An electrochemical aptamer-sensing strategy based on a Ti3C2Tx MXene synergistic Ti-MOF amplification signal for highly sensitive detection of zearalenone.

A refined electrochemical aptamer sensing technique using PEI@Ti-MOF@Ti3C2Tx-MXene was developed for the sensitive detection of ZEN in food samples. A titanium-based metal-organic skeleton (NH2-MIL-125) was synthesized in situ using 2-aminoterephthalic acid as the organic ligand and tetrabutyl titanate as the metal center, followed by the simultaneous hybridization of Ti3C2Tx-MXene to synthesize a Ti-MOF@Ti3C2Tx-MXene composite material. These composites were subsequently functionalized with PEI and covalently linked to form a sensing platform on gold electrodes. Integrating a metal-organic framework (MOF) with MXene materials not only improved the electrochemical properties compared to those of individual elements but also decreased the stacking effect and increased the number of binding sites for the aptamer. The limit of detection (LOD) of this sensor was 1.64 fg mL-1. Additionally, the sensor could efficaciously detect ZEN in cornmeal and beer samples, exhibiting outstanding stability, reproducibility, and selectivity. This highlighted its effectiveness in applications in quality supervision and food safety.

Electrochemical aptamer sensor based on AgNPs@PDANSs and "sandwich" structure guidance for the detection of tobramycin in water samples.

In this study, an ultrasensitive detection platform for tobramycin (TOB) was developed, featuring a "sandwich" structure guided by AgNCs@PDANSs and Thi-AuNCs@ZnONSs. To address the issue of large background current peak signals in tagless sensors, Thi-AuNCs@ZnONSs composites were synthesized as signal tags. Zinc oxide nanosheets (ZnONSs) served as the loading agent, and AuNCs with the electroactive molecule Thi acted as carriers. Furthermore, AgNPs@PDANSs nanocomposites, possessing excellent electrical conductivity and large specific surface areas, were prepared as substrate materials for the modified electrodes. A "sandwich" structure strategy was also introduced to enhance the accuracy of the electrochemical aptasensor. This strategy, utilizing a dual sequence for target labeling and capture, yielded higher sensitivity and simplified the sensor construction compared to methods employing a single sequence. Under optimal conditions, the detection limit for TOB was established at 1.41 pM, with a detection range of 0.05-5000 nM. The aptasensor was effectively applied in the detection of TOB in tap and lake water, demonstrating outstanding reproducibility, selectivity, and stability. These results may serve as a reference for environmental TOB detection.

Conserved moonlighting protein pyruvate dehydrogenase induces robust protection against Staphylococcus aureus infection.

Developing an effective Staphylococcus aureus (S. aureus) vaccine has been a challenging endeavor, as demonstrated by numerous failed clinical trials over the years. In this study, we formulated a vaccine containing a highly conserved moonlighting protein, the pyruvate dehydrogenase complex E2 subunit (PDHC), and showed that it induced strong protective immunity against epidemiologically relevant staphylococcal strains in various murine disease models. While antibody responses contributed to bacterial control, they were not essential for protective immunity in the bloodstream infection model. Conversely, vaccine-induced systemic immunity relied on γδ T cells. It has been suggested that prior S. aureus exposure may contribute to the reduction of vaccine efficacy. However, PDHC-induced protective immunity still facilitated bacterial clearance in mice previously exposed to S. aureus. Collectively, our findings indicate that PDHC is a promising serotype-independent vaccine candidate effective against both methicillin-sensitive and methicillin-resistant S. aureus isolates.

Development of a live attenuated vaccine candidate for equid alphaherpesvirus 1 control: a step towards efficient protection.

