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Oncolytic HSV-1 Encoding IL-12 and Anti-PD-1 Reprograms CNS Immunity: Open-Label Study of MVR-C5252 for Safety, Tolerability, and Efficacy (PuMP) 2259418
Abstract Introduction To overcome immunosuppression in glioblastoma (GBM), we evaluated MVR-C5252, an HSV-1 expressing interleukin-12 (IL-12) and an anti—PD-1 antibody fragment, coupling oncolysis with Th1 polarization and checkpoint blockade. Methods Stage 1 of PuMP (NCT06126744) established safety of a single convection-enhanced intratumoral infusion of MVR-C5252 in adults with recurrent IDH-wildtype GBM (5 × 106 or 1 × 108 PFU; n = 6). A reservoir in the contralateral ventricle collected CSF at baseline, 1 h, and 28 d post-infusion. Pre- and post-treatment tumors were analyzed by Xenium spatial transcriptomics and CODEX proteomics, whole-genome sequencing (WGS) of tumor and CSF quantified ctDNA and viral genome. LC—MS metabolomics targeted IFN-γ—linked tryptophan/kynurenine and arginine/NO pathways. Stage 2, now ongoing, uses an implanted SynchroMed pump for immune priming and boosting via repeat dosing. Results Adverse events were grade 1—2 only with no viral shedding in urine or saliva by PCR. At 28 d, CSF immune cells exhibited sustained IFN-γ—driven transcriptional programs. Dendritic cells upregulated cross-presentation genes (TAP1, TAPBP, CD83). CD8+ T cells expressed increased IFNG, GZMB, and PRF1 with minimal PDCD1 or LAG3 reinduction. NK cells showed transient suppression at 1 h followed by recovery of cytotoxic and IFN-γ—responsive programs (GZMB, PRF1, STAT1), indicating reactivation of antigen-presenting and effector circuits. Conclusion Analyses will relate immune and metabolic remodeling to viral kinetics and ctDNA dynamics. Single-cell and spatial profiling (CODEX, Xenium), LC—MS/MS metabolomics, and CSF—tumor—blood analyses to define immune—metabolic interactions. These findings establish a mechanistic model of localized IL-12/anti—PD-1 virotherapy converting an immunologically quiescent GBM microenvironment into a Th1-polarized, cytotoxic state. Stage 3 will refine dosing and schedule, and Stage 4 will expand at the Phase 2 dose to evaluate efficacy. Funding Source Immvira pharmaceutical grant Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)
Guanosine Modulates Purinergic Signaling to Suppress Astrocyte-Mediated Inflammation 2260527
Abstract Introduction Astrocytes are central regulators of the CNS, integrating metabolic and inflammatory cues to orchestrate neuroimmune responses. During neuroinflammation, they adopt a reactive phenotype with morphological remodeling, altered cytokine release, receptor reorganization, and oxidative signaling. While adenosine’s immunoregulatory roles are well studied, its analog guanosine remains enigmatic. This study investigates how guanosine modulates astrocyte reactivity via purinergic receptor engagement and PI3K/AKT signaling. Methods Primary astrocytes were stimulated with LPS (100 ng/mL) and IFN-γ (20 ng/mL). Guanosine (100 μM) was applied concurrently or post-stimulation. Cytokine secretion (IL-1β, TNF-α, IL-10) was quantified via ELISA. Immunocytochemistry and western blotting assessed adenosine receptor (A1R, A2AR) expression, ENT1/ENT2 regulation, and PI3K/AKT activation. PI3K inhibitors probed pathway dependence. Ongoing FSCV experiments monitor real-time extracellular purine flux in reactive astrocytes. Results Preliminary data suggest guanosine reshapes the cytokine landscape of LPS/IFN-γ—stimulated astrocytes, attenuating proinflammatory signaling while enhancing IL-10 secretion. Purinergic receptor expression appears rebalanced following guanosine treatment, with increased AKT activation, consistent with pathway engagement. FSCV measurements reveal shifts in extracellular purine release between naïve and stimulated astrocytes, supporting guanosine’s regulatory role. Integration with morphological assessments underscores its neuroprotective potential. Conclusion These findings suggest guanosine reprograms reactive astrocytes toward an anti-inflammatory phenotype through purinergic PI3K/AKT dynamics. By modulating cytokine output and recalibrating receptor networks, guanosine fine-tunes CNS immune tone. This work uncovers a novel axis linking purine metabolism to neuroinflammatory regulation, positioning guanosine as a compelling mediator of astrocyte-driven neuroinflammation. Funding Source R01 Guanosine Grant Topic Categories Neuroimmunology (NEUR)
Uncoupling autoreactive B cell receptor expression from autoimmune activation identifies early checkpoints in lupus 2259295
