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Ordered Polar Topological Domains Enabling Giant Second‐Harmonic Generation in Ferroelectric Nematic Liquid Crystals
ABSTRACT Developing high‐performance nonlinear optical materials that simultaneously deliver large effective nonlinearity and device‐level integrability remains a longstanding challenge. Ferroelectric nematic liquid crystals (FNLC) have emerged as a promising platform owing to their intrinsic non‐centrosymmetry, theoretically predicted large second‐order nonlinearity, and solution processability on diverse substrates. However, realizing strong second‐harmonic generation (SHG) in FNLCs has been hindered by pronounced orientational disorder inherent to fluidic systems without lattice constraints. Here, this limitation is overcome through a surface‐anchoring strategy that induces highly ordered polar topological structures within self‐assembled FNLC droplets. The resulting architecture yields a giant effective SHG coefficient of 56.9 pm/V—an order of magnitude higher than previously reported FNLC systems—together with SHG efficiency surpassing that of benchmark LiNbO 3 films of comparable thickness. Moreover, the system exhibits broadband SHG response, while the engineered polar topology enables passive, field‐free spatial optical modulation with a contrast ratio of 330%. The combination of giant nonlinearity, outstanding SHG efficiency, broadband SHG response, spatial optical modulation, and solution processability establishes a new paradigm for integrated nonlinear photonic devices.
Lymphoid Tissue-Activated Donor T cells (LADTs) for Improved Donor Lymphocyte Infusion (DLI) against Blood Cancers 2250463
Abstract Introduction Donor lymphocyte infusion (DLI) is a key strategy for treating relapses after HLA-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT) for blood cancers. In DLI, T cell alloreactivity drives both the highly beneficial graft vs leukemia (GvL) effect and the potentially fatal graft vs host diseases (GvHDs), which severely limits DLI’s application and efficacy. Here, we propose to separate GvL and GvHD by engineering Lymphoid Tissue-Activated Donor T cells (LADTs) that display alloreactivity only in lymphoid tissues where cancer cells accumulate but not in organs involved in GvHD including the skin, liver and guts. Methods To generate LADTs, PBMCs were first used to expand alloreactive T cells from an HLA-mismatched donor. Then, T cell alloreactivity was abrogated by knocking out CD3ε using CRISPR/Cas9, which blocks cell surface expression of the TCR/CD3 complex. Finally, the T cells were engrafted with a B cell-sensing mechanism that includes a CD19-specific synthetic Notch receptor (synNotch) and an expression cassette that drives the expression of an exogenous copy of CD3ε in response to synNotch engagement. This mechanism enables LADTs to restore alloreactivity only in lymphoid organs where B cells are abundant but not in other organs where B cells are scarce. Results We have established a process of generating LADTs with consistent yields. LADTs bearing CD19-specific synNotch receptors restored surface expression of TCR/CD3 in response to CD19+ cells and showed strong cytotoxicity against HLA+ U266 myeloma cells but not against HLA- ones. After removing CD19+ cells, LADTs gradually lost TCR/CD3 cell surface expression. Conclusion Our study demonstrated the feasibility of LADT generation and the proof-of-concept of using LADTs for improved DLI. This cellular immunotherapy approach does not depend on cancer-specific biomarkers thus may be applied to a wide range of hematopoietic malignancies. Funding Source Nemours Foundation Topic Categories Tumor Immunology: Checkpoints, Prevention, and Treatment (TIPT)
Understanding unconventional T cell development to generate an ‘off the shelf’ immunotherapy to treat human diseases 2226160
Abstract Introduction Unconventional T cells represent more than 10% of total T cells within the blood and tissues of humans and play critical roles in cancer and infection. Human unconventional T cells exhibit dual effector functions; they rapidly secrete cytokines upon stimulation, and they express granzymes and perforin that enables them to lyse infected or cancerous cells. Methods Here, we studied the development of unconventional T cells isolated from the postnatal human thymus and from artificial thymus organoids, tracking changes in development using flow cytometry, RNA-sequencing, precursor-product experiments and cytokine expression. Results Our research has deciphered how unconventional T cell subsets including MAIT cells, γδ T cells and NKT cells develop in the human thymus. We reveal that all three subsets develop via a linear pathway defined by the upregulation of the transcription factor PLZF and the Killer cell lectin-like