T cells play a vital role in hepatitis B virus (HBV) infection, mediating both protective immunity and immunopathology. Deep profiling of the T cell receptor (TCR) complementarity-determining region 3 (CDR3) provides critical insights into T cell composition and function. However, the characteristics of the TCR α-chain (TRA) and β-chain (TRB) repertoires in individuals with different HBV infection and vaccination statuses remain incompletely understood. We employed high-throughput sequencing (HTS) to profile the TRA and TRB CDR3 repertoires in peripheral blood mononuclear cells (PBMCs) from 6 inoculation-positive responders (IPR) to hepatitis B (HepB) vaccination, 6 individuals with chronic hepatitis B (CHB), and 5 healthy donors (HD). We systematically compared the frequency, diversity, similarity, and V/J segment usage of TRA and TRB CDR3s across the three groups. The CHB group exhibited significantly fewer unique TRA and TRB CDR3s than both IPR and HD groups. Overlap analysis revealed that the BUB index for TRAV was significantly higher in IPR than in CHB and HD groups (P = 0.0073 and P = 0.0317, respectively), while for TRBV, a significantly higher BUB index was observed only between IPR and CHB groups (P = 0.0002). Additionally, identical TRA CDR3s were preferentially shared among individuals within each group, whereas TRBJ usage remained relatively stable across groups. Collectively, our findings delineate distinct CDR3 repertoire characteristics of circulating T cells in IPR and CHB, which may enhance understanding of the protective response to HepB vaccination and inform future investigations into TRA- and TRB-based strategies for HBV diagnosis and prevention.
Ankylosing spondylitis (AS) is a chronic inflammatory autoimmune disease where T-cell dysregulation plays a pivotal role. A balance of co-stimulatory and co-inhibitory molecules governs the activation of T cells. This study aimed to investigate the mRNA expression profiles of these immune checkpoint signals in AS patients. A cohort of 33 AS patients and 28 healthy controls (HC) from the Taiwanese population was recruited. The mRNA expression levels of five co-stimulatory molecules (CD28, CD40, CD40L, OX40, and OX40L) and one co-inhibitory molecule (TIM-3) were quantified using qRT-PCR. Relative expression was calculated via the 2−△△ct method, normalized to beta-actin. Significant downregulation of several immune checkpoint molecules was observed in AS patients. Compared with HCs, AS patients exhibited a 2.67-fold reduction in CD28 (p < 0.0001), a 2.37-fold reduction in CD40L (p < 0.0001), a 2.20-fold reduction in TIM-3 (p < 0.0001), and a 1.47-fold reduction in CD40 (p = 0.006). No significant differences were found in the expression of OX40 and OX40L between the two groups. The concurrent downregulation of both activating and inhibitory signals suggests a trend toward coordinated downregulation of the immune checkpoint network in the peripheral blood of AS patients. This phenomenon may represent chronic immune exhaustion or the redistribution of activated immune cells to sites of inflammation. These molecules, particularly CD40L and TIM-3, may represent candidates for further biomarkers that warrant further investigation in large, age- and sex-matched cohorts.
Psoriasis is a chronic inflammatory skin disease with highly variable responses to biologic therapies. Predictive tools to guide personalized treatment remain an unmet clinical need. To develop and validate a multiomics-clinical model for predicting a 75
Sepsis remains a critical clinical challenge with high mortality. Uncovering reliable biomarkers and key regulatory mechanisms is crucial for early diagnosis and prognosis improvement. Serum miR-181a-2-3p was detected via RT-qPCR in 167 sepsis patients and 103 healthy controls (HC). The relationship of this miRNA to conventional diagnostic markers for sepsis was evaluated by employing Pearson and Spearman correlation methods. ROC curves evaluated the diagnostic efficacy of this miRNA and TMEM173. The prognostic value of miR-181a-2-3p was evaluated by Kaplan-Meier and Cox regression analyses. The functional targeting effect of miR-181a-2-3p on TMEM173 was confirmed by dual luciferase reporter assay. A sepsis model was established using LPS-stimulated THP-1 cells. Cell proliferation was assessed using the CCK-8 assay. Inflammatory cytokine concentrations were measured via ELISA. Serum miR-181a-2-3p was lower in sepsis patients than in HC. Among septic patients, miR-181a-2-3p expression was inversely associated with PCT, CRP, APACHE II, and SOFA scores. In addition, low expression levels of this miRNA were independently associated with poor prognosis. TMEM173 levels were up-regulated in sepsis patients, effectively distinguishing them from HC. Furthermore, miR-181a-2-3p may contribute to the progression of sepsis by regulating the cGAS-STING and NF-κB signaling pathways via TMEM173. Serum miR-181a-2-3p had diagnostic and prognostic value for sepsis. miR-181a-2-3p may exert a protective effect in sepsis by targeting TMEM173.
Peritonitis is a major complication of peritoneal dialysis and is closely associated with inflammatory injury of peritoneal mesothelial cells (PMCs). Solute carrier family 7 member 11 (SLC7A11), a key component of the cystine/glutamate antiporter system xc−, has been implicated in inflammatory regulation; however, its role in peritonitis remains unclear. In this study, we found that SLC7A11 expression was significantly upregulated in peritoneal tissues from a lipopolysaccharide (LPS)-induced mouse model of peritonitis and in LPS-stimulated PMCs. Knockdown of SLC7A11 attenuated LPS-induced inflammatory responses, as evidenced by decreased expression of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and cluster of differentiation 14 (CD14). In addition, SLC7A11 knockdown reduced LPS-induced phosphorylation of p65, a major subunit of nuclear factor-kappa B (NF-κB), suggesting that SLC7A11 may contribute to inflammatory activation, at least in part, through NF-κB signaling. Mechanistically, chromatin immunoprecipitation sequencing (ChIP-seq) analysis identified SLC7A11 as a potential downstream target of peroxisome proliferator-activated receptor gamma (PPARγ). ChIP-qPCR further confirmed that LPS reduced the binding of PPARγ to the SLC7A11 promoter. Moreover, activation of PPARγ by rosiglitazone suppressed SLC7A11 expression, whereas inhibition of PPARγ by GW9662 enhanced SLC7A11 expression. Collectively, these findings demonstrate that SLC7A11 contributes to LPS-induced peritoneal inflammation and is negatively regulated by PPARγ. The PPARγ–SLC7A11 axis may represent a potential regulatory mechanism and therapeutic target in peritonitis.
This exploratory study aimed to characterize peripheral blood TCRβ CDR3 repertoire features in patients with multidrug-resistant tuberculosis (MDR-TB) compared with drug-sensitive tuberculosis (DS-TB). TCRβ CDR3 diversity and clonotype composition were analyzed in five MDR-TB patients and five DS-TB patients using adaptive immune receptor repertoire sequencing (AIRR-seq). In this small exploratory cohort, MDR-TB patients showed lower overall clonotype numbers, increased dominant clone expansion, and a more focused T-cell response pattