A SARS-CoV-2 vaccine based on conjugation of SARS-CoV-2 RBD with IC28 peptide and mannan.

SARS-CoV-2 is a particularly transmissible virus that causes a severe respiratory disease known as COVID-19. Safe and effective vaccines are urgently needed to combat the COVID-19 pandemic. The receptor-binding domain (RBD) of SARS-CoV-2 spike protein elicits most neutralizing antibodies during viral infection and is an ideal antigen for vaccine development. In particular, RBD expressed by E. coli is amenable to low cost and high-yield manufacturability. The adjuvant is necessitated to improve the immunogenicity of RBD. IC28, a TLR5-dependent adjuvant, is a peptide from bacterial flagellin. Mannan is a ligand of TLR-4 or TLR-2 and a polysaccharide adjuvant. Here, IC28 and mannan were both covalently conjugated with RBD from E. coli. The conjugate (RBD-IC28-M) elicited high RBD-specific IgG titers, and a neutralization antibody titer of 201.4. It induced high levels of Th1-type cytokines (IFN-γ) and Th2-type cytokines (IL-5 and IL-10), along with high antigenicity and no apparent toxicity to the organs. The mouse sera of the RBD-IC28-M group competitively interfered with the interaction of RBD and ACE2. Thus, conjugation with IC28 and mannan additively enhanced the humoral and cellular immunity. Our study was expected to provide the feasibility to develop an affordable, easily scalable, effective vaccine SARS-CoV-2 vaccine.

Conjugation with 8-arm PEG and CRM197 enhances the immunogenicity of SARS-CoV-2 ORF8 protein.

Safe and effective vaccines are urgently needed to combat the COVID-19 pandemic. However, the SARS-CoV-2 variants raise concerns about the effectiveness of vaccines. As a SARS-CoV-2 antigen target, ORF8 strongly inhibits the IFN-ß and NF-κB-responsive promoter, and can be potentially used for the development of SARS-CoV-2 vaccine. However, it is necessary to improve the immunogenicity of ORF8 by adjuvants or delivery systems. CRM197 was a carrier protein with the ability to activate T helper cells for antigens. Eight-arm PEG could conjugate multiple antigen molecules in one entity with inherent adjuvant effect. In the present study, ORF8 was conjugated with CRM197 and 8-arm PEG, respectively. The cellular and humoral immune responses to the conjugates (ORF8-CRM and ORF8-PEG) were evaluated in the BALB/c mice. As compared with ORF8-CRM and ORF8 administrated with aluminum adjuvant (ORF8/AL), ORF8-PEG induced a higher ORF8-specific IgG titer (2.6 × 104), higher levels of cytokines (IFN-γ, TNF-α, IFN-ß, and IL-5), stronger splenocyte proliferation. Thus, conjugation with 8-arm PEG was an effective method to improve the immune response to ORF8. Moreover, ORF8-PEG did not lead to apparent toxicity to the cardiac, liver and renal functions. ORF8-PEG was expected to act as an effective vaccine to provide the immune protection against SARS-CoV-2.

Characteristics of air quality in different climatic zones of China during the COVID-19 lockdown.

The diverse climate types and the complex anthropogenic source emissions in China lead to the great regional differences of air pollution mechanisms. The COVID-19 lockdown has given us a precious opportunity to understand the effect of weather conditions and anthropogenic sources on the distribution of air pollutants in different climate zones. In this study, to understand the impact of meteorological and socio-economic factors on air pollution during COVID-19 lockdown, we divided 358 Chinese cities into eight climate regions. Temporal, spatial and diurnal variations of six major air pollutants from January 1 to April 18, 2020 were analyzed. The differences in the characteristics of air pollutants in different climate zones were obvious. PM2.5 reduced by 59.0%-64.2% in cold regions (North-East China (NEC) and North-Western (NW)), while O3 surged by 99.0%-99.9% in warm regions (Central South (CS) and Southern Coast (SC)). Diurnal variations of atmospheric pollutants were also more prominent in cold regions. Moreover, PM2.5, PM10, CO and SO2 showed more prominent reductions (20.5%-64.2%) in heating regions (NEC, NW, NCP and MG) than no-heating regions (0.8%-48%). Climate has less influence on NO2, which dropped by 41.2%-57.1% countrywide during the lockdown. The influences of weather conditions on the atmospheric pollutants in different climate zones were different. The wind speed was not the primary reason for the differences in air pollutants in different climate zones. Temperature, precipitation, and air pollution emissions led to prominent regional differences in air pollutants throughout the eight climates. The effect of temperature on PM, SO2, CO, and NO2 varied obviously with the latitude, at which condition temperature was negatively correlated to PM, SO2, CO, and NO2 in the north but positively in the south. The temperature was positively correlated to ozone in different climate zones, and the correlation was the highest in NEC and the lowest in SC. The rainfall has a strong removal effect on atmospheric pollutants in the climate regions with more precipitation, but it increases the pollutant concentrations in the climate regions with less precipitation. In regions with more emission sources, air pollutants experienced more significant variations and returned to pre-lockdown levels earlier.

Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli.

SARS-CoV-2 is responsible for a disruptive worldwide viral pandemic, and renders a severe respiratory disease known as COVID-19. Spike protein of SARS-CoV-2 mediates viral entry into host cells by binding ACE2 through the receptor-binding domain (RBD). RBD is an important target for development of virus inhibitors, neutralizing antibodies, and vaccines. RBD expressed in mammalian cells suffers from low expression yield and high cost. E. coli is a popular host for protein expression, which has the advantage of easy scalability with low cost. However, RBD expressed by E. coli (RBD-1) lacks the glycosylation, and its antigenic epitopes may not be sufficiently exposed. In the present study, RBD-1 was expressed by E. coli and purified by a Ni Sepharose Fast Flow column. RBD-1 was structurally characterized and compared with RBD expressed by the HEK293 cells (RBD-2). The secondary structure and tertiary structure of RBD-1 were largely maintained without glycosylation. In particular, the major ß-sheet content of RBD-1 was almost unaltered. RBD-1 could strongly bind ACE2 with a dissociation constant (KD) of 2.98 × 10-8 M. Thus, RBD-1 was expected to apply in the vaccine development, screening drugs and virus test kit.

Engineering a self-navigated MnARK nanovaccine for inducing potent protective immunity against novel coronavirus.

Effective vaccines are vital to fight against the COVID-19 global pandemic. As a critical component of a subunit vaccine, the adjuvant is responsible for strengthening the antigen-induced immune responses. Here, we present a new nanovaccine that comprising the Receptor-Binding Domain (RBD) of spike protein and the manganese nanoadjuvant (MnARK), which induces humoral and cellular responses. Notably, even at a 5-fold lower antigen dose and with fewer injections, the MnARK vaccine immunized mice showed stronger neutralizing abilities against the infection of the pseudovirus (~270-fold) and live coronavirus (>8-fold) in vitro than that of Alum-adsorbed RBD vaccine (Alu-RBD). Furthermore, we found that the effective co-delivery of RBD antigen and MnARK to lymph nodes (LNs) elicited an increased cellular internalization and the activation of immune cells, including DCs, CD4+ and CD8+ T lymphocytes. Our findings highlight the importance of MnARK adjuvant in the design of novel coronavirus vaccines and provide a rationale strategy to design protective vaccines through promoting cellular internalization and the activation of immune-related pathways.

Impact of urbanization on air quality in the Yangtze River Delta during the COVID-19 lockdown in China

The urban agglomeration of Yangtze River Delta (YRD) is symbol of China's rapid urbanization during the past decades. Urbanization can significantly impact land-cover properties, surface heating, and emissions of air pollutants. To control the spread of COVID-19, China imposed very rigorous restrictions, leading to dramatic reductions in air pollutants (except O3) from satellite and ground-based data. As such, inter-transportation of air pollutants was weak during the lockdown, which was conducive to discuss the impacts of urbanization on the air quality. During the lockdown, the rates of surface PM2.5, PM10, SO2, NO2 and CO reductions in different urban types ranged from 6.6% to 62.4% in the YRD. Urbanization exerted great impacts on the pollutant variations in urban agglomerations despite such large decreases in primary pollution in YRD. Lower values of AOD and tropospheric NO2 columns were noticeably observed over large cities during the lockdown. The extents of surface PM, SO2, NO2 and CO reductions in large cities (first-tier and second-tier) were found to be larger (4.7%-10.6%) than those in small-medium cities (third-tier and fourth-tier) during the lockdown, which was also the case for the extent of the increase (33.0% - 53.0%) in O3. PCA analysis revealed that the PM decreases in large cities made greater improvement in the air quality compared with the small cities during lockdown, while the urbanization had non-obvious influence on the photochemical reactions. It is imperative to adopt policies and programs to mitigate the air pollution in urban agglomerations in the fast urbanization process.

Air pollutant variations in Suzhou during the 2019 novel coronavirus (COVID-19) lockdown of 2020: High time-resolution measurements of aerosol chemical compositions and source apportionment.

