Two sequential activation modules control the differentiation of protective T helper-1 (Th1) cells.

Interferon-γ (IFN-γ)-producing CD4+ T helper-1 (Th1) cells are critical for protection from microbes that infect the phagosomes of myeloid cells. Current understanding of Th1 cell differentiation is based largely on reductionist cell culture experiments. We assessed Th1 cell generation in vivo by studying antigen-specific CD4+ T cells during infection with the phagosomal pathogen Salmonella enterica (Se), or influenza A virus (IAV), for which CD4+ T cells are less important. Both microbes induced T follicular helper (Tfh) and interleukin-12 (IL-12)-independent Th1 cells. During Se infection, however, the Th1 cells subsequently outgrew the Tfh cells via an IL-12-dependent process and formed subsets with increased IFN-γ production, ZEB2-transcription factor-dependent cytotoxicity, and capacity to control Se infection. Our results indicate that many infections induce a module that generates Tfh and poorly differentiated Th1 cells, which is followed in phagosomal infections by an IL-12-dependent Th1 cell amplification module that is critical for pathogen control.

Immune tolerance of food is mediated by layers of CD4+ T cell dysfunction.

Gastrointestinal health depends on the adaptive immune system tolerating the foreign proteins in food1,2. This tolerance is paradoxical because the immune system normally attacks foreign substances by generating inflammation. Here we addressed this conundrum by using a sensitive cell enrichment method to show that polyclonal CD4+ T cells responded to food peptides, including a natural one from gliadin, by proliferating weakly in secondary lymphoid organs of the gut-liver axis owing to the action of regulatory T cells. A few food-specific T cells then differentiated into T follicular helper cells that promoted a weak antibody response. Most cells in the expanded population, however, lacked canonical T helper lineage markers and fell into five subsets dominated by naive-like or T follicular helper-like anergic cells with limited capacity to form inflammatory T helper 1 cells. Eventually, many of the T helper lineage-negative cells became regulatory T cells themselves through an interleukin-2-dependent mechanism. Our results indicate that exposure to food antigens causes cognate CD4+ naive T cells to form a complex set of noncanonical hyporesponsive T helper cell subsets that lack the inflammatory functions needed to cause gut pathology and yet have the potential to produce regulatory T cells that may suppress it.

Novel virus-like nanoparticle vaccine effectively protects animal model from SARS-CoV-2 infection.

The key to battling the COVID-19 pandemic and its potential aftermath is to develop a variety of vaccines that are efficacious and safe, elicit lasting immunity, and cover a range of SARS-CoV-2 variants. Recombinant viral receptor-binding domains (RBDs) are safe vaccine candidates but often have limited efficacy due to the lack of virus-like immunogen display pattern. Here we have developed a novel virus-like nanoparticle (VLP) vaccine that displays 120 copies of SARS-CoV-2 RBD on its surface. This VLP-RBD vaccine mimics virus-based vaccines in immunogen display, which boosts its efficacy, while maintaining the safety of protein-based subunit vaccines. Compared to the RBD vaccine, the VLP-RBD vaccine induced five times more neutralizing antibodies in mice that efficiently blocked SARS-CoV-2 from attaching to its host receptor and potently neutralized the cell entry of variant SARS-CoV-2 strains, SARS-CoV-1, and SARS-CoV-1-related bat coronavirus. These neutralizing immune responses induced by the VLP-RBD vaccine did not wane during the two-month study period. Furthermore, the VLP-RBD vaccine effectively protected mice from SARS-CoV-2 challenge, dramatically reducing the development of clinical signs and pathological changes in immunized mice. The VLP-RBD vaccine provides one potentially effective solution to controlling the spread of SARS-CoV-2.

Cutting Edge: Mouse SARS-CoV-2 Epitope Reveals Infection and Vaccine-Elicited CD8 T Cell Responses.

