Browse Articles

Simultaneous Detection of Transcription Factors and RNA Using TotalSeq™ Antibodies with 10x Genomics Chromium Single Cell Flex Gene Expression 2258335

Abstract Introduction Transcription factors (TFs) are key proteins that regulate gene expression by binding specific DNA sequences. They control essential cellular processes such as cell cycle, metabolism, immune responses, and more. Characterizing TFs in defined cell types is crucial to understanding their physiology and role in homeostasis and disease. While other techniques such as intracellular flow cytometry are commonly used, single-cell sequencing offers comparable resolution while providing simultaneous transcriptome characterization and higher throughput protein detection. Methods Here, we profiled human PBMCs and multiple cell lines using the 10x Genomics Chromium Fixed RNA Profiling Kit with our TotalSeq-B Human Universal Cocktail (140 antibodies), plus 13 intracellular effector proteins, and 25 TFs. Cells were stained using our optimized TotalSeq™ Cell Surface and Nuclear Protein Staining protocol, and RNA, surface, and intracellular proteins were simultaneously profiled. Results TFs such as T-BET, FOXP3, TCF1, and THPOK, and signaling proteins like ZAP70 and ITK were reliably detected in T and NK cell clusters as expected. SPI1, NTAL, BCL11B, and SYK were enriched in B cell and monocyte clusters. OCT4, NANOG, and SOX2 were detected in the human embryonal carcinoma cell line NCCIT, and FOXA1 in the luminal epithelial-like breast cancer cell line MCF-7. RNA-protein correlation was confirmed where expected. Conclusion In summary, we demonstrate that our workflow enables sensitive and specific single-cell profiling of both RNA and nuclear proteins while preserving RNA integrity. Funding Source BioLegend Topic Categories Technological Innovations in Immunology (TECH)

Myeloid-Intrinsic Protein Tyrosine Phosphatase Non-Receptor Type 2 (PTPN2) Programs Hematopoietic Stem Cells Towards Pro-Inflammatory Macrophages 2309892

Abstract Introduction Protein tyrosine phosphatase non-receptor type 2 (PTPN2) suppresses inflammatory signaling, and its loss is linked to several inflammatory diseases. PTPN2 helps dampen mucosal inflammation by shifting macrophage balance from pro-inflammatory M1 to protective M2 types. While PTPN2’s role in macrophage activation is known, its impact on myeloid progenitor cells and macrophage differentiation remains unclear, potentially offering new stem cell therapy opportunities. Methods We studied myelopoiesis and macrophage responses in myeloid-specific PTPN2-deficient mice (KO) and wild-type controls (WT), both under naïve conditions and after M2 macrophage polarization via helminth infection. A congenic bone marrow chimera strategy was used to assess cell-intrinsic effects of PTPN2 on monocyte-to-macrophage transition and survival, and Cellular Indexing of Transcriptomes and Epitopes by sequencing (CITE-seq) was performed on the WT and KO donor cells. Results Flow cytometry showed that PTPN2 KO mice had increased Ly6C-Hi P1 inflammatory monocytes and decreased P4 resident macrophages in the colon, with heightened M1 activation post-infection. This led to improved helminth clearance but increased lung and intestinal inflammation. KO donor cells had a reconstitution advantage over WT cells, coupled with a differentiation blockade and accumulation of P1 monocytes. After infection, KO donor cells maintained elevated inflammatory monocytes in the colon, stabilizing local inflammation. CITE-seq revealed increased transcriptional activity and enrichment of antigen presentation and cell survival genes in PTPN2-deficient cells. Conclusion Overall, myeloid-intrinsic PTPN2 deficiency primes the myeloid compartment for inflammation, boosting immunity to helminth infection but sustaining mucosal inflammation. PTPN2 regulates macrophage activation during myelopoiesis, restraining excessive monocyte-to-macrophage transition and limiting cell survival and antigen presentation. Funding Source The National Institutes of Health (RO1AI135195 to DM and MGN) and the UCR Division of Biomedical Sciences (Burden/Pease Fellowship to SA) Topic Categories Mucosal and Regional Immunology (MUC)

