Browse Articles

Acute Fasting Suppresses Neutrophil Metabolism and Effector Function 2254957

Abstract Introduction Acute fasting is common in perioperative care and may create a transient window of vulnerability to hospital-acquired infections. Because Staphylococcus aureus drives significant bacteremia in this setting, we asked how a short-term (12-hour) fast alters neutrophil metabolism and antibacterial function. Methods Healthy adult participants underwent an overnight 12-hour fast. Neutrophils were isolated to assess the fasting state ex vivo, then the same individuals consumed standardized breakfast and lunch, and postprandial neutrophils were isolated for repeat assays. Readouts included glycolytic and mitochondrial activity using extracellular flux analysis and quantifying the oxidative burst, NET release, and bactericidal activity upon S. aureus stimulation. In mice, a single 12-hour fast preceded systemic S. aureus challenge and outcomes included survival, tissue burdens, and in vivo neutrophil function. Results During the fasting state, human neutrophils showed reduced glycolytic and mitochondrial activity and failed to mount an oxidative burst or release NETs in response to S. aureus resulting in impaired bactericidal activity compared to postprandial neutrophils. In mice, a 12-hour fast markedly increased susceptibility to staphylococcal bacteremia. This was not explained by uniformly elevated tissue burdens; rather, in vivo analyses revealed tissue-specific neutrophil dysfunction consistent with impaired early containment and progression to excessive, nonproductive inflammation. Conclusion A brief, clinically relevant fast induces a reversible state that suppresses neutrophil metabolism and effector programs, heightening vulnerability to severe S. aureus infection. These findings position acute fasting as a modifiable perioperative risk factor and motivate practical mitigation strategies around feeding timing. Funding Source 1R35GM154838 Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Therapeutic treatment with a single CD4+ T cell epitope containing PLGA nanoparticles reverses CD8+ T Cell induced Type I Diabetes 2260609

Abstract Introduction T cell-mediated autoimmune diseases, such as type I diabetes (T1D) are mediated by autoantigen-specific CD4+ and CD8+ T cells. We have pioneered CD4+ T cell antigen (Ag)-specific tolerance induction therapy via biodegradable poly(lactide-co-glycolide) (PLGA) nanoparticles (also knowns as TIMPs/CNPs) treatment. While TIMP/CNPs have progressed into Phase I/IIa clinical trials in celiac disease, myasthenia gravis, primary biliary cholangitis, and T1D, less is known about how treatment regulates CD8+ T cells. Methods Therefore, the RIP-mOVA mouse model of T1D was presently used to mechanistically address remaining questions. Following OTI CD8+ T cell-induced early onset T1D. Results Following OTI CD8+ T cell-induced early onset T1D, a single dose of CNP-OVA323, i.e., a CD4+ T cell epitope, reversed dysglycemia/T1D in a regulatory T cell-dependent manner for up to 9 weeks post dosing. Of note, CNP-OVA323 treated mice develop a secondary dysglycemia at this timepoint. An event hypothesized to be driven by early IFN-γ-induced beta cell death resulting in spread epitope-specific CD4+ T cell activation during the initial dysgycemic event. As confirmation, treatment with CNP-T1D (containing Ag for the spread epitope-specific T cells) inhibited the development of secondary dysgycemia. Data presented will track the transferred OTI T cells over time, ex vivo recall responses to beta cell expressed proteins during secondary dysgycemia, and the ability of CD4+ T cells to regulate autoreactive CD8+ T cell responses post Ag-specific TIMP treatment. Conclusion CNP containing CD4 Epitopes from Ag’s are able to regulate CD8 T cell responses to the same Ag’s. Furthermore, inclusion of spread epitopes in a second dose of CNP are able to regulate secondary dysglycemia in the RIP-mOVA mouse model of T1D. Funding Source Breakthrough T1D Topic Categories Therapeutic Approaches to Autoimmunity (THER)

Antibody-Mediated Entry Renders Non-Susceptible Macrophages Permissive to SARS-CoV-2 Replication 2307324

