Browse Articles

Toward a Pan-Arenavirus T-cell Vaccine: Identification and Preclinical Evaluation of Conserved Epitope constructs 2260477

Abstract Introduction Arenaviruses represent a rapidly expanding group of rodent-borne emerging human pathogens with significant pandemic potential. Diseases caused by these viruses, such as the Old-World Arenavirus (OWA) Lassa virus (LASV), which causes Lassa Fever, and the New-World Arenavirus (NWA) Junin virus (JUNV), which causes hemorrhagic fevers, currently lack effective therapeutics or vaccines. Members of the Arenaviridae family possess bi- or tri-segmented genomes encoding three to four viral proteins: glycoprotein (GP), nucleoprotein (NP), RNA polymerase (L) and matrix protein (Z). A robust T-cell immune response is critical for viral clearance and for limiting disease severity during the early stages of the infection. Methods Using a primary in vitro immunogenicity assay, we identified Conserved T cell Epitope Regions (CTERs) derived from conserved sequences within OWA and NWA, using LASV and JUNV as respective prototypes. These CTER epitopes were strongly recognized by human CD4+ T cells in vitro and were predicted to provide broad population coverage across diverse ethnicities. CTER constructs were designed based on the number of viral proteins included (GP+N+L, N+L, or L) and were assembled using AlphaFold into stable and unstable forms. Plasmids for OWA and NWA were codon-optimized and subsequently packaged into mRNA constructs. Results Our ongoing BDF1 mouse studies demonstrate that the CTER-based T-cell vaccine is both safe and immunogenic, as assessed using a combined activation-induced marker (AIM) and intracellular cytokine (ICS) assay. AIM+ T cells isolated from the spleen and lymph nodes show strong cross-reactive potential across the Arenaviridae family. Conclusion With further in vitro and in vivo evaluation, this work represents an initial step toward the development of a pan-arenavirus T-cell vaccine. Funding Source CEPI, NIH Topic Categories Vaccines and Immunotherapy (VAC)

IL-9 at the Maternal-Fetal Interface Drives a Novel Immunological Mechanism of Preterm Labor and Birth 2257559

Abstract Introduction Preterm birth (PTB) — the leading cause of neonatal morbidity and mortality - frequently arises from spontaneous preterm labor (PTL) without an identifiable infectious trigger, which is categorized as idiopathic (iPTL). Here, we uncover a previously unrecognized immunological mechanism driving a major subset of iPTL. Methods We integrated single-cell, spatial, and flow cytometric profiling of human decidual tissues from women with idiopathic or inflammation-associated PTL. A novel ultrasound-guided murine model was developed to deliver IL-9 at the maternal-fetal interface (MFI) and was compared to established models of PTB. In vitro and in vivo assays assessed IL-9’s role in inducing the common pathway of labor, and bulk and single-cell analyses assessed cellular homeostasis. Neutralization of IL-9 was tested to confirm causality, and vitamin D3 supplementation was evaluated for PTB prevention. Results IL-9—producing T cells were enriched at the MFI in women with iPTL but not in inflammation-associated cases. While systemic (maternal) and intra-amniotic (fetal) delivery of IL-9 did not cause adverse outcomes, IL-9 at the MFI induced PTB and neonatal mortality in mice. Mimicking iPTL, IL-9 at the MFI did not cause systemic or intra-amniotic inflammation, progesterone withdrawal, or fetal growth restriction. IL-9 induced myometrial contractility, membrane activation, and cervical shortening. Single-cell analyses revealed that IL-9 disrupted macrophage—stromal signaling and suppressed TGF-β—mediated immune regulation. Neutralization of IL-9 or vitamin D3 supplementation prevented PTB. Conclusion IL-9 is key immune effector driving non-infectious PTL through localized immune dysregulation at the MFI and represents a modifiable pathway for immune-based prevention of prematurity. Funding Source NIAID/NIH (RAI184491A) and Burroughs Wellcome Fund Topic Categories Mucosal and Regional Immunology (MUC)

Embryonic Stem Cell—Derived Signals as Potential Modulators of Prostate Tumor Cell Growth 2258320

