Search Results (1,612)

Background/Objectives: Preclinical studies of head and neck squamous cell carcinoma (HNSCC) commonly use subcutaneous heterotopic (flank) tumor models for simplicity; however, orthotopic models may better reflect the native tumor environment. Direct comparisons of the tumor immune microenvironments (TIME) and tumor-draining lymph nodes (tdLNs) between these models remain limited. Better understanding of site-specific immune differences could improve model selection and interpretation of translational HNSCC studies. Methods: ROC1 tumors were established in murine heterotopic and orthotopic sites, followed by assessment of tumor growth kinetics, survival, and the tumor microenvironment. Immune composition of tumors, blood, tdLNs, and spleen was evaluated at three tumor progression timepoints using multiparameter spectral flow cytometry. Results: Heterotopic and orthotopic tumor models showed similar growth kinetics and survival. Immune profiling revealed increased infiltration of CD3⁺ T-cells, natural killer (NK) cells, and myeloid populations in both models. Heterotopic tumors were enriched in dendritic cells (DCs), plasmacytoid DCs, and monocytic myeloid-derived suppressor cells (M-MDSCs), whereas orthotopic tumors showed increased macrophages, granulocytic MDSCs, and M-MDSCs. Despite temporal variation, both TIMEs were dominated by macrophages, DCs, and CD3⁺ T-cells. Late-stage heterotopic tumors contained more CD4⁺ T-cells. Reduced T-cell cytotoxicity (PD-1, CD107a) and increased immune checkpoint expression across myeloid cells indicated an immunosuppressive TIME. Systemically, effector cells were preserved despite suppressive cell trafficking, and tdLNs in both models exhibited immunosuppressive PD-L1 expression. Conclusions: Heterotopic and orthotopic ROC1 tumors share key immune features, but site-specific differences in the TIME and tdLNs reveal tissue-dependent regulation. These local effects align with systemic changes, supporting global tumor-associated immunosuppression. Full article

Ubiquitination is a key post-translational modification regulating cellular signaling and innate immunity, and its reversal by deubiquitinases (DUBs) represents a critical mechanism exploited by pathogens for immune evasion. While ovarian tumor (OTU) domain-containing DUBs are well characterized in viral systems, their roles in fungal pathogens remain largely unexplored. In this study, we investigated two putative OTU domain-containing proteins derived from the plant pathogenic fungi Melampsora larici-populina (MlpOTU, EGG09943.1) and Taphrina deformans (TdOTU, CCG84064.1). Recombinant MlpOTU and TdOTU proteins were successfully expressed and purified from E. coli and exhibited high solubility and proper folding. Functional analyses in HEK293T cells demonstrated that both proteins significantly reduce global ubiquitination levels, confirming their deubiquitinase activity in vivo. Despite this shared enzymatic function, the two proteins displayed markedly distinct effects on host immune gene expression. MlpOTU selectively suppressed key antiviral effectors, most notably MX1, suggesting a targeted immune evasion strategy. In contrast, TdOTU induced robust upregulation of multiple immune-related genes, including type I interferons, indicating a divergent role. Neither MlpOTU nor TdOTU triggered robust apoptosis, supporting their role as modulators of host signaling rather than cytotoxic effectors. Collectively, these findings provide the first functional evidence that fungal OTU domain-containing proteins act as active deubiquitinases and reveal distinct strategies by which plant pathogens may manipulate host immune responses. This study establishes fungal OTU domains as promising targets for antifungal intervention and broadens our understanding of cross-kingdom evasion mechanisms. Full article

