Browse Articles

The potential application of bioflavonoids in cancer treatment 2309347

Abstract Introduction Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide. The 5-year survival rate of HCC is very low at advanced stages. No effective treatments are available for advanced HCC. Therefore, new therapeutic drugs and treatment methods for HCC remain to be developed. This study aims to investigate the role of bioflavonoids in HCC treatment. Methods Bioinformatic tools and computer-aided techniques advance the processes of drug development. Here, we incorporated bioinformatic analyses and computer-aided drug development processes to investigate the potential application of bioflavonoids as a therapeutic agent for HCC treatment. Results Our results revealed that predicted hub target genes of bioflavonoids displayed functional importance in cancer-related signaling pathways in different cancers, including HCC. Importantly, the mRNA expression levels of the key hub genes, such as vascular endothelial growth factor, were significantly different between normal and HCC tumor samples. Their expression levels were significantly associated with the clinical survival outcomes of HCC patients, as well as the immune cell infiltration in the HCC tumor microenvironment. In addition, these four genes showed significant co-expression correlated with immune checkpoints, including PD-1, PD-L1, and CTLA-4. Furthermore, we investigated the binding affinity and potential binding mechanisms between bioflavonoids and target proteins encoded by the hub genes. Conclusion This study shed light on the potential application of bioactive molecule bioflavonoids in HCC treatment by regulating four key hub genes. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

A unique epigenetic circuitry defines CD8 T cell stemness shared across chronic diseases 2330201

Abstract Introduction In autoimmunity and chronic infection, both settings of persistent (self or foreign) antigen, immune responses are sustained by stem-like CD8 T cells, which self-renew and give rise to differentiated progeny. However, if and how T stemness is epigenetically encoded, which transcription factor(s) regulate the stem-T cells, and whether the stem-T cell state is disease-specific or shared across diseases, is currently not known. Methods We used clinically relevant models of autoimmune type 1 diabetes (T1D) and chronic infection and conducted serial T cell transplantation studies in vivo, combined with single cell paired RNA- and ATAC-sequencing on antigen-specific T cells. We developed CRISPR/Cas9-mediated gene-editing approaches in primary T cells as well as CUT&RUN studies, identifying a novel hierarchy of transcription factors regulating stem T cell identity and function. Results We discovered that a small subset of stem-T cells (TSC) express lymphoid enhancer-binding factor 1 (LEF1), a member of the TCF/LEF TF family. Paired single cell transcriptomic and epigenomic analyses reveal that the LEF1+ TSC harbor a unique epigenetically encoded molecular state enriched in genes and pathways characteristic of embryonic and adult (somatic) stem cells (e.g. neural stem cells). Strikingly, we found that TSC in chronic infection harbor a LEF1+ TSC pool sharing the core stemness epigenetic and molecular program observed in autoimmune LEF1+ TSC. Loss- and gain-of-function studies in both autoimmune T1D and chronic infection confirmed the critical role of LEF1 in maintaining T cell stemness. CUT&RUN analyses provide clues as to how LEF1 instructs the epigenetically encoded program of stem-T cells. Conclusion Here we reveal novel insights into the molecular circuitries of CD8 T cell stemness and differentiation. We discover LEF1 as the master regulator defining T cell stemness and identify novel targets for therapeutic intervention. Funding Source NIH R01AI173249, JDRF SRA-2023-1410-S-B, MSKCC Basic Research Innovation Award, The Hearst Foundation Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)

Integrative Curation of Human and Rhesus Macaque Immunoglobulin Germline Loci for Population-Scale Immunogenomics 2267549

