The Bipartite Sequence Motif in the N and C Termini of gp85 of Subgroup J Avian Leukosis Virus Plays a Crucial Role in Receptor Binding and Viral Entry.

Subgroup J avian leukemia virus (ALV-J), belonging to the genus Alpharetrovirus, enters cells through its envelope surface unit (gp85) via specifically recognizing the cellular receptor chicken Na+/H+ exchanger type I (chNHE1), the 28 to 39 N-terminal residues of which were characterized as the minimal receptor functional domain in our previous studies. In this study, to further clarify the precise organization and properties of the interaction between ALV-J gp85 and chNHE1, we identified the chNHE1-binding domain of ALV-J gp85 using a series of gp85 mutants with segment substitutions and evaluating their effects on chNHE1 binding in protein-cell binding assays. Our results showed that hemagglutinin (HA) substitutions of amino acids (aa) 38 to 131 (N terminus of gp85) and aa 159 to 283 (C terminus of gp85) significantly inhibited the interaction between gp85 and chNHE1/chNHE1 loop 1. In addition, these HA-substituted chimeric gp85 proteins could not effectively block the entry of ALV-J into chNHE1-expressing cells. Furthermore, analysis of various N-linked glycosylation sites and cysteine mutants in gp85 revealed that glycosylation sites (N6 and N11) and cysteines (C3 and C9) were directly involved in receptor-gp85 binding and important for the entry of ALV-J into cells. Taken together, our findings indicated that the bipartite sequence motif, spanning aa 38 to 131 and aa 159 to 283, of ALV-J gp85 was essential for binding to chNHE1, with its two N-linked glycosylation sites and two cysteines being important for its receptor-binding function and subsequent viral infection steps.IMPORTANCE Infection of a cell by retroviruses requires the attachment and fusion of the host and viral membranes. The specific adsorption of envelope (Env) surface proteins to cell receptors is a key step in triggering infections and has been the target of antiviral drug screening. ALV-J is an economically important avian pathogen that belongs to the genus Alpharetrovirus and has a wider host range than other ALV subgroups. Our results showed that the amino acids 38 to 131 of the N terminus and 159 to 283 of the C terminus of ALV-J gp85 controlled the efficiency of gp85 binding to chNHE1 and were critical for viral infection. In addition, the glycosylation sites (N6 and N11) and cysteines (C3 and C9) of gp85 played a crucial role in the receptor binding and viral entry. These findings might help elucidate the mechanism of the entry of ALV-J into host cells and provide antiviral targets for the control of ALV-J.

Strain-Modulated Platinum-Palladium Nanowires for Oxygen Reduction Reaction.

Electrocatalytic activity of alloy nanocatalytsts can be manipulated effectively by tuning their physical properties (ensemble, geometric, and ligand effects) to afford optimal surface structure and compositions for proton exchange membrane fuel cell (PEMFC) application. Herein, highly catalytic platinum-palladium nanowires (PtnPd100-n NWs) with a subtle lattice strain and Boerdijk-Coxeter helix type morphology are synthesized through a surfactant-free, thermal single phase solvent method. X-ray diffraction results show that PtnPd100-n NWs are exposed through the (111) facets and their shrinking or expanding lattice parameters can be modulated by the alloy compositions. Electrochemical results reveal that their high catalytic activity correlates with the lattice shrinking, facets, and bimetallic compositions, showing higher activity when the ratio of Pt and Pd is ∼78:22, which is further supported by DFT results. Compared to the nanoparticle type platinum-palladium alloyed catalysts with similar metal compositions (PtnPd100-n NPs), the PtnPd100-n NWs exhibit significantly improved electrocatalytic activity and stability for the oxygen reduction reaction. These findings open new strategies to design the highly active and stable alloy nanocatalysts with controllable compositions.

Origin of High Activity and Durability of Twisty Nanowire Alloy Catalysts under Oxygen Reduction and Fuel Cell Operating Conditions.

