Intranasal immunization with the recombinant measles virus encoding the spike protein of SARS-CoV-2 confers protective immunity against COVID-19 in hamsters.

BACKGROUND: As the nasal mucosa is the initial site of infection for COVID-19, intranasal vaccines are more favorable than conventional vaccines. In recent clinical studies, intranasal immunization has been shown to generate higher neutralizing antibodies; however, there is a lack of evidence on sterilizing immunity in the upper airway. Previously, we developed a recombinant measles virus encoding the spike protein of SARS-CoV-2 (rMeV-S), eliciting humoral and cellular immune responses against SARS-CoV-2. OBJECTIVES: In this study, we aim to provide an experiment on nasal vaccines focusing on a measles virus platform as well as injection routes. STUDY DESIGN: Recombinant measles viruses expressing rMeV-S were prepared, and 5 × 105 PFUs of rMeV-S were administered to Syrian golden hamsters via intramuscular or intranasal injection. Subsequently, the hamsters were challenged with inoculations of 1 × 105 PFUs of SARS-CoV-2 and euthanized 4 days post-infection. Neutralizing antibodies and RBD-specific IgG in the serum and RBD-specific IgA in the bronchoalveolar lavage fluid (BALF) were measured, and SARS-CoV-2 clearance capacity was determined via quantitative reverse-transcription PCR (qRT-PCR) analysis and viral titer measurement in the upper respiratory tract and lungs. Immunohistochemistry and histopathological examinations of lung samples from experimental hamsters were conducted. RESULTS: The intranasal immunization of rMeV-S elicits protective immune responses and alleviates virus-induced pathophysiology, such as body weight reduction and lung weight increase in hamsters. Furthermore, lung immunohistochemistry demonstrated that intranasal rMeV-S immunization induces effective SARS-CoV-2 clearance that correlates with viral RNA content, as determined by qRT-PCR, in the lung and nasal wash samples, SARS-CoV-2 viral titers in lung, nasal wash, BALF samples, serum RBD-specific IgG concentration, and RBD-specific IgA concentration in the BALF. CONCLUSION: An intranasal vaccine based on the measles virus platform is a promising strategy owing to the typical route of infection of the virus, the ease of administration of the vaccine, and the strong immune response it elicits.

Recombinant measles virus encoding the spike protein of SARS-CoV-2 efficiently induces Th1 responses and neutralizing antibodies that block SARS-CoV-2 variants.

Owing to the rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants, the development of effective and safe vaccines has become a priority. The measles virus (MeV) vaccine is an attractive vaccine platform as it has been administered to children for more than 40 years in over 100 countries. In this study, we developed a recombinant MeV expressing the full-length SARS-CoV-2 spike protein (rMeV-S) and tested its efficacy using mouse and hamster models. In hCD46Tg mice, two-dose rMeV-S vaccination induced higher Th1 secretion and humoral responses than one-dose vaccination. Interestingly, neutralizing antibodies induced by one-dose and two-dose rMeV-S immunization effectively blocked the entry of the α, ß, γ, and δ variants of SARS-CoV-2. Furthermore, two-dose rMeV-S immunization provided complete protection against SARS-CoV-2 in the hamster model. These results suggest the potential of rMeV-S as a vaccine candidate for targeting SARS-CoV-2 and its variants.

Uncertainty in GRACE/GRACE-follow on global ocean mass change estimates due to mis-modeled glacial isostatic adjustment and geocenter motion.

Global mean sea level has increased about 3 mm/yr over several decades due to increases in ocean mass and changes in sea water density. Ocean mass, accounting for about two-thirds of the increase, can be directly measured by the Gravity Recovery and Climate Experiment (GRACE) and GRACE Follow-On (GFO) satellites. An independent measure is obtained by combining satellite altimetry (measuring total sea level change) and Argo float data (measuring steric changes associated with sea water density). Many previous studies have reported that the two estimates of global mean ocean mass (GMOM) change are in good agreement within stated confidence intervals. Recently, particularly since 2016, estimates by the two methods have diverged. A partial explanation appears to be a spurious variation in steric sea level data. An additional contributor may be deficiencies in Glacial Isostatic Adjustment (GIA) corrections and degree-1 spherical harmonic (SH) coefficients. We found that erroneous corrections for GIA contaminate GRACE/GFO estimates as time goes forward. Errors in GIA corrections affect degree-1 SH coefficients, and degree-1 errors may also be associated with ocean dynamics. Poor estimates of degree-1 SH coefficients are likely an important source of discrepancies in the two methods of estimating GMOM change.