Abstract Introduction The immunoglobulin (IG) loci are among the most polymorphic and structurally complex regions of mammalian genomes, yet current germline references remain incomplete and inconsistent across species. We developed unified resources for human and rhesus macaque IG loci by integrating high-fidelity long-read genomic assemblies with matched AIRR-seq data. Our framework curates validated alleles, recombination signal sequences (RSSs), and regulatory elements across 170 human and 300+ macaque samples, establishing a foundation for population-scale and comparative immunogenomics. Methods Targeted long-read assemblies (PacBio HiFi) of IGH, IGK, and IGL loci were analyzed with AIRR-seq repertoires using a unified Nextflow pipeline. Personalized genomic germline sets were inferred and validated by repertoire evidence. Novel alleles were identified through similarity-based clustering and cross-modal validation. RSSs and leader sequences were annotated from flanking regions, and population variation was assessed through allele frequencies, Jaccard similarity, and repertoire concordance. Results For humans, we curated IGH, IGK, and IGL allele sets from 170 individuals, identifying hundreds of novel, population-specific alleles validated by AIRR-seq concordance and annotated with RSSs, leader, and regulatory regions linked to ancestry. In rhesus macaques, we identified 1,643 coding alleles from 300+ animals, including 1,338 novel variants, and cataloged 328 RSS motifs revealing conserved heptamer—nonamer cores but locus-dependent spacer variability. Conclusion The human and macaque unified allele resources establish harmonized, cross-species IG germline references. By integrating genomic and repertoire data, these resources advance immunogenomic annotation, diversity analysis, and modeling of antibody evolution in humans and nonhuman primates. All data and annotations will be available through VDJbase.com, supporting open, standardized immunogenetics research. Funding Source NIAID R24 AI162317 Topic Categories Computational and Systems Immunology (COMP)

Stress‐Lensed Electrochemical Sintering Enables Fast and Stable Lithium‐Silicon Alloy Chemistry in All‐Solid‐State Batteries

ABSTRACT The fundamental challenge in all‐solid‐state batteries (ASSBs) lies in regulating the dynamic reconstruction of solid‐solid interfaces under electro‐chemo‐mechanical conditions. Currently, no mechanism exists to reconcile the conflicting requirements of structural stability and rapid ion transport for high‐capacity silicon (Si) anodes: expansion‐accommodated encapsulation strategies preserve integrity but block interparticle ionic contact, while intrinsic electrochemical sintering restores conduction but creates excessive agglomeration that fractures the electrode. Here, we propose a stress‐lensed electrochemical sintering (SLES) strategy to guide selective interfacial bonding by depositing Si conformally into a porous carbon host, specifically utilizing its high‐curvature pore entrances as geometric constrictions. During cycling, these constrictions act as “stress lenses”, concentrating the volumetric expansion stress of Si precisely at interparticle contacts. This focused mechanical energy locally lowers the atomic diffusion barrier, guiding the formation of a robust, percolating Si network while preserving internal voids to buffer volume changes. The resulting Si‐SLES anode resolves the stability‐transport conflict, achieving ∼100% capacity retention after 100 cycles with superior rate capability and demonstrating practical viability in full cells over 700 cycles.

Structural and biophysical analysis of the four CHRD domains of human chordin reveals a novel binding site for glycosaminoglycans

Progressive degradation-aware distillation for robust indoor object detection

In situ Transglutaminase 2-mediated Crosslinking of Engineered IL-7 to the Tumor Matrix Enhances Immunotherapy and Anti-PD-1 Response 2308334

Abstract Introduction Cytokine therapies in immuno-oncology have been limited by rapid clearance and systemic toxicity. We present a novel strategy to anchor cytokines to the tumor ECM via tissue transglutaminase 2 (TG2)—mediated transamidation, an enzymatic process that covalently crosslinks ECM proteins. Because the enzymatic activity of TG2 is restricted to the tumor microenvironment, our strategy enables tumor-specific conjugation of systemically administered cytokines, thereby enhancing their antitumor activity while limiting systemic exposure. Methods We engineered a TG2 substrate domain—fused interleukin-7 (TSD-IL-7) that undergoes TG2-mediated covalent conjugation to lysine-containing extracellular matrix components. Tumor retention and bioactivity were evaluated following intravenous administration across multiple murine solid tumor models. Therapeutic efficacy was assessed as monotherapy and in combination with αPD-1 immune checkpoint blockade, with immune phenotypes analyzed in tumors and tumor-draining lymph nodes. Results TSD-IL-7 exhibited potent antitumor activity across multiple murine solid tumor models including orthotopic and metastatic disease, and enhanced responses to αPD-1 immune checkpoint inhibition without systemic toxicity. Remarkably, it drove complete tumor clearance in most animals in a checkpoint-refractory ovarian cancer model. Mechanistically, TSD-IL-7 promoted T cell proliferation and antitumor cytokine and chemokine production in the tumor and tumor-draining lymph nodes, while reducing T cell exhaustion and promoting CD8+ central memory T cell development within the tumor. Conclusion We present a novel strategy for covalently anchoring therapeutics to the tumor matrix following systemic administration by harnessing tumor-specific transglutaminase-2 activity, amplifying the antitumor immunity of engineered IL-7 while synergizing with immune checkpoint blockade to clear otherwise therapy-resistant solid tumors. Funding Source National Cancer Institute grant R21-CA277414 (J.A.H.). National Cancer Institute grant R01-CA219304 (M.A.S.). National Institutes of Health Immunology Training Grant T32AI07090 (H.C.). National Research Foundation of Korea Basic Science Research Program, Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Altered composition of peripheral immune subsets in Four Core Genotype mice 2260628

