Browse Articles

Mirtron Overexpression Drives Immune Modulation and Tumor Progression Through Macrophage Polarization and Oncogenic Pathway Regulation 2309835

Abstract Introduction Mirtrons, a specialized subclass of intron-derived microRNAs (miRNAs), bypass canonical Drosha-dependent processing by undergoing splicing and debranching to form pre-miRNA hairpins directly processed by Dicer. This alternative pathway imparts high structural stability and resistance to exonucleolytic degradation, supporting their persistence in the tumor microenvironment. This study investigates the expression of specific human mirtrons and their influence on macrophage proliferation and polarization within cancer-associated conditions. Methods Human cancer cell lines (T47D, MDA-MB-231, PC-3, and MCF-7) and normal HEK cells were cultured to assess mirtron expression. Quantitative RT-PCR was performed to measure levels of human mirtrons such as miR-5010, miR-1228, miR-1224, miR-6756, miR-7110, and uc002mxv.2. Exosomes isolated from mirtron-overexpressing cancer cells were co-cultured with THP-1 and U937 macrophage models. Cell proliferation assays evaluated macrophage growth following exposure, while qRT-PCR quantified polarization markers to distinguish M1 and M2 phenotypes. Results All tested mirtrons showed marked overexpression in cancer cell lines relative to normal HEK controls. Exosome-mediated delivery of these mirtrons to macrophages significantly reduced proliferation rates. Expression analysis revealed downregulation of M1-associated markers (TNFα, IL-6, ARG2) and upregulation of M2-associated markers (CD206, IL-10, TGFβ), indicating polarization toward an immunosuppressive phenotype. Conclusion These results suggest that mirtrons modulate macrophage fate through post-transcriptional regulation of inflammatory pathways, linking RNA processing mechanisms to tumor—immune crosstalk. Their overexpression in cancer cells promotes M2 macrophage polarization and suppresses macrophage proliferation, thereby supporting immune evasion and tumor progression. These findings highlight mirtrons as potential diagnostic biomarkers and RNA-based therapeutic targets in molecular oncology. Funding Source NIGMS (R16GM153648) Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Loss of pressure induced constriction responses in cerebral vessels promotes injury and inflammation in a mouse model of systemic lupus erythematosus 2310241

Abstract Introduction SLE is a chronic autoimmune disease in which up to 75% of patients develop neuropsychiatric symptoms (NPSLE), characterized by cognitive dysfunction, mood disorders and seizures; however, the pathogenesis of NPSLE is unclear. Pressure-induced constriction (PIC) is an intrinsic mechano-dependent response in small arteries in the brain. Because of the prevalent hypertension and vascular dysfunction in SLE, we hypothesized that SLE mice would have a loss of the PIC response as the disease progressed, leading to vascular injury, inflammation, and disruption of the blood brain barrier (BBB), all of which could contribute to the development of NPSLE. Methods Middle cerebral arteries were isolated from control (NZW) and SLE (NZBWF1) mice at 10 (pre disease) and 30 (advanced disease) weeks of age for assessment of PIC responses. In additional subsets of animals, cerebral vessels were isolated for flow cytometric analyses, cerebrovascular cytokine expression, and blood brain barrier integrity analysis. Results In SLE mice, PIC responses were attenuated 50% at 10 weeks of age and totally abolished by 30 weeks of age. In addition, SLE mice had increased expression of MCP-1, MCP-5, and CSF-1, and ICAM-1 in cerebral vessels compared to controls. We also found increased numbers of CD45+ leukocytes and CD11b+Ly6C+ monocytes in cerebral vessels of SLE mice. Finally, SLE mice also had a loss of BBB integrity, as indicated by increased Evans blue extravasation into brain tissue. Conclusion Taken together, these findings indicate progressive PIC impairment in SLE, leading to vascular inflammation and BBB disruption. Loss of the PIC response by 10 weeks of age precedes the onset of overt autoimmune disease, suggesting loss of PIC responses may be a causative factor in cardiovascular consequences in SLE. Future studies will examine mechanisms underlying loss of the PIC response in SLE cerebral vessels and protection against NPSLE symptoms. Funding Source R01 DK137167, P30 GM149404, U54 HL169191 Topic Categories Basic Autoimmunity (BA)