Secular polar motion observed by GRACE.

A long-term drift in polar motion (PM) has been observed for more than a century, and Glacial Isostatic Adjustment (GIA) has been understood as an important cause. However, observed PM includes contributions from other sources, including contemporary climate change and perhaps others associated with Earth's interior dynamics. It has been difficult to separate these effects, because there is considerable scatter among GIA models concerning predicted PM rates. Here we develop a new method to estimate GIA PM using data from the GRACE mission. Changes in GRACE degree 2, order 1 spherical harmonic coefficients are due both to GIA and contemporary surface mass load changes. We estimate the surface mass load contribution to degree 2, order 1 coefficients using GRACE data, relying on higher-degree GRACE coefficients that are dominantly affected by surface loads. Then the GIA PM trend is obtained from the difference between observed PM trend (which includes effects from GIA and surface mass loads) and the estimated PM trend mostly associated with surface mass loads. A previous estimate of the GIA PM trend from PM observations for the period 1900-1978 is toward 79.90° W at a speed of 3.53 mas/year (10.91 cm/year). Our new estimate for the GIA trend is in a direction of 61.77° W at a speed of 2.18 mas/year (6.74 cm/year), similar to the observed PM trend during the early twentieth century. This is consistent with the view that the early twentieth-century trend was dominated by GIA and that more recently there is an increasing contribution from contemporary surface mass load redistribution associated with climate change. Our GIA PM also agrees with the linear mean pole during 1900-2017. Contributions from other solid Earth process such as mantle convection would also produce a linear trend in PM and could be included in our GIA estimate.

Immunization with RBD-P2 and N protects against SARS-CoV-2 in nonhuman primates.

Since the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), various vaccines are being developed, with most vaccine candidates focusing on the viral spike protein. Here, we developed a previously unknown subunit vaccine comprising the receptor binding domain (RBD) of the spike protein fused with the tetanus toxoid epitope P2 (RBD-P2) and tested its efficacy in rodents and nonhuman primates (NHPs). We also investigated whether the SARS-CoV-2 nucleocapsid protein (N) could increase vaccine efficacy. Immunization with N and RBD-P2 (RBDP2/N) + alum increased T cell responses in mice and neutralizing antibody levels in rats compared with those obtained using RBD-P2 + alum. Furthermore, in NHPs, RBD-P2/N + alum induced slightly faster SARS-CoV-2 clearance than that induced by RBD-P2 + alum, albeit without statistical significance. Our study supports further development of RBD-P2 as a vaccine candidate against SARS-CoV-2. Also, it provides insights regarding the use of N in protein-based vaccines against SARS-CoV-2.

Antarctic ice mass variations from 1979 to 2017 driven by anomalous precipitation accumulation.

Antarctic ice mass balance is determined by precipitation and ice discharge, and understanding their relative contributions to contemporary Antarctic ice mass change is important to project future ice mass loss and resulting sea level rise. There has been evidence that anomalous precipitation affects Antarctic ice mass loss estimates, and thus the precipitation contribution should be understood and considered in future projections. In this study, we revisit changes in Antarctic ice mass over recent decades and examine precipitation contributions over this period. We show that accumulated (time-integrated) precipitation explains most inter-annual anomalies of Antarctic ice mass change during the GRACE period (2003-2017). From 1979 to 2017, accumulated Antarctic precipitation contributes to significant ice mass loss acceleration in the Pacific sector and deceleration in the Atlantic-Indian Sectors, forming a bi-polar spatial pattern. Principal component analysis reveals that such a bi-polar pattern is likely modulated by the Southern Annular Mode (SAM). We also find that recent ice mass loss acceleration in 2007 is related to a variation in precipitation accumulation. Overall ice discharge has accelerated at a steady rate since 1992, but has not seen a recent abrupt increase.