The Role of B Cell Specific MHC II Antigen Presentation in Breast Cancer 2247666

Abstract Introduction Breast cancer remains the leading cause of cancer-related mortality in women. Immunotherapies have revolutionized treatment; however, their efficacy relies heavily on immune cell-mediated antigen presentation within the tumor microenvironment (TME). The often-overlooked role of B cell antigen presentation may be important in anti-tumor immunity. B cells are potent antigen presenting cells and the role of B cell antigen presentation in progression of autoimmune diseases in mouse models of systemic lupus erythematous and autoimmune encephalomyelitis has been demonstrated. B cells may have a potent role in breaking immune tolerance towards self-peptides. To this end, we hypothesize that B cell specific MHC II antigen presentation plays a non-redundant role in promoting anti-tumor immunity. Methods To investigate the role of B cell MHC II antigen presentation in tumor progression, we generated B cell-specific MHC II knockout mice (Mb1-Cre+/- MHC II fl/fl) and utilized the E0771-LN26 murine triple-negative breast cancer (TNBC) cell line with a propensity for lymph node and metastases. Tumor growth was assessed in B cell MHC II-deficient and wild-type littermates. To understand the relevance of this study in human TNBCs, we generated co-relation of MHC II expression in breast cancer on patient outcome, we analyzed publicly available TNBC data. Results Our data demonstrate that E0771-LN tumors exhibit accelerated growth in B cell-specific MHC II knockout mice compared to MHC II fl/fl controls, highlighting a critical role for B cell-mediated MHC II antigen presentation in controlling tumor progression. Moreover, T cells from mice lacking B cell specific MHC II expression had were more exhausted. We also found a co-relation of high MHC II expression with better patient outcome. Conclusion These findings suggest that B cell-mediated antigen presentation plays a non-redundant role in modulating anti-tumor immune responses in breast cancer. Funding Source This research project is supported by funding from NCI Topic Categories Classical and Non-Classical Antigen Presenting Cells (APC)

Equol inhibits NLRP3 inflammasome activation in macrophages 2257578

Abstract Introduction The intestinal microbiome produces a vast number of metabolites. Many of these metabolites have immunomodulatory properties including the ability to inhibit the NOD, LRR, and pyrin-domain containing 3 (NLRP3) inflammasome, an established driver of inflammation in many diseases including gout, psoriasis, and Alzheimer’s disease. Methods Therefore, we developed an enzyme-linked immunosorbent assay (ELISA) based screen to identify metabolites with the potential to inhibit NLRP3 activation. Using this screen, we identified equol, a metabolite of the soy isoflavone daidzein, as a strong candidate. Additional ELISAs and caspase-1 immunoblots further validated that equol inhibits ATP dependent NLRP3 activation in murine bone marrow derived macrophages (BMDMs), evidenced by significantly decreased IL-1β secretion and caspase-1 processing after equol treatment. Results Our work indicates equol’s anti-inflammatory effects are specific to the NLRP3 inflammasome; treatment with equol does not alter the secretion of several other inflammatory cytokines/chemokines (TNFα, IL-6, CXCL1, MIP-1α, RANTES) or activation of the AIM2 or NLRC4 inflammasomes in BMDMs. Conclusion These findings lay a strong foundation for future studies examining the immunomodulatory action of equol in mouse models of NLRP3 driven disease. Funding Source This work was supported by funds from the William F. Vilas Trust Estate and the National Institutes of Health P30AG062715, R01AG070973, and R01AG083883 to TKU Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

A 61-Parameter CyTOF Panel for Comprehensive Profiling of Human PBMC to Characterize Activation, Differentiation, Checkpoints and Cytokines 2259198

Abstract Introduction High-parameter cytometric analysis enables discoveries of potential therapeutic targets and informs disease prognoses in translational and clinical research. CyTOF™ technology is a single-cell analysis platform that uses metal-tagged antibodies to resolve 50-plus markers in a single tube. Notably, antibody cocktails and stained samples can be frozen for later use and acquisition, or barcoded and pooled together, minimizing technical variation. Unlike fluorescence-based cytometry, spectral unmixing and single-stain controls are not required. Therefore, it is uniquely possible with CyTOF technology to rapidly design high-parameter panels and use intracellular markers to gain functional insights. The goal of this study was to design a 61-parameter CyTOF panel for in-depth functional immune profiling of human PBMC. Methods The panel contains lineage markers for major immune cell subsets and a diverse array of phenotyping T cell targets focused on activation, differentiation, checkpoint and cytokines and can be used to identify over 60 cell populations. Untreated and stimulated PBMC were barcoded, pooled and stained. Samples were frozen and acquired on a later day using a CyTOF XT PRO system. Results High-dimensional analysis of stimulated PBMC revealed striking cross-lineage immuno-functional diversity at the single-cell level. The expression of over 20 markers spanning the functional landscape of activation, checkpoints and cytokines was revealed across effector, memory, cytotoxic, regulatory and exhausted immune cell populations. Conclusion CyTOF systems enable the highest number of simultaneous measurements in a single panel, allowing for wide immune coverage and high resolution of intracellular targets to interrogate functional potential. Overall, studying functional immunology using CyTOF technology can elucidate the complex nature of immune responses to provide an understanding of how they relate to disease and treatments. For Research Use Only. Not for use in diagnostic procedures., Funding Source n/a Topic Categories Immune Mechanisms of Human Disease (HUM)

