Browse Articles

Fibroblast PIEZO1 Promotes Autoimmune Valvular Carditis 2328060

Abstract Introduction Cardiac valve inflammation accompanies several autoantibody-mediated diseases. Interestingly, this pathology is typically restricted to the higher pressure left side of the heart (mitral and aortic valves), suggesting a link between mechanical stress and inflammation. The K/B.g7 mouse model of autoimmune arthritis and valvular carditis recapitulates key aspects of autoimmune valvular carditis in humans, including attack of the left-sided cardiac valves. We hypothesized that mechanosensing drives this side-specific susceptibility and investigated the role of PIEZO1, a mechanosensitive ion channel, in mediating autoimmune valvular carditis. Methods Using the K/B.g7 mouse model, we conditionally deleted PIEZO1 in endothelial cells (Cdh5-Cre), macrophages (Cx3Cr1-Cre), and fibroblasts (Tcf21-Cre). Cdh5 and Tcf21 Cre-mediated deletion were induced via oral tamoxifen gavage at 4 weeks of age, whereas Cx3cr1 Cre-mediated deletion was constitutive. Hearts were harvested from 8-week-old mice. Valve thickness and pathology were quantified via hematoxylin and eosin (H&E) staining of histological sections. Results Deletion of PIEZO1 in Tcf21+ fibroblasts significantly reduced valve inflammation and thickening (p = 0.0343). In contrast, PIEZO1 deletion in endothelial cells (p = 0.7984) or macrophages (p = 0.9118) did not attenuate pathology. Preliminary experiments show no significant differences in valve adhesion receptor expression, cellular proliferation, or collagen organization following fibroblast-specific loss of PIEZO1. Conclusion These results demonstrate that PIEZO1 expressed by Tcf21+ fibroblasts is a critical contributor to mitral valve inflammation in autoimmune valvular carditis. While specific downstream mechanisms remain to be elucidated, they likely include numerical cell changes, extracellular matrix remodeling, and/or altered intercellular communication. These findings suggest that fibroblast-mediated mechanosensing could underlie the left-sided bias observed in autoimmune valvular carditis. Funding Source 1T32HL166142-01 5T32AI007313-36 5R01HL121093-10 Topic Categories Basic Autoimmunity (BA)

The Inhalation of Deoxynivalenol in a Murine Model of Allergic Fungal Asthma 2310137

Abstract Introduction Deoxynivalenol (DON) is a mycotoxin produced by the fungus Fusarium graminearum. This fungus is commonly found in agricultural fields and can contaminate crops. When ingested, DON can cause changes in the intestinal, nervous, and immune system. However, during harvesting season, Fusarium graminearum and DON can become airborne and inhaled. Minimal research has been done regarding the effects DON has on the respiratory system when inhaled. As DON targets the immune system through intestinal mucosa, it can be hypothesized that DON can also trigger an immune response when inhaled. This study assesses the effect of DON inhalation in an allergic asthma murine model. Methods Allergic mice were exposed to 30 or 60 ng of inhaled DON once daily for six days. On day 7 and 28, serum was analyzed for IgE and IgA, and bronchoalveolar lavage fluid (BALF) were analyzed for IgA levels. Cytokine levels were analyzed in lung homogenates, and goblet cell metaplasia with associated mucus production was visualized by periodic acid Schiff staining of lung histology sections. Results With the inhalation of DON, serum and BALF IgA levels increased. Allergic asthma mediators or allergic asthma symptoms such as serum IgE, cytokine levels, and goblet cell metaplasia were unaffected. Conclusion Inhalation of DON in an experimental allergic asthma murine model did not have an effect on allergic asthma mediators but resulted in increased IgA levels. IgA is an important antibody subtype found in mucosal secretions, and our next step will be to understand the role of pulmonary epithelial cells in mediating this cascade. The results of this study are likely to help inform therapeutics and diagnostics for DON related exposure. Funding Source n/a Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)

The pre-metastatic niche influences neutrophil behaviour, allowing cancer cells to hide in plain site 2309704

Abstract Introduction The fatality rate for metastatic cancers remains high, with the lungs being a common and deadly site of metastasis. Before cancer cells arrive, host tissues undergo changes that prepare them for tumour growth. This process forms the pre-metastatic niche, making the lung environment more receptive to colonization. Neutrophils populate the pre-metastatic niche, but their phenotype and whether they support metastasis at this stage remain unclear. Methods A model of E0771-GFP breast cancer cells that metastasize to the lungs was established, where the lung pre-metastatic niche and arrival of cancer cells can be captured using intravital microscopy. Using this model, we observed that not only does neutrophil behaviour change in the pre-metastatic niche from homeostasis, but their responsiveness to arriving cancer cells is compromised. Results The Kubes Lab has previously demonstrated that under healthy conditions, the majority of resident lung neutrophils crawl along the alveolar capillary wall seeking perturbations. Behavioural analysis of neutrophils imaged in pre-metastatic lungs showed reduced crawling and increased adhesion to the endothelium. Further, pre-metastatic neutrophils were imaged in the presence of both intravenously injected and spontaneously arrived cancer cells. In both cases, neutrophils exhibited limited interactions with cancer cells. In contrast, in healthy lungs, numerous neutrophils maintained prolonged contact with intravenously injected cancer cells. Super-resolution microscopy of healthy resident lung neutrophils showed acquisition of GFP+ particles following interaction with GFP+ cancer cells. Conclusion Our findings reveal that neutrophils in the lung pre-metastatic niche adopt a distinct behavioural state compared to healthy resident neutrophils. Such behaviour may contribute to the permissive environment that enables metastatic seeding in the lungs. These findings lay the foundation for therapeutic targeting of neutrophils to prevent metastatic seeding. Funding Source Canadian Research Excellence Chair (CERC), Infrastructure Operating Fund (IOF), Canadian Foundation for Innovation (CFI), and Bruce Mitchell Research Program Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Lupus Nexus: A Registry, Biorepository, & Data Exchange Platform to Foster Global Collaborations, Accelerate Research, & Advance Precision Medicine 2260377

Abstract Introduction Systemic lupus erythematosus remains a disease of high unmet medical need. The Lupus Nexus (LNx) is a lupus registry, biorepository, and data exchange platform that addresses this by providing a community-wide access point to longitudinal, highly curated, centralized clinical datasets linked to biospecimens and cellular/molecular data. Methods The LNx includes a prospective, longitudinal observational study, the Lupus Landmark Study (LLS), that is currently enrolling up to 3,500 people living with lupus into four cohorts: new-onset, active lupus nephritis, extra-renal flare, and prevalent/quiescent SLE with a 5-year follow-up. The clinical data include parameters such as patient- and clinician-reported outcomes (PRO and CLINRO), full medical history, social history and determinants of health, and environmental exposures, among others. Linked biospecimens include DNA, RNA, urine, plasma, serum, saliva, stool, and tissue, and all are available to the research community. A small portion of the biosamples is currently being used for analyses including genomics, transcriptomics, and proteomics. Results As of October 2025, there are over 570 enrolled participants and over 42,000 stored biospecimens. The registry is reflective of the population affected by lupus. Genetic, transcriptomic, and proteomic assays are underway to generate foundational scientific data, Foundational Analyses (FA), to stimulate research efforts. Conclusion The LNx is a unique, community-wide resource for both researchers and those living with lupus. This biorepository is open to the entire lupus research community and is poised to catalyze advances in understanding disease etiology, genetic drivers and mechanistic heterogeneity in SLE. Funding Source n/a Topic Categories Basic Autoimmunity (BA)

Enabling single-cell multiomics to unravel complex data from human lung cancer tissue preserved in a novel long-term storage buffer 2308753

Abstract Introduction Tumor tissue is highly heterogenous, containing varying cell types with multiple functions and characteristics. Unraveling the complexity of this tissue at a single-cell level can provide critical understanding of tumor microenvironments (TME), setting the foundation for informing disease states, targeted therapies and the discovery of drug targets and biomarkers. However, maintaining the integrity of the tumor from the time of collection to the time of analysis remains a major challenge. Necrosis, cell degradation, and undesired alterations of cellular states can profoundly impact the quality and validity of the data, therefore reliable tumor preservation is critical. Common preservation methods typically use cross-linking agents that can damage protein and nucleic acids and compromise data quality. Methods Here we highlight a novel long-term cryo-preservation buffer that does not contain cross-linking fixatives, that was used to preserve dissociated human lung cancer cells. The tumor was collected in BD®OMICS-Guard and stored for 24 h at 4oC, then processed to obtain single cells that were directly transferred into the novel cryo-preservation buffer for freezing. The samples were then shipped to a location where single-cell multiomic assays were conducted to simultaneously profile mRNA and protein of individual cells. Results Non-supervised clustering revealed 12 distinct cell clusters that included infiltrating lymphocytes (TILs), immune cells, and non-immune tumor cells. Differential gene and protein expression profiles were used to distinguish distinct subpopulations of macrophages., while T cell gene sets of naive, activation, and exhaustion states were used to identify the functional heterogeneity of TILs. Conclusion Together these data demonstrate the use of a novel cryopreservation buffer that enables flexible workflows while yielding high-quality data. For Research Use Only. Not for use in diagnostic or therapeutic procedures. ©2025 BD. All rights reserved. NPM-7268 Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Olfr2 Signaling in Dendritic Cells Enhances Antigen Presentation and T Cell Activation Promoting Atherosclerosis 2309405

