Browse Articles

HSV-2 Infection Induces Lasting Transcriptional and Functional Changes in Vaginal Tissue NK Cells 2258179

Abstract Introduction Mucosal immunity in the vaginal tract (VT) remains poorly defined despite its essential role in defense against sexually transmitted infections. Natural killer (NK) cells are critical for antiviral control, and increasing evidence indicates that they can acquire adaptive-like features following infection. While memory-like NK cells have been established in response to CMV, it is unknown whether other viral infections elicit similar adaptive responses in NK cells, particularly within mucosal tissues. Methods To determine whether HSV-2 infection induces long-term changes in mucosal NK cells, we performed bulk RNA sequencing and high-parameter flow cytometry on vaginal NK cells isolated from naïve mice and from mice 10 days post intravaginal infection with HSV-2 (10dpi). Results Vaginal NK cells exhibited a distinct transcriptional profile following infection, with 10 dpi and naïve populations clustering separately by principal component analysis. Differential gene expression analysis revealed upregulation of Icos (adj. p = 0.22) and Ly6c2 (adj. p = 0.03) at 10 dpi compared to naïve NK cells. Notably, Ly6C has been associated with memory-like NK cells following viral challenge. Analysis by flow cytometry confirmed increased Ly6C expression (p = 0.017) and enhanced IFNγ production (p = 0.006) in NK cells from mice at 10 dpi, suggesting augmented effector capacity. Furthermore, transcripts characteristic of adaptive NK cell populations, including CD3 chain and histocompatibility gene family members, were elevated at 10 dpi. Conclusion Together, these findings demonstrate that local HSV-2 infection induces persistent transcriptional and functional changes in vaginal NK cells, supporting the development of a memory-like NK cell population within mucosal tissue. Funding Source K99 AI180649-02, T32 Ai007509 Topic Categories Viral Immunology (VIR)

Role of Microbial Tryptophan Metabolism in Th17 Activation and Multiple Sclerosis 2260431

Abstract Introduction The relationship between diet, the gut microbiota, and immune cells influences autoimmune diseases such as multiple sclerosis (MS). In a mouse model of MS, a ketogenic diet (KD) reduced disease severity in a microbiota-dependent manner. In further studies, a KD-enriched microbe, Lactobacillus murinus, and its metabolite indole lactate (ILA) both independently conferred disease protection without need for a KD. ILA is a tryptophan metabolite known to inhibit Th17 cells, key immune drivers of MS. However, whether ILA is required for Lactobacillus mediated Th17 inhibition is unclear. Methods To investigate the necessity of microbial ILA production, a microbe deficient in ILA production was tested to determine if it retained the ability to inhibit Th17 activity in vitro and modulate disease in the Experimental Autoimmune Encephalomyelitis (EAE) mouse model of MS. Specifically, a human strain of Lactobacillus reuteri lacking the key enzyme, aromatic aminotransferase (ArAT), involved in tryptophan conversion to ILA was utilized. Naïve mouse CD4+ T cells were treated with cell-free supernatants from wild-type L. murinus, L. reuteri, or L. reuteri ArAT knockout (KO) strains during a Th17 differentiation assay, followed by IL-17A quantification using an ELISA. For EAE studies, mice were orally gavaged with wild-type L. reuteri, L. reuteri ArAT KO, or PBS preceding and throughout disease course. Results ILA producing microbes significantly inhibited IL-17a production in vitro, while the ILA deficient L. reuteri ArAT KO strain did not. Furthermore, L. reuteri ArAT KO treated mice had higher EAE disease scores compared to those receiving wild-type L. reuteri, indicating that ILA is required for disease protection. Conclusion These findings suggest that ILA plays a critical role in microbial-mediated Th17 inhibition and protection against EAE. This work highlights the potential for targeting microbial tryptophan metabolism as a therapeutic for IL-17-driven autoimmune diseases like MS. Funding Source University of Wisconsin-Madison Genetics and Genomics Undergraduate Distinguished Research Fellowship Topic Categories Mucosal and Regional Immunology (MUC)