Unique Transcriptional and Functional Profiles of Neonatal Naïve CD4+ T Cells Expressing KLRG1 2260016

Abstract Introduction Human neonates, particularly those born prematurely, are susceptible to severe infections and inflammatory diseases with long-term complications. The immune factors driving these clinical differences are not well understood. Recent evidence suggests that instead of being merely in a generally naïve state, unique neonatal CD4+ T cell subsets may exist in an effector-like state and differentiate more quickly than their adult counterparts. Killer-like Lectin Receptor G1 (KLRG1), an immune checkpoint receptor found on terminally differentiated T cells in adults, has been observed on ∼30% of CD4+ T cells found in cord blood (CB) of healthy neonates. However, little is known about the functional phenotype and transcriptome of these cells, and their clinical significance in neonates is relatively unexplored. Methods We conducted flow cytometry and CITE-seq on CB mononuclear cells obtained from healthy full-term and premature infants. Additionally, persistent respiratory morbidity (PRD) as a function of KLRG1+ CD4+ T cells was examined in a longitudinal cohort of preterm infants. Results The frequency of KLRG1+ CD4 cells positively correlated with gestational age at birth until term corrected age (TGA), then decreased at one year. Preliminary analysis identified a lower risk of PRD in premature infants with higher KLRG1 frequency by TGA. Compared to KLRG1- CD4+ T cells, cord blood KLRG1+ CD4s were mainly naïve, had lower CD38 surface staining, and higher TNF-α and lower IL-8 production following stimulation. KLRG1+ CD4+ T cells expressed higher ANXA1 and FTH1 genes, which suggests peripheral activation or early differentiation. Conclusion Our results suggest that KLRG1 expression on naïve CD4 T cells identifies a functionally distinct CD4+ T cell population present in neonates, whose frequencies are developmentally regulated. Although preliminary analysis suggests a protective role, further investigation into their role and potential mechanism in protection or immunopathology are warranted. Funding Source NIH NIAID HHSN272201200005C (Respiratory Pathogens Research Center) University of Rochester Department of Pediatrics U24AI152176 Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)

A Rigid‐Flexible Polyinterface Enabling Molecular‐Level Dual‐Ion Regulation for Ultrastable Lithium Metal Batteries

ABSTRACT The unstable lithium metal anode plagued by dendrite growth and parasitic reactions remains a formidable barrier to realizing high‐energy‐density batteries. While artificial solid‐electrolyte interphases offer a protective strategy, most designs are limited to single‐ion regulation and fail to orchestrate the complex, dual‐ion (Li + and anion) chemistry at the molecular level. Here, we report a rationally designed polymeric artificial interphase of poly‐fluorotoluene‐triglycoldimercaptan (PFT) featuring rigid fluorinated benzene rings and flexible sulfur‐oxygen chains for synergistic dual‐ion regulation. The flexible segments enable gradient Li + coordination with differential binding energies, lowering desolvation barrier and facilitating uniform Li + transport, whereas the rigid electron‐deficient fluorinated rings trap TFSI − anions via anion‐π interactions. This dual modulation directs the in‐situ formation of a robust, LiF‐Li 2 S‐rich inorganic composite SEI, as validated by calculations and spectroscopy. The PFT‐based Li anodes exhibit exceptional stability, with symmetric cells operating over 4000 h at 1 mA cm −2 , 1 mAh cm −2 . A high‐loading LiFePO 4 full cell retains 80% capacity after 1000 cycles at 5 C, and an NCM811 pouch cell retains 85% capacity after 160 cycles at 0.5 C, demonstrating practical viability. This work establishes a molecular design principle for dual‐ion regulation via a polyinterface for high‐performance Li metal batteries.

Structural mechanism of 26S proteasome regulation and its pharmacological modulation

Hybrid VAE-GAN generative framework for multi-objective spatial performance optimization of residential landscape morphology

PD-1/PD-L1 enrichment in the IPF lung as a mechanism of senescent cell immune evasion 2266760

Abstract Introduction Idiopathic pulmonary fibrosis (IPF) is an age-related lung disease characterized by the accumulation of senescent cells that deposit extracellular matrix components and soluble proteins, thereby altering the microenvironment. Our previous data indicate a reduced proportion of cytotoxic NK cells in the lungs of patients with IPF. Furthermore, mouse models of lung fibrosis demonstrate that depletion of total NK cells leads to the persistence of senescent cells and exacerbated fibrosis. We postulate that aging and local microenvironmental factors impair the ability of NK cells to eliminate senescent cells. Methods To characterize human IPF cells, single-cell suspensions from lymph nodes and lungs of healthy individuals and IPF patients were analyzed using in vitro cytotoxicity assays, flow cytometry, and single-cell RNA sequencing (scRNA-seq). Results scRNA-seq analysis of lung cell suspensions from healthy and IPF donors revealed that IPF NK cells exhibit reduced expression of cytotoxicity-related genes and increased expression of PD-1, a key regulator of immune cell function. When NK cells were pre-treated with conditioned media from human lung fibroblasts (hLF-CM), only conditioned media derived from IPF fibroblasts induced PD-1 expression in NK cells. Flow cytometry and culture assays further demonstrated that senescent IPF lung fibroblasts express high levels of PD-L1, which in turn suppresses NK cell cytotoxic activity. Conclusion Collectively, our findings suggest that the IPF lung microenvironment modulates NK cell function, promoting the accumulation of pro-fibrotic senescent fibroblasts. These results support the potential use of PD-L1 inhibitors to restore NK cell activity and provide a novel therapeutic strategy for IPF. Funding Source Davis Heart and Lung Research Institute, U01HL145550 Topic Categories Immune Mechanisms of Human Disease (HUM)

Natural Microbial Experience Increases Infiltration of T Cells in the Brains of Alzheimer’s Disease Mice 2258422

Abstract Introduction The immune environment of the brain plays a major role in neurodegenerative conditions such as Alzheimer’s disease. The abnormally clean facilities in which laboratory mice are kept limit immune experience and cause major concerns regarding the translatability of research to humans. These facilities, although not germ-free, lack the normal exposures to diverse commensal and environmental microorganisms experienced by mice in the wild or diverse environments such as a pet store. By breeding inbred lab mice that have previously experienced exposure to the dirty bedding of pet store mice, we can generate “bred dirty” mice with immune systems that mirror those of adult humans more closely. It is unclear whether these immune changes extend to the brain and whether this plays a role in neurodegenerative disease. Methods We used the P301s transgenic mouse model of Alzheimer’s disease that accumulates age-dependent tau pathology. We exposed these mice to dirty bedding obtained from pet store mice, and bred them to establish a line of “bred dirty” tau-P301S mice and appropriate controls. At endpoint, we collected brain and spleen for flow cytometry, immunofluorescence, and bulk RNAseq. Results The proportions of CD3+ T cells, monocytes, and macrophages were significantly increased in the brains of the dirty tau-P301S mice compared to their clean tau-P301S counterparts. The brains of the dirty tau-P301S mice also showed an increase in the proportion of CD8+ T cells, specifically effector differentiated (CD62L-CD44+) CD8+ T cells. The proportion of activated (CD11c+) microglia was increased in tau-P301S mice but unaffected by dirty status. Conclusion Disease-associated infiltration of adaptive immune cells into the brain is amplified by dirty status without grossly affecting microglial activation. Given the increasingly recognized role of T cells in Alzheimer’s disease pathogenesis, our results suggest that more immune educated models might better represent the role of T cells in neurodegenerative disease. Funding Source Mayo Clinic and The Translational Genomics Research Institute Topic Categories Neuroimmunology (NEUR)

Estrogen signaling through estrogen receptor beta in Tregs regulates the differential response to therapy in head and neck squamous cell carcinoma 2310155