Abstract Introduction Prostate cancer progression is influenced by the tumor microenvironment and soluble signaling factors. Mouse embryonic stem cells (mESCs) secrete cytokines such as LIF, BMPs, and TGF-β that regulate cell proliferation and differentiation. Previous reports suggest these signals can alter tumor behavior through paracrine interactions. This study investigates whether mESC-derived factors can modulate the proliferation of MyC-CaP prostate cancer cells and serve as a model for identifying natural inhibitors of tumor growth. Methods MyC-CaP cells were cultured in proximity to mESCs using a transwell co-culture system to allow paracrine signaling without direct contact. Cell viability and morphology were monitored microscopically and quantified using standard viability assays. Future experiments will incorporate a hyaluronic acid hydrogel scaffold to model the extracellular matrix and improve spatial organization for more physiologic signaling assessment. Results Preliminary co-culture data suggest reduced MyC-CaP proliferation when exposed to mESC-secreted factors compared with monoculture controls. Morphologic changes consistent with decreased proliferation and differentiation were observed. Additional assays are underway to quantify cytokine levels and confirm reproducibility under hydrogel conditions. Conclusion mESC-derived signals may suppress MyC-CaP prostate cancer cell proliferation through paracrine cytokine activity. Incorporating a hydrogel microenvironment will further define the mechanisms by which stem cell secretions influence tumor behavior, providing insight into potential translational applications for cancer modulation. Funding Source n/a Topic Categories Translational and Interventional Immunology (TI)

Discovery of Exhausted-like Lung Tissue-Resident Memory T Cells as Key Mediators of Protective Immunity 2257944

Abstract Introduction Respiratory viral infections, especially influenza, remain a major global health challenge, and circulating antibody-based vaccines are limited by frequent surface antigenic drift. In contrast, influenza-specific lung tissue-resident memory CD8+ T cells (TRMs) recognize conserved epitopes and provide both homologous and heterologous protection, serving as a rapid frontline defense during reinfection; however, the mechanisms that sustain and drive their protective functions remain poorly understood. Methods Through analyzing antigen-specific lung TRM from humans and mice, we identified a unique endogenous respiratory TRM population: PD-1HI CD103LO exhausted-like TRMs (TRM-ELs), which differ from PD-1LO CD103HI conventional TRMs (TRM-Cons) observed in most barrier tissues. Results Lung TRM-ELs exhibit unique transcriptional and signaling programs distinct from those of CD103+ TRM-Cons. Unlike previously reported TCR-independent TRM-cons, we demonstrated that local TCR signaling is critical for the maintenance of TRM-ELs. Importantly, the intact TRM-EL is indispensable for heterologous protection, while TRM-Cons don’t exhibit comparable efficiency. Mechanistically, the key molecule mediating TRM degranulation is significantly highly expressed in TRM-ELs, which facilitates the fast recall response and protection against secondary infection. The following experiments reveal that the impaired degranulation of effector molecules in TRM-ELs markedly diminishes the robust protective efficacy and alters the landscape of the anti-viral microenvironment in the lung. Conclusion Taken together, our research provides mechanistic insight into an unrecognized protective lung TRM population and elucidates distinct molecular pathways governing their persistence and antiviral efficacy. These results will lay the foundation for the development of universal respiratory mucosal vaccines to induce sustained local TRM immunity against influenza and related pathogens. Funding Source n/a Topic Categories Mucosal and Regional Immunology (MUC)

Amino-bisphosphonates enhance CD8+ T cell responses to vaccine antigens through an ATRAID-dependent mechanism involving the stromal compartment 2259959