Human Microbiota-Driven Mouse Model Improves Drug Metabolism Prediction 2258986

Abstract Introduction Drug—microbiome interactions are increasingly recognized as critical in pharmacology. The gut microbiome influences drug properties such as bioavailability, absorption, metabolism, excretion, toxicity, and efficacy. Mice are widely used for preclinical studies, but their gut microbiome differs from humans, which can lead to inconsistent results in drug PK/PD and efficacy. To address this limitation, we developed a humanized gut microbiota mouse model. Methods Fecal material from three healthy donors–matched for age and BMI, and with no medication for three months–was pooled and administered by oral gavage twice weekly for four weeks to antibiotic-pretreated C57BL/6 mice. After colonization, a test molecule was given orally, and blood samples were collected at multiple time points for LC-MS/MS analysis. Results At baseline, the mouse microbiome contained ∼40% Muribaculaceae and ∼20% Lachnospiraceae. After FMT, Bacteroidaceae and Lachnospiraceae dominated, while Muribaculaceae dropped to 17% versus 40% in controls. The resulting profile closely resembled the human gut microbiome (∼30% Lachnospiraceae, ∼20% Bacteroidaceae). The pharmacokinetic profile of the tested compound in humanized mice differed markedly from conventional mice, suggesting that human-derived microbes alter enzymatic activity and metabolic pathways, impacting key PK parameters such as Cmax, AUC, and clearance. Conclusion This model provides a relevant platform to study microbiome-driven variability in drug disposition and supports the development of more predictive preclinical approaches for personalized medicine. Funding Source n/a Topic Categories Translational and Interventional Immunology (TI)

Dynamic but Unified Changes in the Perinatal Thymus Promote Distinct Waves of Treg Generation 2254735

Abstract Introduction The thymic environment enforces tolerance by inducing autoreactive T cells to either undergo negative selection or divert to the regulatory T cell (Treg) lineage. We previously found that coordinated transcriptional changes occur in thymic stromal cells and antigen-presenting cells (APCs) as the thymus transitions from neonatal growth to juvenile homeostasis: the first wave (P0-P3) of neonatal growth is accompanied by E2F transcriptional activity, while the second wave, at the “transitional” ages of P7-P14, is characterized by elevated Type I/III interferon signaling and altered antigen presentation. Concomitantly, two distinct waves of Tregs are generated, with the first wave expressing high levels of CTLA-4 and PD-1, and the second wave expressing elevated CD5 and GITR —functional markers of heightened Treg suppressive capacity. These findings raise the question of whether the age of thymocytes and/or the thymus generates phenotypically distinct and uniquely suppressive Tregs in perinates. Methods We performed heterochronic Treg generation assays on live thymic slices, as well as in vitro Treg generation assays to distinguish the contributions of the age of thymocytes versus the thymic environment on Treg selection in perinates. Results We find that both the perinatal age of CD4 single-positive thymocytes (CD4SPs) and the perinatal environment drive expression of key effector markers on Tregs (i.e. PD-1, GITR, and CD5). Perinatal CD4SPs are intrinsically biased towards Treg generation, while in vivo Treg phenotypes are largely driven by the age of the thymus environment. Conclusion Two phenotypically distinct waves of Tregs, first PD-1hi/CTLA-4hi followed by CD5hi/GITRhi, are promoted by both the thymic environment and a CD4SP-intrinsic capacity for Treg generation in perinates. These results provide significant insight into the selection of unique perinatal Treg subsets and reveal more phenotypic heterogeneity than previously described. Funding Source NIH P01 Topic Categories Hematopoiesis and Immune System Development (HEM)

Characterization Of Usutu Virus Replication And Associated Immune Response In Vitro 2308012