Abstract Introduction SARS-CoV-2 entry into epithelial cells via ACE2 is well characterised, but its interactions with alveolar macrophages remain to be fully defined. Macrophages are among the first immune cells to detect infection and initiate protective responses, however, in severe COVID-19 the response is hyperinflammatory. Whether they express ACE2, support viral replication, or directly drive COVID-19 pathology is unresolved. Defining SARS-CoV-2 entry routes and replication competence in macrophages is essential to understanding their dysfunction in severe disease. Methods iPSC-derived macrophages were used as an in vitro model of tissue-resident macrophages. Viral internalisation and intracellular localisation were analysed by imaging flow cytometry. Viral RNA was quantified by smFISH and qRT-PCR, and infectious output virus measured by focus-formation assay. Antigen presentation was assessed in macrophage-T cell co-cultures using intracellular cytokine staining. Results Macrophages efficiently internalise SARS-CoV-2 through a non-specific mechanism and do not express ACE2 or spike-binding receptors. Internalised viral RNA does not replicate and is degraded. Macrophages subsequently present viral antigens and activate CD4+ T cells. Notably, macrophages become both susceptible and permissive to productive infection, when provided with a surrogate entry receptor, such as transgenic ACE2 or specific anti-RBD antibodies. Replication is enhanced by JAK1/2 inhibition, consistent with interferon-mediated restriction. Conclusion These findings show that macrophages are not susceptible to SARS-CoV-2 infection since they do not express ACE2, despite efficiently internalising and degrading the virus. Specific anti-RBD antibodies can act as surrogate receptors that permit productive macrophage infection, suggesting a potential in vivo pathway that may contribute to macrophage dysfunction and hyperinflammation in severe COVID-19. This raises considerations for the therapeutic use of certain monoclonal antibodies. Funding Source University of Oxford, Dunn School of Pathology, EPA Studentship Topic Categories Viral Immunology (VIR)

New‐Era Polymer Thermoelectrics: Material Innovations, Doping Frontiers, Decoupling Strategies, and Unconventional Applications (Adv. Mater. 43/2026)

Macro‐Superlubricity Induced by Synergistic Effect of Molybdenum Carbide/Molybdenum Disulfide Heterojunction and Conformation‐Transformable Ethylene Glycol Molecule

ABSTRACT Macroscopic solid–liquid synergistic superlubricity based on two‐dimensional (2D) materials and liquid molecules integrates the low interlayer shear strength of 2D materials with advantages of liquid superlubricity, significantly reducing friction and wear in industrial applications. However, the low load‐bearing capacity of single 2D material and the unclear synergistic superlubricity mechanism severely constrain the design of long‐lasting superlubricity systems. Herein, through innovatively introducing van der Waals interaction‐induced molybdenum carbide/molybdenum disulfide (Mo 2 CT x /MoS 2 ) heterojunction as additives in ethylene glycol (EG), this Mo 2 CT x /MoS 2 –EG superlubricity system exhibits an ultralow friction coefficient of 0.0044, an extremely low wear rate of 8.17 × 10 −10 mm 3 •N −1 •m −1 , and an ultralong lubrication life of 432 000 cycles (equal to 2160 m). Meanwhile, the maximum average contact pressure reaches 711.9 MPa. The in‐situ formed special tribofilms enriched with heterojunction and homojunction nanosheets effectively reduce the shear strength under boundary lubrication. Notably, the ultralow shear strength at the Mo 2 C‐O/EG solid–liquid interface and the unique pressure‐responsive lubrication properties of EG molecules which leads to the formation of short‐range‐ordered structures are also crucial for achieving superlubricity. This study provides a new perspective for designing novel solid–liquid synergistic superlubricity systems by utilizing heterojunction nanomaterial and has great potential for application in industrial lubrication.

