro, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Zurita, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: González López Ledesma, María Mora. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Bottero, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Malito, Juan Pablo Alfonso. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología e Innovaciones Tecnológicas. Grupo Vinculado Incuinta al IVIT | Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Virología e Innovaciones Tecnológicas. Grupo Vinculado Incuinta al IVIT; ArgentinaFil: Gabrielli, Magali. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Gaillard, María Emilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Stuible, Matthew. National Research Council Canada, Montreal; CanadáFil: Durocher, Yves. National Research Council Canada, Montreal; CanadáFil: Gamarnik, Andrea Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Wigdorovitz, Andrés. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion En Ciencias Veterinarias y Agronomicas. Instituto de Virologia E Innovaciones Tecnologicas. Grupo Vinculado Incuinta Al Ivit | Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Virologia E Innovaciones Tecnologicas. Grupo Vinculado Incuinta Al Ivit.; ArgentinaFil: Hozbor, Daniela Flavia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentin

Ku-0063794 is a specific inhibitor of the mammalian target of rapamycin (mTOR)

mTOR (mammalian target of rapamycin) stimulates cell growth by phosphorylating and promoting activation of AGC (protein kinase A/protein kinase G/protein kinase C) family kinases such as Akt (protein kinase B), S6K (p70 ribosomal S6 kinase) and SGK (serum and glucocorticoid protein kinase). mTORC1 (mTOR complex-1) phosphorylates the hydrophobic motif of S6K, whereas mTORC2 phosphorylates the hydrophobic motif of Akt and SGK. In the present paper we describe the small molecule Ku-0063794, which inhibits both mTORC1 and mTORC2 with an IC50 of ∼10 nM, but does not suppress the activity of 76 other protein kinases or seven lipid kinases, including Class 1 PI3Ks (phosphoinositide 3-kinases) at 1000-fold higher concentrations. Ku-0063794 is cell permeant, suppresses activation and hydrophobic motif phosphorylation of Akt, S6K and SGK, but not RSK (ribosomal S6 kinase), an AGC kinase not regulated by mTOR. Ku-0063794 also inhibited phosphorylation of the T-loop Thr308 residue of Akt phosphorylated by PDK1 (3-phosphoinositide-dependent protein kinase-1). We interpret this as implying phosphorylation of Ser473 promotes phosphorylation of Thr308 and/or induces a conformational change that protects Thr308 from dephosphorylation. In contrast, Ku-0063794 does not affect Thr308 phosphorylation in fibroblasts lacking essential mTORC2 subunits, suggesting that signalling processes have adapted to enable Thr308 phosphorylation to occur in the absence of Ser473 phosphorylation. We found that Ku-0063794 induced a much greater dephosphorylation of the mTORC1 substrate 4E-BP1 (eukaryotic initiation factor 4E-binding protein 1) than rapamycin, even in mTORC2-deficient cells, suggesting a form of mTOR distinct from mTORC1, or mTORC2 phosphorylates 4E-BP1. Ku-0063794 also suppressed cell growth and induced a G1-cell-cycle arrest. Our results indicate that Ku-0063794 will be useful in delineating the physiological roles of mTOR and may have utility in treatment of cancers in which this pathway is inappropriately activated

Search for Decaying Dark Matter in the Virgo Cluster of Galaxies with HAWC

The decay or annihilation of dark matter particles may produce a steady flux of very-high-energy gamma rays detectable above the diffuse background. Nearby clusters of galaxies provide excellent targets to search for the signatures of particle dark matter interactions. In particular, the Virgo cluster spans several degrees across the sky and can be efficiently probed with a wide field-of-view instrument. The High Altitude Water Cherenkov (HAWC) observatory, due to its wide field of view and sensitivity to gamma rays at an energy scale of 300 GeV--100 TeV is well-suited for this search. Using 2141 days of data, we search for gamma-ray emission from the Virgo cluster, assuming well-motivated dark matter sub-structure models. Our results provide some of the strongest constraints on the decay lifetime of dark matter for masses above 10 TeV.Comment: 7 pages, 3 figures. Submitted to PR

Validation of standardized data formats and tools for ground-level particle-based gamma-ray observatories

Ground-based gamma-ray astronomy is still a rather young field of research,with strong historical connections to particle physics. This is why mostobservations are conducted by experiments with proprietary data and analysissoftware, as it is usual in the particle physics field. However in recentyears, this paradigm has been slowly shifting towards the development and useof open-source data formats and tools, driven by upcoming observatories such asthe Cherenkov Telescope Array (CTA). In this context, a community-driven,shared data format (the gamma-astro-data-format or GADF) and analysis toolssuch as Gammapy and ctools have been developed. So far these efforts have beenled by the IACT community, leaving out other types of ground-based gamma-rayinstruments.We aim to show that the data from ground particle arrays, such asthe High-Altitude Water Cherenkov (HAWC) observatory, is also compatible withthe GADF and can thus be fully analysed using the related tools, in this caseGammapy. We reproduce several published HAWC results using Gammapy and dataproducts compliant with GADF standard. We also illustrate the capabilities ofthe shared format and tools by producing a joint fit of the Crab spectrumincluding data from six different gamma-ray experiments. We find excellentagreement with the reference results, a powerful check of both the publishedresults and the tools involved. The data from particle detector arrays such asthe HAWC observatory can be adapted to the GADF and thus analysed with Gammapy.A common data format and shared analysis tools allow multi-instrument jointanalysis and effective data sharing. Given the complementary nature of pointingand wide-field instruments, this synergy will be distinctly beneficial for thejoint scientific exploitation of future observatories such as the SouthernWide-field Gamma-ray Observatory and CTA.<br

Searching for TeV Dark Matter in Irregular dwarf galaxies with HAWC Observatory

We present the results of dark matter (DM) searches in a sample of 31 dwarf irregular (dIrr) galaxies within the field of view of the HAWC Observatory. dIrr galaxies are DM dominated objects, which astrophysical gamma-ray emission is estimated to be negligible with respect to the secondary gamma-ray flux expected by annihilation or decay of Weakly Interacting Massive Particles (WIMPs). While we do not see any statistically significant DM signal in dIrr galaxies, we present the exclusion limits ($95\%~\text{C.L.}$) for annihilation cross-section and decay lifetime for WIMP candidates with masses between $1$ and $100~\text{TeV}$. Exclusion limits from dIrr galaxies are relevant and complementary to benchmark dwarf Spheroidal (dSph) galaxies. In fact, dIrr galaxies are targets kinematically different from benchmark dSph, preserving the footprints of different evolution histories. We compare the limits from dIrr galaxies to those from ultrafaint and classical dSph galaxies previously observed with HAWC. We find that the contraints are comparable to the limits from classical dSph galaxies and $\thicksim2$ orders of magnitude weaker than the ultrafaint dSph limits.Comment: 22 pages, 11 figures, 3 table

HAWC and Fermi-LAT Detection of Extended Emission from the Unidentified Source 2HWC J2006+341

The discovery of the TeV point source 2HWC J2006+341 was reported in the second HAWC gamma-ray catalog. We present a follow-up study of this source here. The TeV emission is best described by an extended source with a soft spectrum. At GeV energies, an extended source is significantly detected in Fermi-LAT data. The matching locations, sizes and spectra suggest that both gamma-ray detections correspond to the same source. Different scenarios for the origin of the emission are considered and we rule out an association to the pulsar PSR J2004+3429 due to extreme energetics required, if located at a distance of 10.8 kpc.Comment: 12 pages, 2 figures. To appear in ApJ

Gamma-ray Emission from Classical Nova V392 Per: Measurements from Fermi and HAWC

