Browse Articles

Immunology and Infection Biology in Cinema: An Interdisciplinary Undergraduate Course 2240912

Abstract Introduction Communicating the complexities of immunology and infection biology to undergraduate students, many of whom may not be science majors, requires innovative pedagogical strategies that balance rigor with accessibility. To this end, we designed a semester long general education course that integrates foundational scientific concepts with cinematic portrayals of infectious disease, immunity, and medical crises. Methods Meeting once weekly, the course provides students with a dual lens: understanding the science of host defense and viral pathogenesis while critically analyzing how cinema– both fiction and documentary-shapes public perceptions of epidemics, vaccines, and biomedical research. The course is structured around thematic modules (e.g., innate and adaptive immunity, HIV/AIDS, vaccines, zoonotic spillover, pandemics, bioterrorism) paired with films such as Contagion, Outbreak, 28 Days Later, and documentaries including Spillover and The First Wave. Students compare narrative cinema with documentary storytelling to interrogate issues of scientific accuracy, cultural framing, and the social dimensions of disease. A guest lecture from a scientist or physician who has collaborated with filmmakers provides a unique opportunity for students to explore the real-world interface between science and media. Results We present a detail overview on this interdisciplinary course which emphasizes science communication, training students to critically evaluate media representations of immunology and to translate complex scientific ideas into accessible language for diverse audiences. Assignments include film critiques, debates, and creative projects that connect cinematic depictions of disease and immunity with real-world public health challenges. Conclusion By bridging the biomedical sciences with the humanities, this course fosters scientific literacy, media critique, and interdisciplinary thinking-skills that are increasingly essential in an era of global pandemics and widespread health misinformation. Funding Source n/a Topic Categories Immunology Education and Communication (EDU)

Oral Bacillus subtilis modulates immune responses and reduces clinical symptoms in a murine model of atopic dermatitis 2260134

Abstract Introduction Allergic diseases like atopic dermatitis (AD) have risen with modern lifestyle changes affecting the microbiome. AD involves skin barrier dysfunction and immune dysregulation. Bacillus subtilis is Gram-positive bacterium that has been shown in a variety of inflammatory diseases to suppress inflammation. This study shows oral Bacillus subtilis protects against House Dust Mite (HDM)-induced AD by reducing clinical symptoms, epidermal thickness, mast cells, and increasing immunoglobulin A (IgA) and intestinal regulatory T cells (Tregs). Methods BALB/c mice received oral Bacillus subtilis or PBS and topical HDM to induce AD. Disease severity was assessed using clinical scoring, H&E staining for epidermal thickness, Toluidine Blue for mast cell infiltration, and ELISA for total and HDM-specific Immunoglobulins. Multiplex and ELISA were used for serum analysis. Flow Cytometry and RT-PCR were used to analyze Tregs. Results Bacillus subtilis treatment reduced clinical symptoms in a murine model of atopic dermatitis. Mice receiving B. subtilis showed lower lesion scores at weeks 3 and 7. While epidermal thickness and Th2-associated antibodies (IgG1, IgE) were unaffected, total IgA increased with B. subtilis treatment. B. subtilis prevented mast cell accumulation in HDM-induced lesions, and elevated intestinal Tregs expressing GATA3 and IL-10. Conclusion Oral administration of B. subtilis reduced clinical symptoms in HDM-induced atopic dermatitis. Protection occurred without lowering IgE or IgG1 but correlated with increased IgA and gut Tregs expressing IL-10 and GATA3. This study supports the value of beneficial microbes for preventing the development of atopic disease, and sheds light on the importance of the gut-skin axis. Funding Source n/a Topic Categories Mucosal and Regional Immunology (MUC)

Type 1 Interferon Restricts mRNA Vaccine Efficacy Through Suppression of Antigen Uptake in Conventional Dendritic Cells 2309444