Advanced exergy and exergoeconomic assessment of a flare-gas-fired GT–HRSG–ST–ORC–LiBr polygeneration system under a locked thermodynamic boundary

Fusion Peptide Priming and Trimer Boosting Strategies for HIV Vaccine Development in Guinea Pigs 2308606

Abstract Introduction Inducing broadly neutralizing antibodies (bnAb) is a central challenge in HIV vaccine development. The HIV envelope (Env) fusion peptide (FP) is a highly conserved bnAb epitope, yet is inefficiently targeted by antibodies elicited by experimental vaccines. Here, we test how Env trimer design and dosing strategies shape the magnitude and specificity of antibody responses by pairing FP priming with boosts using native-like Env trimers. Escalating-dose (ED) immunization regimens are a recently described approach to deliver antigens over a prolonged period, resulting in sustained increased antigen stability in the lymph node and germinal-center activity, improved immune kinetics, and enhanced recruitment and selection of targeted B-cell populations. The impact of ED regimens on boosting HIV-1 FP-specific responses has not been explored. Methods Here, guinea pigs (n = 5 per group) were primed with FP-rTTHC followed by boosts with prefusion trimers from clade A (BG505), clade C (ConC), or a Triple-Tandem-Trimer (three trimeric units of ConC, linked one after another: ConC-DS-14ln-TTT), delivered either as bolus injections or ED regimens. Results All strategies elicited strong Env-specific responses, but the quality of these responses varied among immunogen designs. Bolus boosts using BG505 or ConC generated the most robust FP-specific recall responses, but neutralization levels ultimately converged across regimens. However, the TTT trimer expanded reactivity to non-FP epitopes, suggesting broader epitope engagement during boosting. Notably, cross-reactive BG505-binding antibodies appeared as early as week 6 after two FP primes, highlighting rapid activation of FP-targeting B cells. Conclusion These findings clarify how trimer architecture and dosing influence the trajectory of epitope targeting, strengthening the case for FP priming as a powerful method for engaging a vulnerable HIV Env epitope and demonstrating that the TTT platform may serve as an effective boost to epitopes beyond FP. Funding Source N/A Topic Categories Vaccines and Immunotherapy (VAC)

A rapid IL-9-secreting T cell response mediates uropathogenic Escherichia coli (UPEC) clearance in an acute UTI mouse model 2267492

Abstract Introduction Urinary tract infections (UTIs) are commonly caused by gut-derived pathogenic bacteria, such as Escherichia coli, that infect the bladder. Antibiotic resistance in UTIs is of increasing clinical concern. IL-9 is a cytokine that regulates inflammation at mucosal barrier sites and promotes parasite clearance during helminth infection. Utilizing a uropathogenic E. coli (UPEC) mouse model, we examined IL-9 function in UTI progression, with a focus on epithelial integrity and bacterial clearance. Methods Public datasets and tissue biopsies obtained from the Cooperative Human Tissue Network were assessed for IL9R expression in patients with bacterial UTIs and healthy controls. For in vivo studies, WT, IL-9-deficient, IL-9R-deficient, and CD4-specific Il9 conditional mutant mice were transurethrally administered PBS with or without UPEC (strain UTI89). Mice were euthanized up to 24 hours post-inoculation for tissue processing; bladder bacterial burden, gene expression, immune infiltrate, and intracellular IL-9 protein levels were assessed. Results We found elevated IL-9 receptor expression on the luminal bladder surface in patients with bacterial UTIs compared to healthy controls. In the UPEC model, mice lacking IL-9 (Il9-/-) or its receptor (Il9r-/-) exhibited increased bacterial burden at 24 hours, reduced epithelial sloughing, and impaired immune recruitment, suggesting IL-9 supports rapid physical and immune-mediated bacterial clearance in UPEC infection. Using Il9 conditional mutant mice, we observed T cells as a source of IL-9. IL-9 secretion was detected from multiple T cell subsets, including CD4+ T cells, MAIT, and NKT cells, suggesting the potential involvement of innate T cell subsets. Conclusion T cell IL-9 is important for anti-bacterial immunity in the bladder. Ongoing studies will define the contributions of IL-9-producing T cell subsets in bacterial clearance and explore how these mechanisms may be therapeutically targeted for the treatment of antibiotic-resistant and recurring UTIs. Funding Source NIH NIAID (R01AI180518; R01AI057459; R21AI173929) Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)

A BACH2-STAT5 transcriptional module licenses a short intracellular C3 proteoform in activated CD4+ T cells 2308647

