Browse Articles

Projected region-of-attraction analysis of coupled two-region low-altitude air traffic using microsimulation-calibrated MFDs

Humoral immunity against gut commensals governs their prevalence and influences patient clinical outcome 2267972

Abstract Introduction Evidence continues to link gut microbial dysbiosis with immunotherapy resistance. Patients with reduced responsiveness often lose beneficial bacteria such as Akkermansia muciniphila while gaining taxa detrimental to treatment outcomes. However, whether this dysbiosis is a cause or consequence of altered host immunity remains unresolved. Methods We investigated the immunological drivers of dysbiosis using a combination of flow cytometry, serology, T-cell profiling, and mouse models. Patient samples from NSCLC cohorts were analysed for anti-A. muciniphila IgG titres and recall T-cell responses. Parallel experiments in mice involved serum and splenocyte transfer following allogenic faecal microbiota transplantation (FMT), as well as antibody treatment experiments to evaluate the capacity of IgG to transcytose into the gut lumen and deplete gut commensals. Results High titres of anti-A. muciniphila IgG correlated with the depletion of the bacterium in NSCLC patients. Elevated titres, together with Th17/Th1 recall responses, were associated with poorer survival, increased frequencies of regulatory T-cells, and reduced effector populations. In murine models, serum transfer — but not splenocyte transfer — after allogenic FMT promoted tumour growth. Furthermore, studies using an in-house antibody derived from tumour-infiltrating B cells demonstrated that circulating IgG can cross the gut barrier to bind and eliminate its bacterial target. Conclusion These findings reveal that humoral immunity can directly shape gut microbial composition by promoting the loss of beneficial commensals such as A. muciniphila. This immune-driven dysbiosis may contribute to resistance against immune checkpoint blockade and underscores the need to consider host antibody responses when designing microbiota-centred interventions to enhance cancer immunotherapy outcomes. Funding Source This work was supported by the French Ministry of Health (PIA5, ANR, RHU5 ‘‘ANR-21-RHUS-0017’’ IMMUNOLIFE); the European Union’s Horizon 2020 programme (grant agreements 964590, IHMCSA; 825410, ONCOBIOME — Gut OncoMicrobiome Signatures associated with cancer incidence, prognosis, and treatment response prediction); the Horizon Europe programme (grant 101095604, PREVALUNG EU — Biomarkers affecting the transition from cardiovascular disease to lung cancer for stratified interception); the ANR grant Ileobiome (ANR-19-CE15-0029-01); the European Research Council (ERC) under grant 101052444 (ICD-Cancer — Immunogenic cell death in the cancer-immune dialogue); and by the SIRIC SOCRATE and SIGN’IT ARC Foundation programmes (MICROBIONT-PREDICT, 2021; Made-IT, 2023). Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

Differentiated thymic epithelial cells adopt gene-expression and micro-environmental niches unique to the thymus 2310220

Abstract Introduction The thymus contains a multitude of epithelial cell types that work in concert to educate immature T cells called thymocytes to distinguish self from non-self. A subset of thymic epithelial cells (TECs) express lineage defining transcription factors and differentiate into cell types -- such as muscle, neuroendocrine, and tuft cells -- that are typically found in peripheral tissues. However, the function and organization of these differentiated TECs especially in the human thymus are poorly understood. Specifically, much remains unknown about the gene regulation that enable their function, and their structural morphology and spatial localization in the thymus to ultimately orchestrate thymocyte education. Methods To resolve the diversity of TECs at a single cell resolution, we simultaneously profiled the gene expression and chromatin accessibility of human thymus samples. We integrated our data with all available thymic single cell datasets to establish concordant cell annotations across studies. We leveraged the CELLxGENE database to identify unique gene signatures in the differentiated thymic TECs compared to their peripheral counterparts in 31 tissues. Informed by the gene expression of maturing thymic muscle cells, we used quantitative microscopy to evaluate their cellular morphology and spatial localization in the thymus. Results We show that differentiated TECs share substantial celltype restricted genes, yet take on a celltype specific identity, a phenomenon not shared by their peripheral counterparts. Narrowing in on the maturation of thymic muscle cells, we show that they have distinct cellular morphologies and thymic spatial localization as they differentiate. Conclusion Differentiated thymic celltypes, while sharing similarity to their peripheral counterparts, take on a thymus specific identity. Furthermore, thymic muscle cells have distinct cell morphology and micro-environmental niche as they mature. Funding Source NIH Topic Categories Computational and Systems Immunology (COMP)

