Browse Articles

Unmasking the in vivo role of IGF1R in Chronic lymphocytic leukemia and identifying novel inhibitors for its function 2298900

Abstract Introduction Insulin-like growth factor 1 receptor (IGF1R) signaling is a key regulator of cellular anabolism and is critically implicated in the pathogenesis of various cancers, including Chronic Lymphocytic Leukemia (CLL). Previous in vitro studies have shown that pharmacological inhibition of IGF1R effectively induces CLL cell death. Our study aimed to understand the precise role of IGF1R in CLL development in vivo and to identify novel endogenous inhibitors of its function. Methods We generated a B cell-specific IGF1R knockout mouse model in the Eï•­-TCL1 transgenic background. In parallel, to identify novel endogenous inhibitors of IGF1R, we performed a large-scale screening of a human hemofiltrate peptide library, a valuable resource of bioactive molecules. Results Result: The deletion of IGF1R from B cells in the TCL1 mice led to a significantly reduced disease load compared to their undeleted TCL1 littermates reflecting that IGF1R actively enforces CLL development in vivo. Concurrently, screening of hemofiltrate library led to the identification and purification of a 14 kDa fragment derived from Insulin-like growth factor binding protein 6 (IGFBP6) that strongly inhibit IGF1-IGF1R interaction. Conclusion Our data collectively demonstrate the critical involvement of IGF1R in CLL progression and validate it as a therapeutic target. Furthermore, the identified IGFBP6 fragment represents a novel, endogenous agent for blocking IGF1R function and developing new CLL treatments. Funding Source CRC1279, German Research Foundation (DFG) Topic Categories Immune Mechanisms of Human Disease (HUM)

Electrolyte Engineering for 5 V Lithium Metal Batteries

ABSTRACT Lithium‐metal batteries employing lithium‐rich manganese‐based oxide (LRMO) cathodes offer great potential for achieving gravimetric energy densities exceeding 750 Wh kg −1 . However, their practical deployment is hindered by severe electrolyte decomposition and rapid capacity fade, particularly under high‐voltage conditions. Herein, N ‐methyltrifluoroacetamide (NMTFA) as a novel electrolyte additive is introduced, in combination with lithium difluoro(oxalato)borate (LiDFOB), to modulate the electrolyte solvation structure and enhance interfacial stability. Endowed with lone pair electrons and an N‐H bond, NMTFA acts as an electron donor capable of scavenging oxygen radicals released from the LRMO cathode, thereby mitigating electrolyte degradation and suppressing gas evolution. Additionally, hydrogen‐bonding interactions between NMTFA and the primary solvent, along with the incorporation of DFOB − anions into the primary solvation sheath, weaken Li + ‐solvent interactions, promoting the formation of a nitrogen‐/boron‐rich interphase that facilitates faster Li + desolvation and improves electrode stability. As a result, the Li||LRMO full cell delivers exceptional cycling stability, retaining over 70% of its initial capacity after 800 cycles at a cut‐off voltage of 4.8 V and over 80% retention after 300 cycles at 5.0 V. This study provides critical insights into the rational design of advanced electrolytes for high‐voltage lithium‐metal batteries.

Theoretical calculation method for seismic analysis of twin-arch tunnels under near-fault ground motions: methodology, numerical and physical test validation, parametric analysis

Oncostatin M Limits Lung Injury during Influenza by Modulating Leukocyte Recruitment and Inducing Tissue Repair Factors 2258236

Abstract Introduction Oncostatin M (OSM), an IL-6 family cytokine, is upregulated during pneumonia. We and others have previously shown that OSM is required for protection during pneumonia. However, the exact mechanisms of how OSM mediates this protection remain a knowledge gap. Methods We intratracheally instilled influenza A virus A/HKx31 into the lungs of 6-12-week-old OSM knockout (OSM-/-), OSM receptor knockout (OSMrβ-/-) mice and wild type (WT) controls and measured various outcomes of influenza. To investigate lung cell-type-specific responses to OSM, we performed single-cell RNA sequencing of WT and OSMrβ-/- lungs intratracheally instilled with recombinant mouse OSM or a vehicle control. Results During influenza infection, loss of OSM increases morbidity during peak inflammation, leads to greater recruitment of leukocytes including neutrophils, monocytes and lymphocytes, and increases the number of PD-L1 high immunosuppressive alveolar macrophages. Interestingly, we observed no phenotype in OSMrβ-/- compared to WT mice indicating that these effects are OSMrβ independent. Single-cell RNA sequencing and immunofluorescence analyses revealed that OSM activates STAT3 and ERK pathways in lung endothelial cells and fibroblasts and induces the expression of genes that promote tissue repair including proangiogenic factors and Serpins both in the presence and absence of OSMrβ. Since the related IL-6 family receptor, leukemia inhibitory factor receptor (LIFrβ), is highly expressed by these cells, OSM may be signaling through this receptor in OSMrβ-/- cells. Conclusion Our study indicates that during influenza, OSM limits excessive leukocyte recruitment and thus helps limit lung injury. OSM signaling in endothelial cells and fibroblasts induces the expression of tissue repair factors that may help regenerate the lungs post influenza infection. Furthermore, OSM may signal through LIFrβ to rescue the loss of OSM-OSMrβ signaling during influenza. Funding Source NIH and NHLBI Topic Categories Mucosal and Regional Immunology (MUC)

Functional Identification and Characterization of Human Tissue Resident Memory T Cells Using Ex Vivo Lung Perfusion 2309862

Abstract Introduction Identifying tissue resident memory T cells (TRM) in human lungs has traditionally relied on imprecise surrogate markers of tissue retention, markers that can change based on activation state. Using ex vivo lung perfusion (EVLP), we can identify circulating and tissue resident T cells in a functional manner. Better knowledge of differential gene regulation in TRM will allow for improved analysis of their role in various disease states, as well as identify potential therapeutic targets. Methods Human lungs declined for organ donation were ventilated and attached to a normothermic EVLP system for 2-4 hours (2 experiments with 2 human lungs each). A biotinylated CD45 antibody was instilled into the perfusate to label circulating cells for the final 20 minutes; tissue-resident cells were identified as those cells “protected” from the circulating antibody. Immune cells were isolated and then stained with oligo-tagged streptavidin as well as oligo-tagged antibodies against several traditional TRM markers in order to perform simultaneous single cell RNA sequencing and CITE-seq. Results UMAP clustering of scRNA-seq data identified 13 discrete T cell clusters with differential proportions of labeled vs. protected cells. Clusters corresponding to canonical CD4+ and CD8+ TRM populations contained almost no labeled cells. Differential gene analysis of protected vs labeled CD8+ T cells show that functionally defined CD8+ TRM have a gene expression signature similar, but not identical, to canonical marker-defined TRM. Of particular interest is upregulation of galectin-3 (LGALS3), a member of the lectin family previously implicated in inflammatory and fibrotic disease. Conclusion EVLP allows for effective functional identification of tissue resident T cells in the human lung, which overlap with but are not identical to TRM defined by canonical surface markers. Future directions include kinetics of antibody labeling, validation of differentially regulated gene targets, and analysis of TCR sequencing data. Funding Source T32HL007563 Topic Categories Immune Mechanisms of Human Disease (HUM)

IFN-γ shapes neutrophil function and lung host defense against Aspergillus fumigatus 2220821

Abstract Introduction Aspergillus fumigatus (Af), a ubiquitous airborne fungus, causes >90% of invasive aspergillosis (IA). While triazole antifungals are standard therapy, rising resistance underscores the need to define host-protective mechanisms. Neutrophils are essential for antifungal defense, and we identified them as the predominant source of interferon-γ (IFN-γ) during pulmonary Af infection. Loss of IFN-γ signaling increases mortality and reduces neutrophil effector responses, underscoring its role in host defense. Methods scRNA-seq of lung neutrophils revealed enrichment of immature subsets in Af-infected WT mice versus naïve controls, consistent with infection-driven granulopoiesis. To test whether neutrophil-derived IFN-γ integrates lung cues to influence neutrophil development, we are analyzing Af-infected versus naïve Mrp8CreStat1fl/fl and Mrp8CreIfnlr1fl/fl mice, combining bone marrow and lung analyses with flow maturation markers. In parallel, to capture IFN-γ activity beyond neutrophils, we are performing whole-lung scRNA-seq of Af-infected WT and IFNLR-deficient mice, encompassing both immune (CD45+) and structural (CD45⁻) cells. Ongoing confocal microscopy with labeled spores, EpCAM, and Ly6G further examines neutrophil—epithelial interactions. Results Bulk RNA-seq of lung neutrophils showed induction of interferon-stimulated genes (Oasl1, Ifit1, Rtp4, Zbp1, Cmpk2) in WT mice after Af infection, which was reduced in IFNLR-/- and IFNAR-/-IFNLR-/- mice. RT-qPCR confirmed these findings. Preliminary analyses of conditional knockouts suggest context-dependent contributions of IFN-γ to neutrophil maturation and effector responses. Conclusion Neutrophil-derived IFN-γ is a critical regulator of antifungal defense. Current and future studies will define its dual role: (1) controlling neutrophil development through lung—bone marrow axes, and (2) shaping multicellular pulmonary immunity by acting on immune and structural compartments during Af infection. Funding Source N/A Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Tissue tuned cell death: harnessing death for therapeutic efficacy 2260883