Abstract Introduction Immunotherapies (IO) have largely failed in head and neck squamous cell carcinoma (HNSCC). The subset of patients who respond to therapy tend to be premenopausal women. Estrogen (E2)-based hormone replacement therapy (HRT) has been shown to impart a survival advantage in HNSCC with 22% lower risk for ever-users of HRT, supporting the role of sex hormones as a driver of this advantage. It has been shown that estrogen receptors influence the immunosuppressive activity of regulatory T cells (Tregs), which are known drivers of resistance to IO in HNSCC. Based on these previous findings, we hypothesized that E2 reduces the immunosuppressive functionality of Tregs in the HNSCC tumor microenvironment (TME), thus enhancing effector T cell function. Methods We utilized the MOC2 and LY2 orthotopic models of HNSCC, implanted into C57BL/6 and BALB/C mice, respectively. Using surgical, pharmacologic, and genetic methods, we manipulated the level of sex hormones present to study their role in tumor growth, their impact on the TME using flow cytometry, and their effect on Treg cell state in vitro using qRT-PCR. Results We demonstrate that supplemented E2 leads to immune cell differences within the TME that are associated with improved response to radioimmunotherapy. We show that treatment with an estrogen receptor beta (ERβ) agonist results in a reduction of Treg immunosuppression and an increase in effector T cell functionality, leading to decreased tumor growth. Concordant with these findings, we found that conditional deletion of ERβ in male Tregs blocks the decrease in tumor growth observed with E2 supplementation. We provide further evidence that E2 signaling through ERβ in Tregs is decreasing their suppressive state via a decrease in stability. Conclusion This study reveals ERβ signaling in Tregs as a mediator of the immune response in HNSCC. Our findings provide rationale for therapeutic integration of E2 or ERβ agonists in immunotherapy clinical trials for males and post-menopausal females with HNSCC. Funding Source T32-AI007405, R01 DE028529-01, R01 DE028282-01, R01 CA284651-01 Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Adaptive immune responses in the skull bone marrow for CNS immunosurveillance and brain tumor immunity 2247790

Abstract Introduction Mounting evidence indicates that the central nervous system (CNS) maintains active communication with the peripheral immune system, yet the mechanisms by which adaptive immunity monitors the CNS remain incompletely understood. Recent discoveries have revealed specialized vascular and cellular conduits linking the dura mater with the skull, allowing cerebrospinal fluid and immune cell exchange between the CNS and the skull bone marrow (BM) under both steady-state and pathological conditions. Although the skull BM has been recognized as a reservoir of innate immune cells supporting CNS immunity, its contribution to adaptive immune responses has not been thoroughly defined. Methods We analyzed mouse skull bone marrow using single-cell RNA sequencing, flow cytometry, and confocal imaging. Results In this study, we describe organized lymphoid aggregates within the skull BM that display germinal center-like architecture and contain a distinct subset of follicular helper T cells. These T cells drive B cell activation and humoral responses through their communications. We show that adaptive immune cells within these skull BM niches detect and react to CNS-derived antigens and are indispensable for effective antitumor immunity in mouse models of brain cancer. Conclusion Our identification of these specialized adaptive immune hubs in the skull BM establishes a new paradigm for CNS immunosurveillance and unveils therapeutic opportunities for neurological disorders, including malignancies of the brain. Funding Source The ChadTough Foundation Topic Categories Neuroimmunology (NEUR)

IL-9 secreting CD4+ Trm cells are reciprocally regulated by PU.1 and BCL6 in a recall model of allergic lung inflammation 2257477