The ability to control the surface composition and morphology of alloy catalysts is critical for achieving high activity and durability of catalysts for oxygen reduction reaction (ORR) and fuel cells. This report describes an efficient surfactant-free synthesis route for producing a twisty nanowire (TNW) shaped platinum-iron (PtFe) alloy catalyst (denoted as PtFe TNWs) with controllable bimetallic compositions. PtFe TNWs with an optimal initial composition of ∼24% Pt are shown to exhibit the highest mass activity (3.4 A/mgPt, ∼20 times higher than that of commercial Pt catalyst) and the highest durability (<2% loss of activity after 40 000 cycles and <30% loss after 120 000 cycles) among all PtFe-based nanocatalysts under ORR or fuel cell operating conditions reported so far. Using ex situ and in situ synchrotron X-ray diffraction coupled with atomic pair distribution function (PDF) analysis and 3D modeling, the PtFe TNWs are shown to exhibit mixed face-centered cubic (fcc)-body-centered cubic (bcc) alloy structure and a significant lattice strain. A striking finding is that the activity strongly depends on the composition of the as-synthesized catalysts and this dependence remains unchanged despite the evolution of the composition of the different catalysts and their lattice constants under ORR or fuel cell operating conditions. Notably, dealloying under fuel cell operating condition starts at phase-segregated domain sites leading to a final fcc alloy structure with subtle differences in surface morphology. Due to a subsequent realloying and the morphology of TNWs, the surface lattice strain observed with the as-synthesized catalysts is largely preserved. This strain and the particular facets exhibited by the TNWs are believed to be responsible for the observed activity and durability enhancements. These findings provide new insights into the correlation between the structure, activity, and durability of nanoalloy catalysts and are expected to energize the ongoing effort to develop highly active and durable low-Pt-content nanowire catalysts by controlling their alloy structure and morphology.

Development and evaluation of a gp85 protein-based subgroup-specific indirect enzyme-linked immunosorbent assay for the detection of anti-subgroup J avian leukosis virus antibodies.

Avian leukosis virus subgroup J (ALV-J) is an important pathogen for various neoplasms and causes significant economic losses in the poultry industry. Serological detection of specific antibodies against ALV-J infection is important for successful clinical diagnosis. Here, a 293F stable cell line was established to stably express gp85 protein. In this cell line, gp85 protein was expressed at approximately 30 mg/L. A subgroup-specific indirect enzyme-linked immunosorbent assay (iELISA) was developed using ALV-J gp85 protein as coated antigen to detect antibodies against ALV-J. The sensitivity of the iELISA (1:51200 diluted in serum) was 16 times more than that of indirect immunofluorescence assay (IFA; 1:3200 diluted in serum). Moreover, there was no crossreactivity with antibodies against other common avian viruses and other avian leukosis virus subgroups, such as subgroups A and B. The practicality of the iELISA was further evaluated by experimental infection and clinical samples. The results from experimental infection indicated that anti-ALV-J antibodies were readily detected by iELISA as early as 4 weeks after ALV-J infection, and positive antibodies were detected until 20 weeks, with an antibody-positive rate of 11.1% to 33.3%. Moreover, analysis of clinical samples showed that 9.49% of samples were positive for anti-ALV-J antibodies, and the concordance rate of iELISA and IFA was 99.24%. Overall, these results suggested that the subgroup-specific iELISA developed in this study had good sensitivity, specificity, and feasibility. This iELISA will be very useful for epidemiological surveillance, diagnosis, and eradication of ALV-J in poultry farms.

Development and application of a colloidal gold test strip for detection of avian leukosis virus.