Optimized LNP-encapsulated IVT mRNA as a robust platform for the development of next-generation cancer immunotherapies 2309761

Abstract Introduction Chimeric antigen receptor T cell therapy (CAR-T) and therapeutic cancer vaccines show great promise as next-generation cancer immunotherapies. However, these approaches traditionally rely on plasmid DNA or viral-vector-based gene delivery systems which are limited by potential safety risks and suboptimal antigen expression. Lipid nanoparticle (LNP)-encapsulated in-vitro-transcribed (IVT) mRNA offers a safer alternative while facilitating enhanced targeting, antigen expression, and overall therapeutic efficacy. Methods IVT mRNA vectors were rationally designed and vector components optimized for enhanced expression. Modified nucleotides were incorporated into the RNA during in vitro transcription prior to encapsulation with optimized LNP formulations. To assess efficacy versus other approaches, we compared antigen expression of our optimized LNP-mRNA to a protein-based vaccine in mice. Finally, we tested anti-CD19 CAR-T cells generated using LNP-mRNA in a cytotoxicity assay, and evaluated the anti-tumor effects of an LNP-mRNA anti-TRP2 vaccine in vivo. Results Through considered vector design and optimization, we successfully engineered IVT mRNAs with enhanced stability and translation efficiency, achieving robust transgene expression both in vitro and in vivo. By incorporating modified nucleotides and optimizing LNP formulations, we reduced unwanted host immune responses that might otherwise hamper therapeutic efficacy. Compared with a recombinant protein-based vaccine, optimized LNP-mRNA was more effective, eliciting markedly higher antibody titers against a complex transmembrane antigen. Functionally, anti-CD19 CAR-T cells engineered using LNP-mRNA exhibited potent cytotoxicity against target cells ex vivo, and mRNA-based anti-TRP2 vaccines inhibited melanoma growth in vivo. Conclusion Overall, these findings highlight the potential of optimized LNP-encapsulated IVT mRNA as a platform for engineering safer and more effective cancer immunotherapies. Funding Source n/a Topic Categories Vaccines and Immunotherapy (VAC)

Food Allergen Challenge Elicits Gut-Derived, Migratory iILC2s that Reshape the Immune Profile of Remote Lungs 2309438

Abstract Introduction Allergic progression across the skin-gut-lung axis remains unclear. A subset of migratory, intestine-derived inflammatory type 2 innate lymphoid cells (iILC2s) expand in response to IL-25 and disseminate type 2 immunity along the gut-lung axis (GLA) in helminth infection. Here we tested if food allergen challenge similarly elicits mucosal organ crosstalk by expanding gut-derived, lung-migratory iILC2s, promoting lung IL-13-mediated mucin dysregulation. Methods Wildtype mice sensitized by intraperitoneal (i.p.) injections of ovalbumin (OVA)+alum or PBS+alum (days 0, 14) were challenged by oral gavage (o.g.) with OVA or PBS on days 21-23. Sphingosine-1 phosphate (S1P) modulator FTY720 blocked intestinal egress of iILC2s. Lung, small intestine, and mesenteric lymph nodes (MLNs) were analyzed by RT-PCR and flow cytometry. IL-25 (i.p.) induced lung-migratory iILC2s in an allergen-free system. DdblGATA1-/- (eosinophil deficient) mice were used to parse effects of iILC2s and eosinophils. Results iILC2s (CD45+Lin-CD4-ST2-KLRG1hiIL17Rb1+IL7Ra+) were found in the lung within 1 hour of OVA o.g. challenge. Lung iILC2s positively correlated with Il13 mRNA levels (R2=0.77, p = 0.02), and preceded eosinophil infiltration and mucin dysregulation. O.g. challenge increased duodenal tuft cell signatures (p < 0.05) and IL-25 protein (p < 0.00005), correlating with local iILC2 expansion (R2=0.68, p < 0.0005). FTY720 inhibition of iILC2 intestinal egress reduced iILC2 numbers in MLNs and lungs, indicating intestinal origin of lung-migrated iILC2s. IL-25 treatment (bypassing allergen) elicited gut-derived, lung-migratory iILC2s that correlated with lung Il13 (R2=0.84, p < 0.0005) and preceded eosinophil infiltration and induction of Muc5ac mucins. IL-25 likewise elicited lung-migrated iILC2s and an Il13 signature in eosinophil deficient mice; however, Muc5ac was not increased in the absence of eosinophils. Conclusion Our data implicate migratory iILC2s in food allergen-driven dissemination of type 2 immunity along the GLA. Funding Source This work was supported by NIH R01AI168134 to Lisa A. Spencer. Topic Categories Mucosal and Regional Immunology (MUC)