Abstract Introduction mRNA-based vaccines are a versatile tool reshaping vaccinology. Therefore, it is critical to understand the underlying mechanisms governing their efficacy. These vaccines transiently induce type I interferons (IFN), and studies conflict as to whether IFN is beneficial or detrimental to the vaccine-specific immune response. We sought to determine how IFN shapes the efficacy of mRNA—lipid nanoparticle (mRNA-LNP) vaccines. Methods We hypothesized that IFN inhibits the ability of dendritic cells (DCs) to acquire and express mRNA-LNPs. To test this, we administered mRNA-LNPs in vitro to bone marrow—derived dendritic cells (BMDCs) that were inhibited from signaling through the IFN alpha/beta receptor (IFNαR) using an antagonistic antibody, MAR1-5A3. In addition, we inhibited IFN signaling in vivo using MAR1-5A3 and by genetically deleting IFNαR in DCs via CD11c-specific Cre—mediated deletion of loxP-flanked Ifnar1. These mice were immunized with mRNA-LNPs and assessed for vaccine mRNA uptake and expression, as well as induction of a vaccine-specific immune response. Results We found that IFN reduces vaccine-specific mRNA expression by DCs in vitro and in vivo. Inhibition of IFNαR signaling or genetic ablation of IFNαR in DCs in vivo improved vaccine-specific CD8+ T cell responses. Additionally, IFN induction prior to vaccination reduced vaccine-specific immune responses. Thus, we find that IFN inhibits vaccine uptake by DCs, limiting their ability to mount vaccine-specific immune responses. Conclusion In conclusion, we find that DCs that had not yet acquired vaccine antigen are inhibited from antigen uptake upon IFN signaling. Inhibiting nonessential IFN signaling augmented antigen uptake and downstream vaccine-specific immune responses. These insights provide a compelling rationale for optimizing formulation strategies to transiently modulate IFNαR signaling pathways to maximize antigen acquisition, mRNA expression, T cell priming, and efficacy. Funding Source n/a Topic Categories Vaccines and Immunotherapy (VAC)

Strategy to activate ephrin-Eph signaling pathways and develop a novel therapeutic approach to regenerative wound repair in veterinary medicine 2259503

Abstract Introduction Intact skin is an essential part of innate immunity. In horses, full-thickness distal limb wounds are common and the limited amount of surrounding soft tissue creates forces that prevent primary closure. Repair is also complicated by granulation tissue that forms continually in the absence of epithelial cover, eventually elevating above intact skin. Its irregular surface blocks repair by disrupting keratinocyte proliferation and migration. In other words, excessive granulation tissue blocks re-epithelialization, the main mechanism for distal limb wound closure. While agents that specifically promote re-epithelialization are clinically unavailable, research indicates ephrin-Eph signaling pathways provide targets for strategies that stimulate keratinocyte proliferation and migration. As example, we newly reported topical treatment with a commercially available rEFNA3 protein reversed progestin-mediated vaginal epithelial thinning in mice. Methods We posited ephrin-Eph signaling pathways that stimulate keratinocyte proliferation are an unexplored but effective strategy for accelerating re-epithelialization of full-thickness distal limb horse wounds. To test this hypothesis, we first used rationale design to newly create a potent EFNA3 mimetic agonist peptide (EFNA3-MAP). We then enrolled 10 horses and compared repair in full-thickness distal limb wounds administered EFNA3-MAP in hydrogel vs. hydrogel alone. Results Compared to controls, EFNA3-MAP treatment significantly accelerated keratinocyte proliferation and migration (as measured by gross examination and histology) and this rapid re-epithelialization prevented formation of excessive granulation tissue. Conclusion Our data suggest ephrin-Eph pathways are an important target for agents designed to stimulate regenerative healing. As re-epithelialization is the main mechanism for cutaneous wound closure in horses and humans, our results have the potential to create strategies that enhance cutaneous wound management in equine and human medicine. Funding Source State of Ohio Accelerator Award Topic Categories Veterinary and Comparative Immunology (VET)

Discovery and Characterization of a Potent and Selective Oral Inhibitor of STAT6 for the Treatment of Allergic Diseases 2257118

Abstract Introduction STAT6 is a transcription factor downstream of IL-4 and IL-13 signaling that acts as a key driver of type 2 inflammation, which is associated with atopic dermatitis and asthma. Inhibition of the IL-4/IL-13 pathway is clinically validated for treatment of allergic diseases. Our aim is to develop an oral small molecule that selectively inhibits activation of STAT6 for the treatment of allergic diseases. Methods Binding affinity and mechanism of action were determined using SPR and a nuclear translocation assay, respectively. Compound potency against human STAT6 was assessed in PBMC by measuring phosphorylated STAT6. Selectivity was examined utilizing STAT SH2 scan, KINOMEscan, and cellular assays for STATs 1-5. Inhibition of Th2 biomarkers, TARC and periostin, and cellular proliferation were assessed in cellular assays. Ex vivo target engagement was measured in mouse whole blood. Results EPS-3903 is an orally bioavailable allosteric inhibitor of STAT6 with strong binding affinity (Kd = 0.42 nM) that exhibited robust selectivity against other STAT family members, as well as a broad kinase panel. EPS-3903 suppressed STAT6 nuclear translocation in the human lung cell line A549 with an IC50 of 14 nM. In PBMC, EPS-3903 inhibited IL4-induced STAT6 phosphorylation and TARC production with IC50s of 5 and 11 nM, respectively. EPS-3903 potently prevented IL-13-induced periostin production in primary bronchial cells and STAT6-mediated proliferation in human TF-1 erythroblast cells with IC50s of 3.3 and 7.5 nM, respectively. In mouse whole blood, EPS-3903 suppressed pSTAT6 by > 90% for 24 h. Conclusion EPS-3903 is a potent and selective allosteric inhibitor of STAT6 and demonstrated effectiveness in preventing STAT6 activation and downstream Th2 biomarker production. EPS-3903 suppressed STAT6 activation in vivo by > 90% for 24 h. EPS-3903 may offer an oral therapeutic potential for treating allergic diseases including asthma and atopic dermatitis. Funding Source n/a Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)

Yeast antigens source modulates vaccine potency and immunity mediated by CD8+ T cells in CD4+ T cell-deficient hosts 2305523

Abstract Introduction Given the global rise in fungal infections, which primarily occur in hosts with CD4+ T cell deficiency or dysfunction, and the lack of FDA-approved vaccines, it is imperative to develop novel vaccine platforms. Our previous studies indisputably showed that an attenuated fungal strain vaccine bestowed sterilizing immunity by CD8+ T cells in CD4+ T cell deficiency. Since live vaccines are contraindicated, we delved to identify the cellular source of fungal antigens that impart immunity. Yeast antigens are primarily sourced from the cytoplasm and cell wall, the latter often being glycosylated. Yet, the cross-presentation ability of vaccine and eukaryotic pathogen to confer immunity is not studied. We hypothesize that protective immunity is dictated by the matching of T cell epitopes during vaccine responses with those cross-presented during fungal infection. Methods We vaccinated cohorts of mice subcutaneously with fungal cell wall/membrane (CW/M) and cytosol extract (YCE) antigens, adjuvanted with alum under CD4+ T cell-depleted condition. We assessed CD8+ T cell vaccine responses in lymph nodes and spleen, lung recall responses after pulmonary challenge, and vaccine immunity. Results CW/M antigens elicited superior polycytokine+ CD8+ T cell vaccine responses than YCE antigens. This enhanced immunogenicity of CW/M antigens did not translate into stronger early recall responses. Nevertheless, intriguingly, we found that CW/M Ag vaccination conferred significantly better survival than YCE Ag vaccination. Conclusion Our findings reveal a mismatch between CD8+ T cell vaccine potency and protective efficacy, and immunogenicity readouts do not accurately predict immunity, a factor contributing to vaccine failures in clinical trials. Thus, it is essential to identify matching epitopes in a vaccine that are potentially cross-presented during eukaryotic pathogen infection to impart sterilizing immunity for safer, potent, and efficacious vaccines targeting CD8+ T cells tailored for immunocompromised hosts. Funding Source NIH-NIAID R01153522 (to SGN) Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)

Antibody Repertoire Modulation Alters Alzheimer’s Pathology and Behavior in the 5XFAD Mouse Model 2267871