receptor subfamily B, member 1 (CD161). Further, functional programming of unconventional T cells takes place in the human thymus. We demonstrate that the development of unconventional T cells can be faithfully reproduced using artificial thymus organoids, which allows for detailed interrogation of this process by removal of key factors that influence their development. Furthermore, we demonstrate that our unconventional T cells can be developed as an “off the shelf’ immunotherapy. Conclusion Collectively, our work reveals how understanding unconventional T cell development in the human thymus has led to new opportunities to utilize these cells to treat human diseases such as cancer or microbial resistant infections. Funding Source NHMRC Topic Categories Translational and Interventional Immunology (TI)
Assessment and development of computational strategies for assembling the human FCGR2/3 locus using Nanopore long-read sequencing data 2309911
Abstract Introduction The human FCGR locus encodes Fc gamma receptors (FcγRs) that mediate antibody effector functions and shape responses to infection, autoimmunity, and therapeutic antibodies. Genotyping this region is difficult due to copy number variations (CNVs), small nucleotide variants (SNVs), and highly homologous paralogs in strong linkage disequilibrium. We assessed and developed computational strategies to haplotype the FCGR2/3 locus using Nanopore long-read sequencing. Methods We analyzed 25 individuals from five superpopulations from the 1000 Genome Project (1kGP) Nanopore whole-genome sequencing dataset and assembled FCGR2/3 haplotypes. Performance was evaluated through comparisons to known haplotypes, depth-based copy-number estimates, published assemblies, trio-based validation when available, and independently assembled PacBio HiFi haplotypes from the same individuals. Nucleotide-level assessment was also performed by mapping FcγR genes annotated on human reference genome (T2T-CHM13) to each haplotype assembly to confirm complete coding sequences (CDS). Results We generated contiguous, structurally validated FCGR2/3 haplotypes for 24 individuals. High-copy number FCGR2/3 haplotypes were present in all individuals, with 62% carrying two copies of haplotype 1 (H1) and 20.8% carrying one H1 and one haplotype 5 (H5). Low-copy number haplotypes were rare, observed in 12.5% of individuals carrying one H1 and haplotype 2 (H2). For a subset of individuals, we validated complete, full-length CDSs and generated variant sets relative to T2T-CHM13. Conclusion This work provides a validated computational framework for assembling FCGR2/3 haplotypes, and in some cases improved, germline references for 1kGP individuals. Future studies will focus on applying this framework to efficiently genotype the FCGR2/3 locus in individuals enrolled in human studies. Funding Source NIH/NIAID, R01AI187318 Topic Categories Technological Innovations in Immunology (TECH)
Quantify the age-related decline of CD8+ T cell response to a novel antigen 2302770
Abstract Introduction Aging impairs T cell—mediated immunity. Mechanistically, this decline is thought to result from two major factors: (1) cumulative antigenic stimulation over time driving T cell senescence, and (2) loss of appropriate antigen-specific TCRs due to insufficient usage. Although age-related contraction of the TCR repertoire has been reported, it remains unclear which specific TCRs are lost and how this loss affects responses to novel antigens. To address this question, we examined how aging alters the size, clonality, and functional quality of antigen-specific CD8+ T cell responses. Methods Young (2—3 months) and aged (20—24 months) C57BL/6J mice were infected with Listeria monocytogenes expressing chicken ovalbumin (OVA). OVA257–264 (SIINFEKL)-specific CD8+ T cells were quantified using tetramer staining and profiled by single-cell TCR sequencing. TCR diversity was evaluated using metrics of clonal richness and evenness, while TCR functional quality was assessed based on antigen-binding affinity and downstream signaling capacity. Results Aged mice exhibited significantly delayed clearance of Listeria compared to young mice. OVA-specific CD8+ T cells were markedly reduced in aged mice before infection, at peak response, and during the memory phase. Single-cell TCR sequencing revealed a substantial reduction in OVA-specific TCR repertoire diversity in aged mice, with a 57% decrease in effector cells and an 87% decrease in memory cells. Moreover, clonally expanded OVA-specific CD8+ T cells from aged mice showed impaired tetramer binding and diminished signaling following antigen engagement. Conclusion Our findings demonstrate that aging profoundly reduces CD8+ T cell responses to a novel antigen due to loss of both TCR quantity and functional quality. This deterioration of antigen-specific immunity may contribute to increased infection susceptibility and diminished vaccine efficacy in older individuals. Funding Source Intramural Research Programs of the NIH, National Institute on Aging. Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