Background: Immune effector cell-associated hemophagocytic lymphohistiocytosis-like syndrome (IEC-HS) is a rare, life-threatening complication following CAR T-cell therapy. Diagnosis is challenging due to overlap with severe CRS, sepsis and lack of standardized criteria. Clinical data remain limited. Methods: We retrospectively analyzed 301 patients treated with CD19- or BCMA-directed CAR T-cells for hematologic malignancies at a single center from January 2019 to January 2026. IEC-HS was defined according to American Society for Transplantation and Cellular Therapy criteria. Results: Median follow-up was 31 months. IEC-HS was diagnosed in 14 patients (4.7%), median age 67 years. Underlying diseases included diffuse large B-cell lymphoma (n = 4), multiple myeloma (n = 7), mantle cell lymphoma, Burkitt lymphoma and B-lymphoblastic leukemia (n = 1 each). All patients had hyperferritinemia and cytopenias at baseline; most had high tumor burden (9/14) and elevated LDH (10/14). CRS occurred in all patients and ICANS in 6/14. IEC-HS occurred at median 10 days and was characterized by hyperferritinemia (median 15,321 µg/L), neutropenia, thrombocytopenia, hepatic dysfunction and high CAR-T-cell expansion in peripheral blood. Treatment included corticosteroids and anakinra (12/14). Refractory patients received IVIG (5/14), tocilizumab (3/14), siltuximab, ruxolitinib, emapalumab or etoposide (each n = 1). Infections occurred in 11/14; 4/14 had mixed infections. IEC-HS resolved in 7/14 (median 7 days). Mortality was 79% (11/14), mainly due to IEC-HS (7/14). Three patients were alive at last follow-up. One-year OS was lower vs. the whole cohort (31% vs. 69%, p < 0.0001). Conclusions: IEC-HS was associated with severe cytopenias, hyperferritinemia, hepatic dysfunction and high infection risk. Despite intensive immunosuppressive therapy, outcomes remain poor. Early biomarker-driven identification and multicenter studies are needed. Full article

Mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with potent immunomodulatory properties, making them attractive candidates for treating inflammatory and autoimmune diseases. A key mediator of MSC-induced immunosuppression is programmed death-ligand 1 (PD-L1), a checkpoint molecule that interacts with PD-1 on immune cells to regulate immune responses and promote tolerance. This review synthesizes current evidence on the role of PD-L1 expression in MSCs, emphasizing its effects on both the innate and adaptive immune systems, its therapeutic potential, and its utility as a biomarker for MSC potency and clinical efficacy. We examine how PD-L1 modulates T cell activation, dendritic cell maturation, macrophage polarization, and cytokine profiles, including its role in exosomal contexts. Additionally, we highlight its synergistic interactions with other immune checkpoints and discuss its dual function as both a therapeutic effector and a dynamic biomarker. Finally, we explore its relevance in clinical contexts such as autoimmune diseases, graft-versus-host disease, sepsis, and transplantation and conclude with a discussion of challenges and future directions in harnessing PD-L1 for MSC-based therapies. Full article

Background: Lung cancer is the top cause of cancer-related mortality globally, and chemo-immunotherapy is a core therapeutic strategy for it. The novel bacterial composite vaccine (Neo-BCV) we developed previously can activate anti-tumor immunity. This study explored its synergistic anti-tumor effect with cisplatin (CDDP), along with the underlying immunomodulatory mechanisms and molecular regulatory networks. Methods: A murine Lewis lung cancer (LLC) model was established to evaluate the efficacy of the combination therapy. Flow cytometry and multiplex cytokine assay were used to detect immune cell subsets and functional molecules in the spleen, serum and tumor tissues. RNA-sequencing (RNA-seq) was used to elucidate the molecular regulatory networks following the combination therapy in the tumor tissues. Body weight, blood indexes, serum biochemistry and H&E staining were monitored to verify biosafety. Results: Neo-BCV combined with CDDP achieved an 87.77% tumor growth inhibition rate, showing the most significant anti-tumor effect. The combination promoted DC maturation, enhanced effector immune cell infiltration, reduced immunosuppressive cells, upregulated Th1-type cytokines and downregulated CD8⁺ T cell surface PD-1. RNA-seq confirmed enrichment of multiple immune effector pathways, supporting tumor immune microenvironment remodeling. The combination alleviated CDDP-induced weight loss, had no obvious adverse effects on physiological indicators, and exhibited good biosafety. Conclusions: Neo-BCV combined with CDDP achieves enhanced anti-tumor efficacy and favorable biosafety in murine lung cancer models by regulating immune cell subsets and activating immune-related molecular pathways, providing a solid preclinical basis for its clinical translation in lung cancer treatment. Full article