Abstract Introduction Atherosclerosis is a chronic inflammatory disease driven by dysregulated lipid metabolism and maladaptive interactions between innate and adaptive immune cells. Olfactory receptors (ORs), the largest family of G-protein—coupled receptors, are emerging regulators of vascular inflammation beyond their canonical sensory roles. Olfr2 is expressed in monocytes and macrophages, where it senses the lipid peroxidation product, octanal to activate the NLRP3 inflammasome and promote inflammation; still, its role in other myeloid cells is unclear. Methods Dendritic cells (DCs) are professional antigen-presenting cells (APCs) that coordinate adaptive immune responses; however, whether Olfr2 regulates DC-mediated T cell activation during atherogenesis remains poorly defined. We found that Apoe-/-Olfr2-/- mice fed a high-fat diet (42% calories from fat) for 10 weeks exhibit smaller lymph nodes (LNs) and reduced CD4+ and CD8+ T cell activation, as assessed by CD44 and CD69 expression, compared with Apoe-/- controls. We detected significant Olfr2 expression in CD11c+ CD11b+ DC subsets both in the spleen and LNs. This phenotype was associated with decreased MHC-II and CD86 expression on Olfr2-deficient CD11c+CD11b+ DCs, suggesting impaired APC-T cell signaling. Results To determine whether this defect was APC-intrinsic, we initially assessed T cell activation following anti-CD3/CD28 stimulation and found no differences. Next, we performed co-culture experiments using Nur77-GFP-OT-I CD8+ T cells and WT or Olfr2-/- FLT3 ligand-differentiated bone marrow-derived DCs (BMDCs) under OVA peptide—pulsed and non-pulsed conditions. Conclusion The precise mechanistic role of Olfr2 in antigen presentation, DCs activation, and downstream T cell priming during atherogenesis is poorly elucidated. Our data identify Olfr2 as a possible regulator of DC-driven adaptive immunity in atherosclerosis and suggest that targeting this pathway may limit maladaptive APC-T cell interactions and disease progression. Funding Source n/a Topic Categories Classical and Non-Classical Antigen Presenting Cells (APC)

Perception, Synaptic Plasticity, and Spiking Neuron Function Enabled by a 2D Ferroelectric NbOBr ₂ for Neuromorphic Vision

ABSTRACT 2D ferroelectrics integrate electrically tunable polarization dynamics with anisotropic light–matter interaction, providing a unified platform for co‐localized sensing and in‐memory computing. However, most neuromorphic visual systems still rely on heterogeneous components that exploit only a single material functionality at a time, thereby increasing system complexity and fabrication costs. Here, we demonstrate a device–algorithm co‐design for neuromorphic visual recognition based on multifunctional NbOBr 2 . By leveraging its anisotropic broadband photoresponse, robust in‐plane ferroelectricity, and integration with multilayer graphene, we realize three bio‐inspired core functions: anisotropic photoelectric synaptic preprocessing for retina‐like encoding, ferroelectric synaptic weight modulation for in‐memory weighted operations, and leaky integrate‐and‐fire (LIF) neuronal emulation for spike generation. With BDD100K driving scenes converted to drivable maps and encoded into two orthogonal polarization channels (c‐ and b‐axis; 0°/90°), a device‐aware spiking neural network achieves a recognition accuracy of up to 94.2%, outperforming a standard SNN baseline (91.8%). These results illustrate how the intrinsic multifunctionality of a 2D ferroelectric can be harnessed to create compact, biologically plausible perception–computation hardware for high‐performance neuromorphic vision.