Global sea level change signatures observed by GRACE satellite gravimetry.

Ice mass loss on land results in sea level rise, but its rate varies regionally due to gravitational self-attraction effects. Observing regional sea level rates by ocean mass change using the Gravity Recovery and Climate Experiment (GRACE) gravity solutions is difficult due to GRACE's spatial resolution (~a few hundred km) and other limitations. Here we estimate regional sea level mass change using GRACE data (without contributions from temperature and salinity variations) by addressing these limitations: restoring spatially spread and attenuated signals in post-processed GRACE data; constraining ocean mass distribution to conform to the changing geoid; and judging specific corrections applied to GRACE data including a new geocenter estimate. The estimated global sea level mass trend for 2003-2014 is 2.14 ± 0.12 mm/yr. Regional trends differ considerably among ocean basins, ranging from -0.5 mm/yr in the Arctic to about 2.4 mm/yr in the Indian and South Atlantic Oceans.

Double-Injected Human Stem Cells Enhance Rehabilitation in TBI Mice Via Modulation of Survival and Inflammation.

Traumatic brain injury (TBI), a complicated form of brain damage, is a major cause of mortality in adults. Following mechanical and structural primary insults, a battery of secondary insults, including neurotransmitter-mediated cytotoxicity, dysregulation of calcium and macromolecule homeostasis, and increased oxidative stress, exacerbate brain injury and functional deficits. Although stem cell therapy is considered to be an alternative treatment for brain injuries, such as TBI and stroke, many obstacles remain. In particular, the time window for TBI treatment with either drugs or stem cells and their efficacy is still vague. Human placenta-derived mesenchymal stem cells (hpMSCs) have received extensive attention in stem cell therapy because they can be acquired in large numbers without ethical issues and because of their immune-modulating capacity and effectiveness in several diseases, such as Alzheimer's disease and stroke. Here, we tested the feasibility of hpMSCs for TBI treatment with an animal model and attempted to identify appropriate time points for cell treatments. Double injections at 4 and 24 h post-injury significantly reduced the infarct size and suppressed astrocyte and microglial activation around the injury. With reduced damage, double-injected mice showed enhanced anti-inflammatory- and TNF-α receptor 2 (TNFR2)-associated survival signals and suppressed pro-inflammatory and oxidative responses. In addition, double-treated TBI mice displayed restored sensory motor functions and reduced neurotoxic Aß42 plaque formation around the damaged areas. In this study, we showed the extended therapeutic potentials of hpMSCs and concluded that treatment within an appropriate time window is critical for TBI recovery.

Nasal immunization with M cell-targeting ligand-conjugated ApxIIA toxin fragment induces protective immunity against Actinobacillus pleuropneumoniae infection in a murine model.

Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia and severe economic loss in the swine industry has been caused by the infection. Therefore, the development of an effective vaccine against the bacteria is necessary. ApxII toxin, among several virulence factors expressed by the bacteria, is considered to be a promising vaccine candidate because ApxII toxin not only accompanies cytotoxic and hemolytic activities, but is also expressed in all 15 serotypes of bacteria except serotypes 10 and 14. In this study, we identified the peptide ligand capable of targeting the ligand-conjugated ApxIIA #5 fragment antigen to nasopharynx-associated lymphoid tissue. It was found that nasal immunization with ligand-conjugated ApxIIA #5 induced efficient mucosal and systemic immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. More importantly, the nasal immunization induced protective immunity against nasal challenge infection of the bacteria, which was confirmed by histopathological studies and bacterial clearance after challenge infection. Collectively, we confirmed that the ligand capable of targeting the ligand-conjugated antigen to nasopharynx-associated lymphoid tissue can be used as an effective nasal vaccine adjuvant to induce protective immunity against A. pleuropneumoniae infection.