Abstract Introduction Sex-biased immune differences affect autoimmune disease prevalence, susceptibility to infections and anti-tumor responses. The genetic, hormonal and environmental factors that contribute to these differences are difficult to disentangle. Four Core Genotype (FCG) mice harbor an inactivated Sry gene on the Y chromosome (hereafter Sry* and Y*) and an autosomal Sry transgene (Sry-tg) on chromosome 3, allowing the generation of gonadal females with XX and XY* genotypes, and Sry-tg-bearing XX and XY* gonadal males, thereby separating genetic and hormonal/gonadal influences in sex-biased phenotypes. The FCG model has been widely used to investigate sex-linked genetic vs hormonal factors in developmental, behavioral and immune phenotypes. Recently, a 9-gene translocation, including the Tlr7 and Tlr8 genes, was identified on the Y* chromosome. In addition, several genes with potential immune significance are known to flank the Sry-tg insertion site. Methods To evaluate the effect of these genetic alterations on baseline immunity in FCG mice we carried out flow cytometry on peripheral blood, thymus, spleen and superficial lymph nodes of mice of each of the 4 FCG genotypes and wild-type C57BL/6 male controls. Results Mice bearing the Sry-tg (FCG XX and XY* males) showed a 3- to 5-fold reduction in peripheral CD8+ T cell fractions (p < 0.01 compared to WT males), but not in the thymus, arguing against a CD8 T cell development defect. B cell analysis identified a significant ∼2-fold decrease in splenic marginal zone B cell fractions in FCG XY* males and females, which bear the 9-gene Y* translocation (p < 0.01 compared to WT males and XX females). Conclusion These results indicate that baseline immune subset composition is significantly altered in FCG mice, differentially affecting T and B cell populations depending on the genotype. Transcriptional and immunological effects associated with these differences, and their potential impact on immune responses to tumors and pathogens will be discussed. Funding Source Camden Cancer Research Center Seed Grant Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)

Regulation of Inflammatory Diseases by Solute Carrier Transporters 2266201

Abstract Introduction Cytosolic innate immune sensing is essential for maintaining the integrity of barrier tissues such as the skin and gut. The pattern-recognition receptors NOD1 and NOD2 detect bacterial peptidoglycan fragments (muropeptides) to trigger antimicrobial and inflammatory responses. However, how these muropeptides gain access to the cytosol of epithelial cells has remained unclear. Methods Expression of Slc46a2 and Slc46a3 was analyzed by qRT-PCR, immunofluorescence, and RNA-seq datasets. CRISPR-Cas9—mediated knockout and overexpression studies were performed in epithelial cells to assess NOD1/2 activation using NF-κB reporter, cytokine (IL-8) assays, and labeled muropeptide uptake. Slc46a2⁻/⁻, Slc46a3⁻/⁻, Nod1⁻/⁻, and Nod2⁻/⁻ mice were used in imiquimod-induced psoriasis and DSS or Citrobacter rodentium colitis models. Inflammation and barrier integrity were evaluated by histopathology, cytokine profiling, and immune cell analysis. Results Our study identifies a family of solute carrier transporters, SLC46s, as key mediators of this process. We demonstrate that SLC46A2 and SLC46A3 selectively transport DAP- and MDP-type muropeptides, respectively, thereby triggering cytosolic NOD1 and NOD2 activation in epithelial cells. SLC46A2 is highly expressed in epidermal keratinocytes, where its loss impairs DAP-muropeptide uptake and NOD1-dependent responses, leading to reduced psoriatic inflammation in mice. Conversely, SLC46A3 is expressed in intestinal epithelial cells and is required for MDP transport and NOD2 activation. Slc46a3-deficient mice exhibit increased susceptibility to DSS-induced and Citrobacter rodentium-associated colitis, mirroring the pathology of Nod2-/- animals Conclusion Together, our findings reveal SLC46 transporters as critical gateways for muropeptide entry into epithelial cells, establishing a mechanistic link between microbial sensing and barrier homeostasis, and suggesting novel therapeutic targets for inflammatory skin and intestinal diseases. Funding Source NIH Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