Abstract Introduction Systemic lupus erythematosus arises when tolerance mechanisms fail and autoreactive B cells are activated to produce pathogenic antibodies. While mechanisms sustaining chronic autoimmunity — such as germinal center (GC) persistence and epitope spreading — are well characterized, the checkpoints that link autoreactive B cell receptor (BCR) expression to the initiation of disease remain poorly defined. Methods Lupus progression can be modelled using a murine mixed bone marrow chimera system in which a single ‘rogue’ B cell clone with broad ribonuclear complex reactivity (clone 564) breaches tolerance and recruits wild-type (WT) B cells into activation. To investigate early tolerance checkpoints, we combined BCR knock-in models, adoptive transfer, and bone marrow chimera approaches to examine how antigenic context and T cell interactions influence B cell activation. Results Within mixed chimeras, WT B cells were recruited into spontaneous responses, diversified, and contributed to an expanding autoreactive repertoire. From these responses, we identified two WT-derived BCRs with lupus-associated specificities: M05, reactive to single-stranded DNA and CCL22, and G55, reactive to the Smith D2 protein. M05 and G55 knock-in mice lacked spontaneous GCs and autoantibodies, with tolerance maintained through receptor editing. However, in the presence of the 564 clone, these autoreactive B cells became activated and secreted autoantibodies, revealing that autoreactivity alone is insufficient to trigger disease. We now dissect the antigen- and T cell-dependent cues that enable tolerance breach. Conclusion This work defines the earliest antigen- and T cell-dependent checkpoints that determine whether autoreactive B cells remain silent or become pathogenic. Understanding how these tolerance barriers are breached offers new opportunities to target the initiation phase of lupus rather than its chronic sequelae. Funding Source This work was supported by grants awarded to M.C.C.: National Institutes of Health, National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01 AR074105) and National Institute of Allergy and Infectious Diseases (R01 AI130307); U.S. Department of Defense (HT9425-24-1- 0598); and the Lupus Research Alliance. Topic Categories Basic Autoimmunity (BA)
Deletion of RIAM in myeloid cells accelerates tumor growth by macrophage reprogramming 2248360
Abstract Introduction Myeloid cells within the tumor microenvironment (TME) critically regulate cancer progression. Integrins orchestrate essential myeloid functions, including adhesion, migration, phagocytosis and antigen presentation. Rap1-interacting molecule (RIAM) is an indispensable component of integrin activation machinery. The role of RIAM in tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) is unknown. Methods To investigate this, we generated mice with myeloid-specific RIAM deletion (RIAM fl/fl LysM-Cre ) and implanted MC17-51 fibrosarcoma or B16-F10 melanoma. Results Compared to WT controls, RIAM fl/fl LysM-Cre mice developed significantly larger tumors indicating that RIAM expression in myeloid cells restrains tumor progression. Flow cytometry and RNA sequencing revealed that RIAM-deficient TAMs were skewed toward an M2-like phenotype, with upregulation of CD204, CD206, and CD163, and downregulation of CD86, CD88, and MHC-II. Transcriptomic pathway analysis showed enrichment of IL-4/IL-13 signaling and suppression of NF-κB—mediated inflammatory programs, consistent with impaired pro-inflammatory function. In contrast, RIAM-deficient splenic MDSCs had attenuated immunosuppressive capacity. Thus, despite impaired MDSC activity, RIAM deletion in the myeloid compartment accelerates tumor growth by reprogramming TAMs toward a tumor-promoting state. Conclusion These findings identify RIAM and integrin signaling as novel therapeutic targets for reversing TAM-mediated tumor progression. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Leveraging allelopathy for drug discovery: ocean phytoplankton metabolomes as anti-T. gondii drug candidates: a bioassay strategy 2330921
Abstract Introduction Toxoplasma gondii is an obligate intracellular apicomplexan parasite responsible for fatal foodborne illness (toxoplasmosis) and severe neurological and cognitive defects in vulnerable populations. Current treatments, namely sulfadiazine and pyrimethamine, are ineffective and cause adverse effects. Allelopathy, the process by which phytoplankton species exude compounds to affect competitive behavior, provides a potential strategy as novel inhibitors of T. gondii. The endometabolomic and exometabolomic extracts from several phytoplankton species were screened for anti-parasitic activity. Methods Previous data suggests allelochemicals of candidate species may inhibit another apicomplexan parasite, Plasmodium falciparum. Extracts were collected using solid phase extraction, separating endometabolome and exometabolomes