Abstract Introduction IL-2-STAT5 signaling is vital for CD4+ T cell function, whereas BACH2 restrains activation. How these transcription factors co-operate at the genome remains unclear. Intracellular C3 supports T cell survival and effector programming, but how it is transcriptionally controlled and whether it matches serum C3 is unknown. We hypothesized that a BACH2-STAT5 module coordinates C3 expression and licenses a distinct C3 proteoform in activated CD4+ T cells. Methods We profiled wild-type and Bach2—/— CD4+ T cells by RNA-seq and mapped STAT5 and BACH2 binding using ChIP-seq and H3K27ac Hi-ChIP. STAT5 and BACH2 were re-expressed in Stat5—/— T cells, and shared elements at the C3 locus were deleted by CRISPR. IL-2/STAT5 inhibition, C3 overexpression or knockdown, and complement activation assays defined C3 expression and function. AlphaFold2 modeling and domain analyses characterized novel C3 proteoforms. Results IL-2/STAT5 target genes were enriched in Bach2—/— CD4+ T cells, with overlapping STAT5 and BACH2 binding at differentially expressed loci. Combined activating or repressive control was confirmed by re-expression in Stat5—/— cells. At the C3 locus, BACH2-STAT5 co-occupancy and IL-2/STAT5 signaling were required for C3 transcription; C3 was lost in Bach2—/— cells, after IL-2/STAT5 blockade, or upon CRISPR deletion of shared elements. Resting CD4+ T cells expressed canonical, signal peptide-containing C3, whereas activation induced a shorter, signal peptide-deficient transcript predicted to encode an intracellular proteoform. Purified short C3 failed to seed classical C3 convertase or support hemolysis, indicating altered function. Conclusion We identify a BACH2—STAT5 transcriptional module that is essential for C3 expression in activated CD4+ T cells and redirects C3 output toward a short, intracellular proteoform with noncanonical complement activity. Proteoform-specific, T cell-intrinsic complement suggests new opportunities to target BACH2-STAT5 and C3 in autoimmunity and immunotherapy. Funding Source NA Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

MEX3B as a Potent Regulator of Inflammasome Signaling 2267250

Abstract Introduction Inflammasomes are crucial for innate immune defense to many microbial infections but are also subjected to stringent cellular regulation to avoid persistent, aberrant activation that could lead to lethal sepsis and autoinflammatory conditions. Ubiquitination is one of the most important posttranslational modifications involved in many immune signaling pathways, including the nucleotide-binding and oligomerization domain (NOD)-like receptor 3 (NLRP3) inflammasome. However, its role in the non-canonical caspase-4 inflammasomes is poorly understood. Methods To address this significant gap, we constructed 375 individual ubiquitin E3 ligase knockout lines by CRISPR-Cas9 and performed an unbiased screening. This library represents almost all the currently known definite E3 ligases (total ∼377). Results Our screen identified 15 positive regulators of the caspase-4 inflammasome; one of the top hits was Muscle Excess 3B (MEX3B), an RNA-binding protein with a ubiquitin ligase domain. We found that deletion of MEX3B inhibited caspase-4 and gasdermin D (GSDMD) activation, pyroptosis, and secretion of inflammasome-dependent inflammatory cytokines in human cell lines and murine primary macrophages upon priming with interferon gamma (IFN-γ) and transfection with lipopolysaccharide (LPS). Notably, MEX3B was also crucial for canonical inflammasome signaling such as NLRP3 and NLRC4. Mechanistically, MEX3B was required for caspase-1 activation but not NLRP3 oligomerization. The role of MEX3B in inflammasome signaling was reliant on its RNA-binding, but not E3 ligase activity. However, the expression of caspase-4 and GSDMD, IFN-γ and Toll-like receptor (TLR4) signaling, and apoptosis remained intact in MEX3B-/- cells. Conclusion Our results suggest that MEX3B is a pan-inflammasome regulator targeting inflammatory caspases. Ongoing work is to investigate both the NLRP3 and caspase-4 inflammasome signaling in Mex3b-/- mice and elucidate the molecular mechanism of action of MEX3B. Funding Source National Institute of Allergy and Infectious Diseases, USA Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