Abstract Introduction Usutu virus (USUV), a mosquito-borne flavivirus, is an emerging public health concern capable of causing neurological disease. However, the cellular mechanisms driving USUV neuropathogenesis remain unclear. Different neural and endothelial cell types may vary in their susceptibility to infection and virus-induced damage. This project investigates early cellular responses to USUV in four models: Vero C1008 (E6), SK-N-SH (human neuronal cell line), primary human brain cortical astrocytes (HBCA), and primary human brain microvascular endothelial cells (HBMVE). Determining which cell types support viral replication or exhibit greater cytotoxicity will provide insight into how USUV impacts the central nervous system. Methods Cells were infected with USUV at an MOI of 1, and pellets and supernatants were collected at different time points. Plaque assays quantified infectious titers, and RTq-PCR measured host immune responses. The CellTiter 96 Aqueous One Solution assay assessed USUV-induced cell damage relative to controls. Results USUV infection was successfully established in the four cells lines. Infected cultures showed morphological changes such as rounding, shrinkage, and detachment, with VERO E6 exhibiting the strongest cytopathic effect. Plaque assays confirmed infectious virus in all cell types, with higher titers in VERO E6 and HBCA cells. Cytotoxicity assays showed reduced viability in infected wells, especially in more permissive cells. Infection also induced strong immune activation, marked by increased cytokine, chemokine, and interferon RNA levels. RNA-seq of infected and mock-treated cells provided an unbiased view of virus-induced cellular reprogramming and highlighted pathways that may contribute to cell-type vulnerability and potential therapeutic targets. Conclusion Together, this work establishes a framework for understanding USUV interactions with brain-associated cell types and supports future studies on the mechanisms driving USUV-related neurological injury. Funding Source This work was supported by GSU Institutional Funds. Topic Categories Viral Immunology (VIR)

Strategically leveraging Influenza A induced trained immunity to shape immune profiles to subsequent vaccination with heterologous antigens 2309699

Abstract Introduction Trained immunity, the epigenetic and metabolic reprogramming of innate immune cells upon inflammatory signaling shapes subsequent immune responses to the same as well as unrelated antigens within both local tissues and broadly within the hematopoietic compartment. However, the role trained immunity plays in vaccination remains to be fully elucidated. Here, we demonstrate that vaccination through different routes (mucosal vs. systemic) differentially harnesses trained immune responses established by prior viral respiratory infection Methods Mice were infected with A/New/Caledonia/20/1999 H1N1 influenza virus. 4 weeks post infection, mice were immunized according to a prime/boost/boost schedule. To examine the responses to homologous vs. heterologous antigen and the impact of route, mice were immunized with tetrameric influenza neuraminidase (tetNA) or the spike protein from SARS-CoV-2 (WT-S) given intramuscularly (IM) with Addavax, or intranasally (IN) with our rationally designed adjuvant, NE/IVT, a nanoemulsion combined with IVT, a RIG-I agonist. Results Despite similar serum antibody levels between regimens to respective antigens, the mucosal antibody response to heterologous antigen, WT-S was enhanced by prior IAV infection but only with IN vaccination. Further, while IN NE/IVT/S increased the neutrophil frequencies within the lungs, IM Addavax formulations increased eosinophil frequencies regardless of antigen. However, infection with IAV before IM Addavax immunization enhanced the frequency of CD101+eosinophils. These unique changes were associated with shifts in Th balance upon antigen recall response, with prior-infection increasing Th1 cytokines to heterologous WT-S antigen in the mucosal lymph nodes in both immunization regimens and suppression of Th2 cytokines to homologous tetNA antigen in IM Addavax regimens. Conclusion Respiratory viral infection shapes the local inflammatory cell composition and subsequently alters the Th and mucosal antibody response induced by immunization. Funding Source Supported by National Institutes of Health (NIAID) grants R01AI160706, R21AI176069, R21AI180874 (PW, MS) Topic Categories Vaccines and Immunotherapy (VAC)

S-1117, an engineered pan-IgG cleaving enzyme, reduces IgG BCR mediated B cell activation and removes Fc fragments from tissue-bound anti-AChR IgG 2253830