Abstract Introduction Alzheimer’s Disease (AD) is one of the most prominent neurodegenerative diseases. The disease is associated with amyloid-β (Aβ) accumulation, protein tau neurofibrillary tangles, neuronal loss, and chronic inflammation in the brain. Aβ plaque accumulation leads to an inflammatory response orchestrated by resident microglia and ultimately neurodegeneration. Recent literature suggests that the immune system, particularly antibodies, plays a key role in disease progression. We sought to characterize the role of specific antibodies in AD by manipulating the repertoire of the 5XFAD model system. Methods We crossed MD4 mice, which have a B cell repertoire restricted to an irrelevant antigen, with 5XFAD mice, which model early onset Alzheimer’s disease, to address whether specific antibodies modulate disease progression. These mice underwent a battery of behavioral tests to measure cognitive impairment, followed by tissue collection for analysis of plaque accumulation, microglia activation, and antibody deposition by immunohistochemistry (IHC). Results Analysis of 5XFAD brains using monoclonal antibodies specific for IgG subclasses revealed an increase in microglia-associated IgG1 within the hippocampus compared to non-transgenic littermates. MD4 5XFAD double transgenic mice had sex specific increases in cognitive impairment, including increased mobility in the open field, nighttime hyperactivity, and decreased spontaneous alternation in a cross-maze compared to 5XFAD littermates. By IHC, we detected a modest trending increase in amyloid-β plaque accumulation at 10 months of age in double transgenic mice compared to 5xFAD brain. The wildtype and MD4 mice brains did not display any amyloid-β plaque accumulation, as expected. Conclusion Our results suggest that specific antibodies restrict the progression of cognitive impairment in 5XFAD mice. Ongoing work is aimed at uncovering the specificities and origin of the spontaneous antibodies that can be found within the brains of 5XFAD mice. Funding Source The National Institute of Health (NIH) Topic Categories Neuroimmunology (NEUR)

Differential Image Sensor With Decoupled Static and Dynamic Outputs

ABSTRACT Acquiring and processing full‐motion details in machine vision typically consumes a substantial amount of energy. In contrast, a hierarchical processing architecture, combining a low‐power standby front end with an on‐demand activated back end, provides an optimized energy‐performance tradeoff. To achieve this, the complete acquisition and decoupling of static (brightness) and dynamic (amplitude and polarity) output at the sensory level are essential for activating on‐demand vision function. Here, we report a differential image sensor (DIS) that leverages differential photodiodes with decoupled differential and tunneling modes. These modes can be read out via conventional ROICs, paving the way for the up‐scaled integration (e.g., 640 × 512). With on‐demand activated dynamic and static modes, the DIS implements a hierarchical motion‐processing pipeline—from sparse motion detection to optical flow and depth analysis. This work provides a power‐efficient and scalable strategy for advancing vision‐based AIoT applications.

Integrative analysis of network pharmacology and machine learning suggests potential anti-colorectal cancer mechanisms of Taohong-Siwu decoction

CX3CR1 Signaling Drives Tertiary Lymphoid Structure Formation and Fertility Impairment Following Treg Depletion in the Epididymis 2251411

Abstract Introduction Using an autoimmune model induced by Regulatory T cell (Treg) depletion, we found B and T cells form tertiary lymphoid structures (TLSs) in the epididymis, producing autoantibodies and inflammation that impair male fertility. In this organ, mononuclear phagocytes (MPs) express CX3CR1, while epithelial cells produce CX3CL1. CX3CR1 drives TLS formation in other tissues. We studied whether CX3CR1 deficiency in CX3CR1EGFP/EGFP mice prevents MP-mediated B cell activation and TLS formation in the epididymal mucosa following Treg depletion. Methods CX3CR1EGFP/+ and CX3CR1EGFP/EGFP mice were bred with Foxp3-DTR mice. Treg depletion was induced by diphtheria toxin (DT) injections. Readouts were analyzed at 2 and 8 weeks using confocal microscopy, flow cytometry, ELISA, and fertility assessment. Results Following depletion, CX3CR1+ MPs in CX3CR1EGFP/+/Foxp3-DTR mice exhibited increased projections and phagocytosed antibody-opsonized sperm. We observed fewer B cells and TLSs in DT-treated CX3CR1EGFP/EGFP/Foxp3-DTR mice, indicating that CX3CR1 is critical for B cell recruitment and TLS formation. Anti-sperm antibodies persisted only in CX3CR1EGFP/+ mice at 8 weeks. CX3CR1 deficiency rescued the reduction in sperm count and fertility defects following Treg ablation by impairing MP phagocytosis. Conclusion CX3CR1 is essential for epididymal TLS formation. CX3CR1 represents a promising therapeutic target for mitigating immune-mediated reproductive dysfunction and chronic inflammation in male infertility disorders. Funding Source R01 HD104672 Topic Categories Basic Autoimmunity (BA)