Type I IFN Signaling, via IFNb, is involved in regulating antifungal host responses to Aspergillus fumigatus 2260062

Abstract Introduction Invasive pulmonary aspergillosis caused by Aspergillus fumigatus is increasingly common in influenza and SARS-CoV-2 patients, suggesting that antiviral immune responses create susceptibility to fungal infection. We recently demonstrated that differential type I interferon signaling through IFNAR1 vs. IFNAR2 regulates fungal clearance and host damage, respectively, during A. fumigatus infection. Here, we investigated how IFNβ, the strongest IFNAR1/2 activator, regulates these outcomes during infection. Methods We used a murine lung infection model to determine how IFNβ regulates antifungal responses to A. fumigatus via flow cytometry, biochemical assays, and imaging. Results Our preliminary results demonstrate that Ifnβ-/- mice have increased damage, inflammation, and morbidity in response to A. fumigatus infection compared to WT mice. Further, we found that Ifnβ-/- mice are more susceptible to A. fumigatus, demonstrated by decreased survival and increased invasive disease compared to WT mice. We also found that there was no difference in neutrophil cell recruitment to the lungs between Ifnβ-/- and WT mice, suggesting that their anti-fungal responses may be altered. To determine this, we first examined reactive oxygen species (ROS). We found that Ifnβ-/- neutrophils produced increased ROS compared to WT neutrophils in response to both A. fumigatus conidia and hyphae ex vivo. Currently, we are investigating whether ROS directly regulates damage and fungal clearance of the Ifnβ-/- mice in vivo during A. fumigatus infection. Conclusion Here, we demonstrate that IFNβ, like IFNAR2, regulates tissue damage during A. fumigatus infection. Our results suggest IFNβ-mediated regulation of anti-fungal responses contributes to infection susceptibility through damage control. These findings expand our understanding how type I IFN-mediated damage via IFNβ occurs during Aspergillus infections and how antiviral mechanisms create a permissive environment for fungal infection through host damage. Funding Source NIH/NIAD (K22AI153671), Francis Family Foundation, University of California, Merced Start-Up Topic Categories Microbial, Parasitic, and Fungal Immunology (MPF)

Tissue specific cues govern melanoma specific resident memory CD8+ T cell persistence in skin and tumor draining lymph nodes 2310174

Abstract Introduction CD8+ resident memory (Trm) cells are key sentinels of anti tumor immunity, residing durably in tissues and rapidly responding to cognate antigen. We identified melanoma specific Trm cells in skin and tumor draining lymph nodes (tdLN) that protect against metastasis, yet the mechanisms governing their maintenance, spatial organization and heterogeneity remain unclear. Methods Using a B16 induced vitiligo model, we generated gp100 specific CD69+CD103+ Trm cells in skin and tdLNs. Xenium spatial transcriptomics in wild type and β2m conditional knockout mice mapped endogenous and antigen specific Trm cells niches and tested antigen dependence. Conditional T cell receptor alpha (TCR) knockout mice and IL7/IL15 blocking antibodies probed the roles of TCR signaling and homeostatic cytokines. Results High resolution spatial transcriptomics revealed distinct micro-environments for Trm cells in skin and TdLNs. Skin Trm cells clustered with epidermal and hair follicle keratinocytes, whereas TdLN-Trm cells preferentially contacted dendritic cells, antigen presenting B cells, fibroblastic reticular cells and sub-capsular sinus macrophages. This heterogeneity prompted the hypothesis that tissue specific cues govern Trm cell survival. Consistent with this, conditional loss of MHC-I signaling significantly reduced tdLN-Trm cells but spared skin-Trm, a finding confirmed in inducible TCR knockout mice. Blocking IL7 and IL15 significantly reduced Trm cell proportions in both tissues, indicating that while antigen presentation sustains Trm cells in lymph nodes (LN), IL7/IL15 signaling is critical for their survival in both skin and LNs. Conclusion Our investigations show that melanoma specific Trm cells interact with different cellular partners in skin vs. tdLN niches, underscoring the influence of local microenvironment on their survival cues. We reveal Trm residence in LNs requires ongoing antigen presentation, and that both skin and tdLN Trm populations depend on a shared IL7/IL15 cytokine axis for maintenance. Funding Source R01CA254042 and R01CA225028A Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Longitudinal autoantibody profiling in systemic sclerosis uncovers biomarkers associated with disease progression 2309123