of select species. To determine whether these species will inhibit T. gondii invasion, MRC5 human fibroblast host cells infected with T. gondii were treated and incubated with extract treatments in a dose response curve. The T. gondii plaque assay and the MTT cell proliferation assay were used to quantify the effect of treatment on parasite invasion into MRC5 monolayers and compared to the positive control, sulfadiazine. Results Whole exometabolomes sourced from Prymnesium parvum inhibited plaque formation at a 250 µg/mL concentration, with fewer and smaller plaques compared to untreated monolayers. Preliminary analysis suggests P. parvum and Rhodomonas lens inhibit T. gondii invasion in a modest dose response curve. Conclusion Using an allelopathic inhibition approach, select phytoplankton extracts contain compounds with promising inhibition of T. gondii invasion of MRC5 human fibroblasts. Future studies will evaluate biochemically isolated phytoplankton extracts for inhibition of T. gondii and other apicomplexan parasites. Funding Source University of North Carolina at Wilmington - Center for the Support of Undergraduate Research and Fellowships Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)
Comparing Systemic, Localized, and Focused Ultrasound-Mediated Delivery of Evolocumab (α-PCSK9) in a Murine Model of Glioblastoma 2303312
Abstract Introduction Key challenges in treating glioblastoma are the immunosuppressive tumor microenvironment and the blood-brain barrier (BBB). The PESKe clinical trial (NCT04937413) found that evolocumab (a-PCSK9) treatment increased MHC-I expression and CD8+ T cell infiltration in contrast-enhancing tumors. However, this was not observed in non-enhancing tumors with low concentrations of drug, suggesting that delivery of evolocumab is restricted by the BBB. This study aims to determine effective mechanisms to increase evolocumab delivery to brain tumors, investigating both localized intracranial delivery and targeted focused ultrasound (FUS)-mediated delivery. Methods This study used a MTS assay to optimize in vitro parameters, flow cytometry to evaluate immune effects in vitro and in vivo, and LC-MS to quantify concentrations of intratumoral evolocumab. Results In vitro, it was determined that evolocumab significantly increased MHC-I presentation in CT-2A glioma cells. In vivo, C57Bl/6 mice bearing syngeneic CT-2A tumors were treated with evolocumab either intracranially or intravenously. Flow cytometry at multiple timepoints found no significant difference in MHC-I presentation or T cell infiltration. In the same model, FUS-mediated targeted delivery of evolocumab was compared to systemic delivery, finding that FUS significantly increased both gadolinium contrast-enhancement and intratumoral evolocumab concentrations quantified by LC-MS. There was a significant correlation between contrast-enhancement and concentrations, similar to patient data in the PESKe trial. Conclusion In conclusion, intracranial delivery of evolocumab did not significantly impact immune effects, while FUS-mediated delivery of evolocumab successfully increased intratumoral concentration. Future directions include evaluating the immune effects of FUS-mediated delivery, as well as combining FUS-mediated delivery of evolocumab with anti—PD-1 therapy to improve survival outcomes. Funding Source Duke’s Preston Robert Tisch Brain Tumor Center Topic Categories Neuroimmunology (NEUR)
Development of a mucosal vaccine for pan-Sarbecoviruses: a papilloma pseudovirus expressing SARS-CoV-2 M and N proteins 2259718
Abstract Introduction Over the past two decades, three major coronavirus (CoV) outbreaks–SARS, MERS, and COVID-19–have occurred, all caused by Beta-coronaviruses. Future outbreaks of yet-undiscovered CoVs with pandemic potential are anticipated. This underscores the urgent need for a broadly protective CoV vaccine. We hypothesize that targeting conserved regions of the CoV genome, such as the membrane (M) and nucleocapsid (N) proteins shared across Sarbecoviruses, can provide cross-protective immunity. Methods To induce mucosal immune responses at the site of CoV infection (upper respiratory tract), we used papillomavirus-like particles (VLPs) as a delivery vector. Vaccine constructs were designed using rearranged SARS-CoV-2 Omicron-XEC M and N sequences fused to ubiquitin to enhance antigen processing and T cell activation. These constructs were cloned into a pVax1 vector and expressed in HEK293T cells. Expression was confirmed by RT-PCR and Western blotting. The constructs were then packaged into pseudoviruses using bovine papillomavirus VLPs. Mice were immunized with these pseudoviruses. Results Intracellular cytokine staining revealed antigen-specific T cell responses in lymph nodes and spleens of immunized mice following peptide stimulation. These responses were absent in control animals. Furthermore, vaccinated transgenic K18-hACE2 mice showed protection against heterologous challenge with SARS-CoV-2 Omicron-KP.3, indicating cross-variant efficacy. Conclusion Our findings demonstrate that mucosal delivery of conserved CoV antigens via papillomavirus VLPs induces strong, antigen-specific T cell responses and protects against heterologous SARS-CoV-2 challenge. This strategy shows promise for developing a pan-coronavirus vaccine. Funding Source NIH Topic Categories Vaccines and Immunotherapy (VAC)
