Browse Articles

Relationship between CAR spatial organization and CAR T cell function revealed by imaging flow cytometry and transcriptomic profiling 2260438

Abstract Introduction Chimeric antigen receptor (CAR) T cell therapies have revolutionized treatment for hematologic cancers, yet precise detection and characterization by flow cytometry remain challenging. Using high-sensitivity 1-step CAR detection reagent, we characterized CAR expression in primary T cells. Methods Human T cells were isolated from peripheral blood mononuclear cells, stimulated with anti-CD3/CD28 Dynabeads and transduced with a lentiviral vector encoding a B-cell maturation antigen (BCMA)-specific CAR. The transduced cells were further expanded in culture for 2 weeks prior to cryopreservation. Upon thawing, cells were rested overnight in IL-2-supplemented media before being re-stimulated in co-cultures assays with the U266 target cell line and/or subjected to spectral and imaging flow cytometry. Results Spectral flow cytometry enabled clear discrimination of CAR T cells spiked into donor PBMCs from endogenous T cells, allowing detailed profiling of activation, exhaustion, and memory phenotypes. Imaging flow cytometry with BD FACSDiscover™ S8 and BD CellView™ Technology revealed two distinct CAR spatial patterns: circular and polarized. Functional assays with BCMA-CAR T cells showed antigen-specific degranulation, with CD107a externalization co-localizing with CAR in cells exhibiting the circular pattern. These cells also displayed G1-quiescent transcriptional signatures, indicating readiness for activation and enrichment of immune response pathways. In contrast, polarized CAR+ subsets expressed higher levels of MKI67, HMGB1, and cell cycle genes, consistent with proliferative activity. Conclusion These findings suggest CAR spatial organization correlates with functional states and demonstrate the power of integrating advanced flow and imaging cytometry to resolve CAR T cell phenotypes and spatial dynamics with high precision. BD flow cytometers are Class 1 Laser Products. For Research-Use Only. Not for use in diagnostic or therapeutic procedures. ©2025 BD. All rights reserved. NPM-7267. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

CAR-T with ICOS costimulation lacks therapeutic efficacy in patients due to strong trogocytosis and failure of expansion 2261203

Abstract Introduction CAR-T therapy targeting B-cell antigens is effective for relapsed hematologic malignancies, but antigen escape remains a challenge. We previously showed dual CD19/CD20-4-1BB CAR-T therapy was highly effective (Nat Med, 2020). We further developed trispecific CAR-T cells (CD19/CD20/CD22) incorporating ICOS signaling (Sci Transl Med, 2021). Despite promising preclinical activity, trispecific-ICOS CAR-T cells failed to expand and showed no efficacy in a Phase I trial (NCT05094206; ASH 2024). Here, we investigated mechanisms underlying this failed response. Methods Based on our IRB approved protocol and using patient-derived PBMCs and leftover samples from treated patients, we generated tested bispecific-4-1BB, bispecific-ICOS and trispecific-ICOS CAR-T cells. Results NSG mice receiving bispecific- or trispecific-ICOS CAR-T cells had worse survival than those with 4-1BB CAR-T cells. In vivo, ICOS CAR-T cells failed to expand and showed an exhaustion phenotype. When co-culture with Raji cells, ICOS CAR-T cells significantly enhanced surface-FMC63 loss compared to 4-1BB counterparts. Surface-FMC63 expression was mostly recovered at 40 hours on 4-1BB CAR-T cells, but not on ICOS CAR-T cells. Since trogocytosis can mediate CAR internalization and antigen loss on tumor, we examined CD19 transfer. ICOS CAR-T cells displayed higher and more sustained CD19 uptake and fratricide reflected by apoptosis than 4-1BB cells. Consistently, when co-culture with patient autologous B cells, ICOS CAR-T cells again showed greater CAR loss and apoptosis. Extended co-culture mimicking chronic antigen stimulation revealed expansion of 4-1BB but not ICOS CAR-T cells, the latter showing reduced proliferation (Ki67) and elevated exhaustion markers. Conclusion In conclusion, the ICOS domain promotes trogocytosis and fratricide, explaining the poor persistence and function of ICOS CAR-T cells observed in our trial. Thus, our findings raise significant caution for using CAR constructs that rely on ICOS for CAR activation. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Influenza mRNA-LNP vaccines enhance the immunogenicity of protein-based influenza subunit and inactivated split vaccines after co-administration 2253871