Type 2 conventional dendritic cells mediate FVIII recognition in FVIII immune response 2256433

Abstract Introduction Hemophilia A is an X-linked bleeding disorder caused by mutations in the F8 gene, leading to a deficiency of coagulation factor VIII (FVIII). However, FVIII infusions used for the treatment of bleeding can trigger the development of neutralizing FVIII antibodies. Although several studies have demonstrated that various immune cells, such conventional dendritic cells (cDCs), macrophages, monocytes, and marginal zone B cells, co-localize or interact with FVIII, the primary immune cell population mediating FVIII immune recognition remains poorly explored. We hypothesize that type 2 cDCs (cDC2) mediate FVIII recognition and CD4+ T cell activation that enhances FVIII immunity. Methods Batf3⁻/⁻ mice (cDC1 knockout [KO]), Zeb2-Delta3⁻/⁻ mice (cDC2 KO), and Zbtb46-DTR mice without diphtheria toxin depletion (WT) were utilized to isolate bone marrow-derived dendritic cells (BMDC) for FVIII uptake. cDC1 ko mice received five doses of recombinant human FVIII or normal saline via retro-orbital (RO) injection. CD4+ T cells from immunized mice were co-cultured with BMDCs isolated from naïve cDC1 KO, cDC2 KO, and WT mice and labeled with CellTrace Violet (CTV) to assess CD4+ T cell proliferation. Plasma samples were collected to measure anti-FVIII antibody levels by ELISA. Results Immature BMDC endocytosed more FVIII than mature and intermediate-mature BMDC as expected. However, immature BMDC from cDC2 KO mice exhibited reduced FVIII uptake compared to those from WT and cDC1 KO mice. Although BMDC from cDC2 KO mice enhanced CD4+ T cell proliferation compared to WT mice and cDC1 KO mice, cDC2 KO mice treated with FVIII exhibited reduced anti-FVIII antibodies than WT mice and cDC1 KO mice. Conclusion Together, these data suggest that cDC2 predominantly contributes to FVIII-specific adaptive immunity through promotion of FVIII uptake and anti-FVIII antibodies formation. Funding Source K99 Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)

Induction of germinal centers in human Peyer’s patches shapes robust intestinal adaptive immunity in early life 2260324

Abstract Introduction Early life is a critical period in which the immune system encounters a variety of new antigens including pathogens, food antigens, and vaccines. Many of these initial exposures occur in mucosal sites such as the gut, where Peyer’s patches (PP) house local B cell responses that establish protection or maintain homeostasis. However, the dynamics of intestinal adaptive immunity during development remains poorly defined in humans. Methods We profiled paired PP and mesenteric lymph nodes (MLN) from pediatric human organ donors aged 0-11 years using immunofluorescence microscopy and single-cell RNA-seq (scRNA-seq) to investigate germinal center (GC) dynamics across space and time. Results We found robust induction of GC reactions in PPs and MLNs, significantly greater in PPs, peaking at ∼2 years and declining thereafter. Profiling by scRNA-seq revealed prominent GC, plasma, and memory B cell populations, with GC B cell proportions declining and memory B cells increasing across age. GC B cells consisted of 6 subsets corresponding to distinct light and dark zone B cells, which provides a unique window into human GC formation and somatic hypermutation (SHM). Paired BCR analysis showed large clonal expansion in GC B cells and age-associated higher SHM frequency, consistent with ongoing affinity maturation. Interestingly, clonal overlap was minimal between spatially separated PPs within the same donor, indicating GC responses in the gut are highly compartmentalized. Complementing these B cell dynamics, three transcriptionally distinct follicular helper T cell subsets showed discrete dynamics with age and GC magnitude, suggesting subset specific contributions to GC induction and waning. Conclusion Together, our data demonstrate that the first two years of life is a critical window to establish intestinal adaptive immunity, driven by expansive and spatially compartmentalized GC activity in PPs, with implications for the timing and design of vaccination and immunotherapy strategies in children. Funding Source AI100119 , Helmsley Charitable Trust Topic Categories Mucosal and Regional Immunology (MUC)