Abstract Introduction Bisphosphonates (BPs) are a family of drugs that are widely used to prevent bone resorption. Recent retrospective epidemiologic studies have found a strong association between BP use and a marked reduction in incidence and severity of respiratory viral infections. This association was not observed in patients who had received non-BP medications to prevent bone loss, raising the possibility that BPs may boost anti-viral immunity by a mechanism unrelated to their effect on bone metabolism. Indeed, experiments in mice have shown that BPs can markedly boost humoral immune responses to a variety of antigens. Methods Here, we have examined the effect of alendronate (ALD), a nitrogen containing BP, on the response of CD8+ T cells to immunization with a model vaccine consisting of chicken ovalbumin (OVA) and CpG as an adjuvant. Results Compared to OVA + CpG alone, the vaccine combined with ALD elicited a significantly higher frequency of effector CD8+ T cells in the spleen and brachial lymph nodes, and these effector cells produced more granzyme B, perforin, TNF-α and IFN-γ. To access the cytosol of target cell, highly charged amino-BPs such as ALD, require the endosomal transporter, ATRAID. Indeed, vaccine supplementation with ALD in ATRAID-/- mice failed to boost endogenous effector CD8+ T cells. Moreover, reciprocal bone marrow chimeras between wild-type and ATRAID-deficient mice revealed that the ALD-induced enhancement of effector CD8+ T cell responses depends on non-hematopoietic compartments. Conclusion These data indicate that ALD is capable of enhancing the CD8+ T cell response to exogenous antigens by a mechanism that is critically dependent on the presence of ATRAID in the stromal compartment. Off-label combination of BPs with vaccines may potentiate cytotoxic T cell responses. Funding Source Moderna, Inc.; Ragon Institute of MGH, MIT and Harvard; NIH RO1 AI155865 Topic Categories Vaccines and Immunotherapy (VAC)

Ultra-small Saccharibacteria modulate immunoactivation through type IV pili and TLR2 2259816

Abstract Introduction Saccharibacteria (TM7) are episymbiont, that grow on the surface of host bacteria such as Schaalia and Actinomyces (Actinobacteria). They are positively associated with inflammatory diseases within the human microbiome, yet their mechanisms for interacting with the human host and contributing to diseases remain unknown. This study investigated tripartite interactions between TM7, their host/non-host bacteria and their human host. Methods Numerous oral epithelial cell lines were infected with TM7 strains their host/non-host bacteria, and their co-culture, followed by analysis of pro-inflammatory cytokine responses and downstream interaction with epithelial cells. Results TM7 dampened innate immune responses induced by both their host and non-host Actinobacteria in oral epithelial cells. Follow up transcriptomic analysis revealed host bacteria activated TLR2 pathway, while TM7 inhibited it through close interaction between TM7 Type IV pili (T4P) and TLR2 receptor, leading to TLR2 clustering and endocytosis via caveolin, thereby inhibiting host and non-host Actinobacteria induced epithelial innate immune response. Endocytosed TM7 was processed to lysosome, however, a fraction of TM7 persisted and survived to reinfect its host-bacteria upon mechanical lysis. Conclusion This study highlights the role of TM7 in inflammatory diseases and offer novel mechanistic insights into how TM7 influence immune activation and inflammation, highlighting their potential role in shaping the microbiome and contributing to inflammatory disease dynamics. Funding Source National Institute of Dental and Craniofacial Research (NIDCR) under awards 1R01DE031274 (B.B.) and 1R01DE023810 (X.H., J.S.M.). Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Engineering natural killer cells to express immunoglobulin A Fc receptor CD89 unlocks IgA-mediated cytotoxicity responses 2309809

Abstract Introduction Natural killer (NK) cells are critical effector cells of the innate immune system. Classically, NK cells mediate antibody-dependent responses via CD16, the IgG Fc receptor. However, IgG is the minority of Ig expressed at mucosal sites, whereas IgA is > 90% of Ig in mucosal secretions. Thus, harnessing NK cells for mucosal responses via IgA is an attractive therapeutic target. In this study, we demonstrate highly efficient CD89 knock-in that enables potent IgA- and IgG-mediated responses in NK cells. Methods Primary human NK cells were expanded and activated with irradiated K562-mbIL15 cells for 7 days before nucleofection with a transposon vector including the CD89 open reading frame. Knock-in efficiency was determined 4 days post-nucleofection and CD89+ NK cells were FACS purified and expanded with feeder cells. Functional assays were carried out using CD89+ NK cells and monoclonal rituximab antibodies expressing IgG, IgA1, or IgA2 Fc regions. CD89+ NK cells were co-cultured with Raji cells (10:1 E:T) and combinations of rituximab antibodies for 6-hours before staining and data acquisition via flow cytometry. Results CD89+ NK cells retained baseline phenotypes compared to non-nucleofected NK cells, including natural cytotoxicity and co-stimulatory receptors, intracellular signaling molecules, and markers of activation. Most importantly, CD89+ NK cells also retained CD16 expression, allowing them to respond to both IgG and IgA antibody activation. CD89+ NK cells display similar activation profiles as measured by CD107a, interferon-γ, and MIP-1β when activated with IgG, IgA1, or IgA2. As expected, CD89- NK cell controls only responded to IgG and not to IgA, confirming CD89-specific activation. Conclusion NK cells represent a promising avenue for cell-based therapeutics due to various benefits compared to T cell-based therapies. Through our current study we have shown consistent and robust engineering of NK cells to express CD89 to give them the ability to simultaneously respond to IgA and IgG. Funding Source NIAID grants R21AI184422 and P01AI162242 Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