Abstract Introduction Immune cell death programs are not passive endpoints but active regulators of tissue immunity. Neutrophil extracellular trap (NET) formation is one such program, releasing DNA and granule proteins that can either restrict pathogens or reinforce tumor progression. More broadly, the capacity to induce or redirect death pathways represents a powerful immunotherapeutic tool. We examined how manipulating neutrophil death and NETosis shapes antitumor immunity, situating these findings within the broader context of programmed cell death as a driver of immune reprogramming. Methods We combined genetically engineered pancreatic ductal adenocarcinoma (PDAC) models with thick tissue slice imaging to visualize NET formation in situ. Engineered bacteria were used to selectively induce pyroptosis or apoptosis, while NETosis was modulated through genetic and pharmacologic approaches. Spatial transcriptomics and multiplex immunofluorescence were employed to map neutrophil clusters, death pathway activation, and tumor microenvironmental (TME) remodeling. Results NETosis emerged as a dominant neutrophil fate within PDAC, enriched in clusters that restricted lymphocyte access. Inhibition of NETosis reduced tumor burden and enhanced CD8+ T cell activity. In parallel, bacterial induction of pyroptosis promoted inflammatory myeloid states, whereas apoptosis favored immune resolution. These data highlight that distinct death programs can be selectively manipulated to rewire immune responses. Conclusion Our findings identify NETosis as a targetable suppressive program in cancer and show that engineered manipulation of cell death can redirect the TME toward antitumor immunity. Together, these results position neutrophils as central regulators of immunogenic death and illustrate how programmed cell death pathways can be leveraged as a generalizable strategy for immune intervention. Funding Source NIH R00 Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Fever temperature alters metabolic and cytokine activities in activated macrophages in mice, chickens and fish 2266416

Abstract Introduction Response to fever affects the biological process in animals and often they are conserved in animals. In this study we investigate the role of activated macrophages in different vertebrate animal models- in homeothermic (chickens and mice) versus poikilothermic vertebrates (rainbow trout). Methods We use various cell lines from young adult mice cell line, RAW 264.7, chicken cell line HTC and rainbow trout splenic cells RTS 11; tissue macrophages from all three species and mouse bone marrow-derived macrophages (BMDM) to test metabolic changes, gene expression changes in response to fever using Greiss, qPCR, and Seahorse assays Results Stimulating cells with LPS, and IFN-g showed higher NO production, treatment with LPS and Interleukins (ILs) caused increase in iNOS mRNA levels. Metabolism increased during fever temperatures in all three species when glycolytic rates and oxidative phosphorylation rates were measured. Stimulation caused a reduction in OCR (rate of oxidative phosphorylation) and increase in PER (glycolysis) mice and chicken. For fish cells however, observations were strikingly different where unstimulated cells at fever temperatures showed higher metabolic activity, and with stimulation OXPHOS dominates and showed an early PER increase.ATP levels also showed an increase at fever temperatures when compared to the body temperatures for all three species. Conclusion Metabolism increased with fever temperatures in all three species. For Rainbow trout were much more engaged in OXPHOS. Funding Source Fulbright Immunology Fund Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

Spatially Resolved High-Plex Protein Profiling of Immune States in FFPE Tissue 2334779