Regulation of Neutrophil Heterogeneity and Effector Functions by the NLRP3 Inflammasome in Sepsis 2306743

Abstract Introduction Sepsis is a life-threatening condition marked by dysregulated inflammation and immunosuppression. Bacterial clearance relies on effector functions of neutrophils. Besides classic neutrophils (HDNs), inflammatory low-density neutrophils (LDNs) emerge in disease states, driving tissue damage and morbidity in sepsis. Although NLRP3 modulates neutrophil activity, its influence on neutrophil heterogeneity and effector functions remains elusive. Methods Sepsis model: Sepsis was induced in 8-12-week-old male C57BL/6 wild-type (WT) and Nlrp3-deficient (Nlrp3-/-) mice using the cecal ligation and puncture model. Functional Assays Neutrophil Extracellular Traps (NETosis) were quantified using SYTOX Green dye. Phagocytosis was measured using pHrodo-bacterial bioparticles., Single-cell RNA sequencing: Neutrophils were profiled using the 10X Genomics platform. Data were analyzed using CellRanger and Seurat. Data Analysis Results presented as Mean ± SEM. Data were analyzed using Student’s t-test or one-way ANOVA. Differences were considered statistically significant at p < 0.05. Results We found that Nlrp3 is not required for LDN/HDN recruitment in sepsis. However, the LDNs and HDNs from Nlrp3-deficient mice demonstrated reduced NETosis. In contrast, the Nlrp3-/- LDNs and Nlrp3-/- HDNs from the spleen and peritoneum, respectively, showed increased phagocytosis. Inhibition of NLRP3 in human LDNs and HDNs confirmed our mouse findings. scRNA-Seq analysis revealed that neutrophil subtype clustering is altered in Nlrp3-/- mice during sepsis. Conclusion NLRP3 is dispensable for LDN and HDN recruitment to tissues but essential for NETosis during sepsis. NLRP3 activation impairs bacterial phagocytosis. scRNA-Seq analysis revealed significant changes in LDN/HDN clusters in Nlrp3-/- mice, reflecting tremendous neutrophil heterogeneity and attenuation in pro-inflammatory genes. Human LDNs/HDNs analysis confirmed the findings from mouse neutrophils and suggests NLRP3 as a therapeutic target to improve sepsis outcomes. Funding Source R01AI157353 Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

MAP4K2 promotes regulatory T cell differentiation and attenuates T cell immunity 2256886

Abstract Introduction MAP4K family kinases are key kinases for T-cell-mediated immune responses; however, in vivo roles of MAP4K2 in immune regulation remain unclear. Methods We used T-cell-specific MAP4K2 conditional knockout (T-MAP4K2 cKO) mice, single-cell RNA sequencing (scRNA-seq), and mass spectrometry analysis to study the role of MAP4K2 in T cell function and regulation. Results We found that MAP4K2 protein levels were increased in Treg cells compared to other T helper cells. We found that MAP4K2 interacted with DDX39, induced Foxp3 gene expression, and promoted Treg differentiation. Mechanistically, MAP4K2 directly phosphorylated the DEAD box protein DDX39, leading to DDX39 nuclear translocation and subsequent Foxp3 RNA splicing. MAP4K2-induced Foxp3 mRNA levels were abolished in DDX39 knockout Jurkat T cells. Furthermore, T-MAP4K2 cKO mice displayed the reduction of Treg population and sustained inflammation during remission phase of EAE autoimmune disease model. Remarkably, the anti-PD-1 immunotherapeutic effect on pancreatic cancer was drastically improved in T-MAP4K2 cKO mice, Treg-specific MAP4K2-deficient mice, or MAP4K2-inhibitor-treated mice. Consistently, scRNA-seq analysis of human pancreatic or lung cancer patients showed an increase of MAP4K2 levels in the tumor-infiltrating Treg cells; MAP4K2 mRNA levels were correlated with Foxp3 mRNA levels in the tumor-infiltrating T cells. Conclusion Collectively, MAP4K2 promotes Treg differentiation by inducing DDX39 nuclear translocation, leading to the attenuation of tumor immunity. Funding Source n/a Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