Abstract Introduction Autoreactive IgGs and BCRs trigger tissue damage in autoantibody-mediated diseases. IgG and BCR degradation using S-1117, a novel Fc-fused pan-IgG protease represents a new therapeutic approach. S-1117 cleaves soluble IgG eliminating IgG-mediated effector functions including antibody dependent cellular cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), and immune complex-mediated cell activation. Here, we describe S-1117’s effect on the function of IgG BCR and autoreactive IgG bound to acetylcholine receptor (AChR) on tissue. Methods In vitro, purified human B cells were treated with S-1117 and BCR-mediated activation was assessed by phospho-flow cytometry. S-1117 cleaves IgG BCR on memory B cells preventing BCR-mediated phosphorylation of Erk and Syk. The in vivo effect of S-1117 on complement deposition in tissue was assessed by immunofluorescent staining of mouse diaphragm administered human anti-AChR IgG antibody. A single dose of S-1117 following human anti-AChR IgG injection, reduced IgG Fc presence by > 95%, thereby preventing complement activation in vivo. Results S-1117 is a novel engineered pan-IgG protease that targets multiple pathogenic mechanisms by cleaving soluble IgG, tissue-bound IgG and the IgG BCR on memory B cells. IgG and BCR cleavage results in reduced functional activity of these molecules, including eliminating complement deposition and reduced activation of B cells. Conclusion Collectively, these mechanisms have the potential to drive superior clinical outcomes in autoantibody-driven diseases. Funding Source n/a Topic Categories Therapeutic Approaches to Autoimmunity (THER)

Perivascular macrophage regulates sympathetic innervation density in adipose tissue 2309701

Abstract Introduction Metabolic balance within adipose tissue is tightly controlled by both the peripheral nervous system and the local immune environment. Recent studies highlighted an underappreciated interplay between adipose-innervating peripheral nerves and immune cells, especially the resident tissue macrophages that contribute to proper energy expenditure and metabolic health. However, the mechanistic basis of this bidirectional neuro—macrophage communication in regulating WAT metabolic homeostasis remains largely unknown. Methods To test whether perivascular macrophage reprogramming changes sympathetic innervation, we will quantify sympathetic fiber density/structure in WAT by immunostaining and confocal imaging and assess sympathetic activity by measuring norepinephrine and catecholamine-pathway markers in adipose tissue. To identify macrophage-derived mechanisms, we will transcriptomically profile perivascular macrophages from reprogrammed and control mice, prioritizing secreted/extracellular programs linked to axon outgrowth and guidance. Top candidates will be functionally tested using sympathetic neuron neurite-outgrowth assays and complementary in vivo perturbation approaches. Finally, we will evaluate metabolic impact by measuring thermogenic/lipolytic gene programs and systemic phenotypes. Results Transcriptional reprogramming of adipose perivascular macrophages is associated with higher sympathetic fiber density in white adipose tissue. This remodeling correlates with improved metabolic outcomes, including protection from early-onset diet-induced obesity. Conclusion These findings support a model in which resident perivascular macrophages actively modulate sympathetic remodeling in adipose tissue to promote metabolic homeostasis. Funding Source n/a Topic Categories Neuroimmunology (NEUR)

From Behavior to Dynamics: Decoding Carrier Roles in Cu‐based Photocathodes for Solar‐Driven CO ₂ Reduction

ABSTRACT Cu‐based photocathodes offer unique advantages for photoelectrochemical CO 2 reduction (PEC CO 2 RR) due to their earth abundance, tunable electronic structures and capacity to efficiently produce multi‐carbon (C 2+ ) products. However, their practical performance is fundamentally governed by the dynamics of photogenerated charge carriers, whose generation, separation, accumulation, transport, and extraction directly determine both catalytic efficiency and stability. This review examines how carrier behavior governs key reaction steps, evaluates material design strategies for tuning interfacial charge dynamics, and summarizes emerging techniques for probing these dynamics and interfacial transformations in operando. Looking ahead, integrating precise carrier control with stability and selectivity will be critical, requiring deeper insights into the coupling between carrier physics and catalytic function to guide the development of next‐generation Cu‐based photocathodes for PEC CO 2 RR.

Brain-Only Progression After First-Line Therapy in Extensive-Stage Small Cell Lung Cancer: Treat the Sanctuary, Preserve the Backbone

Ancestry-Informed Transcriptomic Analysis Identifies YY1 Axis Disruption and Stromal Program Loss in African American Prostate Cancer 2258332

