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关键词： IL-33   ST2 knockdown   Breast cancer stem cells   Wnt pathway   Immunotherapy  

摘要： Interleukin-33 (IL-33) plays multifaceted roles in tumor progression, but its autocrine regulation of breast cancer stemness and metastasis via the Wnt pathway remains unclear. Here, we investigated the IL-33/ST2 axis in breast cancer using CRISPR/Cas9, single-cell RNA sequencing, and murine models (orthotopic 4T1 and spontaneous MMTV-PyMT). Elevated IL-33 levels correlated with aggressive subtypes and poor prognosis. IL-33 overexpression enhanced proliferation, migration, and cancer stem cell (CSC) marker expression (CD44, ALDH1) in 4T1 and MDA-MB-231 cells, whereas ST2 knockdown via CRISPR or adeno-associated virus (AAV) attenuated tumor growth and metastasis in vivo, reducing CSC frequency. Mechanistically, IL-33 activated Wnt/beta-catenin signaling to promote stemness, which was reversed by the Wnt inhibitor XAV-939. Single-cell analysis revealed that IL-33 overexpression skewed the immune microenvironment toward immunosuppression, while ST2 knockdown restored antitumor immunity. Our findings establish an IL-33-Wnt axis as a critical driver of breast cancer aggressiveness and propose AAV-mediated ST2 silencing as a novel therapeutic strategy. Targeting this axis may offer dual benefits by suppressing stemness and enhancing immune surveillance, warranting clinical exploration for advanced breast cancer.

关键词： CD4(+) T cells   IL-12p35   immune thrombocytopenia   JAK2/STAT3/HIF-1 alpha   regulatory B cells  

摘要： Primary immune thrombocytopenia (ITP) is a condition characterized by immune-mediated platelet loss due to excessive destruction and/or insufficient production. IL-12p35 is involved in autoimmune uveitis;however, its role in ITP remains unknown. In the study, CD19(+) B and CD4(+) T cells were isolated from peripheral blood mononuclear cells (PBMCs) of patients with ITP and healthy individuals. CD4(+)CD25(-) T cells were cocultured with the CD19(+) B cells treated with anti-IL-12p35 antibody or IL-12p35 recombinant protein. The impact of IL-12p35 on CD19(+) B cells was assessed by enzyme-linked immunosorbent assay, Western blotting, quantitative real-time PCR and flow cytometry. PBMCs from patients with ITP showed lower proportions of IL-10(+)CD19(+) B cells and decreased mRNA levels of IL-12p35-coding gene IL12A than those from healthy individuals. Anti-IL-12p35 antibody-treated CD19(+) B cells could reduce the population of IL-10(+) in CD19(+) B cells and regulate the differentiation of CD4(+)CD25(-) T cells, while recombinant IL-12p35 demonstrated the opposite effects. Moreover, the increased IL-10(+) B-cell population and HIF-1 alpha expression in CD19(+) B cells induced by recombinant IL-12p35 were reversed by HIF-1 alpha and JAK2/STAT3 inhibitors. In conclusion, CD19(+)IL-10(+) B cells induced by IL-12p35 regulate CD4(+) T-cell subsets in patients with ITP via the JAK2/STAT3/HIF-1 alpha signalling pathway.