Equid alphaherpesvirus 1 (EqAHV1) is a viral pathogen known to cause respiratory disease, neurologic syndromes, and abortion storms in horses. Currently, there are no vaccines that provide complete protection against EqAHV1. Marker vaccines and the differentiation of infected and vaccinated animals (DIVA) strategy are effective for preventing and controlling outbreaks but have not been used for the prevention of EqAHV1 infection. Glycoprotein 2 (gp2), located on the envelope of viruses (EqAHV1), exhibits high antigenicity and functions as a molecular marker for DIVA. In this study, a series of EqAHV1 mutants with deletion of gp2 along with other virulence genes (TK, UL24/TK, gI/gE) were engineered. The mutant viruses were studied in vitro and then in an in vivo experiment using Golden Syrian hamsters to assess the extent of viral attenuation and the immune response elicited by the mutant viruses in comparison to the wild-type (WT) virus. Compared with the WT strain, the YM2019 Δgp2, ΔTK/gp2, and ΔUL24/TK/gp2 strains exhibited reduced growth in RK-13 cells, while the ΔgI/gE/gp2 strain exhibited significantly impaired proliferation. The YM2019 Δgp2 strain induced clinical signs and mortality in hamsters. In contrast, the YM2019 ΔTK/gp2 and ΔUL24/TK/gp2 variants displayed diminished pathogenicity, causing no observable clinical signs or fatalities. Immunization with nasal vaccines containing YM2019 ΔTK/gp2 and ΔUL24/TK/gp2 elicited a robust immune response in hamsters. In particular, compared with the vaccine containing the ΔTK/gp2 strain, the vaccine containing the ΔUL24/TK/gp2 strain demonstrated enhanced immune protection upon challenge with the WT virus. Furthermore, an ELISA for gp2 was established and refined to accurately differentiate between infected and vaccinated animals. These results confirm that the ΔUL24/TK/gp2 strain is a safe and effective live attenuated vaccine candidate for controlling EqAHV1 infection.

Exo â ¢-assisted amplification signal strategy synergized with Au@Pt NFs/CoSe2 for sensitive detection of enrofloxacin.

Overuse of enrofloxacin (ENR) has posed a potential threat to ecosystems and public health, so it is critical to sensitive and accurate determination of ENR residues. In this work, a novel ultra-sensitive and specific electrochemical aptasensor was fabricated based on the cobalt diselenide loaded gold and platinum nanoflowers (Au@Pt NFs/ CoSe2) and Exonuclease III (Exo III)-assisted cycle amplification strategy for the detection of ENR. Au@Pt NFs/ CoSe2 nanosheets as the substrate material, with large surface area, accelerate electron transfer and attach more DNA probes on the electrode substrate, have effectively enhanced the electrochemical performance of the electrode. With the existence of Enrofloxacin (ENR), the aptamer recognizes and binds to ENR, thus the signal probe cDNA was released and immobilized onto the electrode surface to hybridized with methylene blue (MB) labelled DNA (MB-DNA), thereby triggering the Exo III-assisted cycle for further signal amplification. As expected, the prepared aptasensor demonstrated excellent sensitivity and selectivity, with a wide linear range from 5.0 × 10-6 ng/mL to 1.0 × 10-2 ng/mL for ENR, a low detection limit of 1.59 × 10-6 ng/mL. Consequently, this strategy provided a promising avenue for ultrasensitive and accurate detection of ENR in milk samples.

EZH2 Promotes Glioma Cell Proliferation, Invasion, and Migration via Mir-142-3p/KCNQ1OT1/HMGB3 Axis : Running Title: EZH2 Promotes Glioma cell Malignant Behaviors.