Nasal anti-CD3 modulates T cell and Microglia activation in aged mice 2267372

Abstract Introduction Cellular senescence is characterized by a permanent arrest of the cell cycle, often accompanied by dysfunctional regulatory T cell (Treg) activity and loss of homeostatic microglia, contributing to neurotoxicity. We previously demonstrated that intranasal administration of anti-CD3 (αCD3) induces several Treg subpopulations that produce anti-inflammatory cytokines such as TGF-β and IL-10, which can reduce microglial inflammation and ameliorate neurodegenerative conditions like multiple sclerosis (MS) and Alzheimer’s disease (AD). Methods In this study, we treated aged female mice with intranasal αCD3 and evaluated the effects on brain immune cell populations and behavior. Mice aged 8, 16, and 70 weeks received 1 μg of nasal αCD3 three times per week for three months. Following treatment, we assessed motor function, memory, anxiety, and depression-like behavior. Brain cell suspensions were sorted into three CD45-defined populations for single-cell enriched analysis, and spleens were analyzed by flow cytometry. Results While no significant differences were observed in behavioral outcomes, single-cell analysis revealed that αCD3 treatment altered the myeloid compartment. We found increased clustering of dendritic cells, oligodendrocytes, macrophages, and monocytes in treated mice compared to controls. Additionally, we detected an upregulation of T cell markers such as CD39 and TIGIT, and reduced expression of proinflammatory cytokines including IL-17, TNF-α, and IFN-γ. Conclusion Collectively, our findings suggest that nasal αCD3 induces a systemic anti-inflammatory immune response and modulates microglial-related pathways, which may confer neuroprotective benefits in the context of aging. Funding Source The ALS Association, NIH-National Institutes of Health Topic Categories Neuroimmunology (NEUR)

Investigating the role of Galectin-3 in a model of lung granuloma formation 2309412

Abstract Introduction Granulomas are structured clusters of macrophages and lymphocytes that form in tissues in response to persistent antigens and, in some diseases, without an identifiable cause. Granulomatous inflammation is central to the pathogenesis of certain infections, for example, those caused by mycobacteria. Mechanisms that drive granuloma formation, and whether they represent an adaptive or maladaptive response, remain poorly defined. Methods We used mice deficient in Tsc2 gene expression in myeloid lineage cells (Tsc2ffLys2Cre+). An inoculum of 5x105 of the mycobacterium Bacille Calmette-Guérin (BCG) was administered intranasally and lung flow cytometry, histology, immunofluorescence and single cell RNA sequencing were performed. Results Following BCG challenge, we observed histologic evidence of progressive granuloma formation in the lungs, mediastinal lymph nodes, spleen, skin, and heart tissues over 11 weeks in Cre-positive mice, but not Cre-negative littermates. The granulomas were composed of large tissue macrophages, some forming multinucleated giant cells, surrounded by lymphocytes. Lung single cell transcriptional profiling identified an atypical histiocyte population in Cre-positive animals. We used these data to develop a flow cytometry gating strategy to identify this histiocyte population, using Galectin-3 as a key marker. Using Immunofluorescence, we observed the presence of Galectin-3 on the cell surface. The atypical histiocytes exhibited cell surface Galectin-3 expression and increased mitochondrial superoxide production. Targeting the carbohydrate-binding domain of galectin-3 with the synthetic small molecule inhibitor, TD139 (olitigaltin), resulted in lower numbers of these atypical histiocytes in the lungs. Conclusion Histiocytes play a critical role in granuloma formation and maintenance. Galectin-3 can serve as a marker for histiocytes and may be a promising target for therapies aimed at treating granulomatous diseases. Funding Source n/a Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Electrophysiological characterization of primary and immortalized immune cells using automated patch-clamp technology 2335008

Abstract Introduction Ion channels and transporters mediate key functions of innate and adaptive immune cells, including the regulation of membrane potential and Ca2+-mediated intracellular signaling pathways. Kv1.3 voltage-gated potassium channels and KCa3.1 intermediate conductance Ca2+-activated K+ channels are particularly important for regulating membrane depolarization and oscillations that facilitate intracellular calcium-release activated calcium (ICRAC) currents and activation of naïve T-cells. Methods Here, we use automated patch-clamp electrophysiology (Sophion QPatch48) to record and characterize K+ currents from both immortalized and primary B-cells and T-cells. Primary murine T-cells were collected and compared with immortalized T- and B-cell lines and stable cell lines expressing Kv1.3 and KCa3.1 channels. Recording solutions and compounds were optimized for each target ion channel of interest. Results Inactivating voltage-dependent outward currents were determined to be Kv1.3 mediated by inhibition with low concentrations of the isoform-specific blocker PAP-1. Intermediate conductance (IK) KCa3.1 channels were measured using different Ca2+ concentrations in the intracellular solutions and applying voltage ramps during the whole-cell break in period. IK currents grew in amplitude with each sweep as Ca2+ diffused into the intracellular environment, and these currents could be either enhanced with riluzole or blocked by TRAM-34, suggesting they were mediated by KCa3.1. Conclusion Our results demonstrate the utility of high-throughput automated patch-clamp for measuring ion channel composition and function across primary and immortalized immune cells. This approach permits rapid, simultaneous recording of many cells under the same conditions, allowing more accurate assessment of ion-mediated processes during immune cell activation. Funding Source n/a/ Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)

TCR affinity mediated T cell clonal type development 2267484

Abstract Introduction T cell receptor (TCR) functional potency is determined by membrane-proximal, two-dimensional (2D) kinetics–on-rate, off-rate, and 2D affinity–which correlate more closely with cellular responses than conventional 3D measures. The CD8 co-receptor enhances bond number and lifetime, and normalized synergy quantifies its contribution across affinity ranges. However, how individual TCR biophysical traits relate to in vivo clonal outcomes–expansion and differentiation (naïve, TCM, TEM, cytotoxic)–remains unclear in human pathogen-specific repertoires. We aim to connect quantitative 2D biophysics with single-cell phenotypes in a SARS-CoV-2 cohort to identify features predicting robust expansion and defined differentiation. Methods SARS-CoV-2 vaccine and infection—annotated donors were analyzed. Antigen-specific CD8+ T cells were identified by peptide—MHC tetramers and single-cell sequencing to recover paired TCRα/β and transcriptomic profiles. Selected TCRs were cloned into Jurkat-76 (J76) derivatives expressing or lacking CD8, including Nur77/NFAT reporter lines, to assess co-receptor dependence. 2D kinetic parameters were measured using micropipette adhesion and bio-membrane force microscopy. Single-cell phenotypes were clustered (Seurat/totalVI), linked to clonotypes, and analyzed for correlations between biophysical features, expansion, and differentiation. Results SARS-CoV-2—specific TCRs exhibited wide 2D affinity and CD8 dependency ranges, indicating diverse activation mechanisms. Integrating biophysical and single-cell data revealed that expansion and differentiation correlate with combined metrics of affinity, stability, and co-receptor cooperation. Highly expanded clones were not always the strongest binders; rather, normalized CD8 synergy identified low-affinity yet functionally potent clones. Conclusion These insights suggest that composite biophysical signatures, not affinity alone, govern clonal fate and may inform TCR-based immunotherapies and vaccine design for durable protection. Funding Source n/a Topic Categories Viral Immunology (VIR)

Direct Visualization and Regulation of Interfacial Ion Concentration Layer at Zinc Metal Interfaces via an Ion‐Buffering Artificial Solid Electrolyte Interphase

ABSTRACT Direct experimental visualization of the electrical double layer (EDL) and its associated layer formation at metal–electrolyte interfaces remains challenging yet is essential for understanding interfacial instability in zinc metal anodes. Herein, we engineer a PEO/PIM‐1 hybrid polymer (PIP) as an artificial solid–electrolyte interphase (SEI) that replaces field‐driven interfacial ion accumulation with confined ion buffering, thereby suppressing EDL amplification and chaotic interfacial dynamics. Liquid‐phase atomic force microscopy (AFM) measurements demonstrate the elimination of voltage‐dependent force amplification upon PIP modification, evidencing effective decoupling of electric‐field‐induced ion crowding at the Zn surface. In situ Raman spectroscopy, electrochemical analysis, and molecular dynamics simulations further reveal reduced interfacial water activity, regulated ion distribution, and homogeneous Zn stripping and deposition. PIP significantly enhances anode stability, enabling Zn|Zn symmetric cells to cycle for 2700 h and delivering a Coulombic efficiency of 98.6% in Zn|Ti cells. When coupled with I 2 cathodes, PIP@Zn supports long‐life 4e − Zn|I 2 batteries, achieving >6000 cycles at 4 A g −1 and >27 000 cycles at 8 A g −1 . For the practical application, 4e − Zn|I 2 pouch cell delivers 500 mAh capacity and 500 cycles. This work offers a mechanistic framework for probing EDL dynamics and a generalizable SEI strategy for stabilizing Zn metal.