Photothermal‐Activable Artificial Macrophage With Amplified Systemic Antibacterial Responses to Combat Primary and Secondary Infection
ABSTRACT The rapid evolution of antibiotic‐resistant bacteria, particularly methicillin‐resistant Staphylococcus aureus ( MRSA ), poses a critical threat to human healthcare, underscoring the need for antibiotic‐free antibacterial strategies that achieve both effective pathogen eradication and sustained protection against reinfection. Here, inspired by the functions of natural macrophages, we report the de novo design of an Ir/TiN‐based photothermal‐activable artificial macrophage (ITN Art‐M) with enhanced systemic antibacterial activity to combat primary and secondary infections caused by antibiotic‐resistant bacteria. The experimental studies and theoretical calculations reveal that the ITN Art‐M displays spiky topology, robust photothermal properties, and electron‐rich Ir‐Ti catalytic sites for reactive oxygen species (ROS) production, which can efficiently capture and kill antibiotic‐resistant bacteria. Meanwhile, the synthesized ITN Art‐M can induce severe bacterial antigen leakage and exposure through synergistic oxidative and hyperthermic stress, thereby potently stimulating systemic antibacterial responses, assisting in the elimination of primary infections and establishing durable defensive surveillance against secondary infections. Notably, ITN Art‐M demonstrates superior MRSA eradication and recurrence prevention, offering a potent, intelligent alternative that may surpass conventional antibiotics in combating bacteria and many other pathogenic cells.
Alox15 diminishes inflammation by inducing STING and Type I Interferon production in response to schistosome eggs in dendritic cells 2259492
Abstract Introduction Schistosomes are trematode helminths that cause widespread. Schistosoma mansoni (S. mansoni) infection causes a spectrum of hepato-intestinal disease driven largely by host CD4+ T cell responses to egg antigens: CBA mice develop severe, Th17-biased hepatic granulomas, while C57BL/6 (BL/6) mice show smaller, Th2-dominant lesions. The host factors that determine this divergent pathology remain unclear. Arachidonate 15-lipoxygenase (ALOX15) produces specialized pro-resolving mediators (SPMs) from polyunsaturated fatty acids that limit inflammation. Separately, STING signaling promotes type I interferon responses and can shape downstream immune programs. We hypothesize that STING-mediated induction of ALOX15 generates SPMs that restrain egg-induced inflammation and thereby modulate disease severity. Methods Alox15 mRNA/protein in bone marrow—derived dendritic cells (BMDCs) from high-pathology CBA and low-pathology BL/6 mice, including BL/6 STING Knockouts were assessed by RT-qPCR and immunoblot analysis. Functional effects were assessed by inhibiting ALOX15 with ML351 and measuring cytokine output after egg stimulation. Results BMDCs from CBA mice showed substantially reduced Alox15 expression at both transcript and protein levels compared with BL/6 controls. Importantly, STING deficiency in BL/6 BMDCs led to a marked drop in Alox15 expression after egg stimulation, supporting a model in which STING activity promotes Alox15 expression. Functionally, pharmacologic blockade of ALOX15 with ML351 potentiated IL-1β production by egg-stimulated DCs, shifting the cytokine milieu toward a pro-inflammatory profile known to favor Th17 differentiation and enhanced immunopathology. Conclusion ALOX15 is a STING-regulated checkpoint in DCs that limits pro-inflammatory cytokine output and may protect against exaggerated egg-driven hepatic pathology. Enhancing ALOX15—SPM signaling or restoring STING-dependent ALOX15 induction warrants exploration as a host-directed strategy to promote resolution in schistosomiasis. Funding Source NIH NIAID R01 AI018919 Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)
Differential B Cell Epitope Recognition in Closely Related Arbovirus Infections 2260486