Changes in lumbar spinal cord Alox12 in doxycycline inducible HIV Tat transgenic mice, a model of HIV neuropathy 2259472

Abstract Introduction HIV neuropathy is the most common neurological complication of HIV infection. We previously demonstrated that doxycycline inducible HIV Tat transgenic (iTat) mice display neuropathy-like behaviors following Tat induction. RNA profiling study via NanoString and qRT-PCR with the lumbar spinal cord of iTat mice indicated a Tat-induced upregulation of arachidonate 12-lipoxygenase (Alox 12) comparing to PBS-treated controls. Alox12 metabolizes arachidonic acid to generate proinflammatory lipid mediators. To further examine the role of Alox12 in Tat neuropathy, we assessed the expression of Alox12 in lumbar spinal cord at the protein level. Methods iTat mice were injected daily with doxycycline for 14 days to induce Tat expression. Lumbar spinal cord tissues were collected at days 0, 3, 7, 14, 21, 28, and 35 following Tat induction and processed for immunohistochemistry (IHC). Results Alox12 was found to be co-localized primarily with neuronal marker NeuN rather than astrocyte marker glial fibrillary acidic protein (GFAP) or microglial marker CD11b. Similar to RNA levels, Tat induced an increase in the number of Alox12+ cell within the spinal cord dorsal horn area that returned to baseline 35 days post the initiation of Tat induction. No sex differences were detected at this time. Conclusion Our current results support the potential contribution of Alox12-mediated inflammation to Tat-associated neuropathy-like behaviors. Funding Source NIH/NINDS 1R01NS132674 (PI Cao) Topic Categories Immune Mechanisms of Human Disease (HUM)

Robust Covalent Organic Frameworks Comprising Accessible Catalytic Sites Enable Fast‐Charging and Long‐Cycling Aluminum–Sulfur Batteries

ABSTRACT Rechargeable aluminum–sulfur (Al–S) batteries are recognized as a promising option for large‐scale energy storage due to high theoretical energy density and cost‐effectiveness. However, the sulfur cathodes suffer from sluggish reaction kinetics and severe shuttle effect during cycling. Here we report robust two‐dimensional covalent organic frameworks (COFs) as sulfur hosts that features accessible catalytic nitrogen sites and confined microporous channels. The fine regulation of different microporous sizes was achieved by controlling different‐length organic ligands of various COFs, finally preparing two kinds of COFs. The sensitive comparation between both COFs demonstrates that smaller microporous channels in COFs possess higher confinement effect for polysulfides due to the stronger capillary forces, facilitating higher Coulombic efficiency and better cycling stability in Al–S batteries. Meanwhile, experimental characterizations and theoretical calculations reveal that accessible catalytic nitrogen sites in COFs promote the multistep conversion kinetics of the sulfur cathode during cycling. Consequently, the small‐sized COF confined sulfur cathode exhibits a reversible capacity of 1120 mAh g −1 at 0.2C and a 93.5% capacity retention after 100 cycles, supporting high capacity and exceptional cycling stability. This work provides a new avenue on rational design of emerging COF materials in Al–S batteries.

Framework for spatiotemporal soil moisture assessment: an application to the integration of model-based clustering with non-parametric change points detection

Respiratory bacterial experience protects from allergic airway eosinophilia 2257259

Abstract Introduction Countries with frequent exposure to respiratory pathobionts report lower incidence of asthma. Despite its implications for global health, unequivocal evidence for- and careful dissection of- a protective mechanism remains scant. Here, we sought to determine if and how microbial experience may alter development of asthma. Methods Using a common human airway colonizing bacteria Streptococcus pneumoniae (Spn), we modeled repeated microbial inhalations experienced by humans, and utilized the common human aeroallergen house dust mites (HDM) to model allergen-induced allergic airway eosinophilia. Results Consistently, naive C57BL/6J mice developed allergic airway eosinophilia following HDM challenge, while their Spn-experienced counterparts did not. Protection by Spn experience was irrespective of atopy, as genetically susceptible FVB/NJ mice were also protected. Transcriptional analysis demonstrated that Spn experience-induced protection was characterized by lower type 2 cytokine production and subsequent reductions in epithelial-derived eotaxins during HDM encounter. This was accompanied by reduced ILC2s, HDM-specific Th2 cells, cDC2s, and eosinophils, and significantly increased cDC1s and Tregs. Using mice lacking IFN-γ, IL-17, or IL-22, we next evaluated necessity of Spn-induced type 1 and type 3 effector T cells in anti-allergic protection and found no compromise in antiallergic protection. Instead, preliminary data suggest a role for IL-10 in this protection. Conclusion Our findings suggest that frequent exposure to respiratory pathobionts may indeed protect against development of allergic airway eosinophilia. Ongoing studies are focused on dissecting the cellular and molecular mechanisms driving this microbe-induced antiallergic protection. Funding Source NIAID 5T32AI007413, NHLBI 4R00HL157555 Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)