Avian leukosis virus (ALV) is an avian oncogenic retrovirus that induces leukemia-like proliferative diseases in chickens. ALV infection can result in the development of immunological tolerance and persistent viremia. Since effective vaccines against ALV are not yet available, its current prevention primarily depends on detection and eradication to establish exogenous ALV-free poultry flocks. In this study, a rapid and simple colloidal gold test strip method, specific for the group-specific antigen, p27 protein, was developed and systematically evaluated for the detection of ALV from different samples. The detection limit of this assay was as low as 6.25 ng/ml for p27 protein and 80 TCID50/ml for different subgroups of ALV. Besides, the test strip showed high specificity in the detection of different subgroups of ALV, including ALV-A, ALV-B, ALV-J, and ALV-K, with no cross-reaction with other avian pathogens. Furthermore, we artificially infected specific pathogen-free (SPF) chickens with ALV-J, collected cloacal swabs, and examined viral shedding using both test strips and ELISA. Results from the test strip were highly consistent with that from ELISA. In addition, 1104 virus isolates from anti-coagulant blood samples, 645 albumen samples, and 4312 meconium samples were tested, and the test strip results agreed with those of ELISA kit up to 97.1%. All the results indicated that the colloidal gold test strip could serve as a simple, rapid, sensitive, and specific diagnostic method for eradication of ALV in poultry farms.

Heterostyly promotes compatible pollination in buckwheats: Comparisons of intraflower, intraplant, and interplant pollen flow in distylous and homostylous Fagopyrum.

PREMISE OF THE STUDY: Heterostyly, the reciprocal positioning of stigmas and anthers in different floral morphs, has long been thought to promote intermorph pollination. However, extensive intramorph pollination occurs commonly in heterostylous species, leading to recurrent questions about the functional and evolutionary significance of heterostyly. METHODS: To identify the sources of stigmatic pollen (autogamous [intraflower], geitonogamous [intraplant], vs. interplant), we emasculated either one flower or entire plants in experimental populations of the two closely related buckwheat species, distylous Fagopyrum esculentum and homostylous F. tataricum. Differences in pollen size allowed unambiguous identification of pollen on stigmas. RESULTS: Only 2.4% of F. tataricum pollen and 1.5% of F. esculentum pollen arrived successfully on compatible stigmas of other plants. In the former (homostylous) species, 71.3% of the pollen load on stigmas was autogamous, 10.8% was geitonogamous, and 17.9% was interplant. In the latter (distylous) species, 37.45% of the pollen on stigmas was autogamous, 13.8% was geitonogamous, 17.0% was intramorph, and 31.75% was intermorph. The amount of incompatible pollen arriving on stigmas was greatly decreased by both one-flower and whole-plant emasculations, and thus, the proportion of compatible pollen deposited increased with one-flower emasculation and increased even more with whole-plant emasculation. CONCLUSIONS: Our quantification of pollen-donor sources in these two species indicated that heterostyly in Fagopyrum esculentum provided a nearly 2-fold fitness advantage (in terms of compatible pollination) over expected (random) pollen transfers between morphs. Because of reduced herkogamy, the homostylous F. tataricum was highly autogamous.

Composition Tunability and (111)-Dominant Facets of Ultrathin Platinum-Gold Alloy Nanowires toward Enhanced Electrocatalysis.

The ability for tuning not only the composition but also the type of surface facets of alloyed nanomaterials is important for the design of catalysts with enhanced activity and stability through optimizing both ensemble and ligand effects. Herein we report the first example of ultrathin platinum-gold alloy nanowires (PtAu NWs) featuring composition-tunable and (111) facet-dominant surface characteristics, and the electrocatalytic enhancement for the oxygen reduction reaction (ORR). PtAu NWs of different bimetallic compositions synthesized by a single-phase and surfactant-free method are shown to display an alloyed, parallel-bundled structure in which the individual nanowires exhibit Boerdijk-Coxeter helix type morphology predominant in (111) facets. Results have revealed intriguing catalytic correlation with the binary composition, exhibiting an activity maximum at a Pt:Au ratio of ∼3:1. As revealed by high-energy synchrotron X-ray diffraction and atomic pair distribution function analysis, NWs of this ratio exhibit a clear shrinkage in interatomic bonding distances. In comparison with PtAu nanoparticles of a similar composition and degree of shrinking of atomic-pair distances, the PtAu NWs display a remarkably higher electrocatalytic activity and stability. The outperformance of NWs over nanoparticles is attributed to the predominant (111)-type facets on the surface balancing the contribution of ensemble and ligand effects, in addition to the composition synergy due to optimal adsorption energies for molecular and atomic oxygen species on the surface as supported by DFT computation of models of the catalysts. The findings open up a new pathway to the design and engineering of alloy nanocatalysts with enhanced activity and durability.