Immunomodulatory Effects of Atrazine in Human A549 Lung Epithelial Cells 2310225
Abstract Introduction The endocrine system is objectively essential for immune functioning and formation. Agents such as Atrazine (ATR) disrupt the ability of the endocrine system to signal the immune system, resulting in an increased risk of autoimmunity, cancer, and neurodegenerative diseases. Atrazine is sprayed on corn cropping systems across the United States, which introduces small particles that can be absorbed into the alveolar sacs, damaging epithelial lung cells and possibly impacting gas exchange. This molecule has been known to inhibit cytokine production, such as IL-6, and cell proliferation. However, there remains a gap in current research as to which pathways ATR inhibits. Methods This research utilized the A549 cell line to evaluate cytotoxic effects post ATR exposure. Atrazine was prepared in DMSO and diluted with media to obtain concentrations of 0, 5, 10, 20, 50, and 100 µM. Cells were exposed for periods of 24, 48, and 72 hours. Following exposure, MTT assay was utilized to confirm treatment with ATR did not result in cell death but remained metabolically active before immunocytochemistry can occur. Results The cell viability (%) mean after 24 hours at 100 μg/mL was significantly lowered to ∼50% when compared to the control of 100%. After both 48 and 72 hours of exposure to 50 and 100 μg/mL of ATR, cells showed significantly less metabolic activity when compared to the control. 48 hours with means of ∼50% and ∼25% for the 50 and 100 μg/mL, respectively, and 72 hours with means of ∼40% and ∼45% of ATR, respectively. Atrazine, particularly at higher concentrations for longer durations, decreased the cells’ metabolic activity, meaning ATR affected cell function. Conclusion Overall, the results indicate that ATR has a statistically significant effect on A549 cells at multiple concentrations over 24-to-72-hour time frames. Further research will dive into specific cytokine signaling pathways, to understand the mechanisms involved in immunomodulation. Funding Source This research project was funded in part by the Judith Strong Scholars research fund. Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
A discovery framework for IBD target discovery utilizing a tissue-derived single cell atlas 2259245
Abstract Introduction Despite advancements for Inflammatory Bowel Disease (IBD), high rates of non-response, relapse, and adverse effects underscore the need for novel therapeutic interventions. This necessity is compounded by the inherent complexity of IBD; wherein, multiple resident cellular components receive and generate aberrant signals within tissues and migrating immune cells. Methods To identify novel therapeutic targets, we constructed a harmonized single-cell tissue atlas (IBD atlas) derived from published clinical single-cell datasets. Machine learning-based methods were employed to extract relevant transcriptional signals and their contributing genes, which were then ranked using orthogonal translation metrics to generate a list of candidate targets for in vitro functional validation. Using this approach, we nominated candidates for cell type-specific functional evaluation in macrophages and fibroblasts. We utilized in vitro models optimized to recapitulate specific transcriptional states of these cells in inflamed IBD tissue, and evaluated the impact of target deletion in the presence or absence of appropriate ligands. Results We highlight a specific target for which deletion reduced the inflammatory proteome and induced an anti-inflammatory transcriptional state similar to that of healthy intestinal myeloid cells with a reduction in IBD associated signatures. Comparison to transcriptional shifts induced by standard-of-care therapies showed that this target’s profile is distinct from anti-TNF and anti-integrin while recapitulating anti-inflammatory effects of JAK inhibitors. We contrast this with a second target for which ablation induced a proinflammatory response in macrophages but a favorable, anti-fibrotic response in intestinal fibroblasts highlighting two novel cell type targeted therapeutic approaches. Conclusion In conclusion, our framework leverages a robust single-cell data foundation to nominate disease-relevant targets in specific cell types optimally poised for desirable clinical outcomes. Funding Source n/a Topic Categories Mucosal and Regional Immunology (MUC)
Tumor extracellular vesicles induce immunosuppressive myeloid cells via TIGIT/CD155 axis, dampening NLRP3-driven inflammatory response in glioblastoma 2257553