Wavelet-derived periodic variability and regional differences of surface ozone in southeastern Nigeria from 2002 to 2023

Prenatal Benzene Exposure Induces Sex-Dimorphic Reprogramming of Offspring Pulmonary Immunity 2256142

Abstract Introduction Air pollution during pregnancy are associated with increased risk of abnormal immune responses in offspring, including susceptibility to respiratory infections, asthma, and allergies. The developing fetal immune system is highly vulnerable to environmental exposures, yet how volatile organic compounds such as benzene affect postnatal pulmonary immunity remains unclear. Methods We exposed pregnant C57BL/6 mice to benzene (5 ppm, full-body inhalation, 24 h/day) from embryonic day 0.5 (E0.5) to E17.5. Offspring were analyzed at three timepoints: fetal (E17.5), postnatal day 10 (PND10), and PND35. Fetal lungs were analyzed by qPCR, western blot, Luminex, flow cytometry, bulk RNAseq, scRNAseq, and proteomics to identify persistent transcriptomic changes. PND10 pups were intraperitoneally infected with MHV68 and evaluated at PND17 for viral susceptibility using molecular, immunological, and histological assays. PND35 offspring were intranasally challenged with LPS to assess bacterial susceptibility; bronchoalveolar lavage fluid (BALF) cytokines and alveolar immune cells were analyzed by flow cytometry. Alveolar macrophages isolated at PND35 were evaluated for inflammatory responses. Results Prenatal benzene exposure induced sex-dimorphic reprogramming of pulmonary immunity, with alveolar macrophages and neutrophils in both females and males adopting a persistently inflammatory state. In females, this was associated with enhanced antiviral responses but increased lung inflammation and tissue damage, linked to A20 suppression. In males, pre-activated immune cells drove exaggerated inflammatory responses upon bacterial challenge. Conclusion These results indicate that gestational benzene exposure programs fetal lung immune cells into a dysregulated, pro-inflammatory state, contributing to long-lasting alterations in pulmonary immunity and heightened infection susceptibility in a sex-dependent manner after birth. Funding Source NIAID 5R01AI145829-05 5P42ES030991, R01HD111146 Topic Categories Hematopoiesis and Immune System Development (HEM)

CD39 regulates cerebrovascular dysfunction during sepsis 2334223

Abstract Introduction Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. Vascular alterations play a central role, promoting endothelial barrier disruption and neuroinflammation, key features of sepsis-associated encephalopathy. Activated endothelial cells release ATP, which acts as an alarmin by activating purinergic receptors or being degraded by ectonucleotidases like CD39, thereby modulating ATP-driven inflammatory responses. This study aimed to evaluate the role of CD39 in sepsis-associated cerebrovascular alterations. Methods In vitro, mouse brain capillary endothelial cells (bEnd.3) were stimulated with LPS (100 or 1000 ng/mL) for 8 h or 24 h. To assess the role of CD39, the enzyme inhibitors POM-1 (10 µM) and AR67156 (100 µM) were used 30 min before LPS stimulation. In vivo, sepsis was induced by cecal ligation and puncture (CLP) in WT and CD39 -/- C57BL/6 mice. Mice were euthanized 24 h after sepsis induction, and brain structures and serum were collected. Brain intravital microscopy experiments were also performed to evaluate leukocyte adhesion in cerebral microcirculation, and coagulation was determined by recalcification tests. Results LPS stimulation reduced CD39 protein expression and enzymatic activity in bEnd.3 without affecting purinergic receptor protein and mRNA levels. CD39 inhibition increased LPS-induced mRNA expression of endothelial markers (ICAM-1, VCAM-1, CXCL-1). Quantification of CXCL-1 by ELISA and immunocytochemical analyses for VCAM and ICAM also showed similar results. Septic CD39 -/- mice showed higher mRNA expression of ICAM-1 and CXCL-1 in the brain as well as elevated levels of CXCL-1 protein in the brain and serum compared to septic WT mice. Septic CD39 -/- mice showed increased leukocyte adhesion and a more procoagulant profile. Conclusion These findings indicate that the enzyme CD39 plays a protective role in modulating endothelial activation, leukocyte adhesion, and cerebral microvascular inflammation during sepsis. Funding Source CAPES; CNPq (INCT Saúde cerebral; INCT Sinalização purinérgica: Desafios para o século 21); FAPERJ. Topic Categories Neuroimmunology (NEUR)

Spatial—glycomic immune signatures in metabolic dysfunction-associated steatotic liver disease (MASLD) progression 2310277