Abstract Introduction To evaluate dynamic changes in autoantibody profiles in naïve systemic sclerosis (nSSc) patients under treatment and to identify biomarkers associated with disease progression. Methods Serum samples from 50 nSSc patients at baseline and follow-up after treatment for 5 years, along with 50 controls, were analyzed. Autoantibody profiles were assessed using an autoantigen microarray targeting 120 autoantigens. Proteomic analysis was conducted via liquid chromatography—mass spectrometry in data-independent acquisition mode. Cox proportional hazards models were used to assess associations with disease progression, and Spearman’s correlation was applied to analyze antibody-protein interactions. Results Baseline autoantibody profiling identified 20 significant elevated autoantibodies which divided the patients into two subgroups: SSc-high and SSc-low, with the SSc-high patients showed more severe clinical characteristics. Longitudinal analysis revealed that 85% of patients with increasing autoantibody titers experienced disease progression, while 65% of those with decreasing titers showed clinical improvement. Notably, increased anti-Topoisomerase I, anti-CENPB and anti-TPO antibody titters were strongly linked to disease progression. Proteomic analysis identified 117 proteins involved in tissue repair and immune modulation in patients with declining titters and clinical improvement, whereas 42 proteins linked to lipid metabolism and oxidative stress were enriched in those with rising titers and disease progression. Antibody-protein correlations highlighted key molecular interactions, including SERPINE1 and muscarinic receptor antibodies, were identified. Conclusion Dynamic changes in autoantibody titers, rather than baseline levels, were more strongly associated with SSc trajectory. Proteomic signatures suggest lipid metabolism and oxidative stress as potential drivers, highlighting novel biomarkers and therapeutic targets for personalized SSc management. Funding Source n/a Topic Categories Immune Mechanisms of Human Disease (HUM)

High Numerical Aperture Metalens for the Deep Ultraviolet

ABSTRACT Deep‐ultraviolet (DUV) radiation is essential for advanced nanofabrication, biomedical imaging, and emerging photonic technologies, yet the absence of compact, high‐performance optical components in this spectral range has impeded progress. We demonstrate an aluminum nitride (AlN) metalens with record‐high numerical aperture (HNA) from 0.9 to 0.7, operating at 266 nm. Optical measurements confirm diffraction‐limited focusing performance. Importantly, we show a direct laser writing microscopy (DLWM) that lithographic direct writing serves microscopy purposes for the HNA metalens: besides enabling nanofabrication with 100 nm level critical dimension, it provides a universal characterization method for HNA DUV metalenses, addressing the challenges of constructing broadband HNA optical characterization systems in this wavelength range. Furthermore, the HNA AlN metalens for the first time integrates into a confocal photoluminescence system, enabling compact sub‐micrometer spectroscopic imaging with DUV excitation and a confocal photoluminescence imaging system using a metalens (NA = 0.7), resolving isolated features down to 300 nm. This work establishes AlN metalenses as a transformative and practical platform for high‐resolution DUV applications.

Quantifying land use trade-offs in Sub-Saharan Africa: spatiotemporal dynamics and drivers of production-living-ecological spaces in Tanzania (2000–2020)

DC Vaccination Against the Vascular Antigen DLK1 Reduces Triple-Negative Breast Cancer Burden and Metastasis 2299544