Abstract Introduction Understanding immune cell states and signaling within intact tissue architecture remains a central challenge in immunology, particularly for formalin-fixed paraffin-embedded (FFPE) specimens that dominate clinical pathology and retrospective studies. While spatial transcriptomics has enabled high-plex mRNA profiling in FFPE tissue, comparable capabilities in protein profiling remain far behind. Methods Here, we describe a high-plex spatial protein profiling approach that integrates DNA-barcoded antibodies with a high-resolution spatial transcriptomics platform (Visium™ HD FFPE), enabling multiplexed protein detection directly within FFPE tissue sections. Tissues are stained with a panel of 200 DNA-barcoded antibodies targeting diverse proteins, including lineage markers, cytokines, phospho-epitopes, and transcription factors. Antibody-derived tags are captured, spatially barcoded, and sequenced, generating integrated spatial maps of protein expression across intact tissue architecture. Results Using human pancreatic cancer FFPE samples, we demonstrate spatially resolved detection of epithelial, immune, and innate immune compartments, including localized expression of T cell markers (CD3, CD8), cytokines (IFNG), chemokine receptors (CXCR4), and lineage-defining transcription factors (T-bet). Spatial patterns of immune activation and exclusion are observed alongside pathway-level protein signals, including Wnt/β-catenin activation, illustrating how immune contexture can be interrogated within its native microenvironment. This sequencing-based protein readout avoids optical spectral limitations, enables scalable plex expansion, and remains compatible with archived FFPE cohorts. Conclusion Together, this approach provides a robust framework for spatially mapping immune cell identity, functional states, and signaling networks at protein resolution in FFPE tissue, enabling retrospective immune profiling across disease states, therapeutic contexts, and experimental systems. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

The VISTA—LRIG1 Axis Integrates Myeloid and T Cell Suppression to Drive Immune Resistance in Triple-Negative Breast Cancer 2303786

Abstract Introduction VISTA is a myeloid-enriched inhibitory ligand strongly linked to poor therapeutic outcomes in triple-negative breast cancer (TNBC). However, the mechanism by which VISTA suppresses CD8+ T cell immunity has remained unclear. We recently identified LRIG1 as a T cell—intrinsic binding partner for VISTA. Here, we investigate how the VISTA—LRIG1 axis contributes to immune resistance in TNBC using genetic mouse models, tumor immunophenotyping, and integration of clinical datasets. Methods We used global VISTA knockout (VISTA KO) mice and Granzyme B—Cre LRIG1 conditional knockout (GzmB-Cre LRIG1 KO) mice in the E0771 TNBC model. Tumors were analyzed by flow cytometry, and cytokine profiling. Myeloid signatures from VISTA-deficient cells were compared to breast cancer patient datasets. Public single-cell TNBC data (Zhang et al., 2021) and a neoadjuvant chemotherapy cohort were used to assess clinical correlations with VISTA expression. Results VISTA KO mice exhibited significantly improved control of E0771 tumors, along with reduced suppressive myeloid programs and increased tumor-infiltrating CD8+ T cells. The VISTA-deficient myeloid signature correlated with improved survival across breast cancer datasets. Clinically, VISTA was enriched in myeloid cells from TNBC non-responders to PD-L1 blockade and associated with higher residual cancer burden following chemotherapy. GzmB-Cre LRIG1 KO mice displayed enhanced tumor control driven by increased CD8+ T cell proliferation, reduced quiescence, greater cytokine production, and expansion of TCF1+ progenitor-like subsets. Conclusion The VISTA—LRIG1 axis functions as an integrated suppressive pathway across myeloid and T cell compartments in TNBC. Genetic disruption of either VISTA or LRIG1 improves CD8+ T cell function and tumor control. These findings establish this axis as a key driver of immune resistance and a promising target for therapeutic intervention Funding Source NIH, CDMRP and MRF Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Dynamic feature pyramid network for real-time gesture recognition

Breast milk-derived signals regulate epithelial cells maturation and γδ-IEL colonization to shape early life intestinal homeostasis 2258727