B cells induce the age-related accumulation of CD39+CD73+ CD8+ T cells 2305676

Abstract Introduction Even in “healthy” aging, adaptive immune cells become dysregulated, increasing age-associated B cells and exhausted and senescent T cells in the circulation. Although the clinical consequence of this dysregulation remains poorly understood, we previously reported that aged B cells, through the induction of cytolytic CD8+ T cells, can be responsible for the surprising suppression of orthotopic B16-F10 melanoma in aged C57BL/6 mice. Despite this, it remains unclear why this pathway fails to prevent the age-associated increase in other cancers. Methods We performed comprehensive flocytometric analysis combined with scRNA-seq and ATC-seq of adaptive immune cells in young (8-20 weeks old) and aged (17-20 months old) mice. Results While the age-related cancer increase can be explained by exhaustion or senescence of T cells, we do not detect significant accumulation of these cells in aged mice. Instead, we find that aged mice markedly increase a previously unknown subset of CD8+ cells expressing CXCR6 and adenosine-generating CD39 and CD73. These cells (termed DP8), which are also induced by aged B cells, appear to play the key role in the increase of cancer in aged mice. Unlike cancers that regress, cancers that progress in aged mice recruit DP8 cells through the production of CXCL16, which then suppress antitumor CD4+ T cells by generating immunoregulatory adenosine using their ectonucleotidases CD39 and CD73. Conclusion Our results underscore the importance of aged B cells in cancer. By targeting B cells, aging elicits both cancer-promoting and cancer-blocking CD8+ T cells. Funding Source This work was supported by the Intramural Research Program, National Institute on Aging Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Unlocking Therapeutic Vulnerabilities in Leukemia with CRISPR-Based Screens 2255583

Abstract Introduction Background: Leukemia is a cancer of hematopoietic stem cells in the bone marrow. It is classified as lymphoid or myeloid, and as acute (rapid onset) or chronic (slow progression). Advances in CRISPR technology enable deeper study of leukemia biology and therapeutic targets. While AML, CML, ALL, and CLL are distinct, comparative studies highlight shared and unique features. Identifying key pathways across subtypes may yield improved treatments. Our lab has established a unique CRISPR Activation (CRISPRa) platform for gain-of-function screening applications. We hypothesize that genome-wide CRISPRa screening will uncover subtype-specific genetic dependencies and drug resistance mechanisms, offering novel therapeutic insights. Methods We engineered leukemia cell lines by nucleofecting a self-selecting CRISPRa PiggyBac plasmid carrying a blasticidin resistance gene, enabling selection of CRISPRa-competent cells. Functionality was validated using lentiviral delivery of sgRNAs targeting cell surface markers. Cells were transduced with our whole-genome CRISPRa library, Sonata, at MOI 0.4, followed by puromycin selection to enrich for sgRNA-expressing cells. Post-transduction, cells were harvested at various timepoints for sequencing to track sgRNA abundance and identify growth-modulating genes. Results We engineered and validated four CRISPRa-competent leukemia lines: K562 (CML), Jurkat (T-ALL), THP-1 (AML), and HL60 (AML). We have completed whole-genome screening campaigns in the CML and T-ALL backgrounds, identifying hundreds of shared and context specific growth modifiers. Functional validation, pathway analysis, and potential clinical significance is ongoing, as is expansion of our screens with the AML contexts. Conclusion Conclusions: Our screens reveal genes that, upon activation, influence leukemia cell growth. These findings support discovery of new therapeutic targets and enhance understanding of leukemia subtype biology, guiding future personalized treatment strategies. Funding Source The Cole Foundation, FHMR Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Colonic immunometabolic host response mechanisms to ionizing radiation in a mouse model of acute radiation syndrome 2260900