Correction: RIPC improves myocardial injury by promoting mitochondrial protection via the PGC-1α/Nrf2 pathway

Two-way THz vehicular communications with mobile RISs: RSMA/NOMA performance analysis and DRL based optimization

Nuclear Localization of IRG1 Modulates COX2 Expression and Prostaglandin Biosynthesis in Primary Human Macrophages 2308479

Abstract Introduction Immunometabolism within macrophages represents a dynamic process modulated by environmental factors and pathological conditions. Immune Responsive Gene 1 (IRG1) catalyzes the conversion of cis-aconitate, an intermediate in the tricarboxylic acid (TCA) cycle, to itaconate, and is robustly upregulated in macrophages following toll-like receptor engagement or type II interferon stimulation. The upregulation of IRG1 results in reduced macrophage cytokine production and influences polarization. To date, IRG1 localization has primarily been associated with mitochondria. In this study, we present evidence that IRG1 is also present in the nucleus of macrophages, where it associates with several promoters, including one controlling expression of PTGS2 (COX2), the rate-limiting enzyme in prostaglandin synthesis. Methods In these studies, we employed a multi-omic strategy incorporating ChIP-seq, RNA-seq, global proteomics, and metabolomics. To establish the biological significance of this novel IRG1 function, we generated CRISPR knockouts in human monocyte-derived macrophages and utilized a DSS-induced colitis model to assess inflammation in vivo. Results Through a multi-omic approach encompassing ChIP-seq, RNA-seq, global differential proteomics, and metabolomics, we have elucidated a nuclear function for IRG1 in primary human macrophages influencing prostaglandin biosynthesis. Additionally, investigations using a CRISPR-generated catalytically inactive IRG1 point mutant in THP1 cells demonstrated significant binding of IRG1 to the PTGS2 promoter, resulting in increased COX2 transcript and protein expression independent of itaconate production. Conclusion Collectively, these results reveal a novel, nuclear role for IRG1 in macrophages that provides a deeper understanding of how IRG1 may regulate macrophage function in the context of inflammation. Funding Source Eli Lilly and Company Topic Categories Cellular Adhesion, Migration, and Inflammation (CAM)

Comparative therapeutic efficacy and immunological mechanisms of LANCL2 therapeutics for treating recurrent Clostridioides difficile infection 2260935

Abstract Introduction Omilancor is an oral, gut-restricted, LANCL2 agonist with proven therapeutic efficacy in acute and recurrent mouse models of Clostridioides difficile infection (CDI). While gut microbiome disruption is the underlying cause of CDI, paradoxically, the infection is treated with antibiotics that cause further gut microbiome imbalance leading to recurrence. Methods We conducted comparative studies of oral omilancor versus current treatments using a recurrent model of CDI. To induce recurrence, mice received a clindamycin injection 12 d post infection. Colonic bacterial burden was evaluated by culture. Colitis was evaluated by histopathology. Gut microbiome composition was evaluated through 16S sequencing. Results During recurrent CDI, omilancor showed non-inferiority to anti-TcdB and rectal fecal microbiome transplantation (FMT), displaying the greatest therapeutic efficacy. Mice treated with vancomycin had the worst outcome during the recurring phase with a 2.5-fold increase in disease severity scores compared to vehicle-treated mice. Oral omilancor outperformed all current treatments in inducing the fastest C. difficile clearance, which was linked to preservation of the gut microbiome diversity. Immunologically, activation of LANCL2 by oral omilancor decreases the expression of proinflammatory cytokines, including IL-17, and infiltration of inflammatory subsets, such as Th17 cells and neutrophils in the gut mucosa while increasing mucosal regulatory T cells (Treg). Conclusion Oral omilancor treatment outperforms current drugs used for treating recurrent CDI. Given omilancor’s placebo-like safety profile and its superior efficacy, LANCL2 therapeutics have the potential to become first-in-class immunoregulatory, antimicrobial-free, gut microbiome-preserving host-targeting treatment modalities for recurrent CDI. Funding Source N/a Topic Categories Mucosal and Regional Immunology (MUC)