This study investigates the role and molecular mechanism of EZH2 in glioma cell proliferation, invasion, and migration. EZH2, miR-142-3p, lncRNA KCNQ1OT1, LIN28B, and HMGB3 expressions in glioma tissues and cells were determined using qRT-PCR or Western blot, followed by CCK-8 assay detection of cell viability, Transwell detection of invasion and migration, ChIP analysis of the enrichment of EZH2 and H3K27me3 on miR-142-3p promoter, dual-luciferase reporter assay and RIP validation of the binding of miR-142-3p-KCNQ1OT1 and KCNQ1OT1-LIN28B, and actinomycin D detection of KCNQ1OT1 and HMGB3 mRNA stability. A nude mouse xenograft model and a lung metastasis model were established. EZH2, KCNQ1OT1, LIN28B, and HMGB3 were highly expressed while miR-142-3p was poorly expressed in gliomas. EZH2 silencing restrained glioma cell proliferation, invasion, and migration. EZH2 repressed miR-142-3p expression by elevating the H3K27me3 level. miR-142-3p targeted KCNQ1OT1 expression, and KCNQ1OT1 bound to LIN28B to stabilize HMGB3 mRNA, thereby promoting its protein expression. EZH2 silencing depressed tumor growth and metastasis in nude mice via the miR-142-3p/KCNQ1OT1/HMGB3 axis. In conclusion, EZH2 curbed miR-142-3p expression, thereby relieving the inhibition of KCNQ1OT1 expression by miR-142-3p, enhancing the binding of KCNQ1OT1 to LIN28B, elevating HMGB3 expression, and ultimately accelerating glioma cell proliferation, invasion, and migration.

Recent strides toward transforming lignin into plastics and aqueous electrolytes for flow batteries.

Lignin is an abundant polyaromatic polymer with a wide range of potential future uses. However, the conversion of lignin into valuable products comes at a cost, and medium- to high-value applications are thus appropriate. Two examples of these are polymers (e.g., as fibers, plasticizers, or additives) and flow batteries (e.g., as redox species). Both of these areas would benefit from lignin-derived molecules with potentially low molecular weight and high (electro)chemical functionality. A promising route to obtain these molecules is oxidative lignin depolymerization, as it enables the formation of targeted compounds with multiple functionalities. An application with high potential in the production of plastics is the synthesis of new sustainable polymers. Employing organic molecules, such as quinones and heterocycles, would constitute an important step toward the sustainability of aqueous flow batteries, and lignin and its derivatives are emerging as redox species, mainly due to their low cost and renewability.

Type-II IFN inhibits SARS-CoV-2 replication in human lung epithelial cells and ex vivo human lung tissues through indoleamine 2,3-dioxygenase-mediated pathways.

Interferons (IFNs) are critical for immune defense against pathogens. While type-I and -III IFNs have been reported to inhibit SARS-CoV-2 replication, the antiviral effect and mechanism of type-II IFN against SARS-CoV-2 remain largely unknown. Here, we evaluate the antiviral activity of type-II IFN (IFNγ) using human lung epithelial cells (Calu3) and ex vivo human lung tissues. In this study, we found that IFNγ suppresses SARS-CoV-2 replication in both Calu3 cells and ex vivo human lung tissues. Moreover, IFNγ treatment does not significantly modulate the expression of SARS-CoV-2 entry-related factors and induces a similar level of pro-inflammatory response in human lung tissues when compared with IFNß treatment. Mechanistically, we show that overexpression of indoleamine 2,3-dioxygenase 1 (IDO1), which is most profoundly induced by IFNγ, substantially restricts the replication of ancestral SARS-CoV-2 and the Alpha and Delta variants. Meanwhile, loss-of-function study reveals that IDO1 knockdown restores SARS-CoV-2 replication restricted by IFNγ in Calu3 cells. We further found that the treatment of l-tryptophan, a substrate of IDO1, partially rescues the IFNγ-mediated inhibitory effect on SARS-CoV-2 replication in both Calu3 cells and ex vivo human lung tissues. Collectively, these results suggest that type-II IFN potently inhibits SARS-CoV-2 replication through IDO1-mediated antiviral response.

Vaccination-Route-Dependent Adjuvanticity of Antigen-Carrying Nanoparticles for Enhanced Vaccine Efficacy.

Vaccination-route-dependent adjuvanticity was identified as being associated with the specific features of antigen-carrying nanoparticles (NPs) in the present work. Here, we demonstrated that the mechanical properties and the decomposability of NP adjuvants play key roles in determining the antigen accessibility and thus the overall vaccine efficacy in the immune system when different vaccination routes were employed. We showed that soft nano-vaccines were associated with more efficient antigen uptake when administering subcutaneous (S.C.) vaccination, while the slow decomposition of hard nano-vaccines promoted antigen uptake when intravenous (I.V.) vaccination was employed. In comparison to the clinically used aluminum (Alum) adjuvant, the NP adjuvants were found to stimulate both humoral and cellular immune responses efficiently, irrespective of the vaccination route. For vaccination via S.C. and I.V. alike, the NP-based vaccines show excellent protection for mice from Staphylococcus aureus (S. aureus) infection, and their survival rates are 100% after lethal challenge, being much superior to the clinically used Alum adjuvant.