Abstract Introduction Metabolic dysfunction—associated steatotic liver disease (MASLD) progresses from chronic inflammation and fibrosis to hepatocellular carcinoma (HCC). Yet the spatial remodeling of immune-cell N-glycosylation during this transition remains poorly understood. Because glycosylation critically regulates immune cell function, trafficking, and antitumor immune responses, defining how immune glycan landscapes are rewired in situ across MASLD stages is essential for identifying actionable biomarkers and therapeutic targets. Methods We performed imaging mass cytometry (IMC) and MALDI N-glycan imaging on human MASLD liver biopsies spanning multiple fibrosis stages, as well as on end-stage MASLD tissues with and without hepatocellular carcinoma. Multimodal spatial datasets were co-registered and jointly analyzed using our newly developed MIMIC pipeline, enabling pixel-level integration of immune cell phenotypes and N-glycan distributions across tissue compartments. Results IMC revealed a progressive reorganization of immune neighborhoods, shifting from Kupffer cell—T cell interactions in early fibrosis toward T cell—dominated networks in advanced disease. Spatial glycomics uncovered domain-specific glycan organization. Integrated pixel-level analysis identified cell type—associated glycan signatures and demonstrated differences between intratumoral, tumor-adjacent, and healthy glycan features consistent with impaired immune surveillance in MASLD-associated HCC. Conclusion These findings establish immune glycan remodeling as a spatially organized hallmark of MASLD progression and MASLD-HCC. The integrated IMC—glycomics framework provides a practical and scalable approach for biomarker discovery and supports therapeutic targeting of glycosylation pathways in metabolically diseased liver. Funding Source N/A Topic Categories Immune Mechanisms of Human Disease (HUM)

Multiplex Made Practical: An Educational Workflow Using Directly Conjugated Primary Antibodies 2309766

Abstract Introduction Spatial biology multiplex panels built from directly conjugated primary antibodies accelerate staining workflows and improve quantification accuracy of immunomarker expression by eliminating amplification bias. These advantages allow scientists to interrogate complex tissues with greater depth and precision. Here, we outline how to create and optimize primary antibody panels to characterize immune and structural features in healthy and malignant colon tissues. Methods Using a practical framework of steps and considerations that we will highlight in detail, we designed a panel comprising directly labeled antibodies to podoplanin, CD4, FoxP3, Ki67, CD8, CD68, PD 1, and CD3. CD3 was conjugated in-house using an antibody labeling kit, and we will describe the optimization of this procedure. We describe an effective method to combat autofluorescence using a combination of white light photobleaching + H2O2 and highlight ways to achieve superior unmixing performance using EVOS S1000. Results Immune cells comprised only about 10% of total cells in both samples, with tumors showing a marked increase in macrophages and a dramatic rise in proliferating Tregs, both associated with tumor progression and poorer outcomes. In the adenocarcinoma sample, we identified a tertiary lymphoid structure enriched primarily in cytotoxic and helper T cells, suggesting a potentially beneficial anti-tumor immune niche. Overall, these findings highlight how high-plex imaging enables detailed characterization of immune cell composition and spatial organization within colorectal tumors. Conclusion These findings show that directly conjugated multiplex panels offer a robust, scalable way to resolve complex tissue features while providing workflows that generate high-quality spatial data to accelerate biological insight and discovery. All reagents are intended for Research Use Only (RUO). Not for use in diagnostic procedures. Funding Source N/A Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

CTLA-4 mediates oral immunotolerance against systemic allergic sensitization to environmental allergens 2260513

