Ticks rely on salivary immunomodulators to sustain prolonged blood feeding and transmit pathogens, yet the protein effectors that mediate these processes remain incompletely defined. Here, we characterize three divergent cysteine protease inhibitors—Amacstatins—from the Gulf Coast tick Amblyomma maculatum and delineate their roles in feeding success and Rickettsia parkeri transmission. Integrated transcriptomic, structural, and biochemical analyses reveal that Amacstatin-1, -2, and -3 possess distinct reactive centers, inhibitory selectivity, and tissue-specific expression patterns that are dynamically regulated by blood feeding and rickettsial infection. Silencing all three Amacstatins via RNA interference significantly impaired tick feeding, reducing engorgement, fecundity, and R. parkeri burden in both midgut and salivary glands. Functional assays demonstrated that Amacstatins potently suppress host inflammatory responses, including pro-inflammatory cytokine production, nitric oxide release, and leukocyte recruitment. Together, these findings identify Amacstatins as critical immunomodulatory effectors that promote tick feeding and rickettsial transmission. This work provides a protein-level mechanistic insight into the A. maculatum–R. parkeri interaction and highlights Amacstatins as promising molecular targets for anti-tick and anti-rickettsial strategies.
Acute kidney injury (AKI) is an unavoidable complication and critical determinant of prognosis in renal transplantation. M1-polarized macrophages are known to exacerbate AKI progression. Preliminary clinical data reveals a positive correlation between elevated E74-like ETS transcription factor 1 (ELF-1) and pro-inflammatory cytokines in macrophages from post-transplant patients. However, the role of ELF-1 in M1 macrophage and AKI pathogenesis remains unexplored. We find that ELF-1 deficiency in murine macrophages significantly decreases inflammatory M1 polarization and attenuated AKI. Mechanistically, ELF-1 directly binds to GBP4 to promote its transcription, thereby promoting the GBP4-NLR-NF-κB axis and finally driving M1 polarization. Recombinant GBP4 protein restores M1 polarization impaired by Elf-1 deletion. Similarly, adoptive transfer of ELF-1-expressing bone marrow monocytes in vivo reverses the protective effects of Elf-1 deficiency on AKI. Collectively, we identify ELF-1 as a novel transcriptional regulator of M1 macrophage polarization in post-transplant acute kidney injury. The transcription factor ELF-1 directly regulates the expression of the GBP4 gene, thereby promoting GBP4-NLR-NF-κB axis and ultimately driving the M1 polarization of renal macrophages, thereby exacerbating acute kidney injury.
Species diversity strongly predicts productivity, yet the processes linking intraspecific diversity to ecosystem functioning remain poorly understood. Through a controlled marsh mesocosm experiment with the salt-marsh foundation species Scirpus mariqueter, we found that increasing genotypic diversity significantly enhanced plant performance (above-ground and total biomass) and reproductive output (corm number, corm biomass, inflorescence and seed production), with increased soil nitrogen and a significant overall effect on soil organic carbon. Intermediate genotypic diversity significantly altered bacterial community composition. Although overall bacterial community assembly remained predominantly stochastic, β-nearest taxon index (βNTI) analysis indicated a stronger deterministic assembly signal in four-genotype mixtures. Notably, trait expression transitioned from additive effects at four genotypes to nonadditive interactions at eight genotypes, indicating increasing complementarity at higher diversity levels. These findings collectively reveal that genotypic diversity enhances plant productivity and soil nitrogen pools while selectively altering bacterial community composition and assembly processes, providing mechanistic insights into how intraspecific variation stabilizes and promotes ecosystem functioning in salt marshes.
We present the Generalized Activating Function (GAF), a biophysics-grounded framework that accelerates neural activation predictions by up to three orders of magnitude relative to NEURON simulations of multi-compartment cable models while maintaining high predictive accuracy (R2 = 0.99). By overcoming computational bottlenecks in patient-specific modeling, the GAF enables practical translation of computational optimization into neuromodulation therapies such as spinal cord stimulation (SCS). We verify the GAF’s physiological validity by demonstrating near-perfect agreement with multi-compartment simulations for activation thresholds and spike initiation location/timing across multiple pulse widths in an anatomically realistic spine model. We validate clinical relevance by reproducing a published SCS optimization study for lower-limb motor recovery, matching NEURON-based prediction accuracy while reducing computation time from hours to seconds. Finally, we exploit the GAF’s speed to explore previously intractable parameter spaces: Pareto optimization over pulse waveforms uncovers solutions with approximately seven-fold increased energy efficiency at matched activation, while optimization of multielectrode stimulation (16 currents) increases the functional selectivity index for right hip flexion from 52% (clinical benchmark) to 82% in under one minute. By eliminating computational bottlenecks and enabling gains in efficiency and selectivity, the GAF transforms model-driven optimization into a clinically viable tool for patient-specific neuromodulation. A linear generalization of the activating function predicts neural responses to electrical stimulation as accurately as detailed biophysical models but up to 1000 times faster, enabling rapid patient-specific optimization of spinal cord stimulation.