Enhancing immune responses of ESC-based TAA cancer vaccines with a novel OMV delivery system.

Embryonic stem cell (ESC)-derived epitopes can act as therapeutic tumor vaccines against different types of tumors Jin (Adv Healthc Mater 2023). However, these epitopes have poor immunogenicity and stimulate insufficient CD8+ T cell responses, which motivated us to develop a new method to deliver and enhance their effectiveness. Bacterial outer membrane vesicles (OMVs) can serve as immunoadjuvants and act as a delivery vector for tumor antigens. In the current study, we engineered a new OMV platform for the co-delivery of ESC-derived tumor antigens and immune checkpoint inhibitors (PD-L1 antibody). An engineered Staphylococcal Protein A (SpA) was created to non-specifically bind to anti-PD-L1 antibody. SpyCatcher (SpC) and SpA were fused into the cell outer membrane protein OmpA to capture SpyTag-attached peptides and PD-L1 antibody, respectively. The modified OMV was able to efficiently conjugate with ESC-derived TAAs and PD-L1 antibody (SpC-OMVs + SpT-peptides + anti-PD-L1), increasing the residence time of TAAs in the body. The results showed that the combination therapy of ESC-based TAAs and PD-L1 antibody delivered by OMV had significant inhibitory effects in mouse tumor model. Specifically, it was effective in reducing tumor growth by enhancing IFN-γ-CD8+ T cell responses and increasing the number of CD8+ memory cells and antigen-specific T cells. Overall, the new OMV delivery system is a versatile platform that can enhance the immune responses of ESC-based TAA cancer vaccines.

Heterogeneity analysis of main driving factors affecting potential evapotranspiration changes across different climate regions.

Exploring the influencing factors of potential evapotranspiration (PET) is of great significance for further understanding the causes of climate change and improving agricultural irrigation efficiency. In this study, modified Mann-Kendall analysis was used to elucidate the temporal variation characteristics of meteorological factors and PET based on a dataset from 710 meteorological stations in China. Furthermore, we revealed the main factors that influence the temporal and climate heterogeneity of PET by combining sensitivity analysis with the contribution analysis method. The results showed that 1) climate factors and PET exhibited trend changes on a yearly scale, with slope variation ranges of temperature (T), relative humidity (RH), net radiation (RN), wind speed (U) and PET of 0.03-0.04 °C/a, 0.03-0.08 %/a, 0.001-0.007[MJ/(m2/day)]/a, -0.005 to -0.012(m/s)/a and -0.30-0.38 mm/a, respectively. 2) The sensitivity coefficient fluctuated greatly inter-annually, but the trend was more pronounced inter-annually. Most sensitive factor for PET was RN in hyperarid (HAR), arid (AR) and semiarid regions (SAR), while it changed to RH in semihumid (SHR) and humid regions (HR). PET was more sensitive to RN in dry and relatively wet hot seasons, while it changed to RH during wet and relatively dry cold seasons. 3) PET changes were determined by the relative changes and the sensitivity coefficient, and significant temporal heterogeneity was observed. In HAR, AR, SAR and SHR, the relative changes in T and U result in higher contributions. In HR, PET changes were primarily caused by its higher sensitivity to RH and RN. 4) In dry region and humid-cold seasons, the bigger relative changes of climate factors were the main drivers affecting PET changes, but in humid region and arid-hot seasons, the they were determined by the strong nonlinear relationship between PET and factors. This finding holds great significance for the scientific understanding of the evolution mechanism of PET under changing environments.

Environmental carrying capacity and ecological risk assessment of pesticides under different soil use types in the Central Plains Urban Agglomeration (CPUA), China.