When gut stem cells talk to the immune system: impacts on remote tumor 2240442

Abstract Introduction Pancreatic cancer is predicted to be the second leading cause of cancer deaths by 2030. Interleukin 17 (IL-17) signaling in pancreatic cancer cells can trigger neutrophil extracellular traps that impair immunotherapy efficacy. IL-17 signaling also maintains gut homeostasis. Intestinal epithelial cells interact with the gut microbiome, participate in injury repair, and are replenished by intestinal stem cells (ISCs). Our lab showed that deletion of IL-17 receptor A (IL-17RA) in the gut epithelium induces pro-tumorigenic IL-17 signaling due to the disruption of gut homeostasis, suggesting that gut IL-17/IL-17RA—mediated immune responses affect distant tumors. However, the role of IL-17 in specific epithelial compartments and its impact on gut homeostasis and cancer risk remain unclear. We hypothesized that disruption of IL-17/IL-17RA signaling within ISCs impacts tumor growth. Methods We utilized a lineage marker of ISCs, Lgr5, to drive Cre enzyme tissue specific expression to knock out Il17ra in gut ISCs (cKO mice). We used implantable pancreatic tumor models and performed immunoprofiling. Results We found that the pancreatic tumor growth was altered by the remote signal from gut ISCs in cKO mice. Furthermore, we identified that major transcriptomic changes were associated with innate and adaptive immune pathways, especially B cell activation. Conclusion We conclude that IL-17/IL-17RA signaling in gut ISCs changes intestinal immune responses locally that can affect distant tumor immunity. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Systems Immunology of Long Covid: Insights from the STOP-PASC Clinical Trial 2265936

Abstract Introduction Post Acute Sequelae of COVID-19 (PASC), also called Long COVID, is an infection-associated chronic syndrome. Despite proposed viral persistence mechanisms, no therapeutic benefit was observed in randomized placebo-controlled trials of nirmatrelvir/ritonavir (NMV/r) in adults with Long COVID, including the Selective Trial of Paxlovid for PASC (STOP-PASC). This systems immunology analysis aimed to characterize immune profiles of participants during clinical trial intervention, identify biomarkers associated with patient-reported outcomes, and investigate potential mechanisms underlying Long COVID. Methods We performed comprehensive immunological profiling of 152 STOP-PASC trial participants using plasma proteomics (Olink® Explore HT 5400 panel), autoantigen arrays, viral serology, and microclot assays at baseline, day 15, and week 10. We assessed associations between immune features and patient-reported outcomes. We also conducted meta-analysis of nine independent Long COVID proteomics cohorts (n = 590 total samples) to identify conserved inflammatory signatures. Results NMV/r treatment at day 15 compared with baseline induced transient changes in plasma proteins that normalized by week 10, primarily impacting myeloid cell/monocyte, lysosome, and complement activation pathways. Cardiovascular symptoms were negatively associated with SARS-CoV-2 antibody levels at baseline. No widespread differences in autoantibody profiles, Epstein-Barr virus (EBV) reactivation, or microclotting were observed between STOP-PASC Long COVID participants, pre-pandemic controls, and individuals without Long COVID. Meta-analysis of publicly available Olink® data from Long COVID cohorts identified a conserved 60-protein Long COVID Signature (LCS) score revealing multi-compartment immune activation involving monocyte, neutrophil, and T/NK cell modules. Conclusion These findings advance our understanding of Long COVID immunology and may help direct future proteomic biomarker endpoints for Long COVID clinical trials. Funding Source Pfizer Topic Categories Computational and Systems Immunology (COMP)

Spatial Multi-Platform Integration Reveals Distinct Tumor States and Microenvironmental Niches in Colitis-Associated Colorectal Cancer 2309487

Abstract Introduction Colitis-associated colorectal cancer (C-CRC) develops in chronic inflammatory settings, where tumor—microenvironment interactions critically influence progression. Spatial transcriptomics enables analysis of these interactions while preserving tissue architecture. We aimed to define tumor cell states and associated microenvironmental features in C-CRC. Methods Seven tissue samples from three C-CRC patients were analyzed using consecutive sections processed by 10x Xenium and 10x Visium. Single-cell spatial coordinates from Xenium were integrated with Visium spots for combined spatial transcriptomic analysis. Results Xenium profiling identified 3,506,705 cells comprising epithelial, fibroblast, and immune populations. Tumor subclustering revealed two states: proliferating tumor cells enriched for cell-cycle genes (MKI67, TOP2A) and invasive tumor cells with elevated epithelial mesenchymal transition (EMT)-associated genes (CD44, CXCL1). Spatial neighborhood analysis identified a niche unique to invasive tumor cells characterized by proximity to cancer-associated fibroblasts (CAF) and SPP1+ macrophages. Invasive regions showed enrichment of angiogenesis, hypoxia, and inflammatory pathways. Receptor—ligand analysis identified SPP1—CD44 signaling between SPP1+ macrophages and invasive tumor cells. A gene signature distinguishing invasive and proliferating tumor cells stratified TCGA sporadic colorectal cancer samples into invasive-like and proliferating-like groups, with invasive-like tumors enriched for consensus molecular subtype 4. Conclusion This integrated spatial transcriptomics study identifies two tumor populations in C-CRC with distinct microenvironmental contexts. The invasive tumor phenotype is associated with CAF and SPP1+ macrophage proximity and SPP1—CD44-mediated signaling, highlighting the tumor microenvironment as a potential therapeutic target in C-CRC. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Purinergic receptor P2X7R regulates neuropsychiatric lupus 2259440