Background: Cytokine-induced JAK–STAT signaling becomes dysregulated in chronic human diseases, including cancer and autoimmunity, and contributes to immune cell dysfunction. A cytokine-independent approach to activating STAT proteins could “hardwire” pro-survival and effector programs in immune cells to sustain function within diseased tissues. Engineered variants of the herpesvirus saimiri tyrosine kinase interacting protein (TIP) can recruit the SRC family kinase (SFK) LCK to drive STAT phosphorylation and activation. Here, we evaluated the interactome of a TIP-derived, cytokine-independent STAT5 activator and determined whether it could induce STAT5 activation in immune cell lines and primary human CD8+ T cells. Methods: A STAT5 activator (aSTAT5) was characterized by proteomics using affinity purification mass spectrometry (AP-MS) to define its interactome and STAT5 binding specificity. STAT5 phosphorylation was assessed in hematopoietic cell lines and primary human CD8+ T cells. Results: Proteomic analysis confirmed preferential association of aSTAT5 with STAT5 relative to other proteins. In cell-based assays, aSTAT5 induced robust STAT5 phosphorylation in LCK-expressing NK-92 and Jurkat T cells, whereas phosphorylation was not observed in Raji B cells or RAW 264.7 macrophages despite expression of closely related SFKs and STAT5. Cytokine-independent STAT5 phosphorylation supported the viability of NK-92 cells and primary human CD8+ T cells during cytokine withdrawal and preserved the cytotoxic function of CAR T cells. Conclusions: We defined the interactome of a cytokine-independent STAT5 activator and demonstrated its capacity to maintain survival and function in human CD8+ T cells and NK-92 cells. These findings underscore the translational potential of engineered, cytokine-independent STAT5 activation for immune cell therapies. Full article

Chimeric antigen receptor (CAR) T-cell therapy represents a major advance in modern immunotherapy. This narrative review summarizes evidence from the past five years, including case reports, case series, and clinical trials, on its application beyond hematologic malignancies, focusing on autoimmune diseases such as systemic lupus erythematosus (SLE), systemic sclerosis (SSc), as well as solid tumors including melanoma and primary cutaneous lymphomas. CD19-directed CAR T-cells have demonstrated clinical benefits in SLE and SSc, with sustained immune reset, reduced autoreactive antibody levels, and clinical improvement. In melanoma, CAR T-cells targeting GD2, cMET, and CD20 have shown in vivo expansion and tumor infiltration; however, clinical efficacy remains limited, with transient stabilization or disease progression in most patients. In primary cutaneous lymphomas, early-phase studies with anti-CD70 and anti-CCR4.30 CAR T-cells indicate partial tumor regression and disease stabilization, often requiring additional therapy. Key challenges include limited durability of immune reset due to persistent plasma cells in autoimmune disorders, tumor heterogeneity, antigen loss or overlap, infiltration barriers, resistance mechanisms, and T-cell depletion in solid tumors, collectively reducing response durability and safety. The main toxicities include grade 1–2 cytokine release syndrome and rare hematologic complications, while immune effector cell-associated neurotoxicity syndrome is uncommon. Clinical translation remains limited and requires larger studies to improve efficacy and define safety profiles. Full article

This study investigated the role of microglia-derived insulin-like growth factor 1 (IGF-1) in modulating host defense and disease progression in a viral model of neuroinflammation and demyelination. Intracranial infection of susceptible mice with the glial-tropic JHM strain of mouse hepatitis virus (JHMV) induces acute encephalomyelitis, followed by an immune-mediated demyelinating disease that mimics many clinical and histologic features of multiple sclerosis (MS). Utilizing an inducible fractalkine receptor (Cx3cr1) promoter-driven Cre-loxP recombinant system, we performed timed ablation of Igf1 in microglia to assess its impact on the central nervous system (CNS) response to JHMV. While the loss of microglial IGF-1 did not impair the control of viral replication, it significantly exacerbated spinal cord demyelination. CyTOF and imaging mass cytometry analysis of spinal cords indicated increased myelin damage was associated with increased accumulation of CD8⁺Ly6C⁺ effector T cells and reduced expression of TREM2 that impaired transition into a disease-associated microglia (DAM) phenotype capable of sensing and potentially mitigating myelin damage. Collectively, these findings argue that microglial IGF-1 is a non-redundant coordinator of the CNS immune responses that occur in response to CNS viral infection. Full article