Soil environmental safety has received much attention during the past few decades due to its significance in agricultural production and human health. Special attention is required for soil pesticide residues and ecological risks. This study examined 197 soil samples from industrial, residential and agricultural areas for the presence of 12 organophosphorus pesticides (OPPs) and 8 synthetic pyrethroids (SYPs) in the 16 cities in Henan Province, and the center of CPUA, based on the Central Plains Urban Agglomeration (CPUA) concept proposed by China. The total average concentrations of ∑12OPPs in industrial, residential and agricultural soils were 194, 217, 267 ng/g dry weight, and those of ∑8SYPs were 26.8, 35.7, 25.5 ng/g dry weight, respectively. The two pollutants with the greatest concentrations in the soils were malathion and fenpropathrin, respectively, the dominant components of OPPs and SYPs. The soil environmental carrying capacity (SECC) analysis, representing the maximum residual load that can be supported, shows that acephate and cyhalothrin were overloaded, with a predicted period of over 500 years. Among the 16 cities of CPUA, a higher frequency of high ecological risk could be observed only in Shangqiu. The OPPs in children had total non-carcinogenic risk values of more than 1.0. Similarly, the non-carcinogenic risks of SYPs in adults and children in the residential areas were more than 1.0. The study provides knowledge on how to effectively manage soil safety in Henan Province, which is the center of the CPUA, with a large population and grain province to protect ecosystems and reduce the risks of soil pesticide residues in humans.

Divergent trajectory of replication and intrinsic pathogenicity of SARS-CoV-2 Omicron post-BA.2/5 subvariants in the upper and lower respiratory tract.

BACKGROUND: Earlier Omicron subvariants including BA.1, BA.2, and BA.5 emerged in waves, with a subvariant replacing the previous one every few months. More recently, the post-BA.2/5 subvariants have acquired convergent substitutions in spike that facilitated their escape from humoral immunity and gained ACE2 binding capacity. However, the intrinsic pathogenicity and replication fitness of the evaluated post-BA.2/5 subvariants are not fully understood. METHODS: We systemically investigated the replication fitness and intrinsic pathogenicity of representative post-BA.2/5 subvariants (BL.1, BQ.1, BQ.1.1, XBB.1, CH.1.1, and XBB.1.5) in weanling (3-4 weeks), adult (8-10 weeks), and aged (10-12 months) mice. In addition, to better model Omicron replication in the human nasal epithelium, we further investigated the replication capacity of the post-BA.2/5 subvariants in human primary nasal epithelial cells. FINDINGS: We found that the evaluated post-BA.2/5 subvariants are consistently attenuated in mouse lungs but not in nasal turbinates when compared with their ancestral subvariants BA.2/5. Further investigations in primary human nasal epithelial cells revealed a gained replication fitness of XBB.1 and XBB.1.5 when compared to BA.2 and BA.5.2. INTERPRETATION: Our study revealed that the post-BA.2/5 subvariants are attenuated in lungs while increased in replication fitness in the nasal epithelium, indicating rapid adaptation of the circulating Omicron subvariants in the human populations. FUNDING: The full list of funding can be found at the Acknowledgements section.

A dual-cycle amplification-based electrochemical platform for sensitive detection of tobramycin.