JARID1C Regulates Macrophage Function in Diabetic Wounds 2299614

Abstract Introduction Non-healing wounds in patients with Type 2 Diabetes (T2D) are a major cause of morbidity and mortality and are increasing at a concerning rate. The injury response is a complex process that must proceed in an orderly and organized manner to restore homeostatic tissue function. Macrophages are highly plastic in tissue and are recruited in wounds following injury. In vivo, macrophages exist along a continuum where they initially exist in a pro-inflammatory state and then transition to an anti-inflammatory state to promote proliferation and remodeling phases. In T2D, this does not occur, and macrophages remain in an inflammatory state and serve to promote chronic inflammation. Methods Experiments were conducted using C57Bl/6 mice (WT) or Macrophage-selective deletion of Jarid1c transgenic mice (Jarid1c f/f Lyz2 Cre). These mice were placed on a normal diet (ND) and on a high fat diet to generate a Diet Induced Obesity (DIO) diabetic model. Mice were on the requisite diet for 12-16 weeks. Wound macrophages were isolated from the wound via EasySep Cdllb+ Isolation. Macrophage phenotype was assessed by qPCR and chromatin immunoprecipitation (ChIP) following DNase/Liberase digestion of wound tissue. Results Here, we identify JARID1C, a histone demethylase that removes the activating trimethyl mark on histone 3 lysine 4, as a critical regulator of inflammatory gene expression in macrophages. In DIO mice, wound macrophages exhibit reduced JARID1C expression. We demonstrate that JARID1C is required for the resolution of inflammation, enabling IL-6—dependent removal of H3K4me3 at promoters of inflammatory genes. Macrophage-specific deletion of JARID1C significantly delayed wound closure in the DIO wound model, establishing its functional importance in tissue repair. Conclusion Collectively, these findings suggest JARID1C acts as critical epigenetic regulator of macrophage function during wound healing and this axis may be a potential therapeutic target for improving wound healing in patients with T2D. Funding Source CTSA T32 TR004371 Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

A High-Throughput, Systems-based Neutralizing Antibody Assay Identifies Humoral Features Linked with Compartmentalized Neutralization 2304404

Abstract Introduction Antibodies leverage both their fragment antigen binding (Fab) and fragment crystallizable (Fc) domains to confer protection against pathogens. Recent work has shown that the Fc-domain can contribute to breadth of antibody-mediated recognition. Because of this, we sought to identify antibody features correlated to neutralization capacity using a systems serology approach. Moreover, it is unclear if signatures of neutralization are conserved across anatomical sites. We thus employed a newly-developed, systems-predicted neutralizing antibody assay to profile how neutralization signatures against influenza virus were shaped by their milleu. Methods We developed a systems-based predictive neutralizing antibody (SNAb) titer assay to simultaneously quantify influenza neutralization across multiple subtypes and isolates using specimens from H1- and H5-vaccinated non-human primates. We incorporated these results into a systems serology framework to assess the contribution of antibody features to neutralization capacity. Results We find that Fcγ-receptor binding features are strongly associated with neutralization capacity against H1 and H5 influenza clades in serum, while IgG, IgM, and IgA were enriched at the lower respiratory tract mucosa. Features correlating with the neutralization response appeared highly influenced by vaccination priming compositions and boosting route. Neutralization titers were not correlated to the targets of vaccination between serum and lower respiratory tract mucosa. Conclusion Influenza virus neutralization can be quantified in a high-throughput manner and correlated with antibody features in distinct tissues in non-human primates through a modified systems serology approach. Our results are in tight agreement with other neutralizing antibody titer assays such as microneutralization assays and hemagglutinin inhibition assays. Funding Source National Institutes of Health Grant CA260476 National Institutes of Health Grant P01AI165072 National Institutes of Health Grant U19AI135995 National Institutes of Health Contract 75N93021C00029 Topic Categories Technological Innovations in Immunology (TECH)