Abstract Introduction The intestinal epithelium is a specialized barrier that balances tolerance and defense against luminal antigens and microbes. Its maturation during early life is shaped by intraepithelial lymphocytes (IELs), the microbiota, and maternal factors. Natural CD8αα+ αβ and Vγ7+ γδ IELs are critical for epithelial homeostasis, repair, and microbial regulation, yet the mechanisms controlling their postnatal maintenance remain poorly understood. Methods Jackson (Jax) and Taconic (Tac) mice were evaluated by flow cytometry after cross-foster, co-house, and adoptive transfer experiments. Bulk RNA sequencing, and milk proteomic analysis were also performed. Results Tac mice showed fewer natural CD8αα+ and Vγ7+ γδ IELs throughout the small intestine (SI), while colonic frequencies remained comparable to Jax mice, indicating a compartmentalized defect. CD8αα+ IEL loss was microbiota-dependent, as co-housing with Jax mice restored their abundance, whereas Vγ7+ IEL loss was microbiota-independent and emerged at weaning despite normal thymic output. Cross-fostering Tac pups onto Jax dams fully rescued Vγ7+ IELs, implicating milk-derived factors. Proteomic profiling of Tac versus Jax milk revealed broad protein reductions, notably in IGF transport and uptake pathways, with decreased IGF-1 and osteopontin. Adoptive transfers showed a tissue-intrinsic defect, as Tac IELs engrafted in Jax RAG⁻/⁻ hosts but not vice versa. SI epithelial transcriptomics revealed downregulation of c-Myc targets after day 28, coinciding with Vγ7+ IEL loss and defective epithelial maturation. Conclusion Together, these findings reveal that postnatal maintenance of natural IELs depends on distinct environmental cues: while CD8αα+ IELs are shaped by the microbiota, Vγ7+ γδ IELs require maternal milk—derived factors that support epithelial maturation and c-Myc—dependent pathways. This identifies a critical early-life window in which deficient milk signals can impair intestinal immune—epithelial development and long-term mucosal homeostasis. Funding Source n/a Topic Categories Mucosal and Regional Immunology (MUC)

Powering health literacy with STEAM: Using an infographic project to teach the public about complex topics in immunology 2265691

Abstract Introduction An infographic is a brief document that emphasizes the use of charts, figures, or diagrams to graphically represent information or data. There are many advantages to using infographics including concise representation of complex concepts or data, visual attractiveness, and low production time. Since infographics prioritize space for imagery, they rarely contain dense text which allows readers to view and process the information quickly. Methods Creating an infographic to illustrate a complex disease or topic in immunology is a useful assignment for an undergraduate course because it requires students to master immunological concepts and disseminate pertinent information to a general audience. Further, this project requires students to create drafts, perform peer reviews, and create an 11 x 17 inch infographic poster, bibliography, and reflective essay. Students follow a detailed rubric to complete the multistep process which is robust enough to be used as an alternative assessment (i.e. in place of a traditional exam grade). Results Alternative assessments offer more inclusive and equitable ways to assess knowledge and skill acquisition. Importantly, the infographic project fosters artistic creativity and encourages students to read and synthesis information from primary literature and share their findings with a diverse audience. Conclusion Undergraduate and graduate programs in science frequently focus on training students to perform scientific writing — writing for a professional audience in a particular area or field. However, mastering the art of science writing — presenting science to a general or lay audience in a non-technical way, is a valuable skill, which should be emphasized in an undergraduate immunology course. Considering a 2019 study by the National Literacy Institute determined that only 12% of the American public is considered proficient regarding health literacy, it is more important than ever to train emerging scientists to master science writing and communication. Funding Source n/a Topic Categories Immunology Education and Communication (EDU)

Expanded IL-22 producing γδ T cells after neonatal respiratory viral infection are required for lung repair 2241785

Abstract Introduction Human metapneumovirus (HMPV) is a respiratory virus that can lead to lower respiratory tract infection (LRTI), causing 500,000 hospitalizations and 11,000 deaths in children under 5 worldwide annually. Early life viral LRTI leads to persistent abnormalities in lung function and increased risk of asthma development. The lungs continue to develop postnatally via alveolarization for the first few years of life in children and the first 4 weeks in mice. The healing mechanisms in the lung following viral LRTI remain largely unknown. Methods We modeled neonatal HMPV infection with mice at day of life 4-6 compared to adult mice with infection at 6-8 weeks. HMPV infected alveolar epithelial cells 1 day post infection (DPI) with total viral clearance by 7 DPI. Results Neonatal mice saw a robust expansion of γδ T cells at 1 DPI that was not present in adult mice. These γδ T cells expressed high levels of IL-22, a cytokine involved in epithelial protection and repair. γδ T cell deficient mice (e.g. Tcrd-/-) infected as neonates did not produce IL-22 at 1 DPI. At 7 DPI, B6 neonates had markers of lung repair as evidenced by clusters of Krt5+ cells in alveolar spaces. Krt5 was significantly decreased in Tcrd-/- mice at the same timepoint. Neonatal Tcrd-/- mice presented with increased pathology in the alveolar spaces 21 DPI compared to B6 mice. Conclusion These data show that γδ T cells upon neonatal infection are critical to initiate epithelial repair pathways that mediate lung damage caused by viral pathogens. Funding Source AAAAI Foundation Faculty Development Award K08AI182486, National Institute of Allergy and Infectious Diseases Topic Categories Mucosal and Regional Immunology (MUC)

Effects of the Antiretroviral Raltegravir on Amyloid-Beta1-42-Induced Neuroinflammation 2310253

Abstract Introduction There is growing evidence supporting the potential role of microbial infections in the aetiology of Alzheimer’s disease (AD). We aim to evaluate the effect of raltegravir, either alone as a single therapy or combined with elacridar, a P-gp/BCRP inhibitor, on AD-related pathways in vitro. Methods Flow cytometry combined was used to quantify the expression of key inflammatory markers, including interleukin-1β (IL-1β), NLRP3 inflammasome components, costimulatory molecules (CD86), and the class B scavenger receptor (SR-B2) CD163, in HMC3 human microglia cells. Furthermore, the effect of raltegravir on Aβ1-42-induced p-tau 181 expression in H4 human neuroglioma cells was assessed. Results Results. Our data indicate that raltegravir significantly (p < 0.01) abolishes the Aβ1-42-stimulating effect on p-tau 181, and that this effect involves the downregulation of PP2Aα+β. The observed reduction of p-tau 181 by raltegravir is potentially mediated by the inhibition of tau-phosphatases. Aβ1-42 significantly upregulated the expression of the surface receptors CD86 and CD163 in HMC3 microglial cells. Raltegravir treatment significantly reduced CD86 expression, while it had no significant effect on Aβ1-42-induced CD163 expression (p > 0.05). Further,raltegravir significantly (p < 0.05) attenuated IL-1β cytokine release, likely via downregulation of NLRP3-inflammasome signalling pathway in microglia. Conclusion Taken together, our findings suggest that raltegravir exerts a neuroprotective effect by targeting pathological events associated with AD, particularly tau phosphorylation and neuroinflammation. Funding Source n/a Topic Categories Neuroimmunology (NEUR)

MAIT17 Cells and IL-17-Driven iBALT Formation Protect Against Pulmonary Tularemia 2267799

Abstract Introduction Here we demonstrate that respiratory infection with Francisella tularensis induces the formation of induced bronchus-associated lymphoid tissue (iBALT) as a protective mechanism against pulmonary tularemia-like disease in mice. While semi-invariant natural killer T (NKT) cells and mucosal-associated invariant T (MAIT) cells are abundant at the lung mucosa, their roles in infectious diseases remain poorly understood. Pulmonary tularemia, the most severe form of tularemia, arises from respiratory infection with F. tularensis. Using C57BL/6 mice infected with the live vaccine strain (LVS) of F. tularensis to model tularemia-like disease, we previously showed that NKT cells exacerbate disease, as NKT-deficient CD1d⁻/⁻ mice were protected. Notably, protected CD1d⁻/⁻ mice developed iBALT structures in the lungs, which were absent in C57BL/6 mice. Methods Molecular immunology methods. Results Respiratory LVS infection in CD1d⁻/⁻ mice induced a robust expansion of IL-17-producing MAIT17 cells and a decrease in regulatory T cells in the lungs compared to C57BL/6 mice. Single-cell RNA sequencing revealed a gene signature in MAIT cells linked to factors that drive iBALT formation. Supporting this notion, adoptive transfer of LVS-activated lung MAIT cells from infected CD1d⁻/⁻ mice conferred protection against tularemia-like disease in RAG2⁻/⁻ recipients, while splenic MAIT cells did not despite the fact that both populations homed to the lungs after transfer. Lung-derived MAIT cells were enriched in IL-17-producing MAIT17 cells, whereas splenic MAIT cells predominantly consisted of interferon-gamma-producing MAIT1 cells. Last, IL-17 was identified as critical for iBALT development during respiratory LVS infection in CD1d⁻/⁻ mice. Conclusion In summary, our findings establish MAIT cells as central mediators of IL-17-dependent protection against pulmonary tularemia-like disease and underscore the potential protective role of iBALT in disease resistance. Funding Source NIH AI137082, VA Merit BX001610, VA SRCS BX004595 Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)