The magnitude of SARS-CoV-2-specific T cell responses correlates inversely with human disease severity, suggesting T cell involvement in primary control. Whereas many COVID-19 vaccines focus on establishing humoral immunity to viral spike protein, vaccine-elicited T cell immunity may bolster durable protection or cross-reactivity with viral variants. To better enable mechanistic and vaccination studies in mice, we identified a dominant CD8 T cell SARS-CoV-2 nucleoprotein epitope. Infection of human ACE2 transgenic mice with SARS-CoV-2 elicited robust responses to H2-Db/N219-227, and 40% of HLA-A*02+ COVID-19 PBMC samples isolated from hospitalized patients responded to this peptide in culture. In mice, i.m. prime-boost nucleoprotein vaccination with heterologous vectors favored systemic CD8 T cell responses, whereas intranasal boosting favored respiratory immunity. In contrast, a single i.v. immunization with recombinant adenovirus established robust CD8 T cell memory both systemically and in the respiratory mucosa.

Initial stage of titanium oxidation in Ti/CuO thermites: a molecular dynamics study using ReaxFF forcefields.

The paper elucidates the main driving mechanisms at play during the early stage of the Ti/CuO thermite reaction using reactive forcefields in the frame of molecular dynamics calculations. Results show that TiO preferentially forms in immediate contact to pure Ti at temperatures as low as 200 K rather than TiO2. Increasing the temperature to 700 K, the 2 nm TiO2 in contact to Ti is found to be homogeneously depleted from half of its oxygen atoms. Also, the first signs of CuO decomposition are observed at 600 K, in correlation with the impoverishment in oxygen atom reaching the titanium oxide layer immediately in contact to CuO. Further quantification of the oxygen and titanium mass transport at temperatures above 700 K suggests that mostly oxygen atoms migrate from and across the titanium oxide interfacial layer to further react with the metallic titanium fuel reservoir. This scenario is opposed to the one of the Al/CuO system, for which the mass transport is dominated by the Al fuel diffusion across alumina. Further comparison of both thermites sheds light on the enhanced reactivity of the Ti-based thermite, for which CuO decomposition is promoted at lower temperature, and offers a novel understanding of thermite initiation at large.

Anisotropic atomistic shock response mechanisms of aramid crystals.

Aramid fibers composed of poly(p-phenylene terephthalamide) (PPTA) polymers are attractive materials due to their high strength, low weight, and high shock resilience. Even though they have widely been utilized as a basic ingredient in Kevlar, Twaron, and other fabrics and applications, their intrinsic behavior under intense shock loading is still to be understood. In this work, we characterize the anisotropic shock response of PPTA crystals by performing reactive molecular dynamics simulations. Results from shock loading along the two perpendicular directions to the polymer backbones, [100] and [010], indicate distinct shock release mechanisms that preserve and destroy the hydrogen bond network. Shocks along the [100] direction for particle velocity Up < 2.46 km/s indicate the formation of a plastic regime composed of shear bands, where the PPTA structure is planarized. Shocks along the [010] direction for particle velocity Up < 2.18 km/s indicate a complex response regime, where elastic compression shifts to amorphization as the shock is intensified. While hydrogen bonds are mostly preserved for shocks along the [100] direction, hydrogen bonds are continuously destroyed with the amorphization of the crystal for shocks along the [010] direction. Decomposition of the polymer chains by cross-linking is triggered at the threshold particle velocity Up = 2.18 km/s for the [010] direction and Up = 2.46 km/s for the [100] direction. These atomistic insights based on large-scale simulations highlight the intricate and anisotropic mechanisms underpinning the shock response of PPTA polymers and are expected to support the enhancement of their applications.

Unregulated antigen-presenting cell activation by T cells breaks self tolerance.

T cells proliferate vigorously following acute depletion of CD4+ Foxp3+ T regulatory cells [natural Tregs (nTregs)] and also when naive T cells are transferred to syngeneic, nTreg-deficient Rag1-/- hosts. Here, using mice raised in an antigen-free (AF) environment, we show that proliferation in these two situations is directed to self ligands rather than food or commensal antigens. In both situations, the absence of nTregs elevates B7 expression on host dendritic cells (DCs) and enables a small subset of naive CD4 T cells with high self affinity to respond overtly to host DCs: bidirectional T/DC interaction ensues, leading to progressive DC activation and reciprocal strong proliferation of T cells accompanied by peripheral Treg (pTreg) formation. Likewise, high-affinity CD4 T cells proliferate vigorously and form pTregs when cultured with autologous DCs in vitro in the absence of nTregs: this anti-self response is MHCII/peptide dependent and elicited by the raised level of B7 on cultured DCs. The data support a model in which self tolerance is imposed via modulation of CD28 signaling and explains the pathological effects of superagonistic CD28 antibodies.