BACKGROUND: Tobramycin (TOB), an essential aminoglycoside antibiotic in human life, poses potential threats due to its residues in the environment. The primary concern is the adverse impact of excessive TOB on human kidneys, hearing, and other organs, significantly affecting human health. Constructing a sensitive electrochemical platform for simple and rapid trace detection is crucial. Herein, to enhance the sensitivity of TOB detection in the environment and mitigate the risks associated with residual antibiotics, an ultrasensitive electrochemical aptasensor was developed. RESULTS: The sensor employs a dual-cycle amplification strategy involving catalytic hairpin assembly (CHA) and exonuclease III (Exo III) for efficient signal amplification. Simultaneously, the electrode performance was optimized by incorporating gold nanowires (AuNWs) onto the surface of reduced graphene oxide (PDA-rGO). Specifically, in the presence of TOB, which binds to the aptamer (Apt), dsDNA dissociates, releasing cDNA to open hairpin 1 (HP1) and initiate the CHA cycle with the participation of hairpin 2 (HP2). Exo III shears HP1 in the HP1/HP2 complex, freeing HP2 to participate in the CHA cycle again. Ultimately, a significant amount of signal label is retained on the electrode by hybridizing with sheared HP1, generating a robust electrical signal. SIGNIFICANCE: Through the signal amplification strategy, the aptasensor design provides a broad linear range of 0.005-500 nM, with a low detection limit of 0.112 pM for TOB. It is worth mentioning that the aptasensor displayed favorable stability, specificity, and reproducibility, and has been successfully applied to practical samples, demonstrating its utility in practical applications.

An indirect competitive assay-based method for the sensitive determination of tetracycline residue using a real-time fluorescence-based quantitative polymerase chain reaction.

Tetracycline (TC) is an effective antibiotic used to treat humans and livestock, but its inappropriate use imposes toxic effects, including pollution, on environmental ecology and food. Currently, sensitive, accurate, and cost-effective methods that can detect lower concentrations of TC residues in environmental and food samples are needed. In this study, a novel indirect competitive assay-based aptamer method was developed for detecting TC residues through signal amplification by real-time fluorescence-based quantitative polymerase chain reaction. The response surface methodology was introduced to optimize the optimal concentrations (influencing factors) of the three types of single-stranded DNA in the competitive assay process. The optimal conditions for the three types of ssDNA were 112 nM for the specific aptamer of TC (Apt40), 115 nM for the signal DNA, and 83 nM for the DNA catcher. As expected, under optimal conditions, the Ct value was linearly related to the logarithm of TC concentration. The calibration curve equation was Ct = -0.34516 log[TC] + 9.9345 (R2 = 0.998) in the range of 10-3-103 ng mL-1, and the limit of detection was 7.02 × 10-5 ng mL-1. The new method was effectively applied to detect TC residues in wastewater, honey, and milk samples. It achieved an average recovery rate of 101.19% with a small variation of 5.16%. The validation was carried out using an enzyme-linked immunosorbent assay. This approach demonstrates high sensitivity and selectivity, making it well suited for detecting leftover antibiotics in food when using suitable aptamers.

Family-Specific Training Improves Linear B Cell Epitope Prediction for Emerging Viruses.

The rational design of vaccines and antibody-based therapeutics against newly emerging viruses relies on B cell epitopes mainly. To predict the B cell epitopes of a novel virus, several algorithms have been developed. While most existing algorithms are trained on a dataset in which B cell epitopes are classified as 'Positive' or 'Negative'. However, we found that training on such data contaminates the target pattern of specific viruses, leading to inaccurate predictions in some cases. In this paper, we introduce a novel framework for predicting linear B cell epitopes of novel viruses by exclusively using highly similar viruses for training data. We employed kernel regression based on seropositive rates, which are the percentages of seropositive samples among the population, to predict the potential epitopes. To assess our method, we conducted simulations and utilized two real-world datasets. Our method significantly outperformed other existing methods on the testing data of four viruses with seropositive rates. Also, our strategy showed a better prediction in a larger dataset from the IEDB. Thus, a novel framework providing better linear B cell prediction of newly emerging viruses is established, which will benefit the rational design of vaccines and antibody-based therapeutics in the future.

Heavy metals pollution of soil in central plains urban agglomeration (CPUA), China: human health risk assessment based on Monte Carlo simulation.