The CD83 axis between Tregs and CCR7+ dendritic cells governs anti-tumor immune responses 2309951

Abstract Introduction Immune checkpoint blockade (ICB) has advanced the treatment of melanoma and other cancer types. Despite its success, only about half of the treated patients respond, with many developing resistances. One contributing factor is the poor infiltration of effector T cells in the solid tumors, along with the immunosuppression caused by regulatory T (Treg) cells. CCR7+ dendritic cells (DC) interact with T cells, by either supporting the survival of CD8 T cells or promoting the migration of Tregs. While they are known to play a role in mediating tumor immune responses, their specific functions in controlling tumors and T cell responses remain poorly understood. Methods We generated innovative mouse models such as the inducible CCR7+DC conditional knock out (cKO), to defined how CCR7+DC use CD83 to restrain CD8 T cell effector responses and promote tumor growth. Results Reducing the expression of the activation maker CD83 on CCR7+DC, either directly or via blocking its soluble form expressed by Tregs, enhanced intratumoral CD8 T cell accumulation and controlled tumor growth in mice. Genetic perturbation of CD83 retains a pro-inflammatory IL-12+ CCR7+DC state, which is associated with the expansion of effector-like CD8 T cells and improved tumor control. Mechanistically, Tregs regulate IL-12 expression on CCR7+DC via STAT signaling in a CD83 dependent manner in vitro. Furthermore, we also show that CD83 levels on CCR7+DC negatively correlate with intratumoral CD8 T cell infiltration in several human cancers. Conclusion This study demonstrates that the ablation of interaction of CD83 molecule between CCR7+DC and Tregs results in increased anti-tumor immune response. Our findings identify CD83 as a cellular and molecular checkpoint that restrains T cell responses and can be targeted therapeutically. Funding Source This work was supported by funding for M.D.P from Cancer Prevention and Research Institute of Texas (CPRIT; RR210017) Robert J. Kleberg, Jr. and Helen C. Foundation Melanoma SPORE Developmental Research Program (DRP; P50CA221703) American Association of I Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Glucocorticoid signaling shapes the differentiation of tumor-infiltrating CD8+ T cells through TXNIP-dependent metabolic rewiring 2310224

Abstract Introduction Glucocorticoids (GCs) are anti-inflammatory steroid hormones derived from cholesterol. Owing to their potent immunosuppressive activity, exogenous GCs are widely used to treat autoimmune/allergic inflammation and to manage immune-related adverse events (irAEs) during cancer immunotherapy. GCs signal through the glucocorticoid receptor (GR), a ligand-activated transcription factor encoded by Nr3c1. GR regulates gene expression both directly, by binding glucocorticoid response elements (GREs) in target loci, and indirectly, by transrepressing the activity of other transcription factors. We previously showed that endogenous GR signaling dynamically shapes the differentiation of CD8+ tumor-infiltrating lymphocytes (TILs) within the tumor microenvironment (TME); however, the mechanisms and key downstream effectors remain poorly defined. Methods To define GR-dependent regulatory networks in CD8+ T cells, we performed in vivo single-cell RNA sequencing of wild-type (WT) and GR-deficient CD8+ TILs. In parallel, we conducted time-resolved, bulk RNA-seq over 72 hours (six early and late time points) in GC-treated WT versus GR-deficient CD8+ T cells to identify GR-regulated candidate effectors. We then used CRISPR-Cas9 perturbation to validate the functions of prioritized targets in the TME. Results These analyses revealed that GR signaling diverts CD8+ T cell differentiation from effector and tissue-resident memory (TRM)-like lineages toward dysfunctional states unresponsive to immune checkpoint blockade (ICB). Mechanistically, GR signaling repressed glycolysis via TXNIP, thereby compromising the metabolic fitness of CD8+ T cells, limiting their clonal expansion, effector differentiation, and instead promoting dysfunctional phenotypes, ultimately attenuating anti-tumor immunity. Conclusion Together, our findings define a GR-TXNIP metabolic axis that directs the fate of CD8+ TILs and has important implications for the clinical use of exogenous glucocorticoids in patients receiving ICB. Funding Source NIH R01 CA229400, CA282794, CA187975;; NMSS FG-2207-40160 Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Neutrophil Elastase Inhibition Prevents Neutrophil extracellular trap (NETs) Driven Autoimmune Glomerulonephritis Without Off Target Immunosuppression 2256502

Abstract Introduction Neutrophils release Neutrophil Extracellular Traps (NETs) as a final defense to trap and kill pathogens. In sterile inflammation, however, NETs become injurious, exposing over 70 autoantigens that drive tissue damage. This study evaluated pharmacologic neutrophil elastase (NE) inhibition using Bay 85-8501 in a NET-driven model of anti-neutrophil cytoplasmic antibody vasculitis (AAV), a major cause of glomerulonephritis and kidney failure. We aimed to determine whether targeted NET inhibition could match the efficacy of standard immunosuppression while preserving host defense. Methods A 20-day murine AAV model was induced in C57BL/6 mice. NE was inhibited using Bay 85-8501 via daily oral gavage (n = 8 vs vehicle) and compared to mice treated with cyclophosphamide or prednisolone. Kidney NETs were visualized by confocal microscopy, while histology, serum ANCA, RT-PCR for inflammatory mediators, and leukocyte infiltration were assessed. In additional experiments, NE-deficient (NE-/-) mice and inhibitor-treated groups were analyzed for off-target and antimicrobial effects. Results Bay 85-8501 significantly reduced renal NET deposition, glomerular injury, and leukocyte infiltration (all P < 0.01 vs WT). RT-PCR showed decreased IL-1β, MCP-1, CXCL2, and IL-4 expression. NE inhibition provided disease protection equivalent to prednisolone and cyclophosphamide but without the immunosuppressive burden. NE-/- mice mirrored the protection seen with pharmacologic inhibition, confirming specificity. Phagocytosis assays showed NE inhibition did not impair neutrophil antimicrobial function. Conclusion Bay 85-8501 precisely targets NET formation, achieving efficacy comparable to standard immunosuppression while preserving innate defense. These findings highlight NE inhibitors as a promising therapeutic strategy for NET-mediated diseases such as AAV, offering targeted disease control without the risks of broad immunosuppression. Funding Source National Health and Research Council of Australia Ideas grant Topic Categories Therapeutic Approaches to Autoimmunity (THER)

A novel microbial-derived metabolite counteracts exhaustion programming in CD8 T cells 2260531

Abstract Introduction Exhaustion of CD8 T cells during cancer or chronic infections remains a significant barrier to T cell immunotherapies. Recent studies showed that distinct epigenetic changes drive exhaustion by silencing effector and memory-related genes, thereby establishing the dysfunctional state of exhausted T cells (TEX). Thus, targeting epigenetic regulation of exhaustion is crucial for restoring TEX cell function. Short-chain fatty acids (SCFAs) are emerging as key mediators linking cellular metabolism to gene regulation. Notably, certain SCFAs naturally produced by human microbiota have been shown to modulate host immune responses through epigenetic mechanisms. Methods To investigate their effects on TEX cell epigenetic programming, we utilized innovative in vitro T cell exhaustion models that generate stable terminal dysfunction in both human and mouse CD8 T cells. By inducing a state of exhaustion that recapitulates key molecular and functional features observed in cancer and chronic infections, these models enabled us to assess how SCFA treatment affects T cell function and memory-associated stemness features. Results We discovered that a specific microbial SCFA triggered a significant recovery of polyfunctionality and memory programs within both human and mouse dysfunctional T cells. SCFA-treated TEX cells exhibited renewed effector capabilities, such as enhanced cytokine production, degranulation, and tumor-killing activity. The enhanced effector functions persisted even following termination of SCFA treatment, suggesting stable reprogramming of TEX cells. Conclusion These findings identify a novel microbial SCFA as a potential metabolic-epigenetic regulator, capable of reactivating effector programs in TEX cells while blocking terminal exhaustion. These results provide insights into developing new therapeutic approaches to reprogram TEX cells and enhance the efficacy of T cell immunotherapy. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Automated C-FREE™ Workflow for Reproducible Memory T-cell and NK-cell Immunophenotyping 2306832