Bacterial bunker-busters: Novel Immunotherapeutics against Pneumonic Plague 2253158

Abstract Introduction Yersinia pestis (Yp) is a gram-negative bacillus that causes plague in humans and animals and is endemic in Africa, Asia, and the Americas. When aerosolized, bacterium can cause an acute, often fatal disease that requires antibiotic intervention within 24hr of exposure. Antibodies have been previously raised against two protective antigens, F1 and LcrV, unfortunately Yp is able to circumvent these countermeasures. Methods To expand the breadth of protective targets we generated cow anti-sera against four novel antigens with potential induction of bovine exclusive ultralong CDR H3 antibodies. Cows were administered six doses of Pla, LptED, Ail and BamA, 3-4 weeks apart and antibody titers were evaluated throughout the duration of the study. Results Functional activity of anti-sera was evaluated utilizing gentamicin protection assay, by enumeration of internalized Yp by RAW cells in the presence or absence of complement. A prominent anti-LptED antibody response developed early during the course of vaccination, followed by anti-Pla, -Ail and -BamA. The number of RAW cells that internalized viable Yp was considerably lower in the presence of fresh or C8 repleted serum relative to heat inactivated or C8 depleted sera. Conclusion The inhibitory properties of cow anti-sera against four Yp antigens will enable identification of novel protective targets that will be used to produce additional countermeasures, especially if they stem from the ultralong CDR H3 repertoire. Funding Source DTRA-JSTO Topic Categories Vaccines and Immunotherapy (VAC)

Amphiphilic Diketopyrrolopyrrole Polymers Enable Intrinsic Selective Perchlorate Transduction in Organic Electrochemical Transistors

ABSTRACT Controlling ion‐specific interactions in organic mixed ionic–electronic conductors (OMIECs) remains a major challenge for high‐gain electrochemical sensors, particularly in aqueous environments where detection of emerging contaminants such as perchlorate (ClO 4 − ) is limited by ion hydration dynamics and sluggish volumetric electrochemical doping. Herein, we report a membrane‐free organic electrochemical transistor (OECT) platform based on a fully methoxylated diketopyrrolopyrrole (DPP)‐based amphiphilic copolymer. Backbone methoxylation promotes preferential volumetric electrochemical doping by perchlorate ions, enabling efficient bulk polaron stabilization. Consequently, the fully methoxylated polymer exhibits a five‐fold increase in volumetric charge ( Q V  = 32 C cm − 3 ) and a four‐fold enhancement in doping level ( y  = 0.38) relative to its non‐methoxylated analogue. Integrated into OECTs, the material delivers a 300‐fold improvement in detectivity, achieving detection limits of 22 ± 1 ppb in pristine electrolyte and 37 ± 2 ppb in a mixed‐ion background containing 5 mM each of F − , Cl − , NO 3 − , SO 4 2− , and ClO 3 − . Mixed‐interference studies demonstrate ∼12‐fold selectivity for perchlorate with 83% signal recovery, while control experiments and impedance spectroscopy reveal an auxiliary glycol effect, in which backbone methoxylation and glycol side chains cooperatively facilitate efficient bulk doping. These findings establish backbone functionalization as an effective strategy for realizing selective, membrane‐free OMIEC‐based chemical sensors.