Abstract Introduction Current commercially available split inactivated (split) influenza vaccines offer suboptimal protection, highlighting the need for broader and more durable influenza vaccination strategies. We hypothesized that cross-protective influenza mRNA vaccines would enhance the immunogenicity of influenza split and protein-based vaccines via an immunostimulatory lipid nanoparticle (LNP) delivery platform after co-administration. Methods To address the current limitations of the influenza vaccine, we developed cross-protective mRNA-LNP vaccines encoding the conserved M2 ectodomain (5xM2e) or a chimeric M2e-stalk hemagglutinin (HA) protein. Results Mice immunized with 5xM2e or M2e-stalk HA mRNA-LNP generated influenza antigen-specific IgG antibody. Injection of mice with mRNA-LNP vaccines exhibited an acute recruitment of monocytes, macrophages, and dendritic cells. Additionally, co-administration of the 5xM2e and M2e-stalk mRNA-LNP with a conventional split or protein (stalk HA, neuraminidase) influenza vaccines at low doses significantly enhanced IgG antibody responses to viral antigens and offered protection against both homologous and heterologous influenza viral strains. This adjuvant effect was consistently observed in Moderna and Pfizer LNP formulations and other mRNA vaccines in combination with inactivated split or protein vaccines. Conclusion These findings underscore the potential of mRNA-LNP vaccines, which exhibit not only mRNA delivery platforms but also adjuvant effects on co-administered protein-based vaccines. This combinatorial immunization approach holds promise toward enhancing the immunogenicity of a broadly protective influenza vaccine. Funding Source This work was supported by NIH NIAID grants AI154656 (S.M.K.) and F31AI186351 (P.G.) Topic Categories Vaccines and Immunotherapy (VAC)

Use of a 50-Marker Mass Cytometry Intracellular Cytokine Staining Panel Reveals Unforeseen Functional Diversity Within Cell Therapy Products 2258158

Abstract Introduction Despite significant investment, most cancer therapeutics, including CAR T cell therapies, fail in the clinic. Functional assessment of CAR T cells is typically limited to Th1 cytokines (for example, IFNγ), overlooking the known heterogeneity of the T cell subsets that make up the products, including populations with immunosuppressive capacity. This diversity introduces potential “saboteur” cells with off-target effects that may undermine therapeutic efficacy. Surface marker profiling is insufficient for functional characterization, yet traditional flow cytometry lacks resolution for certain intracellular targets, particularly immunosuppressive cytokines. Mass cytometry (CyTOF™ technology) overcomes this technical limitation, providing superior intracellular signal resolution in the context of large panel analyses, a prerequisite of deep functional assessment of single cells. We hypothesized that there are sizeable proportions of saboteur T cells in cell therapy products that exert functions after CAR stimulation that act in direct opposition to the therapeutic intention of the drug. Methods To assess this, we developed a 50-marker CyTOF intracellular cytokine staining (ICS) panel with 24 functional readouts, enabling clear detection of individual human T cells producing immunosuppressive cytokines such as IL-10, IL-13 and TGF-b in human PBMC and cell therapy products. Results Our data revealed an unforeseen prevalence of T cells that cross dogmatic lineage paradigms when stimulated. For example, approximately 30% of T cells from both peripheral blood and a cell therapy product produced the TGF-b-inducing cytokine amphiregulin and often did so simultaneously with IFNg within individual T cells. Conclusion From these results, we predict that application of this novel CyTOF ICS panel for cell therapy development protocols could prove transformative in the treatment of many cancers and other diseases. Funding Source Standard Biotools Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Self‐Regulating Interfacial‐Boosted Electrolytes for Fast‐Charging and Long‐Life Aqueous Batteries