Categorization depends on multiple brain regions, including the prefrontal cortex, striatum, and hippocampus. However, it remains unclear how these regions interact during category learning. We recorded activity in the medial prefrontal cortex (mPFC), dorsomedial striatum (DMS), and the dorsal hippocampus (dHPC) as rats (n = 9, 3 females) learned a rule-based categorization task. Category-selectivity emerged early in the mPFC. In the DMS and dHPC, category-selectivity increased after learning and peaked before the rats’ decisions. A linear regression analysis revealed that all regions contained neurons with stimulus-related selectivity, decision-related selectivity, and “mixed selectivity”. From the local field potentials (LFPs), Granger causality revealed a learning-related reconfiguration of network synchrony. DMS-mPFC theta (4-10 Hz) causality was strongest during early training sessions and was directional from the DMS to the mPFC. mPFC-dHPC theta causality peaked during middle training sessions and was directional from the mPFC to dHPC. dHPC-DMS theta causality increased across training and was directional from the dHPC to the DMS. mPFC-dHPC causality and DMS-dHPC causality peaked before the rats’ decisions. The results indicate that early training sessions engaged a frontal-striatal circuit, whereas later training sessions engaged a hippocampal-striatal circuit. The transition to a hippocampal-striatal circuit is potentially mediated by top-down mPFC control. Characterization of the interacting dynamics among the prefrontal cortex, hippocampus, and striatum during rule-based category learning in rats.
Human GalNAc-T1 is a mucin-type O-glycosyltransferase that modulates immunity, extracellular matrix formation, and salivary gland development and function. However, its substrates and regulatory mechanisms remain unclear. Here, we investigate the Drosophila melanogaster GalNAc-T1 ortholog PGANT5 in salivary glands and provide details into its mechanism of O-glycosylation. Loss of pgant5 causes irregular secretory granule morphology, alters mucin packaging, and disrupts secretion. Alternative splicing of pgant5 yields pgant5A and pgant5B, with higher expression of pgant5A in salivary glands. Rescue with pgant5A, but not pgant5B, partially restores secretory granule morphology and secretion. A single residue difference within the substrate binding pockets of PGANT5A and PGANT5B increases the in vitro activity and specificity of PGANT5A towards salivary gland mucins relative to PGANT5B, corroborating the role of pgant5A in salivary glands. Our studies illustrate how alternative splicing of an O-glycosyltransferase resulting in modest amino acid changes can influence substrate specificity to regulate tissue-specific biological functions. Loss of pgant5, which expresses the mucin-type O-glycosyltransferase Drosophila PGANT5, disrupts salivary gland function. Alternative splicing of PGANT5 results in subtle differences between PGANT5A and PGANT5B, which tweaks their specificities towards salivary gland substrates and influences secretory granule morphology and secretion.
The Ediacara biota represents Earth’s earliest complex macroscopic organisms, but their biology and ecology remain poorly understood. Here we report Dengyingia pennata gen. et sp. nov., an exceptionally preserved frondose fossil from the late Ediacaran ( ~550–543 Ma) Shibantan limestone of South China. Dengyingia possesses a unique body plan: a bifoliate petalodium of unconstrained, flexible primary branches with pronounced two-sided differentiation—a smooth obverse surface and a reverse surface covered by secondary and tertiary modules. This innovation represents a key morphological advance among Ediacaran fronds. Its architecture, basal growth pattern, and suspension-feeding strategy align with those of modern sea pens (pennatulacean cnidarians), reflecting adaptations for dynamic filter-feeding in currents. Dengyingia bridges the ecological gap between the Ediacara biota and Phanerozoic suspension-feeding animals, demonstrating that complex, animal-like ecological strategies emerged before the Cambrian explosion. These findings expand Ediacaran diversity and provide insights into early animal functional differentiation. An exceptionally preserved Ediacaran frond, Dengyingia pennata, from South China shows a unique branched body plan and functional convergence with modern sea pens, suggesting complex suspension-feeding strategies evolved prior to the Cambrian diversification.