Hysteresis Inversion Driven by Polar‐Order Competition in van der Waals Ferrielectric

ABSTRACT Emerging artificial intelligence paradigms increasingly rely on hardware that can dynamically adapt to surging computational workloads, yet present memristive technology remains fundamentally constrained by the fixed polarity of the device, meaning that its directionality for electrical response is unchangeable once integrated into a circuit. Consequently, a single device is locked into a specific operating mode, unable to achieve the flexible reconfiguration required for complex neural networks. Here we introduce a new device concept, a “polarity‐reconfigurable memristor” capable of breaking this constraint by exploiting the complex polar‐order competition in the van der Waals ferrielectric CuCrP 2 S 6 . By dynamically tuning the balance among antiferroelectric (AFE), ferrielectric (FiE), and ferroelectric (FE) states near the phase boundary, the device reversibly inverts its resistive hysteresis, switching between clockwise (CW) and counter‐clockwise (CCW) modes. This reversal allows the same device to alternate between potentiation‐ and depression‐type synaptic plasticity. These results establish polar‐order competition as an effective route to achieve intrinsic polarity reconfigurability for memory and neuromorphic hardware.

Hybrid multi-objective framework for constrained mixed-integer optimisation of industrial IoT gateway configuration

A critical role of epithelial MHCII in initiation of autoimmune tumorigenesis and sustaining premalignancy growth in the stomach 2300319

Abstract Introduction Autoimmunity is emerging as a new etiology for early-onset gastric cancer (GC), especially in young women, but it remains unclear what molecular pathways drive the initiation and progression of autoimmune tumorigenesis. Given that Major Histocompatibility Complex Class II (MHCII) is the strongest genetic risk factor for many autoimmune diseases, we hypothesize that MHCII-mediated autoantigen presentation drives tumorigenic differentiation of epithelial cells. Using an autoimmune gastric tumorigenesis model, we examined how MHC II-mediated presentation of autoantigens by hematopoietic or non-hematopoietic compartments may drive the initiation and progression of gastric premalignancy. Methods Rag knockout mice carrying a conditional MHCII deletion or lacking MHCII, or H2DM, were adoptively transferred with splenocytes from CTLA4KD mice, a model of autoimmunity-driven tumorigenesis that mimics human CTLA4 insufficiency and GC risk. Hematopoietic and nonhematopoietic compartments within the stomach were profiled using 40-color spectral flow cytometry. Histopathological evaluation was performed to assess inflammatory and premalignant lesions. Results Epithelial MHCII, rather than MHCII from immune cells, plays an essential role in the initiation of autoimmunity-driven tumorigenic differentiation of gastric epithelial cells, which was characterized by increased expression of cancer-associated markers with immune-evasive and stem-like features that potentiate premalignant progression. In addition, we show that early, but not advanced, gastric premalignancy is reversible upon the removal of epithelial MHCII. Conclusion These results reveal that epithelial MHCII antigen presentation is essential in the early stages of autoimmune-driven gastric tumorigenesis and highlight epithelial MHCII as a potential biomarker or therapeutic target in early interventions of autoimmunity-driven cancer development. Funding Source National Cancer Institute (R01CA245673) and the Sylvester Comprehensive Cancer Center Topic Categories Classical and Non-Classical Antigen Presenting Cells (APC)

Endosomal TLRs govern disease-stage specific B cell responses in Sjogren’s disease 2257833

Abstract Introduction Sjogren’s disease (SD) is a systemic autoimmune disease that occurs primarily in women. While B cells play an essential role in SD, the molecular mechanisms that govern B cell activation and the striking female disease predilection remain unclear. Using a mouse model of SD, termed NOD.B10, we showed that endosomal Tlrs govern disease in a sex-biased manner, as Tlr7 drove disease in females and systemic ablation of Tlr9 resulted in an increased percentage of splenic B cell subsets that are implicated in autoimmunity, such as germinal center and age-associated B cells. This B cell expansion was diminished in age- and sex-matched NOD.B10 females that lacked systemic expression of both Tlr7 and Tlr9. Based on these data, we hypothesized that Tlr7 and Tlr9 orchestrate disease-stage-specific B cell responses in SD. Methods We performed single-cell RNA sequencing on salivary glands from NOD.B10 females at the clinical disease stage, and observed that Tlr7 and Tlr9 expression was enriched in SD B cells. We then used flow cytometry to assess splenic B cell activation in NOD.B10 strains with clinical disease. Results Our data revealed that the percentage of activated B cells (B220+CD69+) was increased in NOD.B10Tlr9-/- females as compared to NOD.B10 mice, and this was dependent on Tlr7. No differences in B cell activation were observed in any of the male strains. To determine if B cells from NOD.B10 females showed differences in sensitivity to Tlr9 agonism with disease progression, we isolated spleens from pre-disease, clinical disease, and advanced disease stage mice and age- and sex-matched controls and cultured them with CpG-B. SD B cells were hyperresponsive to CpG-B at pre-disease and clinical disease time points, while no differences in CD69+ expression were observed at the advanced disease stage. Conclusion Overall, our study revealed that Tlr9 governs B cell activation in SD females in a Tlr7-dependent manner and SD B cells show differential sensitivity to Tlr9 at distinct disease stages. Funding Source R01 DE029472 Topic Categories Basic Autoimmunity (BA)