ABSTRACT Aqueous batteries are promising for large‐scale energy storage due to inherent safety and low cost; however, their long‐term cycling stability is fundamentally limited by interfacial degradation. Although existing strategies such as “water‐in‐salt” and water–organic hybrid electrolytes can widen the electrochemical stability window (ESW), trace water electrolysis still occurs at the interface, leading to gradual pH drift, electrode dissolution, and eventual battery failure. Here, we propose a self‐regulating interfacial‐boosted electrolyte that effectively addresses these challenges. By introducing phosphate‑based components into wide‑ESW electrolytes, this design autonomously ensures pH stability via intrinsic buffering capability, spontaneously forms a protective cathode–electrolyte interphase in response to metal‐ion dissolution, and reconfigures the ion solvation structure to widen the electrochemical stability window and lowers the interfacial impedance. Demonstrated in a Na 1 . 85 Mn[Fe(CN) 6 ] 0 . 97 ·2H 2 O//NaTi 2 (PO 4 ) 3 full cell with an energy density of 81.7 Wh kg −1 , this electrolyte enables fast charging (75% capacity retention from 1 C to 80 C) and ultra‐long cycling stability (71% capacity retention after 20 000 cycles at 80 C). The strategy offers an effective and generalizable pathway toward high‐performance aqueous batteries.

A Porous Interfacial Photothermal Layer of Fused Core–Shell Carbon‐Polymer Nanospheres for Directional Salt Crystallization and Zero‐Liquid‐Discharge

ABSTRACT Interfacial solar evaporation offers a sustainable route for freshwater production; however, effective zero‐liquid‐discharge (ZLD) operation under hypersaline conditions remains fundamentally challenging because high salinity suppresses water activity and vapor pressure while intensifying coupled heat, mass, and salt management. Herein, we report a hierarchical hydrogel evaporator featuring a thin, porous interfacial photothermal layer composed of fused core–shell carbon‐polymer nanospheres, fabricated via interfacial precipitation polymerization. The surface‐confined photothermal layer ensures effective thermal confinement and abundant liquid‐vapor interfaces, while the underlying hydrogel matrix sustains continuous water replenishment, delivering an exceptional evaporation rate of 6.2 kg m −2 h −1 under one‐sun irradiation without requiring bulky 3D architectures. During ZLD operation, direction‐dependent transport within the thin porous layer drives lateral advection of concentrated brine away from the active evaporation interface while suppressing through‐thickness salt accumulation, thereby enabling localized crystallization in the distal region. As a result, near‐complete spatial separation of water evaporation and salt crystallization is achieved from 20 wt.% brine. Outdoor demonstration further delivers 26.6 L m −2 day −1 of freshwater and 7.7 kg m −2 day −1 of recovered salt, underscoring the potential of this platform for practical ZLD desalination and mineral recovery.

Exploring the organismal role of UFMylation in development, stress resilience, and neurological function in Caenorhabditis elegans

Study on the effect of ZrO2 loading on the formation of CuO-ZnO-ZrO2-Al2O3/ZrO2-modified HZSM-5 bifunctional catalysts as potential catalysts for CO2 valorization

