We recently characterized the microenvironment of Anaplastic Thyroid Carcinoma (ATC) as immune-exhausted, rich in TIM-3-expressing exhausted cytotoxic T lymphocytes (CTLs). PD-1 was notably the least frequently expressed immune checkpoint (ICP). The results provided a potential explanation for the limited efficacy of anti-PD-1/PD-L1 immunotherapy, highlighting TIM-3 as another promising target. This pilot study further evaluated the potential efficacy of targeting TIM-3 in ATC as a therapeutic strategy in an in vitro model. A co-culture model of ATC cell line 8505C with exhausted TIM-3+ve/PD-1+ve CTLs was established and treated with anti-TIM-3 and anti-PD-1 antibodies, alone and in combination. Antibody efficacy was evaluated using flow cytometry, measuring Granzyme B and Perforin for CTL cytotoxic activity, and Annexin V and propidium iodide for tumor cell death. Additionally, differential gene expression before and after ICP blockade was analyzed using the NanoString nCounter Immune-oncology panel. Both individual and combined antibody treatments enhanced CTL cytotoxicity and promoted tumor cell death. Interestingly, ICP inhibition induced an upregulation of immune-activating pathways and downregulation of genes associated with tumor progression. These findings support the potential of TIM-3 blockers as an alternative to or in combination with anti-PD-1 therapy, thereby providing a rationale for further clinical investigation.
Anti-amyloid-β monoclonal antibodies have established immunotherapy as a viable approach for Alzheimer's disease, yet all α-synuclein and tau antibodies tested to date have failed clinically. These setbacks offer invaluable immunology lessons reshaping next-generation therapeutic development. Four key lessons emerge: epitope specificity determines outcomes, with mid-domain and microtubule-binding region antibodies showing promise over failed N-terminal approaches; biomarker-driven stratification using p-tau217 and AI algorithms can unmask efficacy in rapid progressors; most patients harbor mixed proteinopathies, rendering monotherapy insufficient and mandating combination immunotherapy; and neuroinflammation, particularly NLRP3 activation and microglial dysfunction, represents a convergent pathway targetable independently of upstream triggers. Emerging approaches include conformation-specific antibodies, protein-protein interaction inhibitors, and immune checkpoint targeting, such as anti-SIGLEC-10, to restore phagocytosis. These insights inform biomarker-enriched trials and combination regimens, although clinical evidence for specific combinations remains limited and requires validation. The systems framework targeting aggregation, neuroinflammation, and immunity offers a promising paradigm yet awaits empirical testing.
Autoantibodies in the plasma demonstrate potential as biomarkers for early detection of breast cancer. Copper transport proteins ATP7A, ATP7B and COPT1, are represent potential diagnostic biomarkers. This study aims to evaluate the diagnostic value of plasma autoantibodies targeting ATP7A, ATP7B and COPT1 in breast cancer. The plasma expression levels of anti-ATP7A/ATP7B/COPT1 autoantibodies were detected in the cohort with 229 patients with breast cancer (BC), 214 patients with benign breast tumor (BBN), and 169 normal controls (NC) using the ELISA method. The expression levels of three autoantibodies were significantly lower in BC compared to BBN and NC groups. The combination of anti-ATP7A and anti-COPT1 could improve the diagnostic efficacy with an increased AUC value (AUC = 0.844, 95% CI, 0.764-0.925) in patients aged ≤50 years with early HER2+ breast cancer. This study demonstrates the clinical value of these three autoantibodies in the detection of BC, suggesting their potential as diagnostic biomarkers for BC.
Multiple sclerosis (MS) is treated with branded or generic disease-modifying therapies (DMTs), with increasing concern for the composition and quality of off-patent products. Our objective was to evaluate differences in disease-specific biomarkers, a potential indicator of suboptimal therapeutic exposure, between individuals treated with branded and generic DMTs. We compared biomarker variability in MS individuals (n=1,124) treated with generic or brand-name fumarates (diroximel fumarate, dimethyl fumarate (DMF)), fingolimod, and teriflunomide. In the generic DMF group, higher coefficients of variation (COV) for CXCL9, CCL20, and TNFSF13B were observed, with a paradoxically lower variability for CXCL13. Higher COVs for CXCL9 were observed with generic fingolimod and for CXCL9 and CXCL13 with generic teriflunomide when compared to brand. Additionally, all generic DMTs demonstrated higher multi-variable variability index values across 18 proteins associated with MS disease activity. Greater biomarker variability with generic formulations may reflect differences in therapeutic exposure and less consistent disease control.