Abstract Introduction Prostate cancer (PCa) shows racial disparities, with African American (AA) men experiencing higher incidence and worse outcomes than European American men. We investigated ancestry-associated molecular programs to better understand these differences. Methods RNA-seq data from high-grade PCa were analyzed using DESeq2 and GSEA to identify differentially expressed genes and pathways. Candidate genes and pathways were then examined through ongoing wet-lab assays in MDA-PCa-2b and LN-CaP cell-line to evaluate their effect in PCa. Results Multiple genes and pathways exhibited ancestry-associated dysregulation linked to tumor progression and aggressiveness. Notably, several noncoding and pseudogene transcripts, traditionally considered transcriptionally inert, showed reproducible overexpression patterns, suggesting potential epigenetic activation and regulatory cross-talk in AA tumors. Pathway-level findings revealed broad depletion of muscle/contractile programs, robust enrichment of olfactory receptor signaling, and downregulation of YY1 target gene sets, consistent with stroma loss and transcriptional reprogramming observed in experimental models. Conclusion Ancestry-informed transcriptomic analysis identifies chemosensory GPCR activation through olfactory receptor signaling enrichment and loss of stromal support programs, providing a framework for biomarker discovery and therapeutic exploration in AA PCa. These findings highlight transcriptional and microenvironmental alterations that may contribute to tumor aggressiveness and provide a foundation for biomarker discovery and ancestry-tailored therapeutic strategies. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Characterization of the brain immune microenvironment in healthy mice and a human infant with AT/RT 2260665

Abstract Introduction Atypical Teratoid/Rhabdoid Tumors (AT/RT) are devastating brain tumors with high mortality. Due to rare occurrence in infants, lack of understanding of brain microenvironment prior to treatment, and majority of studied tissues being from heavily treated individuals, the overall immune response to these tumors are unknown. Methods We obtained autopsy brain tissue and blood within 12 hours of death and performed high parameter flow cytometry. Results We reliably isolated immune cells from brain and blood (PBMC: 92%, brain: 54.4% viable cells). Unlike brain tissue isolated from newly-diagnosed adult glioblastoma, this 8-week-old AT/RT patient had significant infiltration of activated PD1+ CD8 T cells in the brain. While significant numbers of CD8 T cells with high expression of proliferation marker (Ki67) and activation markers (PD1, CD69) were found, no evidence of exhaustion was seen. Few Tregs and suppressive myeloid cells were in the brain. This immune phenotype is conducive to immunotherapies. In the matched blood, few T cells were found, which reflects infant lymphopenia. To elucidate immune microenvironment of a healthy brain in infancy, we performed flow cytometry on 2- and 4-day old mice, an age developmentally similar to 8-week-old human infants. We found various immune cells including proliferating T cells in brains, indicating a potential role in the developing brain. Conclusion Understanding the state of immunity in brains of infants will pave the way for design of effective immunotherapies. Funding Source n/a Topic Categories Neuroimmunology (NEUR)

A library-based examination of T cell allorecognition 2259104

Abstract Introduction Alloreactive T cells drive acute transplant rejection and Graft-versus-Host-Disease (GvHD) by reacting to peptides displayed on non-self MHC (allo-MHC). Despite their critical role, the molecular mechanisms of T cell allorecognition of peptide-allo-MHC complexes are not well understood. It is unclear how peptide-specific or cross-reactive alloreactive T cells are, and how they compare to conventional T cells. Additionally, little is known about the antigens recognized in an alloresponse–whether they are tissue-specific or ubiquitous peptides, and whether there are immunodominant antigens. Here, we use high-throughput T cell receptor (TCR) sequencing, cloning, and antigen profiling tools to conduct a repertoire-scale examination of T cell allorecognition. Methods We investigate the molecular mechanisms of allorecognition in a single-MHC mismatch murine skin graft model. We conduct single-cell TCR sequencing, use a pooled TCR cloning method to recapitulate the TCR repertoire for functional testing, and conduct high-throughput antigen screens to identify the targets of alloreactive TCRs. Results Single-cell TCR sequencing of the alloreactive repertoire in this model reveals dominant usage of a specific TRBV gene segment. Initial testing of select expanded TCRs indicates that they exhibit some degree of specificity for the peptide, and that they may recognize ubiquitously expressed antigens that are not skin-specific. We are further characterizing the specificity of these TCRs and the rest of our dataset by constructing a TCR library of the entire TCR dataset for functional testing. Future work will include high-throughput library-on-library screens to pair alloreactive TCRs with the antigens they recognize. Conclusion This work will provide a mechanistic examination of allorecognition at an unprecedented scale, providing insight into the molecular rules of allorecognition and the structure of an alloresponse, which may inform the development of improved therapies for transplant rejection and GvHD. Funding Source Cancer Grand Challenges partnership financed by CRUK (CGCATF-2023/100001), the National Cancer Institute (OT2CA297463), and The Mark Foundation for Cancer Research; Ludwig Center at MIT’s Koch Institute for Integrative Cancer Research Topic Categories Transplantation Immunology (TRAN)