Evaluating MMP9 Expression Across Cancer Cell Lines to Identify Models for Functional Analysis 2258440

Abstract Introduction Matrix metalloproteinase-9 (MMP9) is an extracellular enzyme that contributes to tumor invasion and metastasis through degradation of the extracellular matrix. Elevated MMP9 expression has been reported in multiple cancers, but its relative activity differs across tumor types. This study aims to compare MMP9 expression among cancer cell lines to identify a model suitable for downstream functional studies on tumor progression. Methods Initial experiments examined MMP9 expression in the U-87MG glioblastoma cell line using quantitative PCR (qPCR). RNA extraction and cDNA synthesis were performed following standard molecular biology protocols. Due to low MMP9 expression in U-87MG (Ct ≈ 33), additional cancer models were selected based on prior literature, breast cancer and lymphoma cell lines. RNA was isolated from these lines, and qPCR analysis is being conducted to quantify relative MMP9 expression levels and identify the most suitable model for subsequent CRISPR knockout experiments. Results qPCR analysis confirmed minimal MMP9 expression in U-87MG glioblastoma cells, inconsistent with its predicted aggressiveness. High-quality RNA and cDNA were successfully prepared from both breast cancer and lymphoma cell lines. Ongoing expression analysis will determine which exhibits higher MMP9 levels for targeted functional studies. Conclusion MMP9 expression varies significantly across cancer cell types, underscoring the need for model selection prior to mechanistic studies. Identifying a cell line with robust MMP9 expression will enable future CRISPR knockout experiments to investigate its role in tumor invasiveness and metastatic potential. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Using dynamic model building and simulation to understand cytokines and adaptive immunity 2258020

Abstract Introduction An immune response is mediated by a complex interconnected system of cells and molecules. Understanding these responses requires systems-level thinking. Traditional static diagrams in textbooks fail to convey the dynamic interactions between immune components over time. To address this gap, we developed a set of interactive activities using the computational modeling platform Cell Collective. The collection enables students to visualize, predict, and simulate how cytokine receptor binding influences CD4+ T cell differentiation and how Th1 and Th2 subsets shape immune responses to distinct pathogens. Methods The lesson uses scaffolded, hands-on exercises that engage students in computational modeling and simulation to explore CD4+ T cell differentiation and cytokine signaling pathways. The module was implemented in three different immunology-focused undergraduate settings. Students completed pre- and post-assessments to measure learning gains and confidence in computational modeling, along with a post-activity survey evaluating user experience. Results Student knowledge content scores increased from an average of 60.4% pre-lesson to 89.0% post-lesson, indicating significant learning gains. Post-assessment responses revealed that 86% (n = 42) of students agreed that the activity increased their understanding of the key immunological concept that the immune system is interconnected and interacting. Additionally, 89% (n = 36) provided positive feedback regarding their overall experience with the module. Conclusion Integrating computational modeling into undergraduate immunology instruction significantly enhances student understanding of complex, systems-level immune mechanisms. The use of Cell Collective allows learners to move beyond static representations and actively explore how cellular interactions influence immune outcomes. The module is adaptable to various course levels and offers a scalable, evidence-based strategy for improving immunology education through active, technology-enhanced learning. Funding Source This work was supported by the National Science Foundation under Grants #1915131 awarded to Tomas Helikar and RCN-UBE #2316260 awarded to ImmunoReach. Topic Categories Immunology Education and Communication (EDU)