Hepatocellular carcinoma (HCC) develops within a chronically inflamed and immunologically dysregulated hepatic microenvironment that contributes to therapeutic resistance and disease progression. Gasdermin-mediated pyroptosis has emerged as a context-dependent inflammatory cell-death program capable of reshaping antitumor immunity, stromal remodeling, and immune checkpoint responsiveness. Acute and localized activation of GSDMD or GSDME may enhance immune-cell recruitment and tumor antigen exposure, whereas persistent pyroptotic signaling promotes fibrosis, angiogenesis, and immunosuppressive remodeling. This review examines the molecular circuitry linking pyroptosis with inflammasomes, metabolic stress, mitochondrial dysfunction, hypoxia, and epigenetic regulation in HCC. We further discuss how single-cell and spatial transcriptomic approaches reveal distinct pyroptotic immune niches associated with macrophage polarization, T-cell infiltration, and therapeutic response. Emerging therapeutic strategies including inflammasome modulation, gasdermin-targeted interventions, metabolic sensitization, and combination immunotherapy are also evaluated. Collectively, regulated pyroptosis may represent a promising immunomodulatory framework for improving precision therapy in HCC.
The adaptive autoimmune process in SLE is complex, and conventional techniques cannot resolve disease-associated T/B lymphocyte clones at a single-cell granularity. Single-cell RNA sequencing combined with single-cell V(D)J sequencing (scRNA+VDJ-seq) enables simultaneous profiling of cellular transcriptomes and TCR/BCR repertoires. This review highlights recent advances in scRNA+VDJ-seq in SLE and related autoimmune disorders, elucidating the link between receptor clonality, gene expression, and effector phenotypes. It delineates clonal expansion, biased V(D)J usage, and aberrant exhaustion in SLE, and compares T/B cell immune phenotypes across lupus nephritis, RA, pSS, and AS. The review also summarizes CD19 CAR-T and PD-1 agonist applications. Furthermore, it discusses AI integration with scRNA+VDJ-seq to analyze autoreactive T/B cells in SLE: from data integration to clinical translation, and assesses the clinical translational potential of this technology, offering novel targets and strategies for SLE diagnosis, personalized treatment, and prognosis.
BACKGROUND:GBM exhibits extensive necrosis and profound immune dysfunction, yet the role of multimodal cell death-associated inflammatory niches remains unclear. METHODS:Bulk and single-cell transcriptomic datasets were integrated with spatial analyses and functional experiments to characterize these niches and evaluate stromal barriers and GZMB+ CTL activity. RESULTS:Apoptosis, pyroptosis, and necroptosis markers formed spatial gradients toward necrotic tumor cores. These niches were associated with stemness, proliferation, epithelial-mesenchymal transition, p53 stress responses, and metabolic adaptation. Despite enhanced immune-recruiting signals and GZMB+ CTL accumulation, CTLs remained concentrated around necrotic and vascular regions rather than infiltrating tumor cell-dense areas. Extracellular matrix remodeling and CAF-like stromal barriers restricted CTL penetration and antitumor cytotoxicity. CONCLUSIONS:Multimodal cell death-associated inflammatory niches paradoxically recruit GZMB+ CTLs while spatially constraining their function, revealing a potential mechanism linking necrosis to immune evasion and GBM progression.