Abstract Introduction Neuropsychiatric lupus (NPSLE) is one of the common manifestations of human systemic lupus erythematosus (SLE), a complex autoimmune disease that affects both peripheral organs and brain. The mechanisms leading to psychiatric manifestations in NPSLE is still not clear. The P2X7R, one of the well-studied purinergic receptors, is activated by high levels of ATP and has broad expression in innate and adaptive immune cells. We hypothesize that causes of neuroinflammation in NPSLE could be due to cell communication from the periphery via ATP and/or P2X7R, as we have shown in other settings of organ injury. Methods We used the MRL/lpr lupus-prone mouse model and tested the effects P2X7R blockade using Brilliant-Blue G (BBG) compound, 45mg/kg injected i.p. every other day for 2 weeks. Tail-suspension test was performed to assess depressive-like behavior, 48 hr before the endpoint of the experiment. Circulating ATP and soluble P2X7R were measured in plasma by ELISA. Gene expression of P2X7R was measured by RT-qPCR using Taqman probes. Results We show increased expression of P2X7R in spleen and brain and increased plasma ATP levels in old mice (diseased) as compared to young mice (pre-disease). Blockade of P2X7R showed decreased circulating ATP, and P2X7R and reduced depressive-like behavior in the MRL/lpr mice. Conclusion Our results, in pre-clinical model using lupus-prone mice, show that reduced peripheral inflammation via P2X7R reduces neuroinflammation. Therapeutic targets attenuating systemic inflammation in SLE could also ameliorate NPSLE. We are continuing to investigate the mechanisms of regulation of NPSLE via P2X7R pathway. Funding Source n/a Topic Categories Therapeutic Approaches to Autoimmunity (THER)

Metabolic Regulation of Alloimmunity: Inhibition of PKM2 Attenuates T Cell Activation and Prolongs Allograft Survival 2257247

Abstract Introduction Organ transplantation is a life-saving therapy, but its success is limited by immune rejection and the toxicity of immunosuppressants. Activation of immune cells depends on metabolic reprogramming, particularly a shift from mitochondrial respiration to aerobic glycolysis (Warburg effect). Pyruvate kinase M2 (PKM2), a glycolytic enzyme upregulated during immune activation, links metabolism to pro-inflammatory gene expression. Although implicated in cancer and autoimmunity, the role of PKM2 in transplant rejection remains unclear. We hypothesized that PKM2 supports alloimmune T-cell responses and that its inhibition could mitigate rejection and improve graft outcomes Methods To assess PKM2 expression during rejection, C57BL/6 (B6) mice received heart grafts from syngeneic B6 or fully mismatched BALB/c (B/c) donors. Heart grafts were harvested and analyzed by FACS on postoperative days (POD) 3 and 7. For T cell alloresponse, CD3+ cells (B/c mice) were treated with PKM2 inhibitors (PKM2i) or vehicle and co-cultured with allogeneic BMDCs (B6 mice). To assess graft survival and immune mechanisms in vivo, heart transplant recipients (B/c → B6) were treated with PKM2i or vehicle (2×/week, i.p.) from POD1—28. Recipient’s spleen and heart grafts were harvested on POD14 for ex vivo analyses (FACS+Histology) Results PKM2 expression was strongly upregulated in graft-infiltrating immune cells, peaking in T cells by POD7, while syngeneic grafts remained at baseline. Strikingly, PKM2 inhibition significantly extended graft survival and broadly suppressed T-cell alloimmune responses including reduced T-cell infiltration, lower IFN-γ production (in vitro/in vivo), and decreased expression of activation/proliferation markers (CD25, Ki67). These changes were accompanied by clear histologic evidence of reduced immune-mediated tissue injury. Conclusion These findings position PKM2 as a central regulator of T-cell—mediated rejection and a promising therapeutic target for metabolic immune modulation in transplantation. Funding Source n/a Topic Categories Transplantation Immunology (TRAN)