Abstract Introduction Exposure to high levels of ionizing radiation (IR) causes severe health effects, including acute radiation syndrome (ARS). While some therapeutic options exist to stimulate hematopoiesis, there is an unmet clinical need to develop effective therapeutics for gastrointestinal-ARS (GI-ARS). Methods A global transcriptomics time course (d 0, 1, 2, 3, 5, 7, 10 and 18) was conducted on colon of mice total body-irradiated with a single dose of sublethal (3 and 6 Gy) or lethal (12 Gy) IR to evaluate the gene expression changes associated with mortality or recovery in GI-ARS. Results In the lethal IR dose group, the day preceding mortality marked a transcriptional shift of over 12,000 genes. By clustering differentially expressed genes, twenty distinct temporal patterns were identified. Three notable gene clusters were defined by temporal patterns associated with mortality on d 10 and displayed significant functional enrichment characterized by dysregulated effector immunoinflammatory responses and metabolic disruption. The first cluster identified the upregulation of early (d 3) and late (d 10) inflammation, with enrichment of innate immunity genes. The second cluster was enriched in adaptive immunity genes and was characterized by initial downregulation, followed by an upregulated peak at d 10. The third cluster was characterized by an early downregulation with a secondary decrease on d 10 with an enrichment of sterol metabolism genes. Interestingly, lipid metabolism and sterol metabolism were downregulated at early timepoints followed by later re-stabilization, yet only sterol metabolism was downregulated again on d 10. Conclusion This comprehensive system-wide transcriptional analysis of colonic samples provides new insights into the host response mechanisms underlying IR-induced intestinal damage and subsequent mortality, along with identification of novel immunometabolic mechanisms with strong potential for therapeutic development in GI-ARS. Funding Source N/a Topic Categories Immune Mechanisms of Human Disease (HUM)

Developmental and epigenetic remodeling of inflammation in the aging human innate immune system 2238939

Abstract Introduction Epigenetic alterations and chronic inflammation are hallmarks of aging, shaped by lifelong exposure to environmental, infectious, and psychosocial stressors in humans. Advances in systems biology approaches enable interrogation of these processes directly in humans. We tested the hypothesis that aging reprograms hematopoietic stem cells (HSCs), producing myeloid progeny epigenetically poised for chronic inflammation. Methods In two human cohorts, we assessed inflammation using in vitro stimulation and spectral flow cytometry. In selected individuals, we performed single-cell multiome (ATAC+RNA, scMultiome) sequencing of myeloid cells. Results were validated in publicly available single-cell multiome datasets. Results Myeloid cells from older adults showed a higher pro-inflammatory response to stimulation, which was associated with enhanced chromatin accessibility at AP-1 transcription factor motifs, reproducible across four independent datasets. Enhanced AP-1 accessibility led to transcriptional responses associated with inflammation. Because circulating myeloid cells are short-lived, we examined whether the epigenetic remodeling originates in myeloid precursors. Pseudotime trajectory analysis of scMultiome data supports this model, showing increased AP-1 accessibility in HSCs from older adults, pointing to an inflammatory origin in stem cells. To directly assess lineage output and clonal fitness of AP-1-poised HSCs, we leveraged mitochondrial DNA variants as natural barcodes for lineage tracing in humans. Clonotype analysis revealed a polyclonal increase, without selective expansion of AP-1hi HSCs. Nonetheless, AP-1hi HSCs produced a higher proportion of AP-1hi monocytes, though the effect was modest, suggesting that circulating inflammatory mediators reinforce epigenetic poising. Conclusion Our results reveal a developmental origin of inflammatory poising in aging myeloid cells, with AP-1-remodeling as a molecular driver of innate immune aging, and potential target to treat inflammaging. Funding Source University of Arizona Start-up Funds Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)

Soft Skins With Reversible Thickness Morphing: Materials, Mechanisms, and Applications