Abstract Introduction Glioblastoma (GBM), the most aggressive primary brain tumor in adults, is marked by profound immune suppression. Tumor-derived extracellular vesicles (TEVs) are increasingly recognized as key mediators of this immunosuppressive environment. Methods In this study, we examined the tumor-immune microenvironment using ultrasonic aspirate samples from GBM patients (n = 14). Results Several immune checkpoint molecules, including PDL1, B7-H3, and VISTA, were notably elevated. TIGIT, a receptor typically associated with T cells, was widely expressed across multiple myeloid subsets, including monocytic myeloid-derived suppressor cells (mMDSCs; 64.27±7.99%) and non-classical monocytes (NCM; 39.8±4.37%). Co-culture of healthy donor CD11b+ cells (n = 10) with TEVs replicated similar effects (mMDSCs; control 11.691.87 vs EV treated 23.322.0). We found CD155, a primary TIGIT ligand, to be heavily enriched in TEVs and co-localized with TIGIT in co-culture, suggesting the TIGIT/CD155 axis may be involved in promoting immune suppression. induced MDSCs markedly suppressed T cell proliferation (30.65 ± 5.97% vs. control 86.47 ± 2.94, p = 0.0001), accompanied by elevated Th2 cytokine production and increased NLRP3 and IL-1β expression. Interestingly, TIGIT knockdown did not affect NLRP3 expression, but did restore T cell proliferation (64.13±1.33%; p = 0.02) and pro-inflammatory cytokine production. However, this effect was lost upon NLRP3 knockdown, indicating its essential role in mediating immune rescue. Furthermore, IL-13 (a Th2 cytokine) blockade, in this model, enhanced pro-inflammatory response by suppressing phosphorylation of STAT-6 protein. Conclusion Overall, our findings highlight a crucial role for the TIGIT/CD155 axis in driving myeloid-mediated immune suppression in GBM. Furthermore, TIGIT inhibition may unmask NLRP3 function, potentially contributing to immune rescue. Funding Source Neurosurgery Research & Education Foundation through the B*CURED & NREF Young Clinician Investigator Award, as well as intramural K12 funding through the Institute for Clinical and Translational Research and the NCI Paul Calabresi Career Development Award (1K12CA279871). Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Strong Enhancement of Room Temperature Ferrimagnetism in Insulating CoFe <sub>2</sub> O <sub>4</sub> by Site‐Specific Hydrogen Intercalation
ABSTRACT Strong enhancement of magnetization is highly desirable for increasing device efficiency, improving temperature stability, enhancing sensitivity, and enabling device miniaturization. Here, we demonstrate a 2.6−4.7 times higher room temperature ferrimagnetic moment (730−770 emu cc −1 ) compared to 170−260 emu cc −1 in as‐grown CoFe 2 O 4 films, achieved by site‐specifically intercalating hydrogen ions near the Fe 3+ in tetrahedral sites. While the spin moments of the Fe 3 + at octahedral and tetrahedral sites cancel each other out in as‐grown films, X‐ray spectroscopy of hydrogenated films reveals that both Fe 3+ and Co 2+ at octahedral sites can contribute to the enhanced total magnetic moment. This robust enhancement in insulating CoFe 2 O 4 , exhibiting high reproducibility over ≈10 cycles and long‐term stability over ≈10 d, suggests that hydrogenic‐ferrimagnetic coupling holds great promise for ultralow power electronics operating at high frequencies.
Sepsis-induced alterations to the microenvironment promotes maladaptive emergency granulopoiesis and susceptibility to Streptococcus pneumoniae 2253166
Abstract Introduction Sepsis survivors exhibit immune dysfunction and profound hematological changes that increase a patient’s risk for recurrent infections and hospital readmission. Methods Using a cecal-ligation and puncture model (CLP; causing 50-70% mortality), followed by Streptococcus pneumoniae (Spn) intranasal challenge, we evaluated mechanisms driving increased susceptibility to secondary infection. Results Sham-surgery controls uniformly survived Spn infection, whereas CLP survivors exhibited 100% morality when challenged day 20 or 40 post-surgery, revealing long-term susceptibility to Spn infection. Neutrophils were increased in the bronchoalveolar lavage fluid (BALF) of both sham and CLP mice, however, only CLP mice exhibited significantly increased neutrophils in the blood, spleen, and liver. Following challenge, CLP mice exhibited maladaptive emergency granulopoiesis evidenced by systemic neutrophilia, expansion of bone marrow (BM) hematopoietic stem and progenitor cells (HSPCs) and marked reduction of mature neutrophils in the BM. Using BM transplantation, we found that sepsis-induced alterations to the microenvironment were sufficient to promote maladaptive emergency granulopoiesis during intranasal Spn challenge. Sepsis