Glioblastoma (GBM) is characterized by a profoundly immunosuppressive tumor microenvironment that limits the efficacy of conventional and immune-based therapies. Oncolytic herpes simplex virus (oHSV) therapy has emerged as a strategy capable of both tumor-selective infection and immune microenvironment modulation. This systematic review aimed to synthesize current evidence on how oHSV therapy reshapes the immunosuppressive microenvironment of GBM. A systematic search of PubMed (MEDLINE) and Embase identified original studies published between 2016 and 2025 investigating immunological or microenvironmental effects of oHSV-based therapies in GBM (final search: 28 January 2026). Preclinical and clinical studies were included, whereas reviews, editorials, conference abstracts, and studies of non-herpes oncolytic viruses were excluded. Study selection and data extraction were performed independently by two reviewers. Due to heterogeneity in models, viral constructs, and outcome measures, a qualitative narrative synthesis was conducted. Of 214 records, 22 studies met the inclusion criteria. Most were preclinical studies using orthotopic immunocompetent murine models, with limited clinical evidence including an early-phase trial in recurrent high-grade glioma. Therapeutic efficacy frequently correlated with tumor microenvironment remodeling rather than viral replication alone. oHSV infection promoted inflammatory signaling, antigen presentation, macrophage polarization, and effector T-cell recruitment but also induced counter-regulatory mechanisms such as myeloid-mediated immunosuppression and vascular or stromal barriers. The clinical significance and durability of these effects remain to be established in larger prospective studies. Despite heterogeneous designs and limited clinical data, current evidence suggests that oHSV may function as a platform for immune microenvironment reprogramming in glioblastoma. Full article

Background/Objectives: Follicular helper T-cell (TFH)-type lymphomas, including angioimmunoblastic T-cell lymphoma (AITL) and TFH lymphoma, not otherwise specified (TFH-NOS), are characterized by marked immune dysregulation in the tumor microenvironment. We investigated whether TFH-type lymphomas are enriched in immunosuppressive cells and explored immunologic changes associated with denileukin diftitox (DD) treatment. Methods: FOXP3-positive mononuclear cells were quantified by immunohistochemistry in lymph node specimens from 10 patients with TFH-type lymphoma and six with non-TFH-type T-cell lymphoma. Paired skin biopsy specimens obtained before and after DD treatment from two patients with AITL were evaluated for CD4, CD8, CD68, CD163, and FOXP3 expression. Longitudinal flow cytometric T-cell receptor Vβ repertoire analysis of CD8-positive T-cell subsets was performed in three patients with AITL treated with DD. Clinical responses were retrospectively assessed in seven patients with relapsed or refractory TFH-type lymphoma treated with DD. Results: TFH-type lymphomas showed significantly higher intra-tumoral FOXP3-positive cell densities than non-TFH-type lymphomas (p = 0.0024). In paired skin biopsies, DD treatment was associated with reductions in CD68- and CD163-positive macrophage-rich infiltrates and a suggested decrease in FOXP3-positive cells. Among seven patients treated with DD, the overall response rate was 86%, including one complete response and five partial responses. Responders showed selective Vβ over-representation patterns in effector and/or memory CD8-positive T-cell subsets, whereas such findings were not convincing in the non-responder. Conclusions: TFH-type lymphomas were associated with higher FOXP3-positive cell density than selected non-TFH-type comparators. DD treatment may be associated with changes in tissue immune infiltrates and peripheral CD8/TCR Vβ skewing in responding cases, although these findings remain exploratory and do not establish causality. Full article