Abstract Introduction Despite treatment advancements for triple-negative breast cancer (TNBC) utilizing immune checkpoint blockade (ICB) in combination with chemotherapy, the lack of druggable targets such as receptors for estrogen, progesterone, and HER2 yields high recurrence and metastasis rates. A promising immunotherapeutic strategy involves exploiting the tumor microenvironment’s (TME) dependence on a functional vasculature for growth and progression. Methods We hypothesize that alpha-type-1 dendritic cell (αDC1) vaccination against the conserved tumor blood vessel antigen (TBVA) DLK1, which we found is expressed by abluminal pericytes in the TME of 75% of TNBC patients, can provide therapeutic protection against TNBC. To assess in vivo tumor inhibition and changes in immunologic milieu in two murine models, strain-specific DLK1 peptides were pulsed with αDC1 cells to generate vaccines (αDC-DLK1) that were administered on days 5 and 7 alongside ICB on days 8, 11, and 14 in pre-clinical E0771 and 4T1 TNBC models. Tumors and draining lymph nodes were harvested after euthanasia (E0771) or survival surgery (4T1) around day 17 for analysis by histology, immunohistochemistry, immunofluorescence, TUNEL assay, immunoblotting, qPCR, and/or T cell co-culture activation studies. Lungs and vital organs were harvested for evaluation of metastasis and toxicity. Results Our results demonstrated the ability of therapeutic αDC-DLK1 vaccination to safely provoke anti-angiogenic T cell activation, increased TME infiltration, and enhanced effector function that resulted in significant reduction of primary and metastatic tumor burden. Interestingly, ex vivo analyses demonstrated that vaccine-mediated DLK1 responses initiated polyclonal T cell activation against TNBC cellular antigens through cross-priming effects that likely fortified tumor protection. Conclusion Overall, our data support the clinical relevance of DLK1 as an immunotherapeutic target and the development of TBVA-based DC therapies for TNBC patient treatment. Funding Source n/a Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)

LRP1-Targeted Polymeric Nanoconjugates for BBB Penetration and Time-Specific Delivery of PD-L1—Blocking Peptides in Melanoma Brain Metastases 2310230

Abstract Introduction MBM show CNS tropism and limited benefit from ICB. While AXL inhibitors can improve responses, toxicity remains a concern. The TiME features few effector T cells, abundant TAMs, and glial PD-L1, promoting immune evasion. BBB-penetrant strategies with Immuno-Nano-Conjugates (INCs) are under development; intrathecal delivery has not improved survival. LRP-1—mediated INCs delivery may enable BBB transport and TiME reprogramming. Here we report a targeted nanoplatform to cross the BBB and deliver PD-L1—blocking peptides to the TiME. Methods INCs were constructed by attaching a malic acid—based polymer to (i) P-12, a PD-L1—blocking peptide, and (ii) Angiopep-2 (AP2), enabling LRP-1—mediated transcytosis. We evaluated efficacy, safety, toxicity, and biodistribution in a BBB-on-a-chip brain tumor model and in vivo in murine MBM and subcutaneous melanoma models. To translate findings, we analyzed metastatic melanoma transcriptomics (TCGA-SKCM, n = 471) and generated Kaplan—Meier survival curves. Results INCs crossed the BBB and penetrated the BBB-on-a-chip tumor model. Delivery produced selective uptake by activated TiME and promoted immune activation. In vivo, INCs achieved higher brain accumulation than ICB agents and demonstrated meaningful efficacy with favorable safety. High AXL expression correlated with poor ICB response and immunosuppressive myeloid signatures, linking to reduced survival. In subcutaneous melanoma, combining P-12—based peptides with Bemcentinib yielded greater tumor reduction than anti-PD-1 alone, suggesting circumvention of ICB resistance. These findings support a path toward safe clinical translation. Conclusion LRP-1—mediated INC delivery enhances PD-L1 blockade exposure in MBM TiME, supporting a strategy to treat resistant MBM. AXL is a key target for rational combination with nanoplatforms. Further validation in MBM models is planned, and ongoing preclinical testing. Funding Source Stanford Brain Metastasis Consortium Seed Grant Program U54 Brain MetNet Seed Grant Program and American Cancer Society (ACS) Postdoctoral Fellowship Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Single-nuclei multiome-seq and modeling of intestinal cell types nominates regulatory mechanisms driving IBD genetic risk and anti-TNF nonresponse 2309915