The rapid emergence of multidrug-resistant Escherichia coli highlights the urgent need for effective, host-compatible antibacterial strategies. Targeting bacterial redox homeostasis represents a promising non-antibiotic paradigm, yet rational synergistic approaches remain poorly defined. We show that zinc combined with vitamin K3 exhibits potent antibacterial activity against pathogenic E. coli, including enterotoxigenic ETEC K88, by selectively disrupting redox regulation. Mechanistically, Zn²⁺ targets the redox-sensitive transcription factor SoxR, inactivating the SoxR-SoxS pathway and suppressing the downstream superoxide dismutase (SOD) antioxidative axis, thereby impairing the bacterial adaptive response to oxidative stress and rendering the pathogen highly susceptible to oxidative damage. This effect is further amplified by vitamin K3-mediated superoxide generation, resulting in synergistic bactericidal activity. In the present study, the Zn²⁺-vitamin K3 combination was validated across multiple infection models, including in vitro systems, a Galleria mellonella infection model, and an ETEC K88-induced murine diarrhea model, where it reduced bacterial burden, improved host survival, and alleviated intestinal pathology. Importantly, gut microbiota analysis revealed partial restoration of infection-associated dysbiosis without broad microbial depletion. These findings establish redox homeostasis disruption as a microbiota compatible antibacterial strategy against multidrug-resistant E. coli.
Despite positive outcomes for novel targeted therapy screens in preclinical studies, the majority of follow on GBM clinical trials fail to meet their primary endpoints. This translational gap is partly due to models that do not accurately recapitulate the human GBM-TME and therefore fail to accurately predict patient treatment response. Here, we investigated the NFPp10a/NF5310 syngeneic mouse model of mesenchymal GBM, to assess its translational relevance for evaluating TME-targeting agents. Young and aging C57BL/6 mice bearing orthotopic NFPp10a/NF5310-Luc2 tumours underwent treatment with temozolomide (TMZ), tumour resection, anti-PD1 checkpoint blockade, and/or regorafenib (REGO). TME composition was evaluated using the murine microenvironment cell population (mMCP) counter method and multiplexed immunohistochemistry (multiple iterative labelling by antibody neodeposition). To assess translational relevance and conserved human response patterns, immune cell composition and gene expression changes were directly compared to primary human mesenchymal GBM tumours via further hypothesis generating MILAN analysis and by studying publically available ICI clinical trial data. NFPp10a/NF5310-Luc2 tumour-bearing mice showed resistance to monotherapy and neoadjuvant anti-PD1, with limited response also observed to REGO treatment. mMCP analysis revealed modest increases in CD8+ T-cells, B cells, and monocytes following anti-PD1 and REGO treatment. MILAN analysis further indicated increased cytotoxic T-cells following anti-PD1 therapy. Comparison to untreated-primary GBM and ICI-treated human GBM suggested similar exhausted CD8+ T-cell phenotypes, suggesting the NFPp10a/NF5310 TME reflects the mesenchymal GBM T-cell compartment. Overall, the NFPp10a/NF5310 model recapitulates the GBM-TME architecture and therapeutic resistance patterns observed in human GBM, supporting its use in evaluating select TME-targeting therapies. The NFPp10a/NF5310 glioblastoma model demonstrates clinically relevant response patterns and key tumour microenvironment features of mesenchymal glioblastoma, supporting its utility as a translational platform for therapeutic development.