Abstract Introduction Manual staining and wash steps introduce variability in memory T-cell and NK-cell immunophenotyping, particularly when different operators prepare cocktails or perform sample preparation. Automated workflows can minimize this variability by standardizing staining, mixing, and washing conditions. This work evaluates whether automated antibody cocktailing and automated sample preparation improve workflow consistency for a multiparameter immunophenotyping panel. The current dataset uses a ∼9-marker T-cell panel, with NK-cell markers to be incorporated in the expanded evaluation. Methods PBMCs were stained under four preparation conditions: (1) automated cocktail + automated sample prep, (2) automated cocktail + manual sample prep, (3) manual cocktail + automated sample prep, (4) manual cocktail + manual sample prep. Cocktails were prepared manually or via the Pluto workstation; samples were processed using the C-FREE automated workflow or centrifuge-based washing. Quantitative endpoints included stain index (SI), retention, population frequencies, and scatter stability to illustrate performance across preparation conditions. Results Using the T-cell panel, stain index (SI), retention, and major population frequencies from automated cocktailing and automated sample preparation were all within ±20% of values obtained using manual cocktailing and manual sample preparation. Automated steps produced consistent replicate performance with stable scatter profiles. These results establish baseline workflow consistency for extending the evaluation to memory T-cell and NK-cell panels. Conclusion The automated C-FREE workflow, combined with automated antibody cocktailing, offers a reproducible approach for multiparameter immunophenotyping. By reducing manual variability and standardizing preparation steps, the workflow establishes a foundation for robust memory T-cell and NK-cell profiling in research and translational applications. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Potent STAT6 Inhibitor EPS-3903 Has Excellent Preclinical Pharmacokinetics Enabling Sustained STAT6 Inhibition with Once-Daily Oral Dosing in Humans 2257175

Abstract Introduction STAT6 plays a pivotal role in driving type 2 inflammation. Here we present the preclinical pharmacokinetics of EPS-3903, a potent, selective, and potentially best-in-class STAT6 inhibitor for the treatment of type 2 inflammatory diseases. Methods Metabolic stability was evaluated in liver microsomes and hepatocytes across preclinical species and humans. The in vivo pharmacokinetic studies were conducted in mice, rats and dogs with an intravenous administration of 2.5 or 5 mg/kg or an oral dose of 25 mg/kg. Human PK prediction was based on allometric scaling and in vitro/in vivo DMPK data. Results After oral administration of EPS-3903 at 25 mg/kg, plasma exposures were 141.7 α¼g-hr/mL in mice, 43.0 α¼g-hr/mL in rats, and 174.0 α¼g-hr/mL in dogs, with an oral bioavailability of 96 - 100% using a simple oral formulation of 0.5% methylcellulose. EPS-3903 exhibits drug-like characteristics with very low clearance and excellent in vitro - in vivo correlation, and was stable in human hepatocytes. These favorable in vitro metabolic properties, combined with an excellent in vivo pharmacokinetic profile, support the efficacious, once-daily oral dose projection of 50 — 300 mg with a long half-life of > 16 hours in humans. Conclusion EPS-3903 demonstrates a highly favorable pharmacokinetic profile across preclinical species, supporting sustained STAT6 inhibition and feasibility of once-daily oral dosing in humans. These attributes underscore its potential for the treatment of type 2 inflammatory diseases including asthma and atopic dermatitis. Funding Source n/a Topic Categories Therapeutic Approaches to Autoimmunity (THER)