MCT4 loss reprograms MC38 tumor environment and potentiates mRNA cancer-vaccine therapy 2305984

Abstract Introduction We elucidated the impact of cancer metabolism components to potentiate the anti-tumoral effects of therapeutic mRNA vaccines. MCT4 facilitates the symport of lactate and protons to the extracellular environment contributing in the acidification of extracellular environment and consequently initiation of immune escape mechanisms by cancer cells and it is highly associated with poor cancer patient prognosis Methods CRISP knock out of MCT4 in MC38 cells; in vitro and ex vivo metabolic assays; tumor growth and survival in mice; mRNA-therapy; histology; flow cytometry; isotype tracing. Results In vivo studies using MC38 Mct-4 ko tumor cells showed reduced tumor growth and increased rejection rates in immunocompetent mice, while minimal differences in immunodeficient mice, suggesting a high immune response within the MC38 Mct4-KO’s tumor microenvironment (TME). We showed that MC38 Mct4-KO TME increased CD8 and CD4 T cell infiltration with higher cytotoxic functionality and a pro-inflammatory environment characterized by enriched nutrients availability, lower level of lactate and reduced hypoxic areas. Therapeutic mRNA vaccinations, conducted in mice with well-engrafted MC38 Mct4-KO tumors, generated a longer survival rate and a more potent cytotoxic intratumoral immune response then MC38 tumor bearing animals. Interestingly, Seahorse® analysis of tumor-infiltrating CD8 T cells from neoAg-mRNA vaccinated mice revealed that cytotoxic lymphocytes within the Mct4-KO TME displayed a modest increase in the maximal respiratory capacity and a significantly elevated spare respiratory capacity then lymphocyte isolated from MC38 tumors, indicating an enhancement of oxidative metabolic program and metabolic stress adaptions in Mct4-KO tumors with reduced lactate. Conclusion These findings provide valuable insights into optimizing mRNA vaccination by blocking the lactate transporter MCT4 in cancer cells for more efficient therapies for cancer patients. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Developing a Label-Free High-Throughput Flow Assay by Imaging Cytometry on the Attune CytPix 2300698

Abstract Introduction Flow cytometry is a primary tool for characterizing immune cell dynamics in high-throughput screening (HTS) assays. However, traditional staining and acquisition procedures demand substantial antibodies, resources, and time. Moreover, HTS assays are often constrained to a minimal set of markers, limiting biological readouts to largely binary outputs (e.g., activated vs. resting) and oversimplifying the complexity of drug responses. Methods To simplify assays while improving biological insights, we leveraged the imaging capabilities of the Attune CytPix to investigate whether imaging flow cytometry can replace activation and viability markers and detect complex phenotypes based on cellular morphology. Results Using brightfield imaging-derived parameters and UMAP dimensionality reduction analysis, we successfully resolved distinct clusters separating human resting Peripheral Blood Mononuclear Cells (PBMC) from activated CD25+CD69+ PBMCs. Additionally, cell debris and morphologically distinct dead cells formed separate populations, suggesting the ability to differentiate modalities of cell death through imaging alone. We then applied an AI-driven Python-based analysis to the CytPix derived images, and successfully reconstructed label-free drug-response curves comparable to those derived via conventional manual gating in standard flow cytometry. Conclusion Building on these findings, we are now integrating additional imaging-derived parameters to uncover phenotypes that may not be captured by conventional marker-based approaches, with the goal of enabling richer, label-free HTS readouts at scale. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Hydrophilic‐Stable Nucleoside‐Based Hydrogen‐Bonded Organic Frameworks (N‐HOF) for Therapeutic Bacterial Hybrid Systems