Rebooting the Immune System: Thymic Rejuvenation Reverses Alzheimer’s Disease Pathogenesis and Cognitive Decline 2255320

Abstract Introduction Alzheimer’s disease (AD), a devastating age-related neurodegenerative disorder, is profoundly linked to immunosenescence. Thymic involution, as a central driver of this process, causes systemic immune collapse. We hypothesized that reversing involution through thymic rejuvenation could mitigate AD by rebooting the systemic immune environment. Methods We longitudinally assessed thymic output in 5XFAD versus wild-type mice, correlating it with brain pathology, and confirmed the age-dependence of thymic decline in human single-cell RNA-seq data. To test our hypothesis, we transplanted neonatal thymic tissue into 6-month-old 5XFAD mice and assessed thymic output, peripheral T cell profiles, Aβ load, microgliosis, and cognitive function. Systemic and neuroimmune changes were profiled by single-cell RNA-seq. Furthermore, we engineered and transplanted T cell—deficient mouse thymic tissue chips (MTTCs) to evaluate their therapeutic efficacy and translational potential. Results Thymic involution was driven by aging, not AD pathology, in both mice and humans. This functional decline strongly correlated with Aβ burden and microgliosis. Neonatal thymic transplantation robustly enhanced thymic output, which was associated with a ∼45% reduction in Aβ plaques, suppressed microgliosis, and rescued cognitive function. Single cell transcriptomics revealed a reversal of immunosenescent signatures in the periphery and a concomitant reprogramming of disease-associated microglia (DAM) in the brain. Crucially, transplantation of MTTCs recapitulated these therapeutic benefits, demonstrating clinical potential. Conclusion We introduce a paradigm-shifting therapeutic strategy for AD: thymic rejuvenation. By rebooting the aged immune landscape, this approach reverses established neuropathology and rescues cognitive decline, moving beyond single molecular targets in the brain. It thereby establishes a novel class of “immunoregenerative” therapeutics that combat neurodegeneration by restoring systemic immune homeostasis. Funding Source National Natural Science Foundation of China (82502227), Science and Technology Innovation Key R&D Program of Chongqing (CSTB2023TIAD-STX0041), Chongqing International Institute for Immunology-Innovative Basic Research Project (2023YJC02), Brain-Gain Topic Categories Neuroimmunology (NEUR)

Coordinated circulating and tissue-based T cell responses precede xenograft rejection 2308959

Abstract Introduction Despite the life-saving successes of solid organ transplantation, organ demand far exceeds availability. Porcine xenotransplantation, the use of pig organs in humans, holds promise, but acute rejection remains a barrier to long-term xenograft survival. Methods To investigate mechanisms of acute xenorejection, we used the first long-term (61-day) human decedent xenotransplantation model at NYU, in which a decedent received a kidney with a thymus autograft (“thymokidney”) from an α-1,3-galactosyltransferase knockout (GGTA1 KO) pig. This unique model allows serial thymokidney biopsies, paired blood draws, lymph node biopsies and exhibited antibody-mediated (AMR) and cellular rejection despite immunosuppression. We performed TCR sequencing on thymokidney biopsies and 5’ CITE-seq of blood. Xenoreactive clonotypes were identified by mixed lymphocyte reaction. We identified clonal families forming over time and across compartments. Results Longitudinal analysis of the thymokidney biopsies revealed human CD4 and CD8 T cell responses that formed clonotypic families. Circulating activated T cells increased 2 weeks before rejection, highlighting the potential as a cellular biomarker of rejection. Strikingly, a single xeno-reactive CD8 clonotype expanded dramatically in the blood before rejection and was also identified in the xenograft during rejection. T cell clonotypes were shared across xenograft, blood and lymph nodes and formed clonal families with known xeno-reactive clonotypes, suggesting a coordinated response to a limited set of antigenic targets. Serum CXCL13 and circulating T follicular helper cells increased before AMR, indicating formation of germinal centers which suggests therapeutic opportunities to treat impending rejection. Conclusion These findings establish a framework to track coordinated immune responses at cellular, clonal, and clonotypic family levels and identify potential biomarkers of xenorejection, offering new opportunities for early prediction and potential intervention. Funding Source National Institutes of Health (NIH) grant R01AI158617 (RSH) National Institutes of Health (NIH) grant U19AI082630 (RSH) National Institutes of Health (NIH) grant U19AI191396 (ADG, BJK, RAM) Topic Categories Transplantation Immunology (TRAN)