Characterization of the mechanical properties of van der Waals heterostructures of stanene adsorbed on graphene, hexagonal boron-nitride and silicon carbide.

Stanene has revealed a new horizon in the field of quantum condensed matter and energy conversion devices but its significantly lower tensile strength limits its further applications and effective operation in these nanodevices. Van der Waals heterostructures have given substantial flexibility to integrate different two-dimensional (2D) layered materials over the past few years and have proven highly functional with exceptional features, appealing applications, and innovative physics. Considerable efforts have been made for the preparation, thorough understanding, and applications of van der Waals heterostructures in the fields of electronics and optoelectronics. In this paper, we have executed Molecular Dynamics (MD) simulations to predict the tensile strength of van der Waals heterostructures of stanene (Sn) adsorbed on graphene (Gr), hexagonal boron nitride (hBN), and silicon carbide (SiC) (Sn/Gr, Sn/hBN, and Sn/SiC, respectively) subjected to both armchair and zigzag directional loading at different strain rates for the first time, which has enticing applications in electronic, optoelectronic, energy storage and bio-engineered devices. Among all the van der Waals heterostructures, the Sn/SiC heterostructure exhibits the lowest tensile strength and tensile strain. Furthermore, it has been found that zigzag directional loading could endure more tensile strain before fracture. Besides, it has been disclosed that though the rule of mixtures may accurately reproduce the Young's modulus of these heterostructures, it has limitations to predict the tensile strength. Fracture analysis suggests that for the Sn/hBN heterostructure the fracture initiates from the stanene layer while for the Sn/Gr and Sn/SiC heterostructures the fracture initiates from the Gr and SiC layer, respectively, for both armchair and zigzag directional loading. Overall, this study would aid in the design and efficient operation of Sn/Gr, Sn/hBN, and Sn/SiC heterostructures when subjected to mechanical force.

Growth Kinetics and Atomistic Mechanisms of Native Oxidation of ZrSxSe2-x and MoS2 Crystals.

A thorough understanding of native oxides is essential for designing semiconductor devices. Here, we report a study of the rate and mechanisms of spontaneous oxidation of bulk single crystals of ZrSxSe2-x alloys and MoS2. ZrSxSe2-x alloys oxidize rapidly, and the oxidation rate increases with Se content. Oxidation of basal surfaces is initiated by favorable O2 adsorption and proceeds by a mechanism of Zr-O bond switching, that collapses the van der Waals gaps, and is facilitated by progressive redox transitions of the chalcogen. The rate-limiting process is the formation and out-diffusion of SO2. In contrast, MoS2 basal surfaces are stable due to unfavorable oxygen adsorption. Our results provide insight and quantitative guidance for designing and processing semiconductor devices based on ZrSxSe2-x and MoS2 and identify the atomistic-scale mechanisms of bonding and phase transformations in layered materials with competing anions.

Cost-effective production of tag-less recombinant protein in Nicotiana benthamiana.

Plants have recently received a great deal of attention as a means of producing recombinant proteins. Despite this, a limited number of recombinant proteins are currently on the market and, if plants are to be more widely used, a cost-effective and efficient purification method is urgently needed. Although affinity tags are convenient tools for protein purification, the presence of a tag on the recombinant protein is undesirable for many applications. A cost-effective method of purification using an affinity tag and the removal of the tag after purification has been developed. The family 3 cellulose-binding domain (CBM3), which binds to microcrystalline cellulose, served as the affinity tag and the small ubiquitin-related modifier (SUMO) and SUMO-specific protease were used to remove it. This method, together with size-exclusion chromatography, enabled purification of human interleukin-6 (hIL6) with a yield of 18.49 mg/kg fresh weight from leaf extracts of Nicotiana benthamiana following Agrobacterium-mediated transient expression. Plant-produced hIL6 (P-hIL6) contained less than 0.2 EU/µg (0.02 ng/mL) endotoxin. P-hIL6 activated the Janus kinase-signal transducer and activator of transcriptional pathways in human LNCaP cells, and induced expression of IL-21 in activated mouse CD4+ T cells. This approach is thus a powerful method for producing recombinant proteins in plants.