Triple-negative breast cancer (TNBC), as the most aggressive subtype of breast cancer, has a poor prognosis because of its heterogeneity and complicated molecular mechanism. RAD21, a key component of cohesin complex, play vital roles in cellular functions and regulation of cell identity gene expression in cancer cells. However, it remains incompletely understood how RAD21 regulates tumorigenesis and metastasis in TNBC. Here, we present data suggesting that RAD21 was commonly highly expressed in TNBC and was correlated with poor outcome in invasive breast cancer. RAD21 depletion induced autophagy and suppress proliferation, invasion and xenograft tumor growth of TNBC cells, while RAD21 overexpression promoted lung metastasis. Mechanistically, RAD21 bound to the promoter region of the YAP and RAD21 knockdown increased YAP transcription level and the YAP nuclear accumulation in TNBC cells. Subsequently, YAP promoted a transcriptional program leading to enhanced autophagy flux to restrain TNBC cells proliferation and invasiveness. These findings uncover RAD21 as a promoter of TNBC tumorigenesis and metastasis and suggest a new function of RAD21 in regulating TNBC cells autophagy driven by YAP. RAD21 is linked to poor breast cancer outcomes. RAD21 is here found to drive TNBC tumor growth and invasion in vitro and in vivo by suppressing YAP transcription and YAP-mediated autophagy, revealing a novel YAP-dependent role.
The success of trypanosomes in transitioning between hosts and life stages depends on their ability to manipulate host metabolism, immunity, and behavior. Pathogen-favoring behaviors often link tsetse flies’ host behavioral plasticity and sensory adaptations, but the underlying mechanisms remain unclear. To uncover the molecular basis regulating neuronal sensitivity and transmission behavior in tsetse flies, we applied transcriptomics, metabolomics and validated by RT-qPCR and behavioral assays. Uninfected Glossina pallidipes were more attracted to Trypanosoma congolense-infected mice and metabolites from infected cow urine than to healthy ones. T. congolense infection alters host metabolic pathways, modifying the composition of volatile organic compounds in mice and cattle, which enhanced fly attraction. Infection modulated cellular signaling across multiple sensory modalities, including olfaction, vision, taste, thermosensation, and immunity in G. pallidipes. Trypanosome associated odors doubled the catch of infected G. pallidipes in the field, reducing infected flies by 75% and diseases prevalence in cattle by 40% based on agent-based model. These findings reveal the molecular interaction between T. congolense, G. pallidipes, and cattle, highlighting how the parasite manipulates multiple pathways to enhance adaptation, multiplication, and transmission. Muti-omics investigation reveals how trypanosomes manipulate their vertebrate host metabolites and tsetse flies’ multiple sensory modalities and immunity to enhance its survival and transmission.
The dorsolateral prefrontal cortex contributes to cognitive–affective pain regulation, but the optimal frequency and individualization strategy for high-definition transcranial alternating current stimulation remain unclear. Here we show, across three double-blind experiments in healthy human participants using a capsaicin-induced tonic pain model, that analgesia depends on stimulation frequency and the brain state used for frequency selection. Study 1 compares 10-Hz stimulation of the dorsolateral prefrontal cortex with 10-Hz stimulation of the primary sensorimotor cortex in 40 participants. Study 2 compares 7-Hz theta and 10-Hz alpha stimulation over the dorsolateral prefrontal cortex in 40 participants. Study 3 compares fixed 7-Hz stimulation with individualized theta frequencies derived from pain-free or pain-persistent oscillatory profiles in 28 participants. Participants receive 30 minutes of high-definition transcranial alternating current stimulation during tonic pain with continuous pain ratings. Alpha-band dorsolateral prefrontal cortex stimulation produces stronger acute and sustained analgesia than primary sensorimotor cortex stimulation. Theta-band stimulation produces greater and longer-lasting analgesia than alpha stimulation and is accompanied by increased prefrontal beta- and low-gamma activity. Individualized theta stimulation derived from pain- persistent states yields greater analgesia than fixed-frequency or pain-free-state-based stimulation. These findings identify frequency and brain state as key determinants of analgesic responses to dorsolateral prefrontal cortex stimulation. Three double-blind experiments test how frequency and brain state influence analgesia during tonic pain. Prefrontal theta stimulation tailored to brain activity during pain produces the strongest and most sustained pain relief.