Mathematical Modeling of Vertebrate Memory-Based Immunity and Autoimmunity: A Computational Study of Memory Dynamics and Innate-Adaptive Feedback 2259645

Abstract Introduction We present a mathematical and computational study of the vertebrate adaptive immune system, focusing on T cell-mediated responses and their interaction with innate immunity. Our framework employs nonlinear ordinary differential equations to model the dynamics of both innate and acquired immune responses to pathogens and cancer. Methods Our model treats immune memory as a dynamic variable separate from circulating B and T cell concentrations, and includes autoimmunity as a factor influencing host fitness. Using computational simulations and stability analysis, we investigate different scenarios that could either support immune homeostasis or cause tolerance breakdown and the development of autoimmunity. Results The results demonstrate how nonlinear feedback between innate and adaptive immunity can influence the dynamics of pathogen, cancer cells, and healthy host cells. This model thus reveals how selection will act on different parts of the vertebrate immune system under challenges from both pathogens and cancer. Conclusion This modeling framework reveals how regulatory feedbacks shape the stability and adaptability of vertebrate immunity. It identifies conditions that promote immune homeostasis or trigger autoimmune dynamics, offering a quantitative basis for understanding how disruptions in immune regulation contribute to disease progression. Funding Source N/A Topic Categories Computational and Systems Immunology (COMP)

Compositional Homogenization and Dielectric Modulation of Pb/Sr Mixed Perovskites for Efficient and Bright Deep‐Blue Light‐Emitting Diodes

ABSTRACT Metal halide perovskites have emerged as promising candidates for next‐generation displays and lighting due to their tunable bandgap, high color purity, and solution processability. However, the performance of deep‐blue perovskite light‐emitting diodes (PeLEDs) lags far behind their red and green counterparts. Conventional strategies using low‐dimensional or mixed‐halide perovskites suffer from severe Auger recombination, phase separation, and chlorine‐vacancy‐induced nonradiative losses. Although Pb/Sr mixed B‐site compositional engineering provides excellent spectral stability, the compositional segregation and heterogeneity occurring during crystallization lead to poor film quality, low luminescence efficiency, and limited device performance. Herein, we introduce barium trifluoromethanesulfonate (Ba(OTf) 2 ) as a bifunctional additive to regulate the crystallization of Pb/Sr mixed perovskites. It synchronizes the crystallization kinetics of Pb and Sr via the formation of a ternary complex, suppresses compositional segregation, and passivates bromine vacancies, yielding highly homogeneous films with low defect densities. Moreover, Ba doping enhances the exciton binding energy through local dielectric modulation. Consequently, we achieve deep‐blue PeLEDs with a peak external quantum efficiency of 13.1% and a maximum luminance of 4821 cd m −2 at 467 nm, along with tunable emission from 450 to 470 nm. This work offers a viable route toward high‐performance alloyed perovskites for optoelectronic devices.

Achieving 1300 Wh/L in Anode‐Free Li Batteries With Integrated 3D Printed Cathode and Electrolyte

ABSTRACT Anode‐free Li batteries employing ultra‐thick cathodes offer exceptional volumetric energy density, yet are hindered by side reactions and limited Li reservoirs. This study develops an anode‐free pouch cell using a 3D‐printed ultra‐thick cathode (120 mg cm ‒2 ) and a dual‐salt/dual‐solvent electrolyte. The electrolyte forms an anion‐rich solvation structure, fostering an elastic‐plastic solid‐electrolyte interphase layer that enables highly reversible Li plating/stripping on bare Cu current collectors and stable prolonged cycling, even under low temperature conditions. The configuration delivers energy densities of 1,308 Wh L ‒1 and 424 Wh kg ‒1 , alongside a remarkable areal capacity of 19.11 mAh cm ‒2 . This work establishes a scalable and cost‐effective technical route for high‐energy, safe anode‐free batteries, which is expected to expedite their commercialization in electric aviation and the fast‐growing low‐altitude economy.

Energy disaster damage prevention strategies for power grid-highway network coupled power supply systems based on shortest path optimization