关键词： TNFA polymorphisms   TNF-alpha mRNA expression   sTNF-alpha   SLE  

摘要： Background/Objectives: A case-control study was conducted to determine the association of the -238 G>A and -308 G>A TNF-alpha (TNFA) promoter polymorphisms with mRNA and protein expression in 180 Systemic Lupus Erythematosus (SLE) patients and 186 control subjects (CS) from western Mexico. Methods: Genotyping was performed using the PCR-RFLP method. TNFA mRNA expression was assessed by real-time quantitative PCR, and soluble TNF-alpha (sTNF-alpha) levels were quantified by ELISA. For comparison groups, Chi-square, Mann-Whitney U, or Kruskal-Wallis tests were used. Spearman's rank correlation coefficient determined the correlation between variables. The Area Under the Curve was used to determine the diagnosing performance of sTNF-alpha. Results: No differences were found in the genotype distribution of -238 G>A and -308 G>A TNFA polymorphisms between SLE patients and CS. However, the -238A allele was associated with increased SLE susceptibility (OR 1.18 CI 95% 1.02-3.50, p = 0.037). Also, logistic regression analysis showed that LN risk was significantly higher in carriers of the -308A allele (OR 3.11 IC95% 1.15-6.43;p = 0.002). On the other hand, the TNFA mRNA expression was 3.3-fold higher in SLE compared to CS. SLE patients with -308 GG genotype showed higher TNFA mRNA expression compared to GA+AA genotype carriers (p < 0.01). Regarding sTNFa levels, SLE patients showed higher concentration than CS, mainly in lupus nephritis (LN), with a weak negative correlation with estimated Glomerular Filtration Rate and an acceptable accuracy for diagnosing SLE and LN, with areas under the curve of 0.61 and 0.65, respectively. Conclusions: The -238 A allele and -308 A allele of the TNFA gene are linked to a higher risk of susceptibility to SLE and LN in the western Mexican population. Additionally, SLE patients exhibited increased TNF-alpha gene expression and sTNF-alpha, particularly in LN, demonstrating acceptable diagnostic performance.

关键词： Immune related adverse event - irAE   Immune Checkpoint Inhibitor   Cardiotoxicity  

摘要： Background Immune checkpoint blockers (ICBs) have revolutionized cancer therapy, yet they remain largely ineffective in treating pancreatic ductal adenocarcinoma (PDAC). Moreover, ICBs can cause severe immune-related adverse events (irAEs), including fatal cardiac toxicity. Finally, obesity is a risk factor in PDAC that may differentially modulate ICB efficacy in a malignancy-dependent *** We investigated the mechanisms underlying irAEs induced by dual ICB therapy and sought to identify strategies to mitigate them while improving ICB efficacy in the obese setting. To this end, we used a clinically relevant mouse model that integrated key features of human PDAC: (1) high-fat diet-induced obesity, (2) an orthotopic PDAC, and (3) a therapeutic regimen combining chemotherapy (FOLFIRINOX) with ICBs (alpha-programmed cell death protein-1 + alpha-cytotoxic T-lymphocyte associated protein-4 antibodies).Results Obese mice developed cardiac irAEs and had elevated serum interleukin (IL)-1 beta levels after chemoimmunotherapy. IL-1 beta blockade not only prevented myocarditis and reduced cardiac fibrosis but also enhanced the antitumor efficacy of the combination of chemotherapy plus dual ICB therapy and significantly improved the overall survival of PDAC-bearing obese *** Our findings provide the rationale and compelling data to test a Food and Drug Administration-approved anti-IL-1 beta antibody in combination with chemotherapy and dual ICB therapy in patients with pancreatic cancer with obesity.

关键词： IL-33   ST2   Superoxide anion   Glial cells   Inflammation   And pain  

摘要： Reactive oxygen species such as superoxide anion have varied roles in inflammation and pain, which can be mimicked by potassium superoxide (KO2), the superoxide anion donor. Interleukin (IL)-33 has pleiotropic functions by activating its receptor suppression of tumorigenicity 2 (ST2). However, the role of IL-33/ST2 signaling in inflammatory pain initiated by reactive oxygen species (ROS) such as superoxide anion has not been investigated, which was the aim of the present study. IL-33 levels were assessed by enzyme-linked immunosorbent assay (ELISA). Mechanical and thermal hyperalgesia and overt pain were evaluated by electronic von Frey, hot plate, and abdominal writhing/paw flinching/licking, respectively. Edema and leukocyte recruitment (myeloperoxidase assay and total/differential cell count), antioxidant capacity, superoxide anion production and lipid peroxidation were assessed. Paw skin and spinal cord messenger ribonucleic acid (mRNA) expression of pro-inflammatory mediators and glial markers in the spinal cord were evaluated. Immunofluorescence was used to detect spinal glial and neuronal c-Fos activation. KO2 injection triggered IL-33 production in the paw skin and spinal cord of mice, induced hyperalgesia, edema, neutrophil recruitment to the paw tissue, overt pain-like behavior, and leukocyte recruitment to the peritoneum that were reduced in ST2 deficient mice. In the paw skin and spinal cord, KO2 triggered IL-33/ST2-dependent oxidative stress, and mRNA expression of inflammatory molecules, which were reduced by ST2 deficiency. KO2 induced spinal cord glial (at mRNA/protein levels) and neuronal activation in IL-33/ST2-dependent manner. IL-33/ST2 signaling mediates, at least in part, superoxide anion-induced inflammatory pain by modulating local and spinal inflammatory events.