Abstract Introduction Infections caused by related viruses challenge rapid identification, impeding therapeutic decisions. The co-circulation of Zika virus (ZIKV), dengue virus (DENV), and chikungunya virus (CHIKV) in endemic regions complicates viral diagnosis due to shared symptoms and antibody cross-reactivity. The identities of viral epitopes that induce antibodies inform B cell responses during natural infection, which are crucial for understanding viral pathogenesis, vaccine design, and viral diagnosis. Methods A cohort of Colombian patient serum samples collected from ZIKV-endemic and non-endemic areas was used to identify ZIKV epitopes recognized by serum antibodies during early (< 12 weeks post-infection) and late convalescence (>12 weeks). Screening was performed with overlapping peptides spanning the entire ZIKV polyprotein, followed by validation using synthetic linear peptides. Results We identified immunoreactive epitopes that were recognized by serum antibodies from individuals infected (or co-infected) with ZIKV, DENV, or CHIKV at early and later times after infection. A unique subset of peptides (n = 14) was identified to differentiate early from late ZIKV infection and to develop virus-specific diagnostic methods. Conclusion Diverse linear ZIKV epitopes from structural and non-structural proteins that elicited antibodies with unique specificities during natural infection were identified. The differential antibody recognition patterns between early and late convalescence reveal the antibody diversification from the early extrafollicular response to the later germinal center response, providing insight to distinguish early from late convalescence. Comparative analysis of paired sera suggests the dynamic antibody development over time. The identified ZIKV-specific linear epitopes are valuable candidates for ZIKV-specific diagnostics. Funding Source n/a Topic Categories Viral Immunology (VIR)
Pharmacological Disruption of Glioblastoma Extracellular Vesicle-Myeloid Cell Interactions Mitigates Tumor-Induced Immunosuppression 2259453
Abstract Introduction Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, characterized by profound immunosuppression that hinders the effect of immunotherapies. Myeloid-derived suppressor cells (MDSCs), which dampen anti-tumor immune responses, are major contributors to this immunosuppression. GBM-derived extracellular vesicles (GBM-EVs) play a critical role in this process by promoting immunosuppressive polarization of myeloid cells. Therefore, targeting EV-myeloid cell interactions represents a novel target to mitigate GBM-mediated immune suppression. However, the precise mechanisms by which GBM-EVs induce an immunosuppressive effect on myeloid cells remain poorly understood. Methods Here, we utilize both in vitro and in vivo assays to determine the immunosuppressive functions of GBM-EVs and the effects of pharmacological inhibitors on disrupting the interaction between GBM-EVs and myeloid cells. Results Our data show that GBM-EV treatment significantly increases the monocytic MDSCs (mMDSCs) population in vitro (mean difference = 5.87 ± 1.77, 95% CI [2.17-9.56], p = 0.0017). In vivo, retro-orbital administration of EVs isolated from syngeneic murine glioma cell lines (SB28 and GL261) into C57BL/6 mice resulted in splenic contraction (p = 0.006, p = 0.026, respectively), and a profound increase in splenic mMDSCs (p = 0.0031) with SB28. To disrupt EV-myeloid interactions, we utilized two pharmacological agents targeting different mechanisms of EV uptake: methyl-β-cyclodextrin (MβCD) and cytochalasin D (CytoD). Our results showed that both treatments significantly reduced mMDSC frequency (CytoD: p = 0.023, MBCD: p = 0.0056) as well as the expression of immunosuppressive cytokines, including IL-10 (p = 0.045, 0.002) and TGF-β (p = 0.0068, 0.05). Conclusion These findings demonstrate that disrupting GBM-EV and myeloid cell interaction can rescue tumor-mediated immune suppression, providing a foundation for developing novel therapeutic strategies targeting EV-mediated immune suppression in GBM. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Distinct murine chronic graft-versus-host disease models enable the delineation of the diverging and converging contributors to fibrotic scleroderma 2250854