Monocytes-to-lymphocytes ratio increases the prognostic value of circulating tumor cells in non-small cell lung cancer: a prospective study.

Background: Circulating tumor cells (CTCs) has shown important prognostic value in non-small cell lung cancer (NSCLC). However, the present low sensitivity of CTC capture technology restricts their clinical application. This study aims to explore the feasibility of combining the peripheral blood cell (PBC)-derived inflammation-based score with CTCs to increase the prognostic value of CTCs in NSCLC. Methods: Sixty volunteers diagnosed with NSCLC were recruited. CTC count and six inflammation-based scores were examined and the association with progression-free survival (PFS) and overall survival (OS) was explored. The changes in the CTC counts before and after the immunotherapy were observed. Results: Multivariate analysis showed that CTCs >7 [hazard ratio (HR) =9.07; 95% confidence interval (CI): 3.68-22.37, P<0.001] and monocytes-to-lymphocytes ratio (MLR) > 0.2 (HR =3.07; 95% CI: 1.21-7.84; P=0.01) were associated with shorter OS and PFS in patients with NSCLC. Patients with CTCs >7 and MLR >0.2 had 12.30 times increased risk of death (P<0.001) and 6.10 times increased risk of disease progression (P=0.002) compared with those with CTCs ≤7 and MLR ≤0.2. Decreased CTC counts after immunotherapy were closely related to disease control (r=0.535, P=0.01). Conclusions: CTCs and MLR are both independent risk factors for prognosis in patients with NSCLC. The combination of CTCs with MLR significantly increased the prognostic value of CTCs, which would contribute to stratification of NSCLC patients and providing precise treatment. Dynamic monitoring of CTCs efficiently shows the immunotherapy response in NSCLC.

Efficient production of γ-aminobutyric acid using engineered Escherichia coli whole-cell catalyst.

γ-Aminobutyric acid (GABA) has been widely used in the food, feed, pharmaceutical, and chemical industry fields. Previously, we developed a whole-cell catalyst capable of converting L-glutamate (L-Glu) into GABA by overexpressing the glutamate decarboxylase gene (gadz11) from Bacillus sp. Z11 in Escherichia coli BL21(DE3). However, to enhance cell permeability, a freeze-thaw treatment is required, and to enhance GADZ11 activity, pyridoxal 5'-phosphate (PLP) must be added to the reaction system. The aim of this study is to provide a more efficient approach for GABA production by engineering the recombinant E. coli above. First, the inducible expression conditions of the gadz11 in E. coli were optimized to 37 °C for 6 h. Next, an ideal engineered strain was produced via increasing cell permeability by overexpressing sulA and eliminating PLP dependence by constructing a self-sufficient system. Furthermore, an efficient whole-cell biocatalytic process was optimized. The optimal substrate concentration, cell density, and reaction temperature were 1.0 mol/L (the molecular ratio of L-Glu to L-monosodium glutamate (L-MSG) was 4:1), 15 and 37 °C, respectively. Finally, a whole-cell bioconversion procedure was performed in a 3-L bioreactor under optimal conditions. The strain could be reused for at least two cycles with GABA yield, productivity and conversion ratio of 206.2 g/L, 117.8 g/L/h and 100.0%, respectively. This is currently the highest GABA productivity from a mixture of L-Glu and L-MSG reported without the addition of cofactors or additional treatment of cells. This work demonstrates that the novel engineered E. coli strain has the potential for application in large-scale industrial GABA production.