ABSTRACT Soft skins with reversible thickness morphing represent a distinct and underexplored class of adaptive material interfaces. Unlike conventional soft actuators that achieve motion through bending, elongation, or twisting, these systems enable out‐of‐plane deformation, producing localized protrusion, retraction, and programmable contact mechanics without rigid support structures. This review reframes thickness modulation not merely as an actuation outcome, but as a material–architecture strategy that couples energy transduction, geometry, and compliance to enable new modes of haptic interaction, morphological adaptation, and operation in confined or unstructured environments. We present a comprehensive synthesis of thickness‐morphing soft skins, covering actuation stimuli, material platforms, structural architectures, fabrication strategies, modeling frameworks, and system‐level integration. Particular emphasis is placed on hierarchical elastomer composites, origami‐ and kirigami‐inspired designs, electrohydraulic and multimodal hybrid systems, and emerging data‐driven control approaches that expand the functional design space. Despite rapid progress, key challenges remain in durability under cyclic loading, energy efficiency and autonomy, scalable manufacturing, and integration of sensing, actuation, and computation. Addressing these challenges will enable self‐powered, fault‐tolerant, and computationally intelligent soft skins capable of embodied perception and safe autonomous operation, positioning thickness morphing as a foundational design axis for next‐generation haptics and soft robotic systems.

Developing AEB control strategy for ADAS vehicles considering vehicles in front and rear

mRNA Vaccines Enhance Tumor Control and Synergize with 4-1BB Costimulatory Therapy 2309043

Abstract Introduction Although mRNA—lipid nanoparticle (mRNA-LNP) vaccines are being explored as cancer immunotherapies, their clinical application is constrained by the need for personalized identification of tumor neoantigens and time-consuming vaccine design. Here, we investigated whether an off-the-shelf, antigen-mismatched mRNA-LNP vaccine could elicit bystander anti-tumoral effects without the need to express any tumor antigen. Methods C57BL/6 mice were challenged subcutaneously with 1x10^6 B16 melanoma cells, and five days later, they were immunized intratumorally with an mRNA-LNP encoding an irrelevant antigen, ovalbumin (mRNA-OVA), either alone or in combination with a tolerable dose of 4-1BB agonistic antibodies (50 μg of clone 3H3, administered intraperitoneally). Results Remarkably, mRNA-OVA vaccination alone resulted in robust tumor control despite the absence of any tumor antigen. Notably, mice mounting the strongest OVA-specific CD8 T cell responses exhibited the greatest tumor control, suggesting a role for bystander CD8 T cell activation in potentiating anti-tumoral activity. Importantly, the anti-tumoral efficacy of mRNA-LNP vaccination was further enhanced when combined with a low dose of costimulatory anti-4-1BB antibodies. This costimulatory anti-4-1BB regimen was well tolerated and did not result in elevated liver enzyme levels in mice. Mechanistically, mRNA vaccination induced potent type I interferon (IFN-I) responses, which increased the expression of costimulatory molecules, rendering tumors more responsive to costimulatory triggering. Conclusion Together, these findings demonstrate that mRNA-LNP vaccines promote anti-tumor immunity and can synergize with 4-1BB costimulation to enhance tumor control, providing a mechanistic rationale for combining mRNA vaccines with costimulatory immunotherapies. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Dengue-specific T cell immunity influenced by various levels of pre-existing immunity 2267557

Abstract Introduction Dengue virus (DENV) is a mosquito-borne flavivirus responsible for significant global morbidity and mortality. Prior exposure to different DENV serotypes can shape subsequent immune responses and influence both protection and risk. This challenges the development of a universally effective and safe dengue vaccine, capable of inducing balanced and durable immunity against all four serotypes. T cells constitute a critical arm of dengue immunity, acting independently of antibodies. In this work, we have evaluated the dengue-specific T-cell responses elicited by an attenuated monovalent DENV3 vaccine in participants with various levels of pre-existing DENV exposure. Methods Peripheral blood mononuclear cells (PBMCs) from vaccine recipients of different dengue serostatus (seronegative, monotypic, and multitypic-exposure) were stimulated with experimentally defined DENV3 epitope megapools for DENV structural and nonstructural epitopes. We profiled T cell activation markers, memory phenotypes, and cytokine secretion using a panel of 30+ markers in high-parameter flow cytometry. Results T cells responses were analyzed longitudinally pre-vaccination, and 15, 28 and 90 days after vaccination. The DENV-3 challenge induced measurable DENV-specific T-cell responses for both structural and non-structural components. We observed individual variability in T cell responses that are influenced by their baseline serostatus and viremia burden. Conclusion Our study provides valuable insights into the factors shaping cellular immunity to a novel DENV challenge Funding Source 2P01AI106695-11 Topic Categories Viral Immunology (VIR)