Novel mouse models for immunology research 2308519

Abstract Introduction The Jackson Laboratory (JAX) is a centralized resource of genetically defined mutant and transgenic mice to support the scientific research community. This poster will feature new models designed to advance the field of immunology research. Methods Targeted mutagenesis, transgenic injection and CRISPR/Cas technologies are used to generate the mouse models. Results The NSG (NOD scid IL2rgc-/-) portfolio of strains for transplantation/engraftment and humanized mouse research continues to grow, with recent humanized alleles (e.g., IL6, TSLP) expanding the options for immunology studies. New mouse lines for cancer immunology and antibody therapeutic studies combine humanized FCGRT and ALB alleles for human preclinical pharmacokinetics (HuPK), as well as humanized PD-1 and PD-L1 alleles. Novel fluorescent strains include Gene Editing Reporters for visualizing homology-directed repair (HDR) and non-homologous end joining (NHEJ) repair events, as well as several CRISPR Editing, Bxb1 Integrase Editing, Base Editor, Prime Editor and DNA repair pathway tools. In addition to cryopreserving each strain, JAX employs a genetic quality control program that confirms mutation identity and genetic background, and screens for common unwanted alleles (via testing for neo, Cre, FLP, GFP, RFP, etc.). Conclusion Immunology researchers are encouraged to query the collection of innovative mouse models using the JAX Mouse Search website (mice.jax.org). This resource includes models created by many generous donating institutions. Researchers are encouraged to donate their mouse lines via a very short Strain Submission form (jax.org/donate-a-mouse). JAX is supported by the NIH, The Howard Hughes Medical Institute and other private charitable foundations. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)

Longitudinal T cell receptor repertoire sequencing identifies specific T cell clones with rare CDR3 residues against malaria 2306928

Abstract Introduction Naturally acquired immunity (NAI) against malaria develops gradually with age and wanes in the absence of continuous antigen exposure. Comparative analysis of T cell receptor (TCR) repertoires provides valuable insight into T cell development and immune responses during infection. Methods In this study, we analyzed longitudinal TCRb repertoire data from a well-characterized cohort of Malian young children with malaria and established a statistical framework to identify TCRb clone proliferation and contraction over time. Results We detected hundreds of malaria-responsive TCRbs per individual, encompassing both memory-like and newly emerged clones. Although these responding TCRs were largely private, analysis of the complementarity-determining region 3 (CDR3) antigen-contacting FG loop revealed distinct amino acid doublet usage patterns. Malaria expanded CD4+ TCRbs preferentially utilized rare residue doublets such as SW and AM, whereas CD8+ TCRbs promoted DV, FQ, RW, and SI doublets. Conclusion As memory T cell responses depend on effective antigen presentation, our approach provides new mechanistic insights into the development of NAI against malaria and offers a potential framework to inform vaccine design. Funding Source NIH intramural research funding Topic Categories Immune Mechanisms of Human Disease (HUM)

Morphologic and immune characterization of threespine stickleback (Gasterosteus aculeatus) gastrointestinal tract for making accurate in vitro models 2258640

Abstract Introduction In vitro intestinal culture systems are powerful models to study mucosal immunity and host-pathogen/ commensal interactions. However, existing cell line-based models are limited as they can only represent one intestinal region, making representative organ modeling difficult. As our aim is to develop an accurate culture model of the entire threespine stickleback (Gasterosteus aculeatus) intestine, an in-depth intestinal characterization of both stromal and immune cell populations is required. Methods H&E and Alcian blue staining was used to visualize intestinal morphology and goblet cell abundance, respectively. Neutrophilic granulocytes were visualized with myeloperoxidase staining. RNA-ISH was also used detect lymphocyte (IgM+ and CD4+ cells) abundance and distribution within the intestine. Quantification was performed using Fiji software. Results Visualization of all intestinal regions (proximal, mid, hind, and rectum) indicated villi length and complexity (villi branching) decreased moving from the proximal to hind intestine. Conversely, goblet cell density increased distally along the intestine. Myloperoxidase positive cells were distributed equally across all regions of the intestine. Additionally, RNA-ISH revealed that lymphocytes did not form lymphoid structures, but were diffusely distributed along the villi in the epithelial layer and lamina propria and primarily consisted of CD4+ cells. Conclusion This work establishes a baseline for normal stickleback intestinal morphology and cell composition for each region, and provides guidance for the generation of region-specific in vitro models. Our ongoing work will establish stickleback intestinal cell lines and transwells systems which represent the heterogeneous composition and physiology of these anatomical zones. Future work will use these tools to model host-microbiota interactions and quantifying morphologic and immune cell population changes during Schistocephalus solidus infection, a natural helminth of threespine stickleback. Funding Source Gordon and Betty Moore Foundation Symbiosis in Aquatic Systems Initiative (GBMF9323) Topic Categories Veterinary and Comparative Immunology (VET)