Telomeric DNA replication involves coordinated action of telomerase and DNA polymerase α-primase (Polα/primase), which synthesize the G-strand and C-strand, respectively. The conserved Cdc13-Stn1-Ten1 (CST) complex critically regulates this process by both terminating telomerase activity and stimulating Polα/primase. While telomerase-mediated G-overhang synthesis is well studied, how CST-Polα/primase is recruited to telomeres for C-strand fill-in remains poorly understood. Structural analysis reveals an evolutionary diversity of CST interaction with Polα/primase across organisms. Here, using a combination of AlphaFold3 structural modeling with genetic assays, we unveil the assembly mechanism of CST-Polα/primase for telomere maintenance in Saccharomyces cerevisiae. We structurally characterize the uniqueness and specific determinants of the Stn1-Pol12 interaction in the S. cerevisiae CST-Polα/primase complex. The Cdc13-Pol1 and Stn1-Pol12 interactions together mediate CST-dependent recruitment of Polα/primase to telomeres. Functional analyses reveal that disruption of the CST-Polα/primase interaction shows distinct telomere phenotypes compared to CST disassembly, supporting the model that CST recruits Polα/primase after telomerase elongation is terminated. Together, our findings not only resolve the longstanding question on the assembly mechanism of the CST and Polα/primase complexes at yeast telomeres, but also hint that CST-Polα/primase-dependent C-strand fill-in likely governs recombination-based telomere structure maintenance via controlling 3’ overhang length.
The human auditory system represents sounds at multiple levels, from low-level acoustic features to abstract category- and object-level information. Although selective attention enables listening in complex natural soundscapes, it remains unclear which representational levels are modulated by attention and how this depends on scene structure. Using functional magnetic resonance imaging, representational similarity analysis, and cross-experiment decoding, we examined attentional modulation of auditory representations under three conditions: 1) listening to isolated sounds, 2) attending to one of three overlapping sounds from different categories, or 3) one of three overlapping sounds from the same category. We show that attentional modulation is task-dependent: when competing sounds could not be distinguished by category, attention enhanced low-level acoustic feature processing, whereas in cross-category scenes it targeted abstract category-level representations. Object-identity representations were modulated by attention across both scene types, with category-specific differences, demonstrating flexible, task-dependent targeting of auditory features. Varis et al. use fMRI to study how attention shapes auditory representations in scenes of overlapping natural sounds. Attention enhances acoustic features when competing sounds share a category and categorical features when they differ.
Dominant individuals often structure group organization, but less is known about how social networks reorganize in their absence and how variation among subordinates contributes to collective outcomes. Bumble bees (Bombus impatiens) provide an ideal system to study these dynamics: queens typically monopolize reproduction, but in some contexts, individual workers can adopt queen-like social roles. Using multi-animal pose tracking, we compare matched queenright and queenless partitions from the same source colonies, quantifying over 80 million social interactions. Queenless colonies exhibit increased behavioral variation and contain a subset of highly influential workers with elevated movement, spatial centrality, and reproductive activity that is absent in queenright conditions. The emergence of these individuals coincides with a shift from centralized to decentralized, efficient network architectures. These results demonstrate that queen presence constrains latent worker variation, revealing how individual behavioral differences can scale up to reshape collective social organization in hierarchical societies. Automated tracking of bumble bee colonies shows that queen loss unmasks latent worker variation, allowing a subset of workers to become highly influential and reorganize colony social networks.
Within host-microbiome-pathogen systems, the host chemical microenvironment is often overlooked despite its inherent role in host physiology. We used a multifaceted experimental approach encompassing culture-dependent and -independent methods, genomic and metagenomic analyses, and deep neural network modeling to assess the impact of host skin chemistry and the bacterial microbiome on the growth of Ophidiomyces ophidiicola (ophidiomycosis, snake fungal disease). Results suggest that host skin lipid chemistry (e.g., oleic acid, squalene) and bacteria isolated from wild snake skins (e.g., Chryseobacterium sp. and Stenotrophomonas maltophilia) suppress O. ophidiicola growth. Notably, the O. ophidiicola genome contains biosynthetic gene clusters (BGCs) that encode metabolites that may suppress host lipid production, facilitating fungal pathogenicity. A contrastive deep neural network produced a near-perfect alignment of snake skin lipid and microbiome profiles for both individual snakes and disease states. BGCs from bacterial genomes isolated from snake skin overlap with metagenome profiles from wild snakes and are associated with disease state. We highlight antifungal activity found in the diverse lipid milieu of snake skin and bacterial-fungal interactions that structure the skin microbiome. Our results illustrate a strong relationship among a fungal pathogen, the microbiome, and host skin lipid chemistry that may underpin disease susceptibility. Deep learning models accurately align multimodal data including snake skin lipid chemistry and microbiome-pathogen interactions in the snake fungal disease system.