Abstract Introduction Chronic graft-versus-host disease (cGVHD) is a debilitating complication of allogenic hematopoietic stem cell transplantation (alloHSTC) characterized by immune dysregulation, autoimmunity, and fibrosis. Particularly devastating is scleroderma (Scl), a systemic sclerotic disease that targets the skin causing stiffness and delayed wound healing. Despite FDA approved therapies, complete responses in Scl-cGVHD are rare, highlighting an unmet need for more targeted therapies. Methods To delineate the inflammatory and profibrotic mechanisms underlying Scl disease, we used two distinct murine alloHSCT model in which hosts were lethally irradiated and given bone marrow (BM) ± T cells to induce cGVHD. We compared multiple minor antigen mismatch, B10.D2 into Balb/c to a semi-allogeneic, parent-into-F1 C57BL/6 (B6) into (B6D2) F1 models. Results Mice developed scleroderma signs at different rates facilitating the study of overlapping and diverging fibrosis mechanisms. Mice in the B10.D2into Balb/c model had visible skin lesions by d19 with immune cell infiltration seen as early as d3. In contrast, mice in the parent-into-F1 model had comparatively delayed Scl onset with visible lesions appearing ∼d35. In the B10.D2 into Balb/c model, a 20-fold increase in the frequency of IFNg producing CD4 conventional T cells (Tcon) as compared to BM controls was apparent by d21 (p = 0.0119) with very little IL-17 production (< 0.04% IL-17+ of Foxp3-CD4+ Tcon). Conversely, mice in the parent-into-F1 Scl-GVHD model had a strong signature of IL-17 production, with 60% of CD4 Tcon producing IL-17 at d21 (p = 0.0395). To begin to uncover Scl mechanisms, Scl and BM control skin biopsies (d3, 6, 9, 13 and 21; n = 3 mice/group/time) in the B10.D2into BALB/c model are being analyzed by Nanostring GeoMx along with multiplex immunofluorescence of skin from both models and will be presented. Conclusion Together, our data highlight the need for comprehensive studies of cGVHD/Scl models to develop improved translational therapeutics. Funding Source National Institutes of Health (grants P01 HL158505, P01 AI056299, and R37AI34495) Topic Categories Transplantation Immunology (TRAN)
Galectin 3 Expression in Gastrointestinal Tumors Identified Through Comprehensive Bioinformatic Mapping 2334800
Abstract Introduction Abstract Background Gastrointestinal (GI) cancers are a major global health burden with high incidence and mortality. Despite advances in detection and treatment, prognosis for advanced disease remains poor, underscoring the need for new biomarkers and therapies. Galectin-3 (GAL-3), encoded by LGALS3, is a multifunctional β-galactoside-binding protein involved in adhesion, migration, apoptosis, angiogenesis, and immune regulation, making it an important modulator of tumor biology. Methods This study used in silico bioinformatic analyses to examine LGALS3 expression and its relationship with immune infiltration across seven GI cancers: cholangiocarcinoma (CHOL), colon adenocarcinoma (COAD), esophageal adenocarcinoma (ESCA), liver hepatocellular carcinoma (LIHC), pancreatic adenocarcinoma (PAAD), rectal adenocarcinoma (READ), and stomach adenocarcinoma (STAD) Results LGALS3 was significantly upregulated in CHOL, ESCA, and LIHC, with higher but non-significant expression in PAAD and STAD, while downregulated in COAD and READ. Elevated GAL-3 expression correlated with poorer survival, particularly in LIHC and PAAD. Immune infiltration analysis indicated a role in fostering an immunosuppressive tumor microenvironment. Protein—protein interaction analysis identified MAPK3 and PTEN as key partners, while Gene Ontology enrichment highlighted functions in T-cell activation and motility regulation. Conclusion These findings suggest LGALS3 as a promising diagnostic and prognostic biomarker, and a potential therapeutic target to enhance immune responses in GI cancers. Funding Source No Topic Categories Computational and Systems Immunology (COMP)
Neonatal exposure to antibiotics induces obesity in adulthood by permanent microbiota dysbiosis and deficiency of adipose Tregs 2301819
Abstract Introduction The neonatal dysbiosis of gut microbiota represents a critical environmental factor promoting obesity, but the underlying immunological mechanisms remain unknown. Methods Neonatal mice (from day1) were treated with broad-spectrum antibiotics for the first 3 weeks of life (NeoATB) only . The mice were then hosted under a normal environment for rest of their life. Results We show here that neonatal treatment of mice with broad-spectrum antibiotics for the first 3 weeks of life (NeoATB) resulted in the development of obesity and metabolic abnormalities in adulthood. The NeoATB mice exhibited a permanent dysbiosis of gut microbiota, decreased CD4+Foxp3+ regulatory T cells (Tregs), and increased proinflammatory Th1 cells in visceral adipose tissue (VAT). Mechanistically, neonatal antibiotic treatment resulted in increased intestinal permeability that allowed bacterial translocation into the VAT and liver and an increase in systemic LPS levels. Consequently, VAT dendritic cells were activated via the TLR4 pathway to increase IL-12-triggered Th1-inflammation. Moreover, the decrease in VAT Tregs in NeoATB mice was attributed to reduced infiltration of Tregs from the periphery, decreased expansion of adipose IL-33-mediated ST2+ Tregs, and reduced conversion of local Tregs from VAT CD4+CD25- Foxp3- T cells. Conclusion Thus, we have revealed a previously unrecognized immunological link between neonatal microbiota dysbiosis and the development of obesity. Funding Source NIH, NIDCR IRP Topic Categories Mucosal and Regional Immunology (MUC)