Abstract Introduction Environmental exposure to food allergens (e.g. peanut antigens) through the skin or airways increases the risk of developing food allergy (e.g. peanut allergy). In contrast, oral antigen exposure (e.g., consumption of peanut products) early in life reduces the risk. The objective of this study was to investigate the immunologic mechanisms by which oral antigen exposure provides systemic immunotolerance and protection from food allergy development using murine models. Methods Naive mice were administered ovalbumin (OVA) through their drinking water to promote oral immunotolerance. Mice were subsequently exposed intranasally (i.n.) to OVA plus peanut flour, as an allergic adjuvant. CD4+ T cells from lung draining lymph nodes (dLNs) were assessed by single-cell RNA sequencing, MHC-II tetramer staining, and flow cytometry. Results Following i.n. exposure to OVA plus peanut flour, mice developed OVA-specific IgE (OVA-IgE) and showed symptoms of acute anaphylaxis upon intraperitoneal OVA challenge. Oral OVA feeding prior to i.n. OVA exposure reduced OVA-IgE and protected mice from anaphylaxis. Protection was associated with suppression of OVA-specific T follicular helper (Tfh) and germinal center B cells in dLNs. Tolerance was reversed by administration of anti-CTLA-4 blocking antibody but was not affected by deletion of Foxp3+Bcl6+ T follicular regulatory (Tfr) cells. Accordingly, conventional Foxp3+T regulatory (Treg) cells were not increased in dLNs of OVA-fed mice. Rather, protection was associated with the emergence of OVA-specific Foxp3-CTLA4+ cells expressing markers of stemness. Conclusion Oral antigen exposure suppresses the development of antigen-specific Tfh cells in remote dLNs thereby preventing systemic allergic sensitization. Systemic oral tolerance is dependent on CTLA-4 and unlikely requires conventional Foxp3+ Treg cells. Funding Source Supported by NIH F31 AI86242, R37 AI71106, and Mayo Clinic Graduate School of Biomedical Sciences. Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)

Cytokine, plasmablast, and clinical biomarker profiles following monovalent dengue vaccination in individuals with distinct immune histories 2308784

Abstract Introduction Pre-existing dengue immunity shapes subsequent immune responses, which can either be protective or pathogenic, though the underlying mechanisms remain poorly defined. Using a live attenuated monovalent DENV3 vaccine, we investigated how baseline immunity shapes cytokine, plasmablast, and clinical biomarker responses. Methods Participants were classified as naïve, heterotypic to a single serotype, or polytypic based on screening neutralizing antibodies, then restratified using area under the curve of viremia by qRT-PCR. We profiled 30 cytokines by Luminex and assessed plasmablast responses by ELISpot alongside standard clinical biomarkers. Results Low-viremia participants, most of whom had polytypic immunity, had early increases in innate cytokines, such as IL-6, IL-7, GM-CSF, and IFN-γ, followed by downregulated TNF-alpha and IL-2R and elevated MCP-1, lymphocyte, and monocyte counts, consistent with efficient viral control. The intermediate-viremia group showed early elevations in IFN-α and Th1-associated cytokines, followed by expansion of DENV3-reactive IgG+ and IgA+ plasmablasts, and IP-10 and IL1-RA induction, indicating a coordinated antiviral response with moderate inflammation. High-viremia participants, who mostly had heterotypic immunity, had more delayed increases in DENV-specific IgG+ plasmablasts and broader and more sustained elevations in proinflammatory and tissue-activating cytokines, such as G-CSF, VEG-F, and MIP-1 chemokines. Conclusion Pre-existing dengue immunity coupled with post-vaccine viremia delineates individuals into distinct profiles. We observed transient innate activation and controlled inflammation in low viremia participants, a coordinated adaptive response in the intermediate viremia group, and low-level but broad and persistent inflammation in the high viremia group. Understanding these profiles can help refine dengue interventions to improve safety and protective efficacy. Funding Source This research was supported by the Intramural Research Program of the National Institutes of Health (NIH) as well as NIH Bench-to-Bedside Program Funds - Award # 994875, the NIH Director’s Challenge Innovation Award, ReVAMPP grant 1 U19 AI181960-01, and P Topic Categories Viral Immunology (VIR)

A Traditional Herbal Formula Attenuates Chronic Pruritus by Inhibiting IgE—FcεRI Signaling and Sensory Neuron Activation 2333596

Abstract Introduction Chronic pruritus is a debilitating symptom accompanying various inflammatory and dermatologic diseases, yet currently available antipruritic therapies show limited efficacy and frequent adverse effects. Growing interest has therefore focused on mechanism-based and safer alternatives, including traditional herbal medicines. Cheonwangbosim-dan (CWB), a classical Korean herbal formula, was identified as a potential antipruritic candidate, although its immunological and sensory neuronal mechanisms remain unclear. Methods We screened 26 traditional formulations using a biochip assay to evaluate inhibition of IgE—FcεRI binding. CWB’s effect on mast cell degranulation was measured via histamine release in MC/9 cells. In vivo efficacy was assessed using an imiquimod-induced zebrafish inflammation model (locomotor activity and qRT-PCR for il-1β, il-6, il-33, and trpa1) and a histamine-induced mouse scratching model. Furthermore, sensory neuronal activity was monitored via in vivo calcium imaging of dorsal root ganglion (DRG) neurons in GCaMP-transgenic mice following oral or topical CWB administration. Results CWB demonstrated the strongest inhibition of IgE—FcεRI binding among the screened candidates and significantly reduced mast cell histamine release. In zebrafish, CWB dose-dependently suppressed imiquimod-induced hyperlocomotion and downregulated key pro-inflammatory and pruritus-related genes. In murine models, both oral and topical CWB markedly attenuated histamine-induced scratching. Notably, in vivo imaging revealed that CWB significantly inhibited histamine-evoked DRG neuronal activation without inducing spontaneous activity. Conclusion CWB exhibits potent antipruritic activity through dual modulation of immune-mediated histamine signaling and sensory neuronal responsiveness. This combined mechanism distinguishes CWB from conventional antipruritic agents and supports its potential as a safe and effective complementary therapy for chronic pruritus. Funding Source Portions of this work was carried out within the Korea Institute of Oriental Medicine, (grant number KSN18384113). Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