Distinct tissue niches contribute to prostate tissue-resident memory T cell differentiation and heterogeneity 2256438

Abstract Introduction The prostate is an exocrine organ, a barrier tissue to the male reproductive system, and a site of malignancy, yet prostate tissue-resident memory T cells and the unique cues in the tissue environment remain unexplored. Methods To study T cell memory formation in the prostate, we utilized acute systemic viral infection with lymphocytic choriomeningitis virus (LCMV) Armstrong, which has been used to study CD8+ tissue-resident memory T cells in a broad range of other tissues. To further characterize this population, we employed genetic mouse models, scRNA-seq and spatial transcriptomics to investigate antigen-specific prostate tissue-resident memory T cell formation, differentiation, and heterogeneity. Results Acute infection generated a population of long-lived CD8+ T cells with characteristics of tissue residency. These prostate tissue-resident memory T cells protect against reinfection and display functional and phenotypic heterogeneity in both mice and humans. Functional interrogation of TGFβ-, IL-7-, and IL-15-derived signals reveal subset-specific prostate tissue-resident memory T cell dependencies according to their niche-dependent phenotypes and distinct cytokine sources. For instance, tissue-resident memory-promoting cytokines IL-15 and TGFβ are highest in the prostate epithelium where CD8+ T cells are most enriched for tissue-resident memory-associated gene expression and analysis of human prostate samples identified similar patterns of both cytokine and CD8+ T cell localization. Conclusion We define a spatial framework of heterogenous T cell residence in the prostate, with prostate tissue-resident memory T cell heterogeneity being spatially segregated based on discrete cytokine and chemokine niches, with divergence of T cell transcriptional states between T cells localized to the stromal versus epithelial regions in both mice and humans Funding Source F31AI176705 Topic Categories Mucosal and Regional Immunology (MUC)

Alterations in Kindlin-3 recruitment to plasma membrane during neutrophil recruitment visualized by intravital imaging 2260610

Abstract Introduction Neutrophils respond rapidly to infection and are highly migratory cells. Migratory behavior is coordinated by integrins which bind to ligands to regulate crawling. Activation of integrins is tightly regulated by proteins including Kindlin-3 (K3). Lack of K3 leads to a failure of integrin activation and neutrophil migration resulting in a susceptibility of the host to life-threatening infections and death, a disease known as leukocyte adhesion deficiency 3 (LAD3). The mechanism by which K3 regulates integrin activation remains unclear. We hypothesize that K3 is recruited early to the membrane prior to integrin activation whereupon it then recruits other proteins to mediate integrin conformational changes. Methods We generated a knock-in mouse by fusing a fluorescent protein, mScarlet-I, to K3 (KI-K3), allowing tracking of KI-K3 subcellularly using live animal intravital imaging (IVM). Results We confirmed that all hemopoietic cells express KI-K3 and the transgenic mouse survives to adulthood and old age. However, this fusion induced allelic hypomorphicity as it reduced total KI-K3 mRNA and protein levels and caused compensatory neutrophilia. We studied neutrophil migration in the cremaster muscle and skin. Imaging revealed that KI-K3 is homogenously distributed during circulation, concentrated to the neutrophil-endothelial interface during luminal adherence, crawling and transmigration and polarized to the cell lamellipodia and uropod after diapedesis and during migration within tissues. We bred KI-K3 mice to mice expressing human β2-integrin (hITGB2) and quantified integrin activation using the antibody mAb24 which binds to the high-affinity conformation of β2-integrin. Activation of β2-integrin was reduced by half in hITGB2/KI-K3 mice as compared to hITGB2 mice alone. Conclusion Our real time in vivo observations demonstrate that localization of K3 within neutrophils is highly dynamic and reorganizes differentially during the recruitment cascade. Funding Source N/A Topic Categories Cellular Adhesion, Migration, and Inflammation (CAM)