Checkpoint Blockade Selects Innate-Programmed Cytotoxic T Cells with Pathogenic TCR Features in Immune-Related Arthritis 2309982

Abstract Introduction T cell immune checkpoint inhibitors (ICIs) can cause immune-related adverse events (irAEs), including inflammatory arthritis. The cellular and molecular mechanisms underlying irAE arthritis are not well understood. We hypothesize that a particular subset of CD8 T cells expands in irAE arthritis and that these clones possess unique TCR sequences. Methods Single-cell RNA, paired TCR sequencing and computational approach was applied to analyze clonal, transcriptional, and CDR3 biophysical features of circulating CD8 T cells before and after anti—PD-1 therapy. Results Among the cells collected from patients with irAE arthritis, we identified a population of clonally expanding PRF1+ CD8 effector T cells displaying an innate-like inflammatory and metabolically active phenotype. This cell population expresses cytotoxic effector genes, innate sensing, and metabolic pathways. Remarkably, within this population, TCR repertoires exhibited significantly greater hydrogen-bonding in the CDR3 hypervariable region than in no-irAE controls, suggesting distinctive antigen specificity and binding stability. This feature was present before the ICI treatment. It was further amplified after the therapy, consistent with dynamic clonal selection under checkpoint blockade. Conversely, expanding CD8 T cell clones from patients without irAEs exhibited lower hydrogen bond propensity both before and after treatment. Furthermore, irAE arthritis-associated clonotypes also showed increased hydrophobicity in the central CDR3 region. These regions consisted of shorter CDR3 sequences, indicating a compact structural architecture that promotes stable, potentially promiscuous peptide-MHC interactions. Conclusion Our findings suggest that checkpoint blockade selectively expands innate-programmed cytotoxic CD8 T cells with structurally optimized TCRs, promoting autoreactive effector responses and defining integrated cellular and TCR biophysical determinants of irAE susceptibility. Funding Source NIH AI125640 Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

TNFR2 signals promote serine metabolism to preserve Regulatory T cell function by regulating SLC7A10 2254289

Abstract Introduction TNFR2 is primarily expressed by CD4+Foxp3+ regulatory T (Treg) cells, which are crucial in maintaining immune homeostasis and preventing autoimmunity, Amino acid metabolism plays an important role in maintaining the activity and function of Treg cells. However, the molecular mechanism by which TNFR2 signaling regulates Treg cells by influencing cellular amino acid metabolism remains unclear. Methods WT and TNFR2-deficient Treg cells were purified by FACS sorting analysis, then RNA-bulk Seq was performed to examine the differential expressed genes. RT-qPCR, and flow cytometry were conducted to validate the potential genes that are targeted by TNFR2. Limitation of cellular Serine in vitro and in vivo were applied to reveal the change of Treg cell proliferation and function. Results In this study, we found that SLC7A10 was a significant upregulated gene in Tregs by TNFR2 signaling. Silence of SLC7A10 decreased the proportion of Treg cells and their expression of CTLA4, PD-1, and consequently inhibited its proliferation and suppressive function. Since SLC7A10 is one of the representative neutral amino acid transporter genes, especially serine transportation. We further confirmed that knockdown of SLC7A10 reduced the uptake of serine in Treg cells, in consistent, the limitation of serine in vitro significantly suppressed the proliferation and suppressive function of Treg cells. Therefore, SLC7A10, upregulated in TNF-TNFR2 signaling, plays critical role in maintaining the serine metabolism of Treg cells. Conclusion Our study for the first time found that TNFR2 promotes serine metabolism by regulating SLC7A10 in Treg cells. The findings suggest that SLC7A10 is not only expressed by Tregs but also exerts specific functions for maintaining Treg activation. Funding Source n/a Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)