Dendritic cell expression of the signaling molecule TRAF6 is required for immune tolerance in the lung.

Immune tolerance in the lung is important for preventing hypersensitivity, such as allergic asthma. Maintenance of tolerance in the lung is established by coordinated activities of poorly understood cellular and molecular mechanisms, including participation of dendritic cells (DCs). We have previously identified DC expression of the signaling molecule TRAF6 as a non-redundant requirement for the maintenance of immune tolerance in the small intestine of mice. Because mucosal tissues share similarities in how they interact with exogenous antigens, we examined the role of DC-expressed TRAF6 in the lung. As with the intestine, we found that the absence TRAF6 expression by DCs led to spontaneous generation of Th2-associated immune responses and increased susceptibility to model antigen-induced asthma. To examine the role of commensal microbiota, mice deficient in TRAF6 in DCs were treated with broad-spectrum antibiotics and/or re-derived on a germ-free (GF) background. Interestingly, we found that antibiotics-treated specific pathogen-free, but not GF, mice showed restored immune tolerance in the absence of DC-expressed TRAF6. We further found that antibiotics mediate microbiota-independent effects on lung T cells to promote immune tolerance in the lung. This work provides both a novel tool for studying immune tolerance in the lung and an advance in our conceptual understanding of potentially common molecular mechanisms of immune tolerance in both the intestine and the lung.

Computational Synthesis of MoS2 Layers by Reactive Molecular Dynamics Simulations: Initial Sulfidation of MoO3 Surfaces.

Transition metal dichalcogenides (TMDC) like MoS2 are promising candidates for next-generation electric and optoelectronic devices. These TMDC monolayers are typically synthesized by chemical vapor deposition (CVD). However, despite significant amount of empirical work on this CVD growth of monolayered crystals, neither experiment nor theory has been able to decipher mechanisms of selection rules for different growth scenarios, or make predictions of optimized environmental parameters and growth factors. Here, we present an atomic-scale mechanistic analysis of the initial sulfidation process on MoO3 surfaces using first-principles-informed ReaxFF reactive molecular dynamics (RMD) simulations. We identify a three-step reaction process associated with synthesis of the MoS2 samples from MoO3 and S2 precursors: O2 evolution and self-reduction of the MoO3 surface; SO/SO2 formation and S2-assisted reduction; and sulfidation of the reduced surface and Mo-S bond formation. These atomic processes occurring during early stage MoS2 synthesis, which are consistent with experimental observations and existing theoretical literature, provide valuable input for guided rational synthesis of MoS2 and other TMDC crystals by the CVD process.

Transfer of fibres on the hands of living subjects and their persistence during hand washing.

Textile fibres were transferred to the hands of ten living subjects and their persistence was determined after hand washing. Average number of fibres transferred was 300 ± 133 (female 288 ± 92, male 311 ± 163) per 100 cm(2) hand area in the 100 experiments. However the number of fibres transferred was not gender dependent but individual dependent. The hand texture of subjects was compared with the number of fibres transferred but the relationship was not observed. The number of fibres transferred varied significantly for the 10 repeated experiments performed under the same conditions for the same subject. The subjects were then asked to wash their hands with water. One test group washed their hands with standing water, and the other with running tap water. Afterwards, the number of fibres remaining on the test subjects' hands were investigated. Migration of the fibres on the surface of the observed hands did occur but total loss of transferred fibre after hand washing did not occur. The average number of fibres remaining per 100 cm(2) hand area was 14 ± 10 (range=3-72) for hand washing with standing water, and 10 ± 12 (range=0-79) for washing with running tap water. The results of this study show the possibility of finding fibres on the hands of a person involved in a criminal case even after hand washing before fibre collection.

Gold-coated iron oxide nanoparticles as a T2 contrast agent in magnetic resonance imaging.