ABSTRACT Hydrogen‐bonded organic frameworks (HOFs) have emerged as promising materials for biomedical applications owing to their metal‐free biocompatibility and recyclability. Notably, most HOFs are synthesized and utilized in organic solvents, limiting their biomedical translation. Although water is a biologically compatible alternative, it competes for hydrogen bonding and disrupts interactions between building blocks, making the construction of stable aqueous HOFs challenging. Inspired by the DNA base pairing structure, the first nucleoside‐based HOF (N‐HOF‐1) was developed using a multi‐hydrogen bonding strategy. This framework is synthesized entirely in water by simply mixing 2‐amino‐2'‐fluoro‐2'‐deoxyadenosine (2FA) and cyanuric acid (CA), enabling grade production while maintaining stability under physiological conditions. Microcrystal electron diffraction (MicroED) and single‐crystal X‐ray diffraction (SCXRD) studies revealed the confinement of M‐shaped water clusters within the channels of N‐HOF‐1, mimicking DNA hydration and preserving the HOF architecture. Notably, the porous and positively charged properties of N‐HOF‐1 enable interaction with bacteria to form the bacteria–nanoparticle biohybrid systems. Leveraging the intrinsic bioactivity of nucleoside building blocks, this system enhances engineered bacterial colonization in the periodontium, periodontal tissue regeneration, and lymphoma therapy. These findings highlighted the potential of nucleosides as versatile building blocks for hydrophilically stable HOFs, offering new possibilities for their biomedical applications.

A self-training approach to improve multispecies school classification models in fisheries acoustics data

Abstract Automatic classification models of pelagic species based on trawl–acoustic survey data link echotrace characteristics observed in echosounder echograms to species caught in pelagic trawl hauls. In multispecies ecosystems, developing such models is particularly challenging because pelagic trawls provide probabilistic, proportion‑based species information rather than unambiguous labels for individual schools, which limits the applicability of strictly supervised training and motivates weakly-supervised (probabilistic) learning approaches. This study aims to improve the performance of a previously developed weakly-supervised multi-output classification model through a conservative self-training technique. Using the model’s probabilistic multispecies outputs, self-training gradually transforms the most confident predictions among the poorly-labelled cases into pseudo-labelled ones, increasing the ratio of labelled data and strengthening both model learning and evaluation for the next iterations. The approach was tested on a multi-output classification model trained on small pelagic species in the Bay of Biscay. Self-training improved overall out-of-sample accuracy from 63.5% to 72% (F1-score from 53.7% to 60.7%), maintaining or improving performance for each individual species. Notably, it substantially enhanced performance for species that are abundant but rarely captured in monospecific trawls, such as sardine, whose accuracy increased from 50% to 86% (F1-score from 27.3% to 52.7%). In contrast, performance for species with enough labelled cases remained stable throughout the process.

Preliminary mapping of genes involved in the parent-into-F1 murine model of lupus nephritis 2251705

Abstract Introduction Using the parent-into-F1 model of lupus-like nephritis, splenocytes from female BXD recombinant inbred donor mice were transferred i.v. into female B6D2F1 hybrid hosts. Methods The F1 splenic phenotype was evaluated at two weeks for 16 outcome variables by flow cytometry. 56 BXD strains were tested and results for each outcome variable mapped using GeneNetwork2.0. Results We detected a strong locus controlling variation in F1 splenocyte numbers on Chr 5 (93—103.5 Mb) with linkage as high as 5.7 (—logP). This locus contains strong candidates linked to lupus including: heparinase, Hnrnpd, and Hnrnpdl. as well as several other positional candidates: Mir703, Antxr2, Ntcctt8, Prdm8, Rasgef1b, Vamp9, Arhgap24, Mapk10, Ptpn13, Brca1a, Helq, and Antxr2. Variation in donor CD8 T cell numbers mapped to Chr 15 at ∼102.3 Mb near the lupus-associated genes Itgb7 and Itga5 (—logP 4.4). Lastly, variation in donor T cell numbers mapped to several suggestive lupus-associated loci on: a) Chr 17 including the H2 locus, early complement genes (C4a,C4b,Cfb) Tnf, Lta, Ltb, and Daxx; b) Chr 9 including Il18 and Pou2af1 (BOB1); c) Chr11 includingTnip1(A20),IL3, IL4, IL5, IL13, Irf1,Th2-lcr and Anxa6; and d) Chr X including Tlr13,Il2rg, Apool and Xist Conclusion These results demonstrate the feasibility of this model to identify novel gene variants relevant to the development of lupus-like nephritis. Follow up studies with larger numbers of BXD strains and including male transfers will be important. Funding Source NIH R21AI146633 Topic Categories Basic Autoimmunity (BA)