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a dense stromal microenvironment and profound hypoxia, which contribute to therapeutic resistance. Using an in vitro system incorporating pancreatic cancer cells and cancer-associated fibroblasts (CAFs), we show that hypoxia suppresses CD8⁺ T cell accumulation and, in combination with cancer cell- and CAF-derived factors, further impairs T cell fitness by increasing cell death and reducing proliferation. Although the combination of hypoxia and cancer cell/CAF-derived factors enhances IFNγ and granzyme B expression in CD8⁺ T cells on a per-cell basis, the overall number of functional effector T cells is markedly reduced. Analysis of human PDAC single-cell RNA sequencing data corroborates these findings, revealing that CD8⁺ T cells enriched for hypoxia signatures exhibit elevated apoptosis and stress-response pathways. Furthermore, hypoxia is associated with downregulation of stemness-related genes and upregulation of terminal differentiation markers. Together, these data suggest that the integration of intrinsic hypoxic responses and extrinsic cues from tumor cells and CAFs impairs CD8⁺ T cell fitness and correlates with a terminally differentiated, dysfunctional T cell state. Full article

Phosphatidylinositol-3-kinases (PI3Ks) are heterodimers of catalytic and regulatory subunits that regulate cell metabolism, activation, and survival. PI3K, particularly the p110α catalytic isoform, is frequently mutated in cancer, and highly specific inhibitors such as alpelisib are currently used in oncology and in PIK3CA-related overgrowth disorders. Given the relevance of macrophages in anti-tumor immunity, we examined the impact of alpelisib on murine monocytes’ intracellular signaling and on in vitro differentiation, polarization, and effector functions of macrophages. Real-time qPCR (RT-qPCR) showed comparable relative expression of PI3K isoforms (p110α, p110β, p110δ, p110γ and p85) in bone marrow monocytes and in macrophages differentiated with macrophage colony-stimulating factor (M-CSF) or granulocyte-macrophage colony-stimulating factor (GM-CSF). However, alpelisib increased p110α, p110β, and p85 relative gene expression (2–3-fold) during M-CSF-dependent differentiation. Functionally, alpelisib-treated M-CSF macrophages displayed enhanced interleukin (IL)-6 and tumor necrosis factor alpha (TNF-α) secretion and reduced IL-10 production after lipopolysaccharide (LPS) plus interferon gamma (IFN-γ) or LPS stimulation. In contrast, GM-CSF macrophages differentiated with alpelisib secreted lower levels of IL-6 and TNF-α and reduced inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) gene expression. Additionally, cytokine profiles (IL-2, IL-6, IFN-γ and IL-10) were altered when alpelisib-treated macrophages were cocultured with CD4⁺ T cells under either antigen-specific or polyclonal activation conditions, indicating that the inhibitor modifies both differentiation and subsequent effector interactions of the macrophages. Thus, alpelisib induces lasting effects on macrophage differentiation and function, with potential implications in tumor-associated macrophages that develop under M-CSF or GM-CSF-rich cancer microenvironments. Full article

This study aimed to develop and validate a transcriptomic risk signature for uterine corpus endometrial carcinoma (UCEC) and to investigate whether the identified prognostic program reflects immune–tumor uncoupling within the tumor microenvironment. Using transcriptomic data from The Cancer Genome Atlas (TCGA) UCEC cohort, we identified a 28-gene transcriptomic signature defining a high-risk state. The derived risk score robustly stratified patients into distinct survival groups and remained an independent predictor of overall survival after adjustment for clinical covariates. Functional analyses revealed that high-risk tumors are characterized by a distinct immune–tumor uncoupling phenotype, in which interferon-gamma (IFNG)-associated inflammatory signaling is preserved but fails to translate into effective antitumor immune activity. Specifically, effector immune programs, including CD8 T cell-related signatures and cytotoxic activity, were consistently reduced despite elevated IFNG-associated signaling, indicating a functional discordance between immune activation and immune execution rather than classical T cell exhaustion. In parallel, high-risk tumors exhibited consistently elevated cell cycle and DNA repair-associated transcriptional programs, suggesting that proliferative and stress-adaptive mechanisms represent dominant drivers of poor prognosis. External assessment in an independent GEO cohort (GSE17025) demonstrated consistent associations between signature activity, tumor status, and histological grade, supporting the reproducibility of the underlying transcriptional program at the biological and clinicopathological level. Collectively, this study provides transcriptomic evidence for a previously underappreciated immune–tumor uncoupling state in UCEC and highlights the importance of integrating immune signaling and tumor-intrinsic programs to understand disease progression. Full article

Aberrant pathogenic immune responses drive autoimmune uveitides; however, comprehensive leukocyte profiling in the conditions remains limited. Here, this exploratory pilot study aimed to elucidate the immunodynamics of ocular sarcoidosis (OS) and Vogt–Koyanagi–Harada disease (VKH) to identify blood diagnostic biomarkers during their acute phases. We performed a prospective observational analysis of ten newly diagnosed, treatment-naïve OS patients and seven VKH patients during their acute phases, along with eight healthy controls (HCs). Mass cytometry was utilized to quantify the proportions of 37 distinct leukocyte subsets. In OS group, the proportion of CD8⁺ naive was lower than in both VKH and control groups. Furthermore, the proportions of CD8⁺ central memory and γδ T cells were decreased compared to HC group. Hierarchical cluster analysis categorized the leukocyte subsets into four principal clusters: Cluster A (Th17-like, monocytes, neutrophils, etc.), Cluster B (Tregs, B cells, NK cells, basophils, etc.), Cluster C (CD8⁺ T cells, Th1-like, Th2-like, DCs, etc.), and Cluster D (CD4⁺ terminal effector, CD8⁺ terminal effector, and CD66b⁻ neutrophils). Compared to HC group, the abundance of Cluster A was relatively high in OS group, and the abundance of cluster B was relatively high in VKH group. In OS group, the proportions of CD8⁺ T cells and CD8⁺ terminal effector correlated negatively with serum ACE and sIL-2R levels. ROC curve analysis estimated that CD4⁺/CD8⁺ ratio (cut-off value: ≥3.46), the proportion of monocytes (≥9.41%), and the decreased proportions of CD3⁺ T cells (≤43.9%) and CD8⁺ T cells (≤10.0%) in peripheral blood may serve as potential blood biomarkers for diagnosing OS. The exploratory pilot study provides a comprehensive and simultaneous data of leukocyte subset proportions in the acute phase of OS and VKH, and our preliminary findings suggest that the proportions of specific leukocyte subsets may represent potential candidates for blood-based biomarkers in the diagnosis of OS. Full article

Bone metastases remain one of the most clinically devastating complications of advanced cancer, particularly in breast, prostate, and lung malignancies, where they drive pain, fractures, hypercalcemia, and progressive functional decline. Their management is further complicated by a highly immunosuppressive bone microenvironment characterized by osteoclast-driven bone destruction, myeloid cell dominance, impaired antigen presentation, and weak effector T-cell infiltration, all of which limit the activity of conventional immunotherapies. In this setting, nanoparticles are emerging not merely as passive drug carriers but as programmable platforms capable of reshaping the metastatic niche. This review discusses how bone-targeted and immune-responsive nanocarriers can improve therapeutic precision through hydroxyapatite-binding ligands, dual-targeting strategies, stealth coatings, enzyme- and pH-responsive release systems, and externally guided platforms. We further examine how these systems modulate key immune compartments within bone metastases, including reprogramming tumor-associated macrophages and myeloid-derived suppressor cells, restoring cytotoxic T-cell activity, enhancing dendritic-cell activation, and enabling in situ vaccination through photothermal or photodynamic immunogenic cell death. Particular attention is given to the delivery of checkpoint inhibitors, cytokines, siRNA/miRNA, mRNA, and clustered regularly interspaced short palindromic repeats (CRISPR)-based payloads, as well as to the rational combination of these with chemotherapy, bone-modifying agents, and radiotherapy. Finally, we highlight major translational barriers, including lesion heterogeneity, limited penetration into mineralized tissue, off-target immune effects, manufacturing complexity, and the continued lack of bone-specific preclinical and clinical validation. Collectively, immunomodulatory nanoparticles represent a promising strategy to convert bone metastases from immune-refractory sites into more therapeutically responsive lesions. Full article