摘要： Importance:Carriage of HLA-B*58:01 has been shown to have a strong association with the development of allopurinol-induced Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) and drug reaction and eosinophilia and systemic symptoms (DRESS) in many populations globally; however, there is a critical need to determine if this is generalizable to varying populations including those in the United States (US). Objective:To perform HLA class I and II association studies in a cohort of US patients diagnosed with allopurinol-induced SJS/TEN or DRESS compared to allopurinol tolerant and population controls. Design Setting and Participants:We enrolled consenting individuals who had specialist adjudicated allopurinol-induced SJS/TEN or DRESS (collectively allopurinol-SCAR). HLA carriage in these cases was compared to allopurinol tolerant and population controls identified through Vanderbilt University Medical Center (VUMC) BioVU, a biobank which includes 94,489 individuals with imputed human leukocyte antigen (HLA) class I and II typing from genotyping array data. Main Outcomes and Measures:We performed HLA class I and II conditional logistic regression case-control analyses between allopurinol-SCAR cases and both population controls and allopurinol tolerant controls matched on age, sex, and self-identified race. We reported odds ratio (OR) and 95% confidence interval (CI) with Bonferroni corrected ( ) <.05. Results:The conditional logistic regression analyses included allopurinol-SCAR cases (n=16) and 10:1 matched allopurinol tolerant controls (n=160). We found two HLA class I alleles independently associated with increased risk of allopurinol-induced SCAR: HLA-B*58:01 (OR 28 [95% CI, 8.6 - 100.6]) and HLA-A*34:02 (OR 20.6 [95% CI, 3.3 - 131.1]). We did not identify any HLA class II alleles meeting the Pc level of significance. Conclusions and Relevance:We found HLA-B*58:01 to be strongly associated with allopurinol-induced SCAR, generalizing findings from prev

关键词： Idiopathic nephrotic syndrome   scRNA-seq   IFN   PBMCs  

摘要： BackgroundIdiopathic nephrotic syndrome (INS) is a prevalent condition whose recurrence leads to multiple adverse effects. Previous studies on INS pathogenesis primarily focused on immune dysregulation, particularly T-cell changes and their correlation with cytokine shifts. Accumulating evidence suggests that B-cell dysfunction also plays a role. Nevertheless, a comprehensive understanding of the mechanisms and effective treatment strategies remains *** study investigates changes in gene expressions and cellular interactions of immune cells at a single-cell level using peripheral blood mononuclear cells (PBMCs). And subsequently validated through quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and flow *** identified seven main clusters using unsupervised clustering of 103,213 high-quality single cells. Through unsupervised clustering, patient-specific T cells (IFI44L + CD4 + T cells) that exhibited a pronounced elevation of interferon-stimulated genes (ISGs) is identified. Activation of ISGs and interferon (IFN)-related pathways are also observed in other clusters. Specifically, this study demonstrates that interferon-gamma (IFN-gamma) plays a crucial role by promoting the interaction between B-cell activating factor (BAFF) and receptors on B cells. This interaction triggers the release of autoantibodies, thereby initiating INS pathogenesis. Furthermore, telitacicept has shown efficacy in treating pediatric patients with frequent relapse NS(FRNS).ConclusionsOverall, our findings underscore the role of interferon and its related pathways in INS pathogenesis, providing novel therapeutic interventions for NS.