Heligmosomoides polygyrus bakeri secretions (HES) promote Treg differentiation and inhibit pathogenesis in a mouse model of vitiligo 2266584

Abstract Introduction Vitiligo is an autoimmune skin disorder characterized by the progressive destruction of melanocytes, leading to patchy depigmentation. Heligmosomoides polygyrus bakeri is a murine intestinal helminth known for potent immunoregulatory secretions (HES) that promote tolerance across several autoimmune models. Given the central role of Tregs in maintaining peripheral tolerance and their deficiency or dysfunction in vitiligo, we hypothesized that HES could suppress anti-melanocyte immunity by promoting Treg differentiation. In this study, we investigated whether HES treatment could (1) promote functional Treg differentiation in vitro, and (2) inhibit vitiligo pathogenesis in vivo. Methods For in vitro studies, naïve mouse CD4+ T cells were cultured for 72h ± HES (10 ug/mL). Foxp3 induction and suppressive capacity were measured by flow cytometry. For in vivo studies, vitiligo was induced in k14*mSCF mice by adoptive transfer of transgenic CD8+ T cells, and animals were treated with purified HES (25 ug 3x per week) or vehicle control. Disease progression was assessed by depigmentation scoring, histology, and flow cytometry of skin, lymph nodes and spleens. Results In vitro, HES induced Foxp3 expression from naïve CD4+ T cells but did not cause expansion of existing Treg populations. In vivo, HES-treated mice displayed markedly reduced depigmentation and preserved epidermal melanocytes compared to controls. Flow cytometric analysis revealed increased Foxp3+CD25+ Tregs and decreased CD8+ effector cells in lymphoid tissues. Conclusion Together, these findings demonstrate that HES suppresses vitiligo in vivo by promoting regulatory T cell differentiation, suggesting a novel therapeutic approach for autoimmune skin disease. Funding Source NIH-NIAID (U01AI176310), NIH-NIAMS (P50AR080593) Topic Categories Therapeutic Approaches to Autoimmunity (THER)

TLR2/TLR7 Conjugated Adjuvant Enhances Anti-Nucleoprotein Immunity and Provides Protection Against Lethal H1N1 Influenza Infection 2253752

Abstract Introduction Influenza A virus (IAV) poses a significant public health threat due to its high mutation rate. Antigenic drift (slight changes in the outer coat proteins) leads to seasonal epidemics, while antigenic shift (new hemagglutinin (HA) or neuraminidase (NA) antigens) can produce novel strains with pandemic potential. Limited population immunity to these new strains increases the risk of severe illness and mortality, underscoring the urgent need for a universal vaccine. Methods To develop a universal influenza A virus (IAV) vaccine that induces robust B and T cell memory responses we take a two-layered approach. First, we utilize toll-like receptor (TLR) agonists as vaccine adjuvants and second, we add a conserved internal protein (nucleoprotein (NP)) to increase the breadth of the immune response and activate T cell responses to provide a second armament of protection. Results We show that a conjugated, dual TLR2/7 agonist CL413 and NP antigen can enhance both antibody and T cell responses after intramuscular (i.m.) injection and provide enhanced heterosubtypic immunity to lethal IAV infection compared to either agonist alone. In determining the utility of this approach across mouse strains we found that BALB/c mice generated a hyperinflammatory response to the initial i.m. vaccination with CL413 and antigen, with the TLR2 agonist being responsible for the higher side effects. In contrast, the i.m. injection of CL413 (or TLR agonist) in C57BL/6 strain showed no adverse outcomes. Interestingly, vaccination of BALB/c mice via the intranasal (i.n.) route did not induce visible side effects, yet provided complete protection against challenge. Conclusion Together, these data indicate that conjugated adjuvants with conserved IAV proteins provide superior heterosubtypic protection compared to antigen alone, or antigen with singular agonists. Investigating the protective mechanisms of this combination strategy, brings us closer to developing a universal vaccine against both seasonal and pandemic IAV strains. Funding Source Biomedical Research Scholars Program, Trudeau Institute Topic Categories Vaccines and Immunotherapy (VAC)