Abstract Introduction Chronic allergic airway inflammation results in the establishment of a tissue resident memory T (Trm) cell population that can respond to subsequent allergen challenge. A subset of multi cytokine-producing CD4+ Trms that express high levels of IL-9 with type 2 cytokines, is critical to mediate this rapid allergen recall responses upon challenge. PU.1 (Spi1), BCL6, PPARγ, BATF, and IRF4 have been shown to be important in IL-9 expression in Th9 cells, but their role in generating a multi-cytokine phenotype in Trm cells has not been investigated. Methods We used a memory-recall model of chronic allergic lung inflammation that mimics seasonal allergies with house dust mite (HDM) sensitization to challenge wild-type, Spi1 fl/fl iCD4-Cre, Spi1 fl/fl CD4-Cre, and Bcl6 fl/fl iCD4-Cre mice to study the reciprocal roles of PU.1 and BCL6. Mice were challenged intranasally three times a week for 6 weeks with HDM followed by a 6-week rest period absent of any allergen challenge. Mice with inducible CD4-Cre were treated with tamoxifen for the last two weeks of the allergen-free period. Mice were then challenged with HDM 2 day before analysis. Results Deletion of PU.1 resulted in decreased Trms and IL-9, IL-13 production and diminished eosinophilic inflammation in the memory response in lung in vivo. However, deletion of BCL6 from CD4+ T cells resulted in increased Trms and increased IL-9 and IL-13 production and eosinophilic inflammation in the lung in vivo. Conclusion Our results show that during allergic airway inflammation, BCL6 and PU.1 play reciprocal roles in regulating tissue resident T cells and multi-cytokine-producing Th9 cell population that mediates the memory-recall in allergic lung inflammation. Funding Source National Institutes of Health (R01 AI057459 and AI129241 to M.H.K.) Topic Categories Cytokines and Chemokines and their Receptors (CCR)

Structural characterization of antibodies binding the conserved central helix and membrane-proximal lower stalk of coronavirus spike glycoprotein 2248624

Abstract Introduction The ongoing threat of zoonotic coronavirus spillover into humans, exemplified by severe acute respiratory syndrome virus 2 (SARS-CoV-2), underscores the urgent need for pan-coronavirus therapeutics that can be deployed to mitigate future pandemics. Methods Here, we identified monoclonal antibodies from COVID-19 convalescent donors that target two conserved epitopes in the S2 domain of the coronavirus Spike glycoprotein: the central helix (CH) and a membrane-proximal epitope in the heptad repeat 2 (HR2), which we designate the lower stalk (LS). Results CH-directed antibodies exhibited broad cross-reactivity across betacoronaviruses, whereas LS-directed antibodies demonstrated reactivity primarily within sarbecoviruses. Using cryogenic electron microscopy (cryo-EM), we determined sub-4 Å structures of three cross-reactive CH antibodies–ch.005, ch.007, and ch.010–bound to the prefusion-stabilized SARS-CoV-2 S2 protein, revealing distinct binding poses and contact residues relative to previously described CH antibodies. In parallel, X-ray crystallography studies yielded a sub-2 Å structure of the ls.019 Fab in complex with the LS peptide, providing, to our knowledge, the first structural visualization of a human monoclonal antibody engaging this epitope. Conclusion Together, these findings advance our knowledge of two conserved and structurally vulnerable sites within the coronavirus S2 subunit—the central helix and the lower stalk—that can guide the development of broad-spectrum antibody therapeutics and vaccines against current and emerging coronaviruses. Funding Source Howard Hughes Medical Institute Topic Categories Vaccines and Immunotherapy (VAC)

Converge and Contrast: Leishmania donovani’s Impacts on Uninfected Bystander Cells by Life Stage 2259952

Abstract Introduction Visceral leishmaniasis (VL) remains the second most lethal parasitic disease in the world. As a neglected tropical disease, treatments are limited. During VL, systemic infection affects primarily phagocytic cells in the liver, spleen, and bone marrow. There is massive modulation of the immune system. Some immune evasion mechanisms are already known for both the promastigote (initiation) and amastigote (chronic) forms of the parasite. Moreover, our prior observations in mouse macrophages have shown NF-κB sensing is blocked for infected and uninfected bystander cells during promastigote infection. Thus, we aimed to verify bystander immunosuppression in a human model and explore further its impact and the effect of chronic infection. Methods This project used two modes of assessing the bystander immune response. First, we used a human transgenic U937 monocyte line with Green Fluorescent Protein on a NF-κB-responsive promoter as a high-throughput detector. A Dsred2 expressing L. donovani was used for promastigote or amastigote (isolated from animals) infections in vitro. Also, we evaluated single cell RNA sequencing (scRNA-seq) data to explore gene expression from in vivo murine chronic VL in infected and bystander bone marrow monocytes. Results In vitro, our reporter cells have identified that the bystander immunosuppression effect occurs through media components < 3kDa, independent of direct contact to the promastigote. Meanwhile, in the amastigote infection, we saw in vitro NF-κB bystander activation rather than suppression. Also, scRNA-seq revealed that TGF-β is highly upregulated in bystander monocytes among other differentially expressed genes (DEGs). Conclusion We have for the first time confirmed NF-κB bystander modulation during promastigote and amastigote infections of the human model cells. We also have identified DEGs of interest in chronic infection. These various effects by life stage highlight the important role for NF-κB in VL progression. Funding Source GR110769 Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)