Ejectosome of Pectobacterium bacteriophage ΦM1.

Podophages that infect gram-negative bacteria, such as Pectobacterium pathogen ΦM1, encode tail assemblies too short to extend across the complex gram-negative cell wall. To overcome this, podophages encode a large protein complex (ejectosome) packaged inside the viral capsid and correspondingly ejected during infection to form a transient channel that spans the periplasmic space. Here, we describe the ejectosome of bacteriophage ΦM1 to a resolution of 3.32 Å by single-particle cryo-electron microscopy (cryo-EM). The core consists of tetrameric and octameric ejection proteins which form a ∼1.5-MDa ejectosome that must transition through the ∼30 Å aperture created by the short tail nozzle assembly that acts as the conduit for the passage of DNA during infection. The ejectosome forms several grooves into which coils of genomic DNA are fit before the DNA sharply turns and goes down the tunnel and into the portal. In addition, we reconstructed the icosahedral capsid and hybrid tail apparatus to resolutions between 3.04 and 3.23 Å, and note an uncommon fold adopted by the dimerized decoration proteins which further emphasize the structural diversity of podophages. These reconstructions have allowed the generation of a complete atomic model of the ΦM1, uncovering two distinct decoration proteins and highlighting the exquisite structural diversity of tailed bacteriophages.

Broad cross neutralizing antibodies against sarbecoviruses generated by SARS-CoV-2 infection and vaccination in humans.

The outbreaks of severe acute respiratory syndrome coronavirus (SARS-CoV-1), Middle East respiratory syndrome coronavirus (MERS-CoV), and SARS-CoV-2 highlight the need for countermeasures to prevent future coronavirus pandemics. Given the unpredictable nature of spillover events, preparing antibodies with broad coronavirus-neutralizing activity is an ideal proactive strategy. Here, we investigated whether SARS-CoV-2 infection and vaccination could provide cross-neutralizing antibodies (nAbs) against zoonotic sarbecoviruses. We evaluated the cross-neutralizing profiles of plasma and monoclonal antibodies constructed from B cells from coronavirus disease 2019 (COVID-19) convalescents and vaccine recipients; against sarbecoviruses originating from bats, civets, and pangolins; and against SARS-CoV-1 and SARS-CoV-2. We found that the majority of individuals with natural infection and vaccination elicited broad nAb responses to most tested sarbecoviruses, particularly to clade 1b viruses, but exhibited very low cross-neutralization to SARS-CoV-1 in both natural infection and vaccination, and vaccination boosters significantly augmented the magnitude and breadth of nAbs to sarbecoviruses. Of the nAbs, several exhibited neutralization activity against multiple sarbecoviruses by targeting the spike receptor-binding domain (RBD) and competing with angiotensin-converting enzyme 2 (ACE2) binding. SCM12-61 demonstrated exceptional potency, with half-maximal inhibitory concentration (IC50) values of 0.001-0.091 µg/mL against tested sarbecoviruses; while VSM9-12 exhibited remarkable cross-neutralizing breadth against sarbecoviruses and SARS-CoV-2 Omicron subvariants, highlighting the potential of these two nAbs in combating sarbecoviruses and SARS-CoV-2 Omicron subvariants. Collectively, our findings suggest that vaccination with an ancestral SARS-CoV-2 vaccine, in combination with broad nAbs against sarbecoviruses, may provide a countermeasure for preventing further sarbecovirus outbreaks in humans.

Influence of the AC field intensity and frequency on composition and growth mechanism of Au-Pd alloy nanowires.