MARCKS Drives Immunosuppressive Signaling in Tumor-Associated Macrophages to Promote Lung Cancer Progression 2258297

Abstract Introduction Lung cancer is the deadliest pulmonary malignancy, driven in part by immune evasion and limited treatment options. Among key immunoregulatory players, tumor-associated macrophages (TAMs) promote angiogenesis, immunosuppression, and metastasis via M2-like polarization. MARCKS (myristoylated alanine-rich C-kinase substrate), an oncogenic regulator, is highly expressed in both tumor cells and TAMs, with elevated levels linked to cancer progression. However, its role in TAM-mediated immune modulation remains poorly understood. Methods We analyzed single-cell RNA sequencing data from lung cancer patients. TAMs were stratified into MARCKS-proficient (MARCKS+) and -deficient (MARCKS⁻) subsets. Differential expression and pathway enrichment analyses were performed, and CellChat was used to assess intercellular communication. Results MARCKS+ TAMs were enriched for immunosuppressive pathways, including IL-10 family cytokines, efferocytosis, and CD163-associated M2 programs. CCR5-related pathways implicated proliferative and migratory signaling, while suppression of T cell immunity suggested immune evasion. CellChat analysis showed significantly increased communication by MARCKS+ TAMs. Key outgoing ligands included SPP1, CCL, MIF, VEGF, TNF, and TGF-β, whereas incoming signals featured TGF-β, TNF, ITGB2, and SPP1. MARCKS+ TAMs uniquely received TGF-β, a potent M2 inducer, and secreted SPP1, linked to metastasis, therapy resistance, and fibroblast differentiation into cancer-associated fibroblasts. MIF—CD74—CXCR4 signaling further contributed to Treg recruitment and NK cell suppression. Conclusion MARCKS+ TAMs amplify immunosuppressive signaling that enhances macrophage polarization, immune evasion, and tumor-stromal crosstalk. By driving TGF-β- and SPP1-mediated interactions with tumor and immune compartments, MARCKS emerges as a key regulator of TAM-driven immunosuppression and a potential therapeutic target in lung cancer. Funding Source This work was supported by the NIH grant R01HL146802, the UCOP TRDRP grant T32IR5347, and the DoD PRMRP grant W81XWH2110086 (#PR202411). Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Interfacial Self‐Healing Polymer Electrolytes With Gradient Covalent–Noncovalent Dynamic Bonds for 4.6 V‐Class Lithium Metal Batteries

ABSTRACT Solid‐state lithium batteries (SSLBs) have attracted significant attention due to their high energy density and safety. However, the volumetric expansion/contraction of electrodes, along with persistent interfacial side reactions, leads to mechanical stress damage and failure of interfacial ion transport. Herein, we report an interfacial self‐healing polymer electrolyte (SHPE) based on gradient covalent–noncovalent dynamic bonds, enabling multi‐interfacial dynamic self‐healing throughout the lifecycle of SSLBs. Via molecular engineering, high‐strength boronic ester bonds are combined with ultrafast dynamic responsive hydrogen bonds to enable efficient room‐temperature self‐repair of interfacial defects in SSLBs. The rapid dynamic exchange between polymer chains further promoted fast lithium‐ion conduction, achieving an ionic conductivity of 1.6 × 10 −3 S cm −1 (25°C). The Li|SHPE|Li cells demonstrated stable cycling for >3000 h. The incorporation of oxidation‐resistant boron‐ and fluorine‐containing functional groups extended the electrochemical window to 5.2 V. LiFePO 4 |SHPE|Li and LiCoO 2 (LCO)|SHPE|Li cells delivered >1000 and 700 cycles, respectively, while being compatible with LCO under high voltage (4.6 V) and loading (14 mg cm −2 ). Furthermore, the LCO|SHPE|Li (1.2 Ah) and LCO|SHPE|Si‐C (2.5 Ah) pouch cells exhibited 200 and 400 cycles, respectively, and passed nail penetration safety tests. This study provides an interfacial self‐healing strategy for developing high‐energy‐density and high‐safety SSLBs.