IDO-dependent metabolic reprogramming promotes K. pneumoniae airway infection 2255572
Abstract Introduction Klebsiella pneumoniae (Kp) is a leading cause of pneumonia-associated morbidity and mortality in healthcare settings. One strategy by which Kp evades immune clearance is through its modulation of host metabolic pathways. We hypothesize that Kp exploits the host tryptophan catabolic pathway, initiated by the enzymes indoleamine 2,3-dioxygenase 1 and 2 (Ido1/2), to produce immunosuppressive molecules known collectively as kynurenines, thereby impairing effective immune clearance in the airway Methods Using a murine pneumonia model in wild-type BL/6 and Ido1/2⁻/⁻ mice, we compared bacterial burden and altered immune landscape. We also investigated the metabolic response within lung tissues by spatial metabolite imaging (DESI-mass spectrometry). Results We showed that the Ido1/2⁻/⁻ animals exhibited improved survival, lower airway bacterial burden, and enhanced recruitment of neutrophils and T cells. Neutrophil depletion worsened the infection, particularly in the Ido1/2⁻/⁻ mice, suggesting a key role for Ido1/2 in suppressing an effective neutrophil recruitment and response at the site of infection. Notably, lungs from the infected Ido1/2⁻/⁻ mice showed elevated levels of AMP, a metabolite indicative of AMP-activated protein kinase (AMPK) activity. These findings suggest that Ido1/2 may limit neutrophilic immunity at least in part through the suppression of AMPK signaling. Conclusion Targeting tryptophan metabolism or AMPK may thus represent a promising host-directed approach to enhance immunity against recalcitrant multidrug-resistant bacterial infections. Funding Source NIH K99/R00 HL157550 (NHLBI); NIH R35 GM160393 (NIGMS) Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
Stabilized Bi(III) Sites Direct *NH <sub>2</sub> OH Pathway for Efficient Cyclohexanone Oxime Electrosynthesis
ABSTRACT Coupling electrocatalytic nitrite reduction reaction (NO 2 − RR) with cyclohexanone conversion enables a sustainable route to cyclohexanone oxime (CHO) electrosynthesis, a key feedstock for the nylon‐6 industry. However, this approach is fundamentally constrained by the difficulty of sustaining selective *NH 2 OH intermediate formation. Here, we reveal for the first time that the Bi(III) sites can enable the catalytic system to efficiently follow the *NH 2 OH pathway, avoiding the undesired *N pathway. Accordingly, a BiPO 4 /SiO x interface was designed in which amorphous SiO x functions as an electron‐buffer to stabilize Bi(III) active sites. As a result, the BiPO 4 /SiO x catalyst exhibits a high faradaic efficiency (FE CHO ) of 77.0 ± 3.4% and a CHO yield rate of 0.64 ± 0.01 mmol h −1 cm −2 , surpassing all previously reported catalysts in H‐cellMoreover, the BiPO 4 /SiO x catalyst delivers a nearly 100% carbon and nitrogen selectivity to CHO and retains 91.8% of its initial efficiency after extended cycling, substantially outperforming pristine BiPO 4 . Combined experimental and theoretical analyses reveal that the stabilized Bi(III) site suppresses the formation of surface K + H 2 O, effectively suppressing competing hydrogen evolution and over hydrogenation, thereby enabling efficient CHO electrosynthesis.
Additive Manufacturing of Ordered Polymer Nanostructures
ABSTRACT Additive manufacturing (3D printing) allows the fabrication of complex 3D geometries, yet the integration of long‐range ordered nanostructures within printed materials remains a fundamental challenge. In vat photopolymerization, rapid crosslinking kinetics typically arrest block copolymers in kinetically trapped, disordered morphologies. Here, we introduce Polymerization‐Induced Arrangement of Nanostructures with Order‐tunability (PIANO), a strategy that overcomes this kinetic mismatch by decoupling nanoscale ordering from network formation. PIANO utilizes a mobility mediator, ethylene glycol, to enhance polymer chain mobility, enabling rapid in situ ordering, while maintaining a hydrogen‐bonding network capable of sustaining 3D printing stresses. This approach yields tunable lamellar and hexagonally packed cylindrical morphologies with domain spacings of 20–60 nm. Furthermore, ethylene glycol acts as a latent crosslinker during post‐printing annealing, locking the ordered nanostructure while enhancing macroscopic mechanical strength. By reconciling the divergent timescales of molecular self‐assembly and additive manufacturing, this strategy provides a robust platform for the hierarchical design of functional systems.