To control the spread of the 2019 novel coronavirus (COVID-19), China imposed rigorous restrictions, which resulted in great reductions in pollutant emissions. This study examines the characteristics of air pollutants, including PM2.5 (particles with aerodynamic diameters < 2.5 µm), gas pollutants, water-soluble ions (WSIs), black carbon (BC) and elements, as well as the source apportionment of PM2.5 in Suzhou before, during and after the Chinese New Year (CNY) holiday of 2020 (when China was under an unprecedented state of lockdown to restrict the COVID-19 outbreak). Compared to those before CNY, PM2.5, BC, SNA (sulfate, nitrate and ammonium), other ions, elements, and NO2 and CO mass concentrations decreased by 9.9%-64.0% during CNY. The lockdown policy had strong (weak) effects on the diurnal variations in aerosol chemical compositions (gas pollutants). Compared to those before CNY, source concentrations and contributions of vehicle exhaust during CNY decreased by 72.9% and 21.7%, respectively. In contrast, increased contributions from coal combustion and industry were observed during CNY, which were recorded to be 2.9 and 1.7 times higher than those before CNY, respectively. This study highlights that the lockdown policy that was imposed in Suzhou during CNY not only reduced the mass concentrations of air pollutants but also modified their diurnal variations and the source contributions of PM2.5, which revealed the complex responses of PM2.5 sources to the rare, low emissions of anthropogenic pollutants that occurred during the COVID-19 lockdown.

Characterization of the aerosol chemical composition during the COVID-19 lockdown period in Suzhou in the Yangtze River Delta, China.

To control the spread of COVID-19, rigorous restrictions have been implemented in China, resulting in a great reduction in pollutant emissions. In this study, we evaluated the air quality in the Yangtze River Delta during the COVID-19 lockdown period using satellite and ground-based data, including particle matter (PM), trace gases, water-soluble ions (WSIs) and black carbon (BC). We found that the impacts of lockdown policy on air quality cannot be accurately assessed using MODIS aerosol optical depth (AOD) data, whereas the tropospheric nitrogen dioxide (NO2) vertical column density can well reflect the influences of these restrictions on human activities. Compared to the pre-COVID period, the PM2.5, PM10, NO2, carbon monoxide (CO), BC and WSIs during the lockdown in Suzhou were observed to decrease by 37.2%, 38.3%, 64.5%, 26.1%, 53.3% and 58.6%, respectively, while the sulfur dioxide (SO2) and ozone (O3) increased by 1.5% and 104.7%. The WSIs ranked in the order of NO3- > NH4+ > SO42- > Cl- > Ca2+ > K+ > Mg2+ > Na+ during the lockdown period. By comparisons with the ion concentrations during the pre-COVID period, we found that the ions NO3-, NH4+, SO42-, Cl-, Ca2+, K+ and Na+ decreased by 66.3%, 48.8%, 52.9%, 56.9%, 57.9% and 76.3%, respectively, during the lockdown, in contrast to Mg2+, which increased by 30.2%. The lockdown policy was found to have great impacts on the diurnal variations of Cl-, SO42-, Na+ and Ca2+.

Importance of meteorology in air pollution events during the city lockdown for COVID-19 in Hubei Province, Central China.

Compared with the 21-year climatological mean over the same period during 2000-2020, the aerosol optical depth (AOD) and Angstrom exponent (AE) during the COVID-19 lockdown (January 24-February 29, 2020) decreased and increased, respectively, in most regions of Central-Eastern China (CEC). The AOD (AE) values decreased (increased) by 39.2% (29.4%) and 31.0% (45.3%) in Hubei and Wuhan, respectively, because of the rigorous restrictions. These inverse changes reflected the reduction of total aerosols in the air and the contribution of the increase in fine-mode particles during the lockdown. The surface PM2.5 had a distinct spatial distribution over CEC during the lockdown, with high concentrations in North China and East China. In particular, relatively high PM2.5 concentrations were notable in the lower flatlands of Hubei Province in Central China, where six PM2.5 pollution events were identified during the lockdown. Using the observation data and model simulations, we found that 50% of the pollution episodes were associated with the long-range transport of air pollutants from upstream CEC source regions, which then converged in the downstream Hubei receptor region. However, local pollution was dominant for the remaining episodes because of stagnant meteorological conditions. The long-range transport of air pollutants substantially contributed to PM2.5 pollution in Hubei, reflecting the exceptional importance of meteorology in regional air quality in China.