Aberrant NK Cell Migration and Differentiation Underlie Immune Dysregulation in Endometriosis 2259942

Abstract Introduction Endometriosis affects approximately 10% of women of reproductive age and remains a major clinical challenge due to its association with infertility and reduced quality of life. The disease is difficult both to diagnose and to treat, as current approaches–including laparoscopic removal and hormonal therapy–are time-consuming and often fail to achieve a complete cure. Previous studies have demonstrated impaired NK cell function in patients with endometriosis, while murine experiments have suggested a potential role for NK cells in disease regulation; however, the mechanisms underlying this dysfunction remain unclear. This study aims to investigate whether and how aberrant NK cell migration and differentiation contribute to immune dysregulation in endometriosis. Methods A murine endometriosis model induced by intraperitoneal injection of dissociated uterine horn tissue and spatial transcriptomic profiling of patient-derived lesions were utilized to characterize NK cell subsets and their transcriptional signatures within endometriotic tissues in mice and humans. Results Antibody-mediated NK cell depletion using an NK1.1 monoclonal antibody resulted in significantly larger lesions, indicating that NK cells contribute to lesion control. Moreover, lesion weight was negatively correlated with the abundance of tissue-resident NK cells. Using a congenic mouse model, we confirmed that CD49a+ tissue-resident NK cells were not derived from donor uteri, but originated from the host, suggesting that NK cell migration and differentiation are essential for their function in regulating lesion growth. Conclusion Ongoing studies are expanding on these preliminary findings to elucidate how NK cell migration and differentiation shape the immune landscape of endometriosis. A deeper understanding of these mechanisms may provide a foundation for developing novel diagnostic and therapeutic strategies targeting NK cell—mediated immune regulation. Funding Source This research was supported by a grant of the Boston-Korea Innovative Research Project through the Korea Health Industry Development Institute(KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant no. : RS-2024-00403047) Topic Categories Immune Mechanisms of Human Disease (HUM)

Single-cell RNA and CITE-seq analysis of human CD4+ T cells reveals age-associated upregulation of IL-32 2259951

Abstract Introduction Aging profoundly alters the human immune system, leading to increased vulnerability to infection, autoimmunity, and cardiovascular disease. Methods To define how aging reshapes CD4+ T-cell phenotypes and transcriptional programs, we performed single-cell RNA sequencing combined with CITE-seq (51 surface proteins) on peripheral blood mononuclear cells from 61 patients with angiographically confirmed presence or absence of coronary artery disease (CAD). Results We identified 139,343 high-quality single cells, including 40,821 CD4+ T cells distributed across 16 transcriptionally and phenotypically distinct clusters. The proportions of a central memory (TCM) subset and a cluster of TEMRA cells increased significantly with age, particularly in CAD patients, whereas a MMP9+ CD4+ subset declined with age. Transcriptomic analysis revealed 56 genes that were significantly upregulated with aging, many shared among clusters. Notably, IL32 was consistently increased with age, especially in individuals with low CAD burden. IL32 expression was highest in T cells and NK cells, with regulatory T cells (Tregs) showing the strongest IL32 signal among CD4+ T cells. Conclusion These results suggest that aging drives selective expansion of effector-memory and IL32-expressing subsets. Together, our study provides a single-cell framework for understanding age-associated remodeling of human CD4+ T-cell immunity in the context of atherosclerotic disease. Funding Source N/A Topic Categories Computational and Systems Immunology (COMP)

Open resources and data from the Allen Institute: exploring immune health, rheumatoid arthritis, tissue immunity, and cytokine perturbations 2309948

Abstract Introduction A core goal of the Allen Institute is to openly share our immunological data and enable the immunology community to explore and re-use data from our studies of human disease cohorts. We have profiled healthy adults and children as well as patients at risk for rheumatoid arthritis, with multiple myeloma, under treatment for melanoma, and with COVID-19 longitudinally for up to two years, then performed immune profiling using scRNA-seq, plasma or serum proteomics, and clinical lab tests. Methods To process, analyze, and distribute this data, we developed the Human Immune System Explorer (HISE) platform, a flexible, scalable, cloud-based framework to enable storage, interactive analysis, visualization, and generation of Certificates of Reproducibility that enable inspection and replay of any step of an analysis workflow. This platform enables us to openly provide data, interactive visualization tools, scientific context, and analysis methods with the broader immunology community. Results We have applied our tools to both our own studies and large publicly available data resources, including a dataset of ∼10 million PBMCs from a study with 90 cytokine perturbations. These tools and data are freely available to the public, including interactive differential expression, UMAP, clinical data, and longitudinal trend visualizations. Conclusion We invite immunologists to explore this resource and our expanding library of immunology data, insights, and tools at https://explore.allenimmunology.org/. Funding Source n/a Topic Categories Computational and Systems Immunology (COMP)