Cells are the basic unit of life. In multicellular organisms, cells are organized into tissues. This enables a division of labor where tissues perform complex tasks via coordinated actions of specialized cell types. Accordingly, the cellular collective determines tissue function. A comprehensive overview of which cell types exist in solid tissues is lacking. Using skeletal muscle as a model, we discuss basic principles for cell type classification, summarize 62 unified definitions of cell types in terms of lineage, molecular signatures, and function from the literature, and discuss how these cell types contribute to muscle function. For cell types for which quantitative data was available, we compare abundances in immunohistology and single cell and single-nucleus RNA sequencing data, revealing cell types that are commonly over- or underrepresented in each method. The result is a cell type resource that will serve as a benchmark for single-cell studies of skeletal muscle.
Magnetic susceptibility in brain white matter is anisotropic because of the ordered arrangement of myelin lipids, whereas paramagnetic iron is generally not considered a source of susceptibility anisotropy. Whether spatially organized paramagnetic sources also contribute to tissue-level susceptibility anisotropy remains unclear. Here we analyze multi-orientation gradient-echo MRI and diffusion MRI data from a paraformaldehyde-fixed postmortem adult male western chimpanzee (Pan troglodytes verus) brain and use DECOMPOSE-QSM to separate paramagnetic and diamagnetic susceptibility components before tensor reconstruction. The diamagnetic component shows the expected anisotropy in white matter and aligns with major diffusion-based fiber orientations. The paramagnetic component also shows coherent anisotropy, most prominently in deep gray matter and in parts of white matter. Numerical modeling further supports that an ordered spatial arrangement of otherwise isotropic paramagnetic sources can produce orientation-dependent susceptibility. These results suggest that source-separated susceptibility tensor imaging provides complementary information about tissue-specific susceptibility organization beyond conventional susceptibility tensor imaging. Source-separated susceptibility tensor MRI in a postmortem chimpanzee brain reveals distinct paramagnetic and diamagnetic anisotropy patterns, providing complementary views of primate brain microstructure.
Many clinically important bacterial pathogens, including Pseudomonas, Vibrio, Neisseria, and Acinetobacter species, employ dynamic extracellular appendages called type IV pili (T4P) to facilitate virulence through cyclical extension and retraction of pilus filaments. To dissect how T4P dynamics govern pathogenesis, we engineered a genetic system to precisely tune pilus length across a continuum. We demonstrate that pilus length determines four major T4P-dependent virulence traits in P. aeruginosa (motility, surface sensing, biofilm formation, and phage infection) and reveal a hidden subpopulation of pili that are unable to interact with environmental substrates or host cells, rendering them non-contributing to T4P-mediated functions. Integrating molecular dynamics simulations, we show that low inner-membrane abundance of the major pilin forces the extension mechanism into transient idle states, restricting both velocity and final length. Molecularly, this finding reveals how two key biophysical parameters, pilin abundance and diffusion, impose a fundamental physical constraint on T4P assembly, and that regulating pilin abundance presents a strong lever over regulating pilus count for controlling the amount of functionally contributing filaments. Contrary to the view that retraction force dictates T4P-mediated behaviors, our results establish extension dynamics as the overlooked bottleneck constraining retraction-enabled virulence traits, with population heterogeneity in length possibly enabling risk mitigation. The availability of the major pilin PilA limits extension of type IV pili and directly impacts pilus-dependent virulence behaviors.
The sustained human-to-human transmission of Clade IIb monkeypox virus (MPXV) during the 2022 outbreak underscores the need to define host determinants of viral fitness across species. ISG15 encodes a ubiquitin-like modifier with antiviral activity, yet its role in MPXV infection and host adaptation remains unclear. Using representative strains from Clades Ib and II, we identify a striking species-specific difference in ISG15-mediated restriction. In murine cells, ISG15 deficiency primarily increases viral protein accumulation with minimal effects on infectious viral production, indicating limited antiviral restriction. In contrast, in human cells, ISG15 restricts replication of recent epidemic strains since ISG15 deficiency enhances viral growth, establishing ISG15 as a restriction factor in humans. ISGylome profiling identifies candidate viral and host substrates underlying this effect. Together, these findings reveal species-specific ISG15-dependent control of MPXV infection and support a model in which rodents serve as permissive reservoirs while humans represent a more restrictive host due at least in part to the antiviral activity of human ISG15. We study ISG15 during MPXV infection in human and murine cells. ISG15 deficiency increases infectious virus production in human cells but alters viral protein accumulation in murine cells, and ISGylome analysis identifies ISGylated viral proteins.