Multiscale Magnetic Field Engineering for Advanced Lithium‐Based Batteries

ABSTRACT Lithium battery performance remains limited by sluggish kinetics, interfacial instability, and mass transport constraints. The application of magnetic fields as a non‐invasive physical modulation tool offers a promising paradigm to fundamentally circumvent these pervasive bottlenecks. This review first outlines the fundamental physical principles of magnetic fields to establish the theoretical basis for their functionality in electrochemical systems. Building upon this foundation, we systematically elucidate the role of the magnetic field in addressing multiscale scientific challenges within lithium‐based batteries, specifically regarding interfacial dynamics, bulk phase transport, and chemical reactions. Regarding the electrode/electrolyte interface, the magnetic field effectively inhibits lithium dendrite proliferation and stabilizes the solid electrolyte interphase through the homogenization of ionic flux. Within the bulk phase, macroscopic convection induced by the magnetohydrodynamic effect mitigates concentration polarization, while magnetic force facilitates the directional transport of paramagnetic molecules such as oxygen and the targeted capture of detrimental ions. At the reaction level, magnetic modulation inhibits cation mixing via spin‐state regulation and accelerates sluggish multielectron reactions. The objective of this review is to clarify the transformative potential of magnetic fields as multiscale and multifunctional regulatory tool for advancing the development of next‐generation lithium‐based batteries, including lithium–sulfur, lithium–oxygen, and all‐solid‐state batteries.

Structural insights into histone mimicry by the small hepatitis delta antigen

Network analysis highlights socio-demographic patterns of stone tool-using primates

Abstract While the nutritional benefits and social learning of tool use in primates have been widely studied, far less is known about how tool use is associated with social life. We investigated the social aspects of tool use in a hybrid population of long-tailed macaques ( Macaca fascicularis aurea  ×  M. f. fascicularis ) on Koram Island, Thailand, comprising both stone tool users and non-users. To assess potential socio-demographic patterns, we examined whether tool use differed between sexes and whether it was related to variation in dominance rank. Using social network analyses, we assessed whether tool users differed in their associations outside foraging contexts by analyzing their grooming interactions and positions within the grooming network. Males were more likely to use tools than females, whereas rank was unrelated to tool use. Tool users and non-users showed similar social connectivity, grooming rates, and numbers of grooming partners. However, grooming networks indicated that non-tool users preferentially associated with other non-users, whereas tool users showed no affiliative bias. By integrating individual characteristics with social network data, this study provides insight into how tool-use behavior integrates into primate social systems, particularly in populations with substantial variation in technological behavior.

Noncanonical role for Caspase-5 in the human intestinal epithelium 2267092

Abstract Introduction Caspase-5 (CASP5) is a member of the inflammatory caspase family of cysteine proteases involved in inflammation and cell death. CASP5 shares highest homology with inflammatory caspase-4, but while caspase-4 is essential for noncanonical inflammasome activation, CASP5 is generally dispensable. This conundrum has left a knowledge gap in understanding CASP5 function. Methods We mapped CASP5 expression across human tissues and epithelial cell types, then used proteomics and molecular biology to define interacting partners and pathways. We validated function in human intestinal organoids, and localized isoforms in human tissues by immunohistochemistry and RNAscope. Results CASP5 expression was largely restricted to human intestinal epithelium and resolved into three isoforms. Proteomics and interaction mapping suggested that CASP5 interacts with an important developmental signaling scaffold in epithelial cells. Strikingly, during human tissue analysis and organoid differentiation, we noted differential expression of CASP5 isoforms along the crypt-to-lumen axis with specific enrichment of the developmental isoform in crypt regions. Functional studies demonstrated that the developmental isoform augmented the growth of colonic and small-intestinal organoids, which together with supporting data from biochemical studies provided robust evidence for a novel function for CASP5 in sustaining proliferative programs and epithelial renewal. Conclusion Our findings reveal that a select developmental isoform of CASP5 amplifies a key epithelial growth program, peaking in specific cell subsets in the intestinal epithelium to sustain their proliferation during stages critical of intestinal epithelial differentiation and regeneration. Our work broadens the role of inflammatory caspases beyond innate immunity, highlighting their contribution to tissue homeostasis. Funding Source n/a Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)