Electric Breakdown of Dielectric Polymers is Strongly Correlated to Mechanical Toughness

ABSTRACT Dielectric energy storage is critical for technologies demanding ultrafast discharging and pulse power, and preventing electric failure is paramount to the design and engineering of dielectric materials. Under high voltages current traverses the dielectric leading to electric breakdown, which is the primary failure mode of dielectrics. While intrinsic breakdown strength in perfect dielectrics can be predicted from quantum mechanics, failures in dielectric materials often occur under an electric field well below their intrinsic theoretical strengths, and the mechanism is still poorly understood. Here we show that electric breakdown of dielectric polymers is strongly correlated to their mechanical toughness. We reveal through in situ experiments that breakdown of polymers proceeds through nucleation and growth of cracking channels, suggesting a close connection between electric and mechanical failures. We demonstrate that the breakdown strength of polymers can be modulated by their tensile toughness, and there is a universal relationship between electric toughness and mechanical toughness. As a result, the breakdown strength for a wide range of polymers with mechanical and dielectric properties spanning two orders of magnitude can be reliably predicted, uncovering a dominant electromechanical breakdown mode that unifies mechanical and electric failures. Toughening strategies in enhancing dielectric strength can therefore be explored.

A bilateral US-UK airplane wastewater monitoring pilot demonstrates the feasibility of an international wastewater surveillance network

Abstract We present findings from the first known pilot study of transatlantic airplane wastewater monitoring, conducted over six months at two connected international airports in the United States and the United Kingdom. This study demonstrates the feasibility of implementing bilateral wastewater-based pathogen surveillance at international travel hubs. We outline the operational and analytical methodologies employed, highlight key challenges encountered in transnational coordination, and provide recommendations for the design and implementation of future surveillance programs at points of entry.

Double-Negative T Cells Induce Ferroptosis-Dependent, MHC-Independent Cytotoxicity of CD8+ T Cells Against AML 2256323

Abstract Introduction Acute myeloid leukemia (AML) is a blood cancer originating from myeloid cells in the bone marrow with poor prognosis. CD3+CD4-CD8-double negative T cells (DNTs) is a rare subset of immune cells that have shown strong anti-AML properties. Our previous work demonstrated that allogeneic DNTs from healthy donors effectively targeted AML in vitro and in vivo without off-tumor toxicity in preclinical models and early phase clinical trials. However, the mechanisms by which DNTs influence host immune cells’ anti-leukemic activity remain unclear. Methods We cultured patient or healthy donor CD8+ T cells with autologous AML blasts or AML cell lines and characterized their immunophenotype and anti-leukemic activity. We performed proteomic analysis comparing CD8+ T cells activated by DNT+AML, AML alone, or CD3/CD28 beads, and differentially expressed proteins and pathways were identified and validated using functional assays. We conducted blocking experiments targeting ferroptosis and generated MHC-I knockout AML cells to test whether DNT-activated CD8+ T cells can kill AML cells in an MHC-independent manner. Results CD8+ T cells co-cultured with DNTs mediated cytotoxicity toward AML cells. Proteomic profiling revealed that CD8+ T cells activated by DNT+AML upregulated innate immune response and ferroptosis-pathways. Blocking ferroptosis markedly inhibited their killing capacity, indicating that CD8+ T cell cytotoxicity depends on ferroptosis. Notably, these CD8+ T cells could kill MHC-I—deficient AML cells, suggesting an antigen-independent mechanism. Conclusion These findings show that, beyond direct cytotoxicity, DNTs promote a non-conventional anti-leukemic activity of CD8+ T cells through ferroptosis-dependent mechanisms. This synergy between DNTs and CD8+ T cells represents a novel immune axis that could be leveraged to enhance adoptive cell therapy efficacy against AML. Funding Source Alberta Cancer Foundation, Kids Cancer Care, CIHR Project Grant, Cancer Research Society Operating Grant Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Ultra-Sensitive Cytokine Quantification Using Binding Oligo Ladder Detection: A Universal Signal-Amplification Strategy for Standard Immunoassays 2308422