Immune Cell Derived 1,25(OH)2D3: A Critical Defense Against Influenza Infection 2257145

Abstract Introduction Immune cells have vitamin D receptors (VDR) making them direct targets of vitamin D. CYP27B1 is the critical enzyme that hydroxylates circulating, inactive 25(OH)D to form the active, high affinity VDR binding, 1,25(OH)2D. CYP27B1 is expressed primarily in the kidney; however, immune cells can also express CYP27B1. Activated macrophages and T cells have been shown to produce 1,25(OH)2D. The role of immune cell derived 1,25(OH)2D in host resistance to influenza infection was examined here. Methods To investigate the role of CYP27B1 in host resistance to respiratory infection, whole body CYP27B1 knockout (KO) mice and mice with a deletion in the ability of the kidney to make 1,25(OH)2D (double intronic CYP27B1 renal deletion, DIKO) were infected with mouse adapted H1N1 influenza A. DIKO mice only express extra renal CYP27B1 that includes immune cell produced 1,25(OH)2D. Results Influenza infected CYP27B1 KO mice developed severe respiratory symptoms and significantly more lethality compared to wildtype (WT) littermates. Influenza infection of DIKO mice was not different than in WT mice, indicating that immune cell production of 1,25(OH)2D is protective during influenza. CD8+ T cells from CYP27B1 KO mice had impaired granzyme B responses and lack of PD-1 activation as compared to WT littermates. This data supports a role for immune cell production of 1,25(OH)2D which is critical for the cytotoxic function of CD8+ T cells. Experiments are underway to determine the effect of T cell specific CYP27B1 deletion in response to influenza infection. Conclusion Immune cell derived 1,25(OH)2D is a critical regulator of influenza pathogenesis and is required for granzyme B production by CD8+ T cells. Understanding the mechanisms that regulate local 1,25(OH)2D production in T cells or other immune cell types is needed for future recommendations that might include vitamin D supplementation for the prevention or treatment of viral respiratory infections. Funding Source This work was supported by the National Institutes of Health (NIH) grants R01AT005378 to M.T.C., T32GM154124 to M.T and T32DK120509 to N.E.F. And USDA National Institute of Food and Agriculture and Hatch Appropriations Projects #PEN04963 Accession #700657 Topic Categories Viral Immunology (VIR)

Multifunctional Injectable Hydrogel as a Biomimic Lens: Optical–Mechanical Restoration and Dynamic Postoperative Microenvironment Regulation

ABSTRACT Phacoemulsification followed by intraocular lens (IOL) implantation is currently the only effective treatment for cataracts. However, conventional IOLs exhibit limited biological adaptability and fixed or discontinuous focal properties, preventing restoration of the natural lens's continuous accommodation and often causing postoperative visual disturbances and complications. Herein, we report an injectable multifunctional hydrogel‐based biomimic lens that simultaneously reconstructs optical–mechanical properties and dynamically modulates the postoperative lenticular microenvironment. Through multilayered dynamic crosslinking and micro‐capsulorhexis‐assisted in situ gelation within the capsular bag, the hydrogel forms a biomimic lens with continuous refractive characteristics. Meanwhile, its drug‐loading capability enables the incorporation of multifunctional components for early postoperative antibacterial, anti‐inflammatory, and antifibrotic effects. In addition, after inflammation subsides, the loaded photocatalytic elements induce the orderly clearance of residual lens epithelial cells, enabling comprehensive restoration of microenvironmental homeostasis within the capsular bag. In a rabbit model, this hydrogel lens demonstrates effective refractive reconstruction and sustained suppression of postoperative complications, highlighting its potential as a next‐generation intraocular lens.