Space‐Confined Chemical Vapor Deposition of 2D FeS With Crossover Magnetoresistance and Ultra‐High Conductivity

ABSTRACT 2D iron chalcogenides (FeX, X = S, Se, Te) are emerging as an appealing class of materials, owing to their intriguing physical properties (e.g., 2D magnetism, interface superconductivity), and great potentials in next‐generation applications (e.g., spintronics, quantum computing). Recently, FeS has attracted particular attentions due to the discovery of altermagnetism in bulk state. However, the intrinsic physical properties of 2D FeS are still unclear, since the syntheses of high‐quality FeS with thicknesses down to 2D limit remain challenging. Herein, we report the direct syntheses of thickness‐tunable, high‐quality 2D FeS flakes with low defect density and thus strict stoichiometry, via a space‐confined chemical vapor deposition (CVD) strategy. Significantly, we observe an unsaturated magnetoresistance with a crossover behavior (positive‐negative‐positive with decreasing the temperature from ≈350 to ≈2 K), and an ultra‐high conductivity (≈2.8 × 10 7 S m −1 at 2 K) in 2D FeS. Leveraging such high conductivity and robust air stability, we demonstrate 2D FeS as ideal contact electrodes in monolayer‐MoS 2 ‐based field effect transistor, achieving a mobility more than 3 times higher than that of Ti/Au‐contacted device. This work should hereby promote the fundamental property explorations of 2D nonlayered materials, and their applications in next‐generation electronic/spintronic devices.

The C-terminal domain of RD3 enables accumulation of retinal membrane guanylyl cyclase (RetGC) in photoreceptor outer segment

Temporal-spatial reasoning over hypergraph knowledge structures for multimodal retrieval-augmented generation

Abstract Existing graph-based retrieval-augmented generation (RAG) systems represent knowledge with binary relations and rely primarily on semantic similarity for retrieval. This design struggles with multimodal queries requiring temporal constraints, spatial relationships, or higher-order interactions among entities. We present HyperTSRAG, a multimodal RAG retrieval algorithm that models knowledge as a hypergraph and performs explicit temporal-spatial reasoning during traversal. HyperTSRAG performs a bipartite-alternating best-first traversal that alternates between entity nodes and hyperedge nodes to capture higher-order connectivity and ranks candidate evidence using a scoring function applied within traversal that integrates semantic similarity, temporal coherence, spatial overlap, and structural importance. We evaluate HyperTSRAG on a benchmark corpus of 1,000 multimodal documents (text, images, audio, and video) with 500 queries spanning simple lookups through complex multi-hop reasoning. For complex queries, HyperTSRAG achieves 78.3% Recall@10, improving by 12.3% over GraphRAG and 18.3% over LightRAG, while maintaining a 95th-percentile latency of 1.83 s. On temporal-spatial subsets, HyperTSRAG attains 85.2% accuracy on temporal queries and 81.6% on spatial queries. Ablation studies show that individual scoring components contribute 2–5% gains. Relative to a matched Projected-Binary-RAG control derived from the same extracted hyperedges, HyperTSRAG improves Recall@10 by 9.1%, while a separate binary-graph traversal ablation yields a 13.4% decrease, underscoring the value of native n-ary structure. These results support hypergraph-native traversal with temporal-spatial-aware scoring as an effective retrieval paradigm for the evaluated multimodal RAG setting, particularly for constraint-driven queries that semantic-only retrieval cannot address.