Au-Pd alloy nanowires with controllable morphology and composition are useful sensing materials for chemical and biological sensors. This report describes the preparation of such Au-Pd alloy nanowires from an aqueous solution by alternating current (AC) varied-frequency method, focusing on determining the dependence of the composition and morphology of the alloy nanowires on the electric field intensity and frequency. An electric field varied from 0.1 V x m(-1) to 0.4 x 10(6)V x m(-1) at 300 Hz frequency was used for the nucleation, followed by variation of the frequency between 1 and 20 MHz for the growth of the nanowires. The results showed that the Pd content in the alloy nanowires increased with the field intensity and frequency. The nanowire morphology with a less branching and better alignment was obtained at the increased frequency. XRD results showed that the phase structure of the alloy nanowires was face-centered cubic lattice. The nanowire compositions were shown controllable by changing the AC field intensity, frequency, as well as the metal ion ratio in the solution. The growth of the nanowires was shown to obey the Maxwell-Wanger (M-W) law.

Platinum-Cobalt Nanowires for Efficient Alcohol Oxidation Electrocatalysis.

The compositions and surface facets of platinum (Pt)-based electrocatalysts are of great significance for the development of direct alcohol fuel cells (DAFCs). We reported an approach for preparing ultrathin PtnCo100-n nanowire (NW) catalysts with high activity. The PtnCo100-n NW alloy catalysts synthesized by single-phase surfactant-free synthesis have adjustable compositions and (111) plane and strain lattices. X-ray diffraction (XRD) results indicate that the alloy composition can adjust the lattice shrinkage or expansion of PtnCo100-n NWs. X-ray photoelectron spectroscopy (XPS) results show that the electron structure of Pt is changed by the alloying effect caused by electron modulation in the d band, and the chemical adsorption strength of Pt is decreased, thus the catalytic activity of Pt is increased. The experimental results show that the activity of PtnCo100-n for the oxidation of methanol and ethanol is related to the exposed crystal surface, strain lattice and composition of catalysts. The PtnCo100-n NWs exhibit stronger electrocatalytic performance for both methanol oxidation reaction (MOR) and ethanol oxidation reaction (EOR). The dominant (111) plane Pt53Co47 exhibits the highest electrocatalytic activity in MOR, which is supported by the results of XPS. This discovery provides a new pathway to design high activity, stability nanocatalysts to enhance direct alcohol fuel cells.

Co-Fe3C pair sites catalyst with heterometallic dual active sites for efficient oxygen reduction reaction.

Newly emerging metal-based pair sites catalysts show great potential because they can provide more metal active centers with synergistic effect for green catalysis, compared with single site catalysts. However, both the synthesis and catalytic mechanisms of the pair sites catalyst with new structural features need to be developed vigorously to promote the desired chemical reactions, especially carbon-based metal catalysts for green energy storage and conversion devices. Herein, we constructed highly active Co-Fe3C pair sites on N-doped graphite catalyst (CNCo-Fe3C) by a two-step strategy, which have electron interactions of heterometallic atoms and can play better synergistic effect. X-ray absorption spectra and density functional theory (DFT) calculation further identify the presence of heterometallic active sites in the pair sites catalyst, resulting in electron redistribution and positive d-band center due to the electron interactions. The more positive d-band center model predicts the optimization of the adsorption energy of oxygen-containing intermediates, and reduces the energy barrier of the determining step. This further results in superior oxygen reduction reaction (ORR) performance with a half-wave potential of 0.90 V versus reversible hydrogen electrode (vs.RHE) and superior long-term stability for about 20 h with only 2.3 % decrease at 0.75 V vs.RHE in 0.1 M KOH solution. Additionally, it also shows significant peak power density of 124 mW cm-2 and prominent cycling stability performance exceeding 400 h at 5 mA cm-2 in the Zn-air battery (ZAB) test, which is higher than that of Pt/C catalyst. This work provides a new idea for the regulation of intrinsic activity of non-noble metal ORR catalysts through the synergistic effect of the pair sites.

Surface Modification of Hollow Structure TiO2 Nanospheres for Enhanced Photocatalytic Hydrogen Evolution.