survivors challenged with Spn exhibited increased stromal cell populations and osteoblastic progenitors, suggesting profound BM niche remodeling, that correlated with robust splenic extramedullary granulopoiesis. Susceptibility to pneumonia was not due to lung-specific damage as CLP survivors were unable to control intravenous Spn infection. Conclusion Therefore, despite robust granulopoiesis and increased neutrophils, particularly splenic neutrophils, sepsis results in long-term neutrophil dysfunction. Our data demonstrate that sepsis drives durable changes to the microenvironment that support maladaptive granulopoieses programs and increased susceptibility to Spn and pneumonia. Funding Source 5R35GM131842 Topic Categories Innate Immune Responses and Host Defense: Cellular Mechanisms (INC)
Metabolic checkpoint AhR prevents pathogenic B cell expansion in murine lupus 2239969
Abstract Introduction The aryl hydrocarbon receptor (AHR) integrates metabolic and immune cues, but its role in B-cell regulation during lupus remains unclear. We investigated whether AHR restrains pathogenic B-cell development under lupus-mimicking conditions using toll-like receptor 7 (TLR7) stimulation. Methods B-cell-specific Ahr knockout (Ahrf/f Cd19.Cre+/- or Ahr B-KO) mice were generated on B6 and lupus-prone BXD2 backgrounds and compared with Ahr B-wild type (Ahr B-WT) controls. TLR7 was activated using R848. AHR was activated using tryptophan (Trp) metabolites 6-formylindolo[3,2-b]carbazole (FICZ) and kynurenine (KYN). B cell subpopulations were determined by flow cytometry. In vitro B cell responses to FICZ were analyzed for transcriptomics using the 10x Chromium single cell RNA-sequencing (scRNA-seq) method while the metabolomics of B cells was analyzed by untargeted LC-MS/MS. Results In vivo, FICZ reduced R848-induced T-bet+CXCR3+ B cells, germinal center B cells, and CXCR3+CD138+ plasma cells in BXD2 mice. Loss of Ahr in B cells resulted in an increase in R848-induced anti-DNA IgM⁻IgD⁻ and human DN2-equivalents (T-bet+FCRL5+, T-bet+CXCR3+) B cells in B6 Ahr B-KO, compared to Ahr B-WT mice. In vitro, AHR activation preserved naïve B cells and blocked their conversion into T-bet+CD11c+CXCR3+ cells, but this protection was absent in BXD2 Ahr B-KO B cells. AHR activation increased KYN- and indole-pyruvic acid (IPyA) derived AHR agonistic ligand metabolites in BXD2 Ahr intact but not Ahr deficient B cells. scRNA-seq confirmed upregulation of IPyA (Il4i1) and KYN (Kynu, Kmo, Haao) catabolic enzyme genes in BXD2 Ahr B-WT but not Ahr B-KO B cells. Conclusion These findings identify AHR as a metabolic-immune checkpoint that restrains lupus-related autoantibody production. By uncovering a B-cell intrinsic self-reinforcing pathway that links Trp metabolism to AHR signaling, this work reveals AHR as a promising therapeutic target in lupus and other autoimmune diseases. Funding Source VA Merit Review grant (1I01BX006099), NIH R01 AI134023, and 1R01AI182085 to J.D.M. Topic Categories Therapeutic Approaches to Autoimmunity (THER)
VISTA Regulates Proinflammatory Function by Tissue Resident Peritoneal Macrophages in Response to Hemorrhagic Shock and Septic Challenge 2310024
Abstract Introduction Traumatic injury, like hemorrhagic shock, serves as an immune predisposition ‘priming’ event, where upon a secondary challenge a heightened inflammatory response can occur, culminating in clinical complications such as septic multiple organ failure. Large (LPM) and small (SPM) peritoneal tissue resident macrophages play a significant role in peritoneal homeostatic functions and in initiating the local immune response upon injury/infection. V-domain Ig suppressor of T-cell Activation (VISTA) has a protective effect in response to shock and septic challenge able to suppress T-cell effector function and promoting an anti-inflammatory phenotype in monocyte-derived macrophages (Mo-Macs). However, VISTA function in tissue resident peritoneal macrophages in response to shock and sepsis has yet to be studied. Methods Using a ‘double hit’ shock and sepsis induced multiple organ injury mouse model, consisting of fixed pressure hemorrhagic shock (Hem) insult followed by sepsis through the cecal ligation & puncture (CLP) method, the effects of VISTA function on the immune contribution by resident peritoneal macrophages was characterized through flow cytometry. Results Our initial data shows that following Hem/CLP, VISTA expression is upregulated in LPMs but remains constant in SPMs and Mo-Macs. However, both LPMs and SPMs are significantly depleted from the peritoneum after Hem/CLP challenge. Lastly, pro-inflammatory markers are increased in SPMs and Mo-Macs, and VISTA deficiency leads to downregulation of MHCII expression in SPMs. Conclusion These results indicate that VISTA regulates the inflammatory SPM population through its constitutive expression in peritoneal macrophages as a mechanism that promotes an anti-inflammatory phenotype and through VISTA and MHCII mediated interaction with T cells. VISTA plays a novel role as a negative regulator of innate inflammation following shock and septic insult by regulating macrophage mediated inflammation through the activation of anti-inflammatory pathways. Funding Source R35 GM118097 Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)