Scaling measurements of peptide-HLA complex stability using user-defined libraries and mass spectrometry 2310029

Abstract Introduction Human leukocyte antigen (HLA) class I presents intracellular peptides to the immune system on the cell surface. Since this process is crucial for the recognition of cancer cells and the initiation of anti-tumor immunity, peptides presented by HLA are valuable immunotherapy targets. More stable peptide HLA (pHLA) complexes provoke superior immune responses. However, how peptide sequence motifs contribute to pHLA stability is not well understood. Methods We developed a high-throughput assay to quantify stability of thousands of user-defined pHLA produced in E. coli. Peptide libraries and the desired HLA are produced and form pHLA complexes in E. coli. pHLA are purified and stability is evaluated by treating pHLA with a thermal gradient and recovering only the peptides which remain HLA-bound after heat treatment. Peptide depletion over the temperature range is monitored by quantitative tandem mass tag (TMT) enabled mass spectrometry. Results Our new E. coli-based method is reliable for assessing pHLA stability. Detected HLA-binding peptides have the expected binding motifs, and stability data strongly correlates with current gold-standard data. We are able to generate large peptide stability datasets (1,800+ peptides) in one scaled experiment — five times larger than currently available datasets. We show that peptide motifs and anchor residue combinations potentially drive pHLA stability. Additionally, peptides were included in user-defined libraries with public immunogenicity annotations. We observed that immunogenic peptides were significantly more stable than non-immunogenic peptides. Conclusion We generated customizable pHLA stability datasets which show how peptide sequence motifs affect pHLA stability, and may be helpful for improving our mechanistic understanding of pHLA stability. Further, since peptide stability is related to immunogenicity, these large-scale pHLA stability datasets will be useful for improving peptide immunogenicity predictions for the development of immunotherapeutics. Funding Source NIH R01CA155010, Mark Foundation for Cancer Research, Moderna Topic Categories Classical and Non-Classical Antigen Presenting Cells (APC)

Role of EHF in regulating B cell checkpoint molecule TIM-1 and development of CD11c+TIM-1+ Age-associated B cells 2301855

Abstract Introduction Aging is associated with an increased risk of cancer and immunosenescence, coinciding with the expansion of Age-associated B cells (ABCs). While T cell checkpoints are well-established targets, the landscape of inhibitory pathways in B cells, how they regulate anti-tumor immunity remain unclear. This study investigates the transcriptional regulation of the checkpoint molecule TIM-1 on ABCs and its impact on anti-tumor immunity. Methods We combined multi-tissue single-cell RNA-seq across aging, bulk RNA-seq, ATAC-seq, ChIP-qPCR, and a transcription factor CRISPR screen to identify regulators of TIM-1+ ABCs. We generated mice with B cell-specific deletion of the ETS-homologous factor (EHF) and assessed tumor growth, immune infiltration using B16-OVA melanoma and AOM/DSS colon adenocarcinoma models in both young and aged cohorts. Results We identified a progressive age-related expansion of TIM-1+ ABCs that co-express multiple inhibitory checkpoints, a phenotype mirrored by B cells in tumor-draining lymph nodes. We found that TLR9 signaling drives this phenotype and identified EHF as the pivotal transcriptional driver. EHF directly binds the Havcr1 (TIM-1) locus to induce expression. B cell-specific EHF deletion selectively eliminated TIM-1+ ABCs, unleashed Th1-skewed immunity, and enhanced CD8+ T cell activity. Consequently, EHF-deficient mice exhibited robust control of colorectal and melanoma tumors. Crucially, this heightened anti-tumor immunity persisted in aged mice, reversing age-associated immune suppression. Conclusion These findings define a previously unrecognized TLR9-EHF-TIM-1 checkpoint axis in B cells that emerges with age constraining anti-tumor immunity and age-associated inflammation. Targeting the regulatory function of EHF-driven ABCs provides a mechanistic rationale for novel therapeutic strategies to potentiate cancer immunotherapy, particularly in the elderly population where these cells accumulate. Funding Source This work was supported by the Gene Lay Institute of Immunology and Inflammation and grants from the National Institutes of Health (R01 AG080992, AI169075, AI185514, AI184980, P01 AI039671, P01 AI056299 to V.K.K; R01CA187975 to A.C.A.; R01CA282794 to V.K.K. and A.C.A.), GSK Award (“Role of B cells in regulating autoimmunity and anti-tumor immunity”) to V.K.K. and the Melanoma Research Alliance (926682, “Harnessing B cell checkpoints in melanoma” to V.K.K. and A.C.A). Y.-C.K. was supported by the postdoctoral fellowship from National Multiple Sclerosis Society (FG-2007-36929). Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Copolymerized Plasticizer Enables Halogen‐Free Processing Toward 20.78% and 17.83% Efficient Organic Solar Cells and Modules