关键词： MMUD   PTCy   donor age   haploidentical   mismatched unrelated donor   post-transplantation cyclophosphamide  

摘要： BACKGROUND:Haploidentical donors and HLA-mismatched unrelated donors (MMUDs) are increasingly utilized for hematopoietic cell transplantation (HCT), with post-transplantation cyclophosphamide (PTCy) emerging as an effective graft-versus-host disease (GVHD) prophylaxis strategy. Despite the growing use of these donor types, comparative data to guide donor selection remain limited. Donor age is a known predictor of HCT outcomes, yet its specific impact when choosing between haploidentical and MMUD donors with PTCy-based prophylaxis has not been thoroughly explored. OBJECTIVES:This study aimed to evaluate the influence of donor age on HCT outcomes in patients receiving haploidentical or MMUD HCT with PTCy-based GVHD prophylaxis, hypothesizing that younger donors (<30 years) would be associated with improved outcomes compared to older donors (≥30 years) regardless of donor type. STUDY DESIGN:We conducted a retrospective analysis of 7,116 patients with hematologic malignancies from the Center for International Blood and Marrow Transplant Research (CIBMTR) database, transplanted between 2013 and 2021. Donors were categorized into four groups: younger haploidentical (<30 years), older haploidentical (≥30 years), younger MMUD (<30 years), and older MMUD (≥30 years). The primary outcome was GVHD-free relapse-free survival (GRFS), defined as the absence of grade III-IV acute GVHD, chronic GVHD requiring systemic immunosuppressive therapy (IST), relapse, or death. Secondary outcomes included overall survival (OS), treatment-related mortality (TRM), relapse, grade III-IV acute GVHD, overall chronic GVHD, and chronic GVHD requiring IST. Comparisons were made between (a) younger MMUD vs. older haploidentical and (b) younger haploidentical vs. older MMUD groups using multivariable Cox proportional hazards models. RESULTS:In multivariable analysis, the older MMUD group exhibited inferior GRFS (Hazard Ratio [HR] 1.20; 95% confidence interval [CI], 1.06-1.36; p=0.003), higher TRM (HR

关键词： Inflammatory bowel disease   Crohn's disease   T cells   4-1BB   mucosal immunology   PD-1   anti-TNF alpha treatment  

摘要： Objective: Pro-inflammatory T-cell responses dominate in Crohn's disease (CD). This may result from a dysbalanced expression of co-stimulatory and inhibitory T-cell receptors. The present study investigated if a dysbalanced co-stimulatory and inhibitory T-cell receptor expression are present in CD and can be rebalanced by anti-TNF alpha treatment. Methods: Mucosal biopsies from 27 patients with active CD receiving anti-TNF treatment were examined for the mRNA levels of the co-stimulatory 4-1BB and inhibitory PD-1 T-cell receptor. Levels of mRNA were compared between inflamed and noninflamed tissue, and before and after treatment. Peripheral T cells from 12 healthy controls (HC) and 11 active CD patients were evaluated for their expression of 4-1BB and PD-1 by flow cytometry. Results: The 4-1BB mRNA levels in inflamed mucosa were upregulated (> 2-fold) compared with uninflamed mucosa (p < 0.05). Anti-TNF alpha treatment reduced the 4-1BB and PD-1 mRNA levels in the inflamed gut tissue (p < 0.05). In in vitro activated T cells, the percentage of both 4-1BB and PD-1 positive CD4+ and CD8+ T cells increased more than 1.5 fold compared with HC (p < 0.05). The 4-1BB/PD-1 ratio on activated peripheral T cells was significantly reduced in CD after anti-TNF therapy (p < 0.05). Conclusions: A dysbalanced mucosal proinflammatory co-stimulatory T cell receptor expression was present in active CD and modified by anti-TNF alpha treatment. However, anti-TNF alpha treatment did not normalize the expression of 4-1BB or PD-1 on peripheral T cells although a modest increased immunoregulatory capacity could be demonstrated.