Engineering the surface structure of semiconductor is one of the most promising strategies for improving the separation and transfer efficiency of charge, which is a key issue in photocatalysis. Here, we designed and fabricated the C decorated hollow TiO2 photocatalysts (C-TiO2), in which 3-aminophenol-formaldehyde resin (APF) spheres were used as template and carbon precursor. It was determined that the C content can be easily controlled by calcinating the APF spheres with different time. Moreover, the synergetic effort between the optimal C content and the formed Ti-O-C bonds in C-TiO2 were determined to increase the light absorption and greatly promote the separation and transfer of charge in the photocatalytic reaction, which is verified from UV-vis, PL, photocurrent, and EIS characterizations. Remarkably, the activity of the C-TiO2 is 5.5-fold higher than that of TiO2 in H2 evolution. A feasible strategy for rational design and construction of surface-engineered hollow photocatalysts to improve the photocatalytic performance was provided in this study.

Production of N-acetylglucosamine from carbon dioxide by engineering Cupriavidus necator H16.

The conversion of CO2 into valuable bioactive substances using synthetic biological techniques is a potential approach for mitigating the greenhouse effect. Here, the engineering of C. necator H16 to produce N-acetylglucosamine (GlcNAc) from CO2 is reported. First, GlcNAc importation and intracellular metabolic pathways were disrupted by the deletion of nagF, nagE, nagC, nagA and nagB genes. Second, the GlcNAc-6-phosphate N-acetyltransferase gene (gna1) was screened. A GlcNAc-producing strain was constructed by overexpressing a mutant gna1 from Caenorhabditis elegans. A further increase in GlcNAc production was achieved by disrupting poly(3-hydroxybutyrate) biosynthesis and the Entner-Doudoroff pathways. The maximum GlcNAc titers were 199.9 and 566.3 mg/L for fructose and glycerol, respectively. Finally, the best strain achieved a GlcNAc titer of 75.3 mg/L in autotrophic fermentation. This study demonstrated a conversion of CO2 to GlcNAc, thereby providing a feasible approach for the biosynthesis of various bioactive chemicals from CO2 under normal conditions..

SARS-CoV-2 spike-specific TFH cells exhibit unique responses in infected and vaccinated individuals.

Long-term humoral immunity to SARS-CoV-2 is essential for preventing reinfection. The production of neutralizing antibody (nAb) and B cell differentiation are tightly regulated by T follicular help (TFH) cells. However, the longevity and functional role of TFH cell subsets in COVID-19 convalescents and vaccine recipients remain poorly defined. Here, we show that SARS-CoV-2 infection and inactivated vaccine elicited both spike-specific CXCR3+ TFH cell and CXCR3- TFH cell responses, which showed distinct response patterns. Spike-specific CXCR3+ TFH cells exhibit a dominant and more durable response than CXCR3- TFH cells that positively correlated with antibody responses. A third booster dose preferentially expands the spike-specific CXCR3+ TFH cell subset induced by two doses of inactivated vaccine, contributing to antibody maturation and potency. Functionally, spike-specific CXCR3+ TFH cells have a greater ability to induce spike-specific antibody secreting cells (ASCs) differentiation compared to spike-specific CXCR3- TFH cells. In conclusion, the persistent and functional role of spike-specific CXCR3+ TFH cells following SARS-CoV-2 infection and vaccination may play an important role in antibody maintenance and recall response, thereby conferring long-term protection. The findings from this study will inform the development of SARS-CoV-2 vaccines aiming to induce long-term protective immune memory.

Strained Lattice Gold-Copper Alloy Nanoparticles for Efficient Carbon Dioxide Electroreduction.

Electrocatalytic conversion of carbon dioxide (CO2) into specific renewable fuels is an attractive way to mitigate the greenhouse effect and solve the energy crisis. AunCu100-n/C alloy nanoparticles (AunCu100-n/C NPs) with tunable compositions, a highly active crystal plane and a strained lattice were synthesized by the thermal solvent co-reduction method. Transmission electron microscopy (TEM) and X-ray diffraction (XRD) results show that AunCu100-n/C catalysts display a subtle lattice strain and dominant (111) crystal plane, which can be adjusted by the alloy composition. Electrochemical results show that AunCu100-n/C alloy catalysts for CO2 reduction display high catalytic activity; in particular, the Faradaic efficiency of Au75Cu25/C is up to 92.6% for CO at -0.7 V (vs. the reversible hydrogen electrode), which is related to lattice shrinkage and the active facet. This research provides a new strategy with which to design strong and active nanoalloy catalysts with lattice mismatch and main active surfaces for CO2 reduction reaction.