Surface mass balance contributions to acceleration of Antarctic ice mass loss during 2003-2013.

Recent observations from satellite gravimetry (the Gravity Recovery and Climate Experiment (GRACE) mission) suggest an acceleration of ice mass loss from the Antarctic Ice Sheet (AIS). The contribution of surface mass balance changes (due to variable precipitation) is compared with GRACE-derived mass loss acceleration by assessing the estimated contribution of snow mass from meteorological reanalysis data. We find that over much of the continent, the acceleration can be explained by precipitation anomalies. However, on the Antarctic Peninsula and other parts of West Antarctica, mass changes are not explained by precipitation and are likely associated with ice discharge rate increases. The total apparent GRACE acceleration over all of the AIS between 2003 and 2013 is -13.6 ± 7.2 Gt/yr2. Of this total, we find that the surface mass balance component is -8.2 ± 2.0 Gt/yr2. However, the GRACE estimate appears to contain errors arising from the atmospheric pressure fields used to remove air mass effects. The estimated acceleration error from this effect is about 9.8 ± 5.8 Gt/yr2. Correcting for this yields an ice discharge acceleration of -15.1 ± 6.5 Gt/yr2.

Nasal immunization with major epitope-containing ApxIIA toxin fragment induces protective immunity against challenge infection with Actinobacillus pleuropneumoniae in a murine model.

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439-801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.

Characterization of antigenic determinants in ApxIIA exotoxin capable of inducing protective immunity to Actinobacillus pleuropneumoniae challenge.

Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia. Among the virulence factors of the pathogen, ApxIIA, a bacterial exotoxin, is expressed by many serotypes and presents a plausible target for vaccine development. We characterized the region within ApxIIA that induces a protective immune response against bacterial infection using mouse challenge model. Recombinant proteins spanning the length of ApxIIA were produced and antiserum to the full-length ApxIIA was induced in mice. This antiserum recognized fragments #2, #3 and #5 with high binding specificity, but showed poor recognition for fragments #1 and #4. Of the antisera induced in mice by injection of each fragments, only the antiserum to fragment #4 failed to efficiently recognize the full-length antigen, although the individual antisera recognized their cognate antigens with almost equal efficiency. The protective potency of the immunogenic proteins against a challenge injection of bacteria in vivo correlated well with the antibody titer. Fragment #5 induced the highest level of protective activity, comparable to that by the full-length protein. These results support the use of fragment #5 to produce a vaccine against A. pleuropneumoniae challenge, since the small antigen peptide is easier to handle than is the full-length protein and can be expressed efficiently in heterologous expression systems.

The M cell-targeting ligand promotes antigen delivery and induces antigen-specific immune responses in mucosal vaccination.

Oral mucosal immunization can induce protective immunity in both systemic compartments and the mucosa. Successful mucosal immunization depends on Ag delivery to the mucosal immune induction site. The high transcytotic activity of M cells within the mucosa makes these cells attractive targets for mucosal Ag delivery, although it remains unclear whether delivery of Ag to M cells only can guarantee the induction of effective immune responses. In this study, we evaluated the ability of an M cell-targeting ligand with adjuvant activity to induce immunity against ligand-fused Ag. We selected M cell-targeting ligands through biopanning of a phage display library against differentiated in vitro M-like cells and produced the recombinant Ags fused to the selected ligands using the model Ag. One of the selected peptide ligands, Co1, promoted the binding of ligand-fused Ag to mouse Peyer's patch M cells and human M-like cells that had been defined by binding with the M cell-specific and anti-GP2 Abs. In addition, Co1 ligand enhanced the uptake of fused Ag by immunogenic tissue in an ex vivo loop assay and in vivo oral administration experiments. After oral administration, the ligand-fused Ag enhanced immune responses against the fused Ag compared with those of the control Ag without ligand. In addition, this use of the ligand supported a skewed Th2-type immune response against the fused Ag. Collectively, these results suggest that the ligand selected through biopanning against cultured M-like cells could be used as an adjuvant for targeted Ag delivery into the mucosal immune system to enhance immune induction.