ABSTRACT Nonhalogenated solvents are promising for scalable fabrication of organic solar cells (OSCs). However, the intrinsically low solubility of high‐molecular‐weight donor polymers in nonhalogenated solvents, particularly those with tightly packed, highly crystalline backbones such as the pinnacle of donor materials (D18), disrupts multiscale morphology evolution and severely limits device performance. Here, without compromising the polymer's molecular weight or its intrinsic propensity for ordered packing, we establish a copolymerized “plasticizer” strategy that enhances the solvation interaction between a nonhalogenated solvent (toluene) and an appended plasticizing comonomer, thereby improving the processability. By integrating a plasticizing unit (bis(2‐(2‐methoxyethoxy)ethyl) thieno[3,2‐ b ]thiophene‐3,6‐dicarboxylate) into the D18 backbone, the resulting D18‐O exhibits markedly enhanced solubility through dipolar interactions with toluene. These interactions suppress excessive pre‐aggregation, slow down crystallization kinetics of the donor, and ultimately guide the formation of well‐defined nanocrystals and a hierarchy of optimized morphologies during nonhalogenated processing. This approach enables both OSCs and modules processed from toluene to achieve record power conversion efficiencies of 20.78% (certified 20.40%) and 17.83% (certified 17.29%), respectively, along with exceptional stability.

Effects of hydrogen cyanamide and garlic extract on dormancy release, yield, and fruit quality of low-chill apple cultivars at Debre Tabor, northwestern Ethiopia

Abstract Incomplete dormancy release due to insufficient winter chilling is a major constraint to apple ( Malus domestica Borkh.) production in tropical highland environments. This study evaluated the effects of hydrogen cyanamide (1.5%), garlic extract (15%), and their combined application on bud phenology, reproductive performance, yield, and fruit quality of three low-chill apple cultivars (‘Anna’, ‘Dorsett Golden’, and ‘Princesa’) under Ethiopian highland conditions over two consecutive seasons (2023–2024). Data were analyzed using a linear mixed-effects model with repeated measures across the two growing seasons. Dormancy-breaking treatments significantly advanced bud break by 15–24 days and shortened flowering duration from 7 to 8 weeks in the control to 3–5 weeks in treated trees. The combined hydrogen cyanamide + garlic extract treatment produced the earliest bud break and the highest flowering percentage (up to 67%), fruit set (up to 19%), fruit number (up to 63 fruits tree⁻¹), and yield (5.0–6.0 kg tree⁻¹) across cultivars. Yield improvement was associated with increases in fruit number and fruit weight. Significant cultivar × treatment interactions ( P  < 0.05) were observed for most phenological and reproductive traits, demonstrating differential cultivar responsiveness to dormancy-breaking agents. Conversely, total soluble solids and fruit firmness were not significantly affected by treatment, cultivar, or their interaction ( P  > 0.05). These findings indicate that combining low-chill cultivars with dormancy-breaking treatments partially compensates for inadequate winter chilling and improves apple productivity under tropical highland conditions.