Gold-coated iron oxide (Fe3O4) nanoparticles were synthesized for use as a T2 contrast agent in magnetic resonance imaging (MRI). The coated nanoparticles were spherical in shape with an average diameter of 20 nm. The gold shell was about 2 nm thick. The bonding status of the gold on the nanoparticle surfaces was checked using a Fourier transform infrared spectrometer (FTIR). The FTIR spectra confirmed the attachment of homocysteine, in the form of thiolates, to the Au shell of the Au-Fe3O4 nanoparticles. The relaxivity ratio, R2/R1, for the coated nanoparticles was 3-fold higher than that of a commercial contrast agent, Resovist, which showed the potential for their use as a T2 contrast agent with high efficacy. In animal experiments, the presence of the nanoparticles in rat liver resulted in a 71% decrease in signal intensity in T2-weighted MR images, indicating that our gold-coated iron oxide nanoparticles are suitable for use as a T2 contrast agent in MRI.

Photoluminescence of blood by acidic hydrogen peroxide-A preliminary test.

In this study, the authors found that treating blood with 1 M HCl and 2% (w/v) 5-sulfosalicylic acid (SSA) in 1% (v/v) hydrogen peroxide mixture can produce photoluminescence of blood. SSA was added as a blood fixer. The photoluminescence was induced by irradiation of a forensic light source at 505 nm, which was detected using a 550 nm barrier filter. In this experiment, various level of acid and hydrogen peroxide were tested to find the optimal formulation of reagents, spot tests were conducted with diluted blood to test the sensitivity of this reagent, and impressions in blood left on porous/nonporous surfaces were enhanced. The sensitivity of this solution was slightly lower than Bluestar and was similar to leucocrystal violet or leucomalachite green on both porous/non-porous surfaces. The photoluminescence of blood treated with this reagent has been observed over 2 months. Using this reagent, it was possible to observe fingermarks or footwear impressions in blood on a black porous/non-porous surface. Through this, it was found that using this reagent could enhance bloodstains regardless of the porosity or color of the surface.

Atomic-level investigation on the oxidation efficiency and corrosion resistance of lithium enhanced by the addition of two dimensional materials.

Understanding the oxidation and corrosion characteristics of Lithium (Li)-based systems is critical to their successful use as a solid fuel in spacecraft, powerplants, rechargeable batteries, submarines, and many other aquatic and corrosive environments. This study offers a systematic roadmap for engineering the oxidation efficiency and corrosion resistance of Li-based systems using ReaxFF-based Reactive Molecular Dynamics (RMD) simulations for the first time. First, we explored the oxidation mechanism of bare Li (Li/O2) at 1200 K, noticing that the oxidation process quickly ceases due to the creation of a passive oxide film on the Li surface. Afterward, we examined the effect of introducing graphene-oxide (GO) to the oxidation process of Li/O2. Interestingly, the inclusion of GO establishes a new reaction pathway between Li and O2, thus significantly improving oxidation efficiency. Additionally, we realized that when the concentration of GO increases in the system, the oxidation rate of Li/O2 increases considerably. As exposed to O2 and H2O, bare Li is observed to be highly corrosion-prone, while graphene (Gr)-coated Li exhibits excellent corrosion resistance, suggesting that Gr might be used as a promising corrosion-protective shield. Overall, this study is intended to serve as a reference for experimental investigations and assist researchers and engineers in designing more efficient Li-based functional systems.

Dietary antigens suppress the proliferation of type 2 innate lymphoid cells by restraining homeostatic IL-25 production.

Dietary antigens affect the adaptive immunity of the host by inducing regulatory T cells and IgE-producing B cells. However, their roles in innate immune compartments such as innate lymphoid cells (ILCs) and intestinal epithelial cells (IECs) are unclear. Here, using antigen-free (AF) mice, which are germ-free (GF) mice fed with amino-acid-based diet, we found dietary proteins suppress the development of GATA-3-expressing ILC2s independent of the adaptive immune cells. These cells produce more type 2 cytokines and upregulated proliferation and activation markers such as Ki-67, CD69, and CD25. With this, AF mice had increased expressions of tuft cell-specific transcripts such as Il25, Il33, Dclk1, Trpm5, and Pou2f3 in IECs. Accordingly, expanded ILC2s upregulated IL-17RB, a receptor of IL-25, and their proliferation was blocked by IL-25 neutralizing or IL-17RB blocking antibodies. These results suggest a new dialogue between dietary antigens, IECs, and ILCs in which dietary antigens suppress ILC2 activation and proliferation by restraining homeostatic IL-25 production, potentially limiting type 2 immunity by food antigens.

Vascular endothelial growth factor is a key mediator in the development of T cell priming and its polarization to type 1 and type 17 T helper cells in the airways.

Chronic inflammatory airway diseases including asthma are characterized by immune dysfunction to inhaled allergens. Our previous studies demonstrated that T cell priming to inhaled allergens requires LPS, which is ubiquitously present in household dust allergens. In this study, we evaluated the role of vascular endothelial growth factor (VEGF) in the development of T cell priming and its polarization to Th1 or Th17 cells when exposed to LPS-contaminated allergens. An asthma mouse model was induced by airway sensitization with LPS-contaminated allergens and then challenged with allergens alone. Therapeutic intervention was performed during allergen sensitization. The present study showed that lung inflammation induced by sensitization with LPS-contaminated allergens was decreased in mice with homozygous disruption of the IL-17 gene; in addition, allergen-specific Th17 immune response was abolished in IL-6 knockout mice. Meanwhile, in vivo production of VEGF was up-regulated by airway exposure of LPS. In addition, airway sensitization of allergen plus recombinant VEGF induced both type 1 and type 17 Th cell (Th1 and Th17) responses. Th1 and Th17 responses induced by airway sensitization with LPS-contaminated allergens were blocked by treatment with a pan-VEGF receptor (VEGFR; VEGFR-1 plus VEGFR-2) inhibitor during sensitization. These effects were accompanied by inhibition of the production of Th1 and Th17 polarizing cytokines, IL-12p70 and IL-6, respectively. These findings indicate that VEGF produced by LPS plays a key role in activation of naive T cells and subsequent polarization to Th1 and Th17 cells.

Ni-Fe2O4 nanoparticles as contrast agents for magnetic resonance imaging.

Reported herein is the synthesis of a dextran coating on nickel ferrite (Ni-Fe2O4) nanoparticles via chemical coprecipitation. The aqueous solution of the synthesized nanoparticles showed good colloidal stability, and no precipitate was observed 20 months after the synthesis. The coated nanoparticles were found to be cylindrical in shape in the TEM images, and showed a uniform size distribution with an average length and diameter of 17 and 4 nm, respectively. The coated particles were evaluated as potential T1 and T2 contrast agents for MRI. The T1 and T2 relaxations of the hydrogen protons in the water molecules in an aqueous solution of dextran-coated Ni-Fe2O4 nanoparticles were studied. It was found that the T1 relaxivity for the aqueous solution of dextran-coated nanoparticles was slightly greater than that of a commercial Gd-DTPA-BMA contrast agent. The T2 relaxivity, however, was almost twice that of the commercial Gd-DTPA-BMA contrast agent. Animal experimentation also demonstrated that the dextran-coated Ni-Fe2O4 nanoparticles are suitable for use as either T1 or T2 contrast agents in MRI.

Listeria monocytogenes Establishes Commensalism in Germ-Free Mice Through the Reversible Downregulation of Virulence Gene Expression.

The intestine harbors a complex community of bacterial species collectively known as commensal microbiota. Specific species of resident bacteria, as known as pathobiont, have pathogenic potential and can induce apparent damage to the host and intestinal inflammation in a certain condition. However, the host immune factors that permit its commensalism under steady state conditions are not clearly understood. Here, we studied the gut fitness of Listeria monocytogenes by using germ-free (GF) mice orally infected with this food-borne pathogen. L. monocytogenes persistently exists in the gut of GF mice without inducing chronic immunopathology. L. monocytogenes at the late phase of infection is not capable of infiltrating through the intestinal barrier. L. monocytogenes established the commensalism through the reversible down regulation of virulence gene expression. CD8+ T cells were found to be sufficient for the commensalism of L. monocytogenes. CD8+ T cells responding to L. monocytogenes contributed to the down-regulation of virulence gene expression. Our data provide important insights into the host-microbe interaction and have implications for developing therapeutics against immune disorders induced by intestinal pathogens or pathobionts.