Rejuvenating the Aged Hematopoietic System: IL-4 Signaling Restores Lymphopoiesis and Systemic Function 2302788
Abstract Introduction Chronic inflammation and aging disrupt hematopoietic homeostasis, creating a persistent “myeloid bias” that perpetuates immune dysfunction. While the drivers of this skew are well-characterized, strategies to reverse it remain elusive. We hypothesize that anti-inflammatory cytokines act as a fundamental “reset” mechanism, capable of reprogramming hematopoietic stem and progenitor cells (HSPCs) to counteract myeloid skewing and actively restore lymphoid commitment. Methods We utilized in vitro differentiation assays and in vivo models, including LPS-induced inflammation and aging. Hematopoietic stem and progenitor cell (HSPC) subsets were analyzed via flow cytometry and the transcriptomic change in STAT6-KO or IL-4 injected mice. Cellular specificity was dissected using lineage-tracing and bone marrow transplantation. Functional rejuvenation was assessed via cognitive, motor, and metabolic profiling following IL-4 administration. Results We found that IL-4 signaling effectively resolved the inflammation- and aging-driven myeloid bias by promoting lymphoid differentiation in multipotent progenitors (MPPs), but not in hematopoietic stem cells (HSCs). Mechanistically, FLT3 functionally cooperated with the IL-4 receptor to potentiate STAT6 phosphorylation and activation in a cell-intrinsic manner within MPPs, upregulating lymphoid-specific genes while suppressing myeloid programs. Strikingly, systemic IL-4 treatment rejuvenated the aged hematopoietic system, restoring B and T cell output to youthful levels. This hematopoietic restoration translated into systemic benefits, including measurable improvements in cognitive performance, motor coordination, and metabolic parameters in aged mice. Conclusion Our findings identify the IL-4-FLT3-STAT6 signaling axis as a critical regulator of MPP lineage plasticity. This study positions IL-4 as a promising therapeutic agent for correcting hematopoietic dysregulation and mitigating the systemic functional decline associated with inflammaging. Funding Source the Milky Way Research Foundation and the Howard Hughes Medical Institute (HHMI) Topic Categories Hematopoiesis and Immune System Development (HEM)
Effect of Aptamin C on NK cell activity and cytotoxicity in vivo and in vitro: a comparison study with the effect of vitamin C 2330057
Abstract Introduction NK cells are crucial for innate immunity and rapidly target abnormal cells through ligand-receptor signaling, without prior sensitization. Vitamin C is a bioenhancer of NK cells; however, its susceptibility to oxidation limits its efficacy in NK cell activation. This study evaluated the efficacy of Aptamin C, a stable conjugate of vitamin C and an aptamer, in enhancing NK cell activation. Methods For the in vivo study, 109 participants were administered Aptamin C (36.057 mg/day) or a placebo for 4 weeks. NK cell cytotoxicity and serum levels of NK cell related cytokines and cytotoxic granules were assessed. In parallel, in vitro experiments were conducted using NK-92 cells to compare the effects of Aptamin C with those of conventional vitamin C on NK cell proliferation, survival, and cytotoxic activity. Results In vivo, Aptamin C administration resulted in significant increases in NK cell cytotoxicity at weeks 2 and 4. Additionally, cytokine and granule levels associated with NK cell activity peaked in the serum 4 weeks after Aptamin C intake. Consistently, in vitro experiments revealed that Aptamin C treatment led to superior proliferation, survival, and cytotoxic granule production in NK-92 cells relative to vitamin C-treated controls. Conclusion These results demonstrate that Aptamin C more effectively enhances NK cell activity than vitamin C, highlighting its potential as a novel immunomodulatory supplement for NK cell based immune enhancement strategies. Funding Source This work was supported by N Therapeutics Co., Ltd. and Nexmos. Topic Categories Immune Mechanisms of Human Disease (HUM)
Investigating the Role of 7-Ketocholesterol in Alleviating Inflammatory Bowel Disease (IBD) using an Acute Mouse Model of Chemically Induced IBD 2259324