This paper reports on the $\gamma$-ray properties of the 2018 Galactic novaV392 Per, spanning photon energies $\sim$0.1 GeV to 100 TeV by combiningobservations from the Fermi Gamma-ray Space Telescope and the HAWC Observatory.In one of the most rapidly evolving $\gamma$-ray signals yet observed for anova, GeV $\gamma$ rays with a power law spectrum with index $\Gamma = 2.0 \pm0.1$ were detected over eight days following V392 Per's optical maximum. HAWCobservations constrain the TeV $\gamma$-ray signal during this time and alsobefore and after. We observe no statistically significant evidence of TeV$\gamma$-ray emission from V392 Per, but present flux limits. Tests of theextension of the Fermi/LAT spectrum to energies above 5 TeV are disfavored by 2standard deviations (95\%) or more. We fit V392 Per's GeV $\gamma$ rays withhadronic acceleration models, incorporating optical observations, and comparethe calculations with HAWC limits.<br

Constraints on the very high energy gamma-ray emission from short GRBs with HAWC

Many gamma-ray bursts (GRBs) have been observed from radio wavelengths, and afew at very-high energies (VHEs, > 100GeV). The HAWC gamma-ray observatory iswell suited to study transient phenomena at VHEs due to its large field of viewand duty cycle. These features allow for searches of VHE emission and can probedifferent model assumptions of duration and spectra. In this paper, we use datacollected by HAWC between December 2014 and May 2020 to search for emission inthe energy range from 80 to 800 GeV coming from a sample 47 short GRBs thattriggered the Fermi, Swift and Konus satellites during this period. Thisanalysis is optimized to search for delayed and extended VHE emission withinthe first 20 s of each burst. We find no evidence of VHE emission, eithersimultaneous or delayed, with respect to the prompt emission. Upper limits (90%confidence level) derived on the GRB fluence are used to constrain thesynchrotron self-Compton forward-shock model. Constraints for the interstellardensity as low as $10^{-2}$ cm$^{-3}$ are obtained when assuming z=0.3 forbursts with the highest keV-fluences such as GRB 170206A and GRB 181222841.Such a low density makes observing VHE emission mainly from the fast coolingregime challenging.<br

Gamma/Hadron Separation with the HAWC Observatory

The High Altitude Water Cherenkov (HAWC) gamma-ray observatory observesatmospheric showers produced by incident gamma rays and cosmic rays with energyfrom 300 GeV to more than 100 TeV. A crucial phase in analyzing gamma-raysources using ground-based gamma-ray detectors like HAWC is to identify theshowers produced by gamma rays or hadrons. The HAWC observatory records roughly25,000 events per second, with hadrons representing the vast majority($>99.9\%$) of these events. The standard gamma/hadron separation technique inHAWC uses a simple rectangular cut involving only two parameters. This workdescribes the implementation of more sophisticated gamma/hadron separationtechniques, via machine learning methods (boosted decision trees and neuralnetworks), and summarizes the resulting improvements in gamma/hadron separationobtained in HAWC.<br

Galactic Gamma-Ray Diffuse Emission at TeV energies with HAWC Data

The Galactic gamma-ray diffuse emission (GDE) is emitted by cosmic rays (CRs), ultra-relativistic protons and electrons, interacting with gas and electromagnetic radiation fields in the interstellar medium. Here we present the analysis of TeV diffuse emission from a region of the Galactic Plane over the range in longitude of $l\in[43^\circ,73^\circ]$, using data collected with the High Altitude Water Cherenkov (HAWC) detector. Spectral, longitudinal and latitudinal distributions of the TeV diffuse emission are shown. The radiation spectrum is compatible with the spectrum of the emission arising from a CR population with an "index" similar to that of the observed CRs. When comparing with the \texttt{DRAGON} \textit{base model}, the HAWC GDE flux is higher by about a factor of two. Unresolved sources such as pulsar wind nebulae and TeV halos could explain the excess emission. Finally, deviations of the Galactic CR flux from the locally measured CR flux may additionally explain the difference between the predicted and measured diffuse fluxes