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease in which loss of immune tolerance, chronic inflammation, and metabolic reprogramming are closely interconnected. Fatty acid amides (FAAs) are endogenous lipid mediators that include N-acylethanolamines (NAEs), such as anandamide (AEA), palmitoylethanolamide (PEA), oleoylethanolamide (OEA), palmitoleoylethanolamide, and linoleoylethanolamide, as well as primary fatty acid amides such as palmitamide, octadecanamide, and oleamide. The evidence base for FAA metabolism in SLE should be interpreted at two levels. First, direct human multi-omics evidence now supports disease-associated alteration of several FAA-class serum metabolites: a serum proteome-metabolome study identified palmitoleoylethanolamide, linoleoylethanolamide, palmitamide, and octadecanamide among candidate metabolite biomarkers for SLE classification. Second, a targeted endocannabinoid study found increased 2-arachidonoylglycerol (2-AG) and enhanced diacylglycerol lipase (DAGL) activity in peripheral blood mononuclear cells, whereas AEA, PEA, and OEA were not significantly different from healthy controls. Because 2-AG is an endocannabinoid but not an FAA, these data support broader endocannabinoidome dysregulation rather than universal NAE dysregulation. Mechanistic evidence is primarily derived from lupus mouse models: PEA is reduced in serum and spleen of MRL/lpr mice and suppresses TLR9-induced IL-6 production, dendritic-cell and B-cell activation, IgM production, and B-cell proliferation; FAAH is upregulated in B cells from a lupus-prone Sle2z model and FAAH inhibition reduces receptor revision, RAG expression, and polyreactive autoantibody production; nano-encapsulated AEA reduces inflammatory cytokines and lesion severity in a murine model of cutaneous lupus erythematosus. Collectively, these findings indicate that FAA-related pathways are relevant to SLE, although current evidence remains heterogeneous, species-specific, and insufficient to establish causality in patients. Future work should combine targeted lipidomics, cell-type-resolved enzyme profiling, immune perturbation assays, and organ-specific phenotyping to determine whether FAA metabolism contributes to disease pathogenesis, biomarker development, or adjunctive therapy in defined SLE subsets.
Glioma stem cells (GSCs) drive tumor heterogeneity, therapy resistance, and immunosuppression. This study identified molecular subtypes of glioma based on stemness-related genes and validated a key hub gene through bioinformatics and experiments. A stemness gene set was compiled from single-cell data, GeneCards, and MSigDB. Consensus clustering (NMF) on 157 glioma samples from GSE4290 revealed two subtypes (C1 and C2), with C1 showing higher stemness scores. C1 exhibited enriched DNA repair, proliferation, and metabolic pathways, plus greater temozolomide (TMZ) resistance and M2 macrophage infiltration. WGCNA and differential expression identified 43 genes; machine learning (LASSO, SVM-RFE, RF) selected TNFRSF12A as the core hub gene. TNFRSF12A knockdown in GSCs inhibited proliferation and stemness markers (CD133, OLIG2, SOX2), enhanced TMZ sensitivity, and suppressed M2 macrophage polarization. In vivo, TNFRSF12A silencing reduced tumor growth and synergized with TMZ. TNFRSF12A represents a potential therapeutic target for aggressive gliomas.
This study investigated the role of Krüppel-like factor 4 (KLF4) in macrophage-mediated inflammation in type 2 diabetes mellitus (T2DM) with psoriasis. KLF4, tissue inhibitor of metalloproteinases 3 (TIMP3), and a disintegrin and a metalloproteinase domain 17 (ADAM17) levels were measured in skin samples from patients. A diabetic psoriasis-like dermatitis mouse model was established using imiquimod (IMQ) and treated with KLF4-overexpressing lentiviruses. Glucose/lipid metabolism, skin pathology, mast cell counts, and inflammatory markers were assessed. Macrophage status was evaluated by F4/80 staining. KLF4 was downregulated in patients with T2DM with psoriasis. In mice with diabetic psoriasis-like dermatitis. KLF4 overexpression improved glucose/lipid metabolism and reduced skin erythema, scaling, thickening, psoriasis area severity index scores, and mast cell infiltration. KLF4 also decreased inflammation and modulated the TIMP3/ADAM17 pathway. These findings indicate that KLF4 ameliorates IMQ-induced inflammation via the TIMP3/ADAM17 pathway in diabetic psoriasis-like dermatitis.
Mucosal immunity constitutes the first line of defense against respiratory viruses. We characterized serum and salivary SARS-CoV-2-specific and polyreactive antibodies in 784 Dutch healthcare workers at two timepoints (2020-2021). SARS-CoV-2-specific antibodies and salivary polyreactive IgA were quantified using multiplex immunoassays. Associations with infection, vaccination, and symptom burden were analyzed using multivariable regression, Cox proportional hazard models, and mixed-effects models. SARS-CoV-2 infection induced systemic SARS-CoV-2-specific IgG and salivary IgA responses, whereas vaccination increased serum and salivary anti-Spike-1 IgG. Salivary polyreactive IgA increased over time, was associated with smoking (p = 0.022) and asthma (p = 0.021), and higher levels were associated with an increased hazard of subsequent SARS-CoV-2 infection (HR 1.15 [1.01-1.31]). Polyreactive IgA levels declined more rapidly in participants with a higher symptom burden (p = 0.081). SARS-CoV-2 infection, but not vaccination, induces distinct changes in the salivary IgA compartment. Salivary polyreactive IgA levels changed over time, which may reflect repeated mucosal immune stimulation, and were associated with greater symptom burden.
Life-threatening hypereosinophilic syndrome (HES) is a rare medical emergency with limited therapeutic options in corticosteroid-refractory cases. Preliminary reports suggest that Janus kinase inhibitors may be beneficial in eosinophil-associated disorders, including HES. We conducted a nationwide multicenter retrospective study to assess ruxolitinib. Thirteen patients with severe acute HES and organ- or life-threatening involvement were included, most requiring intensive care, with massive baseline absolute eosinophil counts (AEC, median 45 × 109/L) and major involvement including eosinophilic myocarditis, ischemic strokes, and vascular thromboses. Ruxolitinib was initiated after a median of 7 days of corticosteroids. A rapid decline in absolute eosinophil counts was observed after initiation of ruxolitinib, with hematologic response rates of 54%, 85%, and 92% at Days 7, 14, and 30. Clinical improvement was observed across organ systems. Adverse events were manageable, although three deaths occurred during follow-up, only one of which occurred on ruxolitinib.
Previous studies have demonstrated strong links between circulating microRNAs (miRNA), lymphocytes, and immune markers suggesting that miRNAs are key regulators of immune function and immune related diseases. We used the population-based SHIP-TREND cohort (N = 804) to investigate how specific plasma-based miRNAs relate to lymphocytes, cardiometabolic diseases, and clinical parameters. In linear regression, we identified miR-150-5p to be strongly lymphocyte-associated in both TREND waves. Beyond this, miR-150-5p showed significant associations with kidney disease and also with several clinical markers of renal function. Data indicate that these effects mark the transition from healthy to subclinical disease status and were independent of lymphocytes. We further validated the impact of miR-150-5p in independent publically available clinical datasets. Our findings suggest that the pathological effects of lymphocytes on kidney disease may be mediated through miR-150-5p, by regulating the expression of target genes and modulating inflammatory pathways. This highlights miR-150-5p as a potential early marker of subclinical kidney disease.
BACKGROUND:Acute myeloid leukemia (AML) remains an aggressive hematologic malignancy with heterogeneous responses to induction chemotherapy. Reliable biomarkers to distinguish patients achieving complete remission (CR) from those with refractory disease are needed to support immunological assessment of treatment response. OBJECTIVE:To evaluate whether serum soluble CD25 (sCD25), galectin-9 (Gal-9), and CXCL9 differ between AML patients in CR, refractory AML patients, and healthy controls, and to assess their discriminative performance for refractory versus CR status. METHODS:In this single-center case-control study, 155 participants were enrolled (50 healthy controls, 55 AMLCR, 50 AML-Refractory). Serum sCD25, Gal-9, and CXCL9 were quantified by sandwich ELISA. Between-group comparisons used the Kruskal-Wallis test with Dunn post-hoc pairwise contrasts; p-values were adjusted for multiple comparisons using the Benjamini-Hochberg false discovery rate (FDR). Discriminative performance was evaluated by receiver operating characteristic (ROC) analysis with bootstrap-derived 95% confidence intervals (1000 resamples), and pairwise AUC comparisons were performed using DeLong's test. RESULTS:Serum sCD25 and Gal-9 were significantly elevated in refractory patients compared with CR and controls (adjusted p < 0.001 for both), whereas CXCL9 was highest in the CR group and lowest in refractory patients (adjusted p < 0.001). For discrimination between refractory and CR status, CXCL9 showed the highest single-marker performance (AUC = 0.94, 95% CI: 0.90-0.98), followed by sCD25 (AUC = 0.91, 95% CI: 0.86-0.96) and Gal-9 (AUC = 0.89, 95% CI: 0.83-0.94). A combined logistic regression model incorporating all three markers yielded cross-validated AUC = 0.96 (95% CI: 0.93-0.99); the improvement over CXCL9 alone did not reach statistical significance (DeLong p = 0.31). CONCLUSION:sCD25, Gal-9, and CXCL9 collectively describe two complementary immunological axes in AML chronic activation/exhaustion (sCD25, Gal-9) and effector-cell trafficking (CXCL9) and robustly discriminate refractory from remission status in this cross-sectional cohort. Prospective longitudinal studies are required before these markers can be recommended for prognostic use or clinical decision-making. CLINICAL SIGNIFICANCE:The present study identifies a reproducible serum biomarker panel (sCD25, Galectin-9, and CXCL9) that reflects the dynamic balance between immune activation and exhaustion in acute myeloid leukemia (AML). The combined measurement of these markers can distinguish patients in complete remission from those with refractory disease, correlating with treatment response and residual immune dysregulation. These findings highlight a practical, non-invasive immunological tool that could support early detection of relapses, guide therapeutic monitoring, and enhance risk stratification in AML management.
We investigated cytokine expression patterns and B cell-helper functions of T peripheral helper (Tph) cell subsets. CXCR3+CCR6- Tph1 cells have ability to co-produce interleukin (IL)-21 and interferon-γ while CXCR3-CCR6+ Tph17 can co-produce IL-21 and IL-17A. These Tph subsets show almost comparable activity to induce plasma cell differentiation from CD19+ B cells. Noticeably, Tph1 cells can drive plasma cell differentiation from CD11chi double negative (DN) B cells more effectively than classical memory B cells via IL-21. On the other hand, Tph17 cells appear to have low activity to induce plasma cell differentiation from CD11chi DN B cells. Furthermore, in new-onset patients with systemic lupus erythematosus (SLE), Tph1 cells are expanded in the blood and positively correlated with disease activity, autoantibody levels, and CD11chi B cells. These results suggest that extrafollicular interaction between Tph1 cells and atypical CD11chi DN B cells plays a pathogenic role in new-onset SLE.
BACKGROUND:AD involves barrier dysfunction and immune imbalance. Roles of LCE3D and TGF-β1 remain unclear. METHODS:Bioinformatics analysis (GEO: GSE130588), RT-qPCR, ELISA, and Western blot were performed in 30 CE patients and 30 controls, including 15 serum pairs. TNF-α-stimulated HaCaT and NHEK cells were used to establish an AD-like model. LCE3D knockdown and overexpression were conducted, and TGF-β1 silencing was used to explore mechanistic interaction. RESULTS:LCE3D was upregulated in AD tissues and serum, negatively correlated with TGF-β1, and associated with disease severity and IL-4/IFN-γ imbalance. In vitro, TNF-α increased LCE3D and IL-4 while decreasing IFN-γ and TGF-β1. LCE3D knockdown increased TGF-β1, IFN-γ, and barrier proteins (involucrin, loricrin) and reduced IL-4, whereas overexpression showed opposite effects. TGF-β1 silencing reversed these effects. CONCLUSIONS:LCE3D promotes AD progression by suppressing TGF-β1, driving Th2/Th1 imbalance and epidermal barrier dysfunction. The LCE3D/TGF-β1 axis is a potential therapeutic target.
JAK1 gain-of-function (GoF) mutations were considered ultra-rare causes of immune dysregulation, but recent studies suggest a broader clinical spectrum. We report the first Chinese case of a novel likely JAK1 GoF mutation (p.L783F) in a child with early-onset severe atopic dermatitis, eosinophilia, elevated IgE, and growth retardation. Functional assays showed enhanced IL-6-induced STAT1 phosphorylation among tested cytokines; single-cell RNA-sequencing revealed JAK-STAT hyperactivation predominantly in CD16+ monocytes. Mass and flow cytometry revealed expansion of a pro-inflammatory CD4+ effector subset (CD45RA+CCR7-IFN-γ+IL-2+TNF-α+IL-23+CD4+ T cells) and Th2-polarized CD4+ T cells with increased CD25, CD27, and CRTH2. Given the patient's young age, growth retardation, and dermatitis-predominant clinical phenotype, she was treated with dupilumab, improving skin lesions, pruritus, eosinophilia, and hyper-IgE. This case expands the clinical and mutational spectrum of JAK1 GoF-associated disease (JAACD) and demonstrates that targeting downstream signaling such as dupilumab offers a safe, effective precision therapy for pediatric patients with JAK1 GoF mutations.