NETosis is present and influences tumors in Lymphangioleiomyomatosis models 2309907

Abstract Introduction Lymphangioleiomyomatosis (LAM) is a cystic lung disease primarily found in women whereby, over time, small thin-walled cysts take over normal alveolar space, leading to progressive lung function loss. Cysts are caused by estrogen-sensitive smooth muscle LAM tumor cells with mutations in the tuberous sclerosis gene complex (TSC), which leads to constitutive activation of mTORC1 signaling and increased LAM cell proliferation. Our lab proposed that LAM cells originate from the estrogen sensitive myometrium of the uterus and created a mouse model whereby TSC2 knockout in the uterus leads to uterine LAM tumors that metastasize to the lungs. With this model, we demonstrated that neutrophils promote this tumor growth in part through release of neutrophil elastase. Here we examined the potential role of NETosis, a process of releasing modified chromatin and neutrophils serine proteases like neutrophil elastase, in promoting LAM tumor cell growth. Methods We analyzed RNAseq on neutrophils treated with and without estradiol to understand the role of estrogen in neutrophil function. We also generated NETs from bone marrow derive murine neutrophils and stimulated mouse uterine cells. Finally, we used immunofluorescence looking for citrullinated histones and myeloperoxidase in TSC2 null mouse uteri versus littermate control uteri, in xenografts generated from TSC2 null ELT3 rat myometrial cells, and in human LAM lung samples versus controls. Results Neutrophil degranulation and immunity signaling pathways were upregulated in neutrophils treated with estrogen. Mouse uterine cells responded to NETs by upregulating expression of genes associated with cell survival and EMT. NETosis was detected in both the uterine tumor of our mouse model and in human lung samples from patients with LAM in comparison to controls. Conclusion Evidence suggests that NETosis could be contributing to the release of neutrophil elastase in TSC2 null tumor microenvironments, exacerbating tumor growth. Funding Source LAM Foundation DOD NIH Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

High‐Performance Weavable Piezoelectret Fibers via Scalable In Situ Poling Melt‐Spinning for Real‐Time Knee Joint Rehabilitation Monitoring

ABSTRACT The scalable production of high‐performance piezoelectric fibers remains a major hurdle for smart textile applications. Here, we report an in situ polarization strategy integrated into an industrially viable melt‐spinning process for the continuous fabrication of piezoelectric polypropylene/barium titanate (PP/BTO) composite fibers. This approach simultaneously induces interfacial cavitation to form electret pore structures and accomplishes rapid dipole polarization under a high electric field during fiber drawing, eliminating the need for post‐processing. The optimized PP/BTO fibers exhibit a high piezoelectric coefficient ( d 33 ) of 1.8 pC/N and a surface potential of −3.4 V, achieved with an ultralow poling time of 0.3 s. The fibers demonstrate exceptional flexibility and weavability, enabling their integration into large‐scale textiles. As a proof‐of‐concept, a sensor‐woven insole is constructed for real‐time gait monitoring. Combined with machine learning, the system successfully recognizes different gait patterns with over 84% accuracy, showcasing significant potential for personalized rehabilitation diagnostics. This work provides an efficient and scalable pathway for manufacturing functional fibers, bridging the gap between laboratory innovation and industrial production.

Treatment of Multiple Myeloma: ASCO Living Guideline, Version 2026.1.1

This guideline has been updated. This ASCO Living Guideline is reviewed on an eight-week cycle. Please see the online dynamic version for the latest updates. Living guidelines are developed for selected topic areas with rapidly evolving evidence that drives frequent change in recommended clinical practice. Living guidelines are updated on a regular schedule by a standing expert panel that systematically reviews the health literature on a continuous basis, as described in the ASCO Guidelines Methodology Manual . ASCO Living Guidelines follow the ASCO Conflict of Interest Policy Implementation for Clinical Practice Guidelines . Living Guidelines and updates are not intended to substitute for independent professional judgment of the treating clinician and do not account for individual variation among patients. See appendix for disclaimers and other important information ( Appendix 1 and Appendix 2 ). Updates are published regularly and can be found on the ASCO Publications website .