Monoclonal antibodies constructed from COVID-19 convalescent memory B cells exhibit potent binding activity to MERS-CoV spike S2 subunit and other human coronaviruses.

Introduction: The Middle East respiratory syndrome coronavirus (MERS-CoV) and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are two highly contagious coronaviruses causing MERS and COVID-19, respectively, without an effective antiviral drug and a long-lasting vaccine. Approaches for diagnosis, therapeutics, prevention, etc., particularly for SARS-CoV-2 that is continually spreading and evolving, are urgently needed. Our previous study discovered that >60% of sera from convalescent COVID-19 individuals, but <8% from general population, showed binding activity against the MERS-CoV spike protein, indicating that SARS-CoV-2 infection boosted antibodies cross-reactive with MERS-CoV. Methods: To generate antibodies specific to both SARS-CoV-2 and MERS-CoV, here we screened 60 COVID-19 convalescent sera against MERS-CoV spike extracellular domain and S1 and S2 subunits. We constructed and characterized monoclonal antibodies (mAbs) from COVID-19 convalescent memory B cells and examined their binding and neutralizing activities against human coronaviruses. Results and Discussion: Of 60 convalescent serum samples, 34 showed binding activity against MERS-CoV S2, with endpoint titers positively correlated with the titers to SARS-CoV-2 S2. By sorting single memory B cells from COVID-19 convalescents, we constructed 38 mAbs and found that 11 mAbs showed binding activity with MERS-CoV S2, of which 9 mAbs showed potent cross-reactivity with all or a proportion of spike proteins of alphacoronaviruses (229E and NL63) and betacoronaviruses (SARS-CoV-1, SARS-CoV-2, OC43, and HKU1). Moreover, 5 mAbs also showed weak neutralization efficiency against MERS-CoV spike pseudovirus. Epitope analysis revealed that 3 and 8 mAbs bound to linear and conformational epitopes in MERS-CoV S2, respectively. In summary, we have constructed a panel of antibodies with broad-spectrum reactivity against all seven human coronaviruses, thus facilitating the development of diagnosis methods and vaccine design for multiple coronaviruses.

Molecular characteristic and pathogenicity analysis of a novel multiple recombinant ALV-K strain.

Avian leukosis virus (ALV) can induce various tumors and cause serious production problems. ALVs isolated from chickens were divided into six subgroups (A-J). In 2012, a strain of a putative novel subgroup of ALVs was isolated from Chinese native chickens in Jiangsu Province and named as ALV-K. In this study, three ALV-K strains (JS14LH01, JS13LH14, and JS15SG01) were isolated from chickens with suspected ALV infection in Jiangsu Province. Their complete genomes were amplified, sequenced, and analyzed systematically. The results showed that JS14LH01 and JS13LH14 were ALV-K and ALV-E recombinant strains. Whereas JS15SG01 is an ALV-K, ALV-E, and ALV-J multiple recombinant strain containing the U3 region of ALV-J. The pathogenicity test of JS15SG01 revealed that, compared with previous ALV-K strains, the viremia and viral shedding level of JS15SG01-infected chickens were significantly increased, reaching 100 % and 59 %, respectively. More important, JS15SG01 induced significant proliferation of gliocytes in the cerebral cortex of infected chickens, accompanied by the neurotropic phenomenon. This is the first report about a multiple recombinant ALV-K strain that could invade and injure the brain tissue of chickens in China. Our findings enriched the epidemiologic data of ALV and helped to reveal the evolution of ALV strains prevalent in chicken fields.