Abstract Introduction Inflammatory Bowel Disease (IBD) is an inflammatory condition involving complex interactions among the intestinal epithelium, immune system, and gut microbiome. T-helper (Th) cells, particularly CD4+ effector cells, play a significant role in sustained inflammation and tissue damage. This study explores a novel compound, 7-Ketocholesterol (7-KC), an oxysterol, caged in methyl-β-Cyclodextrins (mβCD) for delivery to the cell membrane to disrupt lipid raft-based membrane order. Methods The approach targets CD4+ T cells and inflamed gut tissues in Dextran Sulfate Sodium (DSS)-induced acute IBD mouse model. Disease severity in the 7-KC-treated test and control mice was monitored daily using weight loss, water intake, and stool hemoccult scoring. At the experimental endpoint, mice were euthanized to measure colon length. The proximal, medial, and distal parts of the colon were prepared for histological and immunohistochemical analyses. Results The DSS-only group (disease control) developed severe colitis, confirmed by a significant mean weight loss of 13.92% ± 3.58%, substantial colon shortening (6.97 ± 0.46 cm), and high stool hemoccult scores (2.75±0.46). The 7-KC treatment showed disease mitigation in weight loss, colon length, and stool hemoccult scores. Conclusion These findings show that the treatment with 7-KC alleviated the clinical symptoms in the acute DSS-induced IBD mouse model, suggesting therapeutic benefits of 7-KC. We will present these data along with images of colon histology and assessment of inflammation in the colon. Funding Source Villanova University Topic Categories Translational and Interventional Immunology (TI)
Optimization and Clinical Correlation of the Basophil Activation Test for Drug-Induced Anaphylaxis Diagnosis in Bolivia 2257595
Abstract Introduction Diagnosis of drug-induced anaphylaxis (DIA) remains challenging, especially when in vivo testing is unsafe or contraindicated. The Basophil Activation Test (BAT) offers a promising in vitro alternative using flow cytometry markers CD63 and CD203c, but methodological variability limits its diagnostic reliability. Objective To optimize and standardize key technical parameters of the BAT and evaluate its clinical correlation in patients with suspected drug allergy in Bolivia. Methods This is a diagnostic test study. Peripheral blood samples from patients with confirmed or suspected drug hypersensitivity and healthy controls are being analyzed. Basophil activation is assessed after stimulation with antibiotics, NSAIDs, and anesthetics. Optimization includes gating strategies and stimulation conditions. Results Preliminary Results (the study is ongoing): Early analyses show that methodological refinement improves assay reproducibility. A positive correlation is observed between basophil activation and clinical symptoms. Evaluation of sensitivity, specificity, and predictive values is ongoing. Conclusion This ongoing study integrates methodological optimization with clinical assessment to enhance the diagnostic performance of the BAT. Preliminary findings support its potential as a safe and effective diagnostic tool for drug-induced allergy. Final validation data will be presented at the meeting. Funding Source n/a Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)
Sub‐Diffraction Nanolithography of Halide Perovskite via Reversible All‐Optical Crystallization–Decomposition
ABSTRACT Halide perovskites exhibit exceptional optoelectronic properties, yet their intrinsic chemical fragility and ionic nature pose fundamental challenges for high‐resolution patterning and nanoscale integration. Here we report a reversible, all‐optical structural modulation strategy that enables chemistry‐free, sub‐diffraction patterning of halide perovskite thin films through light‐driven crystallization‐decomposition dynamics. Localized femtosecond laser excitation induces controlled crystallization and an orthorhombic‐to‐cubic phase transition in CsPbBr 3 thin films, markedly enhancing crystallinity and optoelectronic response. In contrast, ultraviolet illumination promotes defect formation and partial decomposition into CsBr and PbBr 2 , reversibly suppressing crystallinity and photocurrent. This optically driven crystallization‐decomposition cycle is repeatable over multiple iterations with more than 85% photocurrent recovery, establishing a robust platform for reversible material‐state control. Leveraging the nonlinear competition between laser‐induced crystallization and UV‐induced inhibition, we further demonstrate resist‐free, sub‐diffraction nanolithography with feature sizes down to 93.5 nm, well beyond the conventional optical diffraction limit. This work reveals a light‐programmable structural degree of freedom in halide perovskites and provides a general materials framework for reconfigurable perovskite architectures, adaptive photonics, and dynamic optoelectronic systems.