Abstract Introduction Inflammatory Bowel Disease (IBD) is an autoinflammatory condition, with increasing incidence in children. TNF inhibitor (TNFi) therapy is the only FDA-approved biologic in pediatric IBD, yet only a third of patients achieve mucosal healing with TNFi. We recently identified polygenic risk scores for IBD as a top predictor of TNFi response in an ensemble model integrating the PRS with clinical measurements. The majority of IBD risk variants are noncoding. If causal, they likely alter transcription factor (TF) binding and downstream gene expression in particular cell types and contexts. Methods Using single-nuclei multiome sequencing (snRNA-seq and snATAC-seq) of pediatric IBD intestine from TNFi responders and nonresponders, we generated accessible chromatin maps for 27 cell types, resolved by TNFi response status. CD4 T cells and macrophages were top-ranked mediators of IBD genetic risk, especially in the TNFi non-response context (PMID:38405748). We curated ligand-response gene signatures to identify cell-cell signaling responses (1) targeted by IBD genetic risk variants and (2) altered in TNFi nonresponders. In CD4 T cells, we constructed genome-scale gene regulatory networks (GRNs) (PMID:36945549) to predict TF mediators of IBD genetic risk and TNFi response. Results Genetic risk colocalization identified Th17 cells and inflammatory monocytes/macrophages as key mediators of IBD risk. In Th17 cells, GRN modeling revealed TF activities associated with TNFi nonresponse, including IBD risk enhancer targets (e.g., CREM, GFI1) and TFs downstream of signaling pathways targeted by next-gen IBD therapeutics (e.g., STAT4 downstream of IL23/IL12). While, in macrophage, IBD risk variant targets and TNFi-nonresponse gene signatures were linked to TNF and IL1 signaling. Conclusion Our integrated modeling nominates cell-type-specific regulatory mechanisms linking IBD genetic risk to TNFi non-response and prioritizes next-gen IBD biologic targets. Funding Source Cincinnati Children’s Research Foundation by Academic Research Committee (ARC) grant #53671 (ERM, LAD, AGM, LCK, MTW); National Institute of Health (NIH) funding: R01AI153442 (ERM), the CCRF Digestive Health Center grant, P30DK078392 (LAD) Topic Categories Computational and Systems Immunology (COMP)

A Multidonor Plasma Antibody Class with Broad Activity Against H5N1 Neuraminidase 2309455

Abstract Introduction Highly pathogenic H5N1 influenza viruses remain a significant pandemic threat, underscoring the need to define antibody responses that target conserved viral epitopes. Here, we characterize a panel of plasma IgG—derived human neuraminidase (NA)—specific monoclonal antibodies (mAbs) that share a convergent CDR-H3 “DR motif,” previously implicated in substrate mimicry at the NA active site. Methods Using IgG proteomics discovery, ELISA, immunofluorescence assays, enzymatic inhibition, and microneutralization assays, we show that multiple DR-motif mAbs bind H5N1 NA with high potency and directly inhibit NA activity. Results Several antibodies exhibit cross-reactivity with divergent influenza A strains and target overlapping epitopes within the NA catalytic site. Importantly, two DR-motif mAbs (41508 and 125308) confer protection in C57BL/6 mice against lethal challenge with A/Texas/37/2024 (H5N1; clade 2.3.4.4b, genotype B3.13). Conclusion Together, these data define a multidonor class of circulating, functional human NA-directed antibodies with broad antiviral activity and highlight the NA active site as an accessible and vulnerable target for next-generation influenza vaccines and antibody-based countermeasures. Funding Source n/a Topic Categories Vaccines and Immunotherapy (VAC)

Neutrophil NETosis and cell free DNA suppress T cell responses in glioblastoma 2260418