The present study conducted the concentration evaluation, pollution assessment, source analysis, and risk assessment of heavy metals in the soil of the CPUA, China, to contribute to the smooth construction of urban agglomeration. Elevated levels of mean concentrations of cadmium (Cd), chromium (Cr), and copper (Cu) in the soils were shown compared to background values. Cu and zinc (Zn) and also lead (Pb) and Cd exhibited spatial similarity. Manganese (Mn) and Cr exhibited point source characteristics such as the concentrations at a point much higher than the surrounding area. The potential ecological risk in the northern region belonged to the moderate risk level category. Cd contributed over 90% to the potential ecological risk. The health risk among children was higher than that among adults. The major exposure pathways were different for adults and children. Exposure, as shown using Hazard Index (HI), to adults was mainly through the skin contact route, while to children was through both the skin contact and ingestion route. The primary CR (carcinogenic risk) to adults was through the inhalation route, while that to children was through the ingestion route. In both children and adults, Cr was the main contributor to HI and CR. According to the Monte Carlo simulation results, the cumulative probability of exceeding the critical value of HI for children was approximately 2.8-3.0 times that for adults. According to the sensitivity analysis results, non-carcinogenic risk prevention should begin mainly by reducing exposure duration and skin contact. The cancer risk may be reduced primarily by decreasing the exposure duration and controlling ingestion. The PMF (Positive Matrix Factorization) source analysis revealed that Pb mainly came from transportation sources. In addition, Cu, Pb, and Mn were derived mainly from agricultural sources. Cr was derived mostly from a natural source, and Cd originated mainly from an industrial source.

The viral fitness and intrinsic pathogenicity of dominant SARS-CoV-2 Omicron sublineages BA.1, BA.2, and BA.5.

BACKGROUND: Among the Omicron sublineages that have emerged, BA.1, BA.2, BA.5, and their related sublineages have resulted in the largest number of infections. While recent studies demonstrated that all Omicron sublineages robustly escape neutralizing antibody response, it remains unclear on whether these Omicron sublineages share any pattern of evolutionary trajectory on their replication efficiency and intrinsic pathogenicity along the respiratory tract. METHODS: We compared the virological features, replication capacity of dominant Omicron sublineages BA.1, BA.2 and BA.5 in the human nasal epithelium, and characterized their pathogenicity in K18-hACE2, A129, young C57BL/6, and aged C57BL/6 mice. FINDINGS: We found that BA.5 replicated most robustly, followed by BA.2 and BA.1, in the differentiated human nasal epithelium. Consistently, BA.5 infection resulted in higher viral gene copies, infectious viral titres and more abundant viral antigen expression in the nasal turbinates of the infected K18-hACE2 transgenic mice. In contrast, the Omicron sublineages are continuously attenuated in lungs of infected K18-hACE2 and C57BL/6 mice, leading to decreased pathogenicity. Nevertheless, lung manifestations remain severe in Omicron sublineages-infected A129 and aged C57BL/6 mice. INTERPRETATION: Our results suggested that the Omicron sublineages might be gaining intrinsic replication fitness in the upper respiratory tract, therefore highlighting the importance of global surveillance of the emergence of hyper-transmissive Omicron sublineages. On the contrary, replication and intrinsic pathogenicity of Omicron is suggested to be further attenuated in the lower respiratory tract. Effective vaccination and other precautions should be in place to prevent severe infections in the immunocompromised populations at risk. FUNDING: A full list of funding bodies that contributed to this study can be found in the Acknowledgements section.

Rational design of a booster vaccine against COVID-19 based on antigenic distance.

Current COVID-19 vaccines are highly effective against symptomatic disease, but repeated booster doses using vaccines based on the ancestral strain offer limited additional protection against SARS-CoV-2 variants of concern (VOCs). To address this, we used antigenic distance to in silico select optimized booster vaccine seed strains effective against both current and future VOCs. Our model suggests that a SARS-CoV-1-based booster vaccine has the potential to cover a broader range of VOCs. Candidate vaccines including the spike protein from ancestral SARS-CoV-2, Delta, Omicron (BA.1), SARS-CoV-1, or MERS-CoV were experimentally evaluated in mice following two doses of the BNT162b2 vaccine. The SARS-CoV-1-based booster vaccine outperformed other candidates in terms of neutralizing antibody breadth and duration, as well as protective activity against Omicron (BA.2) challenge. This study suggests a unique strategy for selecting booster vaccines based on antigenic distance, which may be useful in designing future booster vaccines as new SARS-CoV-2 variants emerge.