Mechanically Spatio‐Chimeric Fibrin Assembly Enables Vascular‐Integrated Muscle Reconstruction for Volumetric Muscle Loss Repair
ABSTRACT Volumetric muscle loss (VML), a severe injury involving irreversible loss of both muscle tissue and vasculature, poses a major barrier to the development of clinically viable muscle grafts. Functional restoration requires engineered constructs capable of reconstructing both contractile and vascular components that can functionally integrate with the host vasculature. Here, we introduce SPARC (spatio‐chimeric, plasma‐based, anisotropic, and shear‐responsive construct), a mechanically bimodal fibrin hydrogel engineered via shear‐guided assembly of plasma fibrin to recreate the structural and mechanical heterogeneity of native muscle. Controlled microfluidic shear generates aligned fibrillar bundles and a spatially graded bimodal stiffness architecture, establishing stiff, bundle‐dense regions that favor myogenic differentiation and compliant regions that promote endothelial morphogenesis. When co‐cultured with myoblasts and endothelial cells, the resulting anisotropic matrix directs spatially organized myogenic maturation and endothelial morphogenesis. In vivo evaluation in a murine VML model shows that vascularized muscle SPARC grafts restore muscle architecture and function, promoting neovascularization, myofiber regeneration, and enhanced motor recovery. Through its spatially mechano‐programmed design, SPARC enables coordinated myogenic and endothelial organization within a single construct, establishing a scalable biofabrication strategy for functional repair of extensive muscle defects.
IRF8 is a Key Transcriptional Regulator of IL-2—Induced NK Cell Activation and Antitumor activity 2253333
Abstract Introduction Interleukin-2 (IL-2) promotes Natural killer (NK) cell survival, proliferation, and activation; however its downstream signaling and transcriptional regulators remain incompletely defined. Interferon regulatory factor 8 (IRF8), known for its essential role in dendritic cell and macrophage development, also has not been fully characterized in NK cell—mediated antitumor responses. In this study, we demonstrate that IRF-8 functions is a key transcriptional regulator of IL-2—induced NK cell activation. Methods We generated NK cell—specific IRF-8 knockout mice and evaluated transcriptional and functional changes upon IL-2 stimulation. Results As a result, we observed that IL-2 stimulation translocated IRF8 from the cytoplasm to the nucleus in wild-type NK cells, and also increased the cytotoxicity of NK cells. On the other hand, treatment with IL-2 in IRF-8-deficient NK cells did not induce cytotoxic activation, and the antitumor effect of IL-2 was significantly reduced in a tumor model. Transcriptome analysis confirmed IRF-8-dependent regulation of gene expression in NK cells. Conclusion This study demonstrates that IRF-8 is an essential downstream effector of IL-2 signaling and a central transcriptional regulator of NK cell activation. Funding Source N/A Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)
Total respiratory tract immunization induces optimal systemic and regional antigen-specific immune responses 2244163
Abstract Introduction Respiratory infections represent a major global health challenge. Different intranasal vaccine strategies can target specific areas of the respiratory tract, which may influence local and systemic immune responses. Methods Here, we combined peptide-major histocompatibility complex class I and II (pMHCI and pMHCII) tetramers with flow cytometry and single cell RNA sequencing (scRNAseq) to comprehensively study spike-specific CD8+ and CD4+ T cells following immunization with SARS-CoV2 spike protein. Following intramuscular (IM) priming with spike protein and poly:IC, we compared different boosting strategies - upper respiratory tract (URT), total respiratory tract (TRT), and IM. Results At 4 weeks post-boost, the number of spike-specific CD4+ and CD8+ T cells was higher in the lungs of mice receiving TRT boosting compared to URT or IM boosting. We also observed higher frequencies and numbers of CD4+ and CD8+ resident memory T cells (TRM) in the lungs of TRT versus URT or IM groups. In the nasal mucosa (NM), both TRT and URT boosting resulted in higher frequencies of spike-specific CD4+ and CD8+ TRM cells than IM. Notably, TRT was the only boost strategy that induced serum and lung spike-specific IgA. However, despite generating fewer spike-specific T cells and antibodies than TRT, URT boosting still protected mice against viral challenge. Moreover, both spike-specific CD4+ and CD8+ T cells obtained from the NM expressed higher levels of IFN-g, and spike-specific CD8+ T cells in the NM produced higher levels of granzyme B compared to those cells recovered from lung and spleen. Finally, scRNAseq analysis with barcoded tetramers revealed that epitope specificity influenced CD4+ T cell differentiation resulting in epitope-specific T helper cell bias. Conclusion In sum, TRT boosting enhanced lung immunity and epitope-driven CD4+ T cell phenotypes offer a promising approach to guide optimized vaccine design. Funding Source NIH P01 AI165072 AND AMERICAN LUNG ASSOCIATION CA-1252074 Topic Categories Mucosal and Regional Immunology (MUC)