Buildable linkers: Site-Specific Antibody-Drug Conjugates with Well-Defined Payloads 2308812

Abstract Introduction Immunotherapy is experiencing rapid growth by increasingly its use of antibody drug conjugates (ADC). Antibodies are a desired vehicle for immunotherapy because they can deliver a drug to a precise cellular location. Heterogenous conjugates have been shown to affect stability, efficacy and binding to intended target. Successful cellular internalization of an ADC with the controlled release of the cytotoxic drug is required for an effective ADC. The creation of a site-specific, tailor-made ADC with a buildable scaffold is expected to increase an ADC’s functional efficacy and its sensitivity in immunoassays. Methods The well-established Trastuzumab is conjugated to various linker structures and using NHS chemistry with 4 molar excesses to generate conjugates that range in degree of labeling (DOL) from 3-14. Trastuzumab site specific conjugates have a buildable scaffold that corresponds to the NHS DOLs. The performance of the antibody conjugates is compared by immunoassay results, including immunofluorescence (IF) and cell viability assay. Results Antibody drug conjugates that are generated using NHS and site-specific chemistry methods show that lower degrees of labeling for NHS and site specific are comparable in binding results. However, as the DOL increases with high labeling and size of the scaffold grows, there’s a distinction between the effectiveness of NHS and site-specific conjugates with expanding drug scaffolds. Conclusion Antibody drug conjugates (ADC) provide the specificity of the monoclonal antibody with the precise delivery of a covalently labeled cytotoxic drug. Therefor generating an ADC that is homogenous will mitigate the undesirable effects of a heterogenous conjugate. Here we show the benefits of creating a buildable drug scaffold that’s accommodating for achieving exact payload conjugated at a specific site on Trastuzumab. This approach will enable all therapeutic antibodies to be conjugated in a uniform manner, accelerating the advancing field of ADC therapeutics. Funding Source National Science Foundation EPSCoR Award #2119237 Topic Categories Technological Innovations in Immunology (TECH)

Evaluation of intrinsic allergenicity of amaranth seed, pinto bean and kidney bean using an adjuvant free mouse model of food allergy 2299638

Abstract Introduction Food allergy is a growing public health problem of critical significance because of the heightened risk of potential for fatal systemic anaphylaxis. Most food allergies in the United States are caused by 9 common foods: Milk, egg, wheat, soy, peanut, tree nut, fish, shellfish, and sesame. Currently, the allergenicity potential of other foods besides these common foods is incompletely understood. In this study, we assessed the intrinsic allergenicity of protein extracts obtained from 3 commonly used foods: Amaranth seed (AS), pinto bean (PB), and kidney bean (KB). Methods Using an adjuvant-free transdermal sensitization (TS)/ oral elicitation (OE) mouse model of food allergy, we tested the hypothesis that these 3 foods will be intrinsically non-allergenic in this model. Groups of Balb/cJ mice (n = 10/group) were exposed to respective food proteins or saline via skin (once a week for 4 weeks) to test their sensitization potentials. Blood samples collected before and after skin exposures were used to measure food specific IgE antibody titers using an ELISA. Sensitized vs. control mice were orally challenged with the respective proteins to elicit anaphylaxis that were quantified by rectal thermometry and clinically assessed using symptom scoring method. Results Specific IgE was not detectable for any of the foods before skin exposures. However, after 4 transdermal exposures, significant specific IgE levels were elicited by all 3 food proteins. However, the oral challenge with PB or KB to sensitized mice or saline control mice elicited weak or no anaphylaxis. In contrast, oral challenge with AS to sensitized, but not control mice, elicited life-threatening anaphylaxis. Conclusion In summary: i) in contrast to PB, and KB, AS--that is becoming a popular food ingredient, exhibits high intrinsic potential to clinically sensitize mice for life-threatening oral anaphylaxis; and ii) TS/OE model is a useful preclinical testing tool to evaluate intrinsic allergenicity of food proteins. Funding Source US EPA STAR Grant RD83482201; USDA/NIFA: MICL02023, 02486, 02796 Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)