4F2hc mediated amino acid transport drives metabolic reprogramming and orchestrate NK cell effector functions 2259423
Abstract Introduction Natural killer (NK) cells are innate cytotoxic lymphocytes that provide defense against infections. However, the mechanisms by which NK cells adapt metabolically during infection remain poorly understood. Here, we investigated metabolic regulation in NK cells during Mycobacterium tuberculosis (Mtb) infection. Methods Peripheral blood mononuclear cells were stimulated with live Mtb H37Rv or γ-irradiated Mtb, and expression of nutrient transporters was analyzed. Functional and metabolic properties of 4F2hc+ versus 4F2hc⁻ NK cells were evaluated by measuring cytokine expression, cytotoxic markers, and glycolytic gene signatures. Pharmacological blockade of 4F2hc-mediated amino acid transport was performed during co-culture of NK cells with Mtb-infected macrophages. Transcriptomic analysis was conducted to identify gene signatures. Results Stimulation with live or γ-irradiated Mtb upregulated the amino acid transporter 4F2hc, but not CD71, Glut1, or CD36. 4F2hc+ NK cells exhibited higher expression of IFN-γ, TNF-α, IL-18R, granzyme B, and glycolytic genes compared with 4F2hc⁻ NK cells. Inhibition of 4F2hc-mediated amino acid transport reduced glycolytic flux, impaired IFN-γ production, and decreased NK cell-mediated control of Mtb growth. NK cells cultured in the presence of plasma from TB patients displayed diminished 4F2hc expression, spare respiratory capacity, glycolytic ATP production, and basal glycolytic activity. Importantly, treatment with 4-hydroxypyridine increased expression of 4F2hc, LAMP-1, and granzyme A. Transcriptomic profiling revealed upregulation of amino acid metabolic pathways, including phenylalanine, tyrosine, glycine, serine, and threonine metabolism. Conclusion These findings identify 4F2hc as a regulator of NK cell metabolic adaptation and effector function during Mtb infection. Moreover, 4-hydroxypyridine emerges as a potential immunometabolic modulator capable of enhancing NK cell activity through the upregulation of amino acid transport and metabolic reprogramming. Funding Source N/A Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
Treatment with male gut microbiota or metabolite downregulates gene expression associated with inflammation in kidneys of lupus-prone mice 2260396
Abstract Introduction Lupus nephritis (LN) is a serious complication of lupus. We have found that male gut microbiota transplants (M-F) suppress LN and improve survival in lupus-prone female NZBxNZWF1 (BWF1) mice, and have identified a microbial metabolite, phytanic acid (PA) as a major player in protection. Previously, we found that these treatments increase peripheral Treg induction and macrophage efferocytosis, but did not evaluate the effects of these treatments directly on kidneys. Here, we examined gene expression in kidneys of female BWF1 mice that received either M-F or PA vs their respective controls. Methods Kidneys were collected from control female BWF1 mice when proteinuria scores indicated severe LN (4; scale of 0-5; 5=death) and age-matched treated BWF1 female mice (scores ≤1). For treatments, female BWF1 were fed with either M-F shortly after weaning then monthly through the end of the experiment, or PA in PBS 3x per week beginning at 12 weeks of age through the end of the experiment. Kidneys were processed for bulk RNAseq and data analyzed using DESeq2 and Ingenuity Pathway Analysis. Results In kidneys of M-F vs control mice, 585 genes were downregulated and 13 genes upregulated. In kidneys of PA-treated vs control mice, 191 genes were downregulated and 21 genes were upregulated. 76% of the genes that were downregulated in PA-treated mice were also downregulated in M-F treated mice; &gt;73 of those genes were immune/inflammation-associated including many genes associated with immune cell trafficking pathways (e.g., chemokines/chemokine receptors, integrins) but also IL-1B, STING, TNFSF1B as well as genes associated with other immune/inflammation-associated pathways. Conclusion These data suggest that male microbiota and PA have similar effects and may downregulate kidney responses to the immune assaults that propagate pathogenesis in LN. Funding Source DOD Topic Categories Therapeutic Approaches to Autoimmunity (THER)
Development of a sensitive assay for detection of pre-existing antibodies to de-risk biotherapeutic molecules for immunogenicity 2259345
Abstract Introduction Using proprietary Invisi-B and Invisi-T machine learning tools, our IMPACT platform allows us to remove B and T cell epitopes from novel drugs to make them invisible to the immune system while retaining optimal drug-like properties. Our platform enables parallelization, thereby eliminating the need for numerous cycles of protein engineering and significantly reducing the discovery time to identify top candidates. Assessment of pre-existing antibodies to novel bacterial derived biotherapeutics is a key aspect of our platform that helps to inform our engineering and is an important consideration for understanding pre-existing immunity. Methods Here, we describe the development and optimization of a bridging assay for highly sensitive and specific detection of pre-existing antibodies to a bacterial derived Protease-Fc fusion. Initially, we optimized the biotinylation and Sulfo-Tag labeling of our biotherapeutic molecule to adapt it for routine use in our assay. Results Human serum samples were assessed and the ratio of signal responses compared to negative controls was used to identify positive or negative samples. Titration of serum samples was performed to understand levels and prevalence of pre-existing antibodies to different biotherapeutics. Conclusion This type of optimized analytical assay allows for sensitive and reliable detection of pre-existing antibodies to novel biotherapeutics, which can also be used to assess the removal of B cell epitopes for novel molecules. Funding Source n/a Topic Categories Basic Autoimmunity (BA)