Abstract Introduction Accurate quantification of low-abundance cytokines is essential for resolving early immune activation, subtle inflammatory shifts, & cytokine dynamics in limited-volume samples. However, many cytokines circulate at concentrations near or below the analytical limits of conventional assays. Here we describe the application of Binding Oligo Ladder Detection (BOLD)–a molecular amplification chemistry that integrates directly into existing immunoassay workflows to achieve sub-pg/mL sensitivity across multiple cytokines. Methods BOLD functions by conjugating an oligo-dT primer to the detector antibody, enabling enzymatic generation of a DNA/RNA hybrid strand that recruits multiple anti-hybrid antibodies for amplified signal generation. The method requires no specialized instrumentation & is compatible with standard detection methods such as colorimetry, fluorescence and ECL. Results Across four cytokine systems, BOLD consistently improved sensitivity as compared to standard assays. For IL-4, the limit of detection (LOD) improved to 0.040 pg/mL (∼50×) under optimized conditions. TNF-α detection improved ∼6×, with LOD reduced to 0.24 pg/mL and with reproducible performance across replicate runs. Newly developed BOLD-enhanced assays for IL-6 and IL-23 demonstrated 7× and 8× improvements in sensitivity, respectively. Across all cytokines, the concentration of primer-modified detectors was significantly reduced compared to standard ELISA detector levels, while maintaining low background & favorable signal-to-noise ratios. Assay robustness was further supported by minimal dependence on tertiary antibody concentration & stable performance across polymerase reaction conditions. Conclusion These results demonstrate that BOLD is a versatile, platform-agnostic technology capable of transforming standard assays into ultra-sensitive detection systems. Its applicability across diverse cytokines highlights its potential for advancing immune monitoring, translational research, & high-sensitivity biomarker discovery. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Linking zygosity and allelic balance to phenotype with programmable heterozygous editing and allele-specific single-cell RNA sequencing 2307550

Abstract Introduction Heterozygous single-nucleotide variants cause many inborn errors of immunity (IEI), yet most functional genomics tools generate homozygous edits, limiting insight into dominant-negative (DN) and dosage-dependent variant effects. Expression imbalance of the mutant and wild-type alleles can further complicate genotype—phenotype relationships, and has been nearly impossible to interrogate experimentally. To overcome this, we built a scalable platform to engineer heterozygous variants in primary immune cells and directly link allelic expression bias to phenotype. Methods We harnessed an innovative CRISPR-based editing system, helicase-assisted continuous editing (HACE), in primary human T cells to engineer heterozygous variants across CARD11, a key regulator of immune signaling, in which heterozygous variants cause diverse IEI with variable penetrance. In parallel, we developed single-cell Allele-Integrated Multi-omics sequencing (sc-AIMseq), which integrates transcriptomics, surface proteomics, and quantification of mutant and wild-type allele expression at single cell resolution. Results HACE screens recovered known pathogenic DN variants in CARD11 and revealed novel DN and haploinsufficient variants not evident in homozygous models. sc-AIMseq of HACE-edited T cells uncovered marked cell-to-cell variability in CARD11 allelic expression bias, which dramatically influenced T cell phenotype. We also performed sc-AIMseq on PBMCs from multiple patients with DN CARD11 mutations, which revealed that CARD11 allelic expression bias is cell-type dependent, providing a mechanistic basis for variable penetrance. Conclusion This work establishes a scalable strategy to model heterozygous genetic disorders in primary and patient-derived immune cells, and dissect their variable penetrance. By directly coupling heterozygous variant engineering, allelic expression bias, and phenotype at single-cell resolution, our approach enables unprecedented functional variant interpretation and advances precision immunology. Funding Source Z.H.W. has received research funding from NCI F30CA298572 and the Melanoma Research Foundation. Topic Categories Technological Innovations in Immunology (TECH)