Abstract Introduction Glioblastoma is an aggressive brain cancer with poor prognosis for which immunotherapies have fared poorly. In part, the failure in immunotherapies is attributed to the existence of severe local and systemic immunosuppression in GBM. We determined that majority of hallmark features of systemic immunosuppression were linked to immunosuppressive factors in serum. Our data indicate that serum of glioma-bearing mice harbors heightened levels of cell-free DNA (CF-DNA). Methods We evaluated the extent to which CF-DNA contributes to systemic T cell dysfunction in mouse models of GBM and GBM patient plasma samples. Results We determined that majority of hallmark features of systemic immunosuppression were linked to immunosuppressive factors in serum. Our data indicate that serum of glioma-bearing mice harbors heightened levels of cell-free DNA (CF-DNA). Removing CF-DNA from serum improves T cell responses while adding CF-DNA derived from serum of glioma-bearing subjects to healthy T cells directly inhibits their functions. We have also verified that CF-DNA is detectible in sera isolated from patients with GBM. When compared to healthy controls, GBM patients have higher quantities of CF-DNA is their plasma. Interestingly, our preliminary data implicates neutrophil NETosis as the likely source of CF-DNA in sera of GBM patients. Moreover, we stained GBM tumor sections with citrullinated histone 3 (cit-H3) to evaluate the extent of NETosis in GBM patients. We determined evidence of significant NETosis within patient brain tumors. Systemic immunosuppression is detrimental to the success of immune-modulating therapies and patient survival. By identifying and removing the source of systemic immunosuppression, we will enhance immune responses against the tumor likely improving patient survival. Conclusion Glioblastoma causes immune suppression outside of the brain via the released CF-DNA. Our studies in GBM patients and mouse models help develop strategies to reverse immune dysfunction and improve outcomes. Funding Source NIH K99/R00 Topic Categories Immune Mechanisms of Human Disease (HUM)

Mpox virus can suppress virus sensing and antiviral Responses in testicular cells: potential role of Sirtuins 2310247

Abstract Introduction The global outbreak of Mpox virus (MPXV) clade IIb, comprising ∼106,000 cases predominantly in men, has raised serious concerns regarding sexual transmission. Pox viruses evade immune detection by blocking pathogen sensing and inhibiting antiviral responses. We here characterized MPXV infection in human testicular cells to gain insights into the landscape of cell-type-specific testicular response to infection. Methods Human primary Sertoli (SC) and Leydig (LC) cells were infected with MPXV clade IIb (MOI 0.005). Infection was confirmed by quantitative PCR and plaque assays. RNA-seq analysis was conducted using RNA samples from infected SC and LC at days 1, 3, and 5 post-infection (dpi). Differentially expressed genes (DEGs) were identified based on strict cutoffs (log2 fold-change >|1.5|; adj. p-value < 0.05) and analyzed for enrichment in virus-response and steroidogenesis pathways using Ingenuity Pathway Analysis. Results Productive infection was confirmed in both SC and LC with peak virus titers at 5 dpi without significant cell death. Although viral replication was evident by 1 dpi, the number of DEGs remained low (86 in LC and 51 in SC), while they were above 3000 at 5 dpi in both cell types. Notably, canonical viral sensing, cell death, and inflammatory pathways showed no significant activation in either cell type at any time point. Sirtuin family genes (SIRTs) were upregulated at 3 and 5 dpi, correlating with downregulation of antiviral response genes, including DNA virus-sensing gene IFI16. Conclusion This is the first evidence that MPXV can infect key human testicular types and suppress key antiviral pathways similar to vaccinia virus. In addition to functioning as post-virus sensing immune brakes, SIRTs are also crucial for spermatogenesis. Our data suggest that SIRT upregulation favors viral replication by promoting cell survival via autophagy and apoptosis inhibition, while simultaneously dampening innate immunity through suppression of NF-κB and interferon signaling. Funding Source R03AI188484 Topic Categories Viral Immunology (VIR)

Conserved Human CD4 T Cell Targets Inform Group A Streptococcus Vaccine Design 2309249

Abstract Introduction Group A Streptococcus (GAS) is a globally prevalent bacterial pathogen responsible for over 500,000 deaths annually and a leading cause of post-infectious autoimmune diseases, yet no vaccine is available. Current vaccine development focuses primarily on antibody responses to a limited set of antigens, while targets of T cell immunity remain largely undefined. Methods In this study, we performed comprehensive mapping of human CD4 T cell responses to GAS through high-throughput screening of 2,661 peptides spanning 196 literature-curated antigens in donors with prior GAS exposure. Results We identified 392 epitopes mapping to 85 antigens, revealing an unexpectedly broad T cell repertoire. Notably, 20 immunodominant antigens, recognized by ≥ 20% of donors, accounted for 75% of total GAS-specific CD4 T cell responses. These included known vaccine candidates (SCPA, streptolysin O, SpyCEP) as well as multiple novel targets with comparable or superior immunogenicity (SibA, Isp and NADase). Functional enrichment analysis revealed that dominant antigens exhibited hydrolase activity (70%) and were predominantly extracellular or cell wall-associated (60%). Conservation analysis across 2,083 globally sampled GAS genomes revealed strong sequence conservation among dominant antigens, supporting universal vaccine potential. T cell response magnitude decreased with donor age, mirroring serological patterns and suggesting reduced recent infection frequency in older individuals. Lastly, we leveraged epitope mapping to develop optimized peptide pools enabling efficient detection of GAS-specific T cells for future mechanistic and clinical studies. Conclusion Together, this study defines the most comprehensive mapping of human GAS-specific CD4+ T cell targets to date. These findings provide a roadmap for next-generation GAS vaccines incorporating robust T cell targets and establish tools to investigate immunopathology in GAS-associated autoimmune sequelae. Funding Source Leducq Foundation Network Research Grant (RHD23VAC02) and NIH Ruth L. Kirschstein National Research Service postdoctoral fellow Award (NRSA) T32HL069766. Topic Categories Vaccines and Immunotherapy (VAC)

Localized Potential Regulation on Polymer Donor Backbone Suppresses Energetic Disorder for Efficient, Stable and Scalable Organic Solar Cells

ABSTRACT High energetic disorder and short exciton diffusion lengths in low‐crystallinity polymer donors ( P D s) critically limit exciton dissociation, charge transport, efficiency, stability, and scalability of organic solar cells (OSCs). Here we report a localized potential regulation strategy to intrinsically suppress energetic disorder of P D through precise backbone engineering. By introducing a strongly electron‐deficient unit into the structurally disordered backbone of the DP1, we developed a P D DP10 that exhibits enhanced backbone rigidity, optimized local electronic polarization, and favorable miscibility with the acceptor L8‐BO. These features synergistically reduce the exciton binding energy and suppress exciton–phonon coupling, thereby extending the exciton diffusion length from 15.5 to 19.9 nm. Consequently, DP10 enables balanced, trap‐tolerant charge transport and reduced non‐radiative recombination in devices. The DP10:L8‐BO binary system achieves a device efficiency of 19.51%, together with exceptional thermal stability and outstanding thickness‐tolerant performance. Additionally, a DP10:L8‐BO:BTP‐eC9 ternary device reaches an efficiency of 20.57% and a 15.1 cm 2 solar module delivers 17.20% efficiency, with impressive fill factors of 81.04% and 77.39%, respectively. This work establishes localized potential regulation as a powerful molecular design principle of P D s for simultaneously achieving high efficiency, thermal stability, and processing robustness in next‐generation OSCs.

Integrated computational and in vitro analysis reveals the toxic mechanisms of PFAS in female diminished ovarian reserve

NLRP3 confers regulatory T cell-mediated control of intestinal inflammation 2256029

Abstract Introduction Inflammasome activation is an essential component of innate immunity. Recently, its role in regulating various T cell responses has also become increasingly recognized. Interestingly, we found increased expression of Nlrp3 and many genes associated with the NLRP3 inflammasome pathway in intestinal regulatory T cells (Tregs) from mice with autoimmune-mediated inflammation. This unexpected finding of a pro-inflammatory pathway upregulated in a cell type typically known for suppressing immune responses called for further investigation. Methods We developed a new mouse model with a Treg-specific deletion of Nlrp3 and subjected the mice to a variety of disease models of intestinal inflammation. We then performed RNA-seq on NLRP3-sufficient and NLRP3-deficient intestinal Tregs isolated from a similar inflammatory environment to gain mechanistic insights. To further characterize the subset of intestinal Tregs that upregulated Nlrp3, we used the novel PrimeFlow RNA Assay. Results In multiple disease models, loss of NLRP3 in Tregs led to elevated Th17 responses accompanied by reduced Th1 responses despite similar Treg frequencies and comparable levels of Foxp3 expression. Mechanistically, NLRP3 likely confers Treg suppressor function against Th17 cells through driving the production of molecules that antagonize IL-1 signaling. Conclusion Collectively, we demonstrate a previously underappreciated anti-inflammatory role of NLRP3 in Tregs in controlling Th17 responses in the intestines. Ultimately, our work should guide effective therapies for intestinal disorders where the anti-inflammatory vs. proinflammatory role of NLRP3 is still debated. Funding Source NIDDK F31 2023-2025 and Biolegend Fellow in Immunology 2021-2022 Topic Categories Immune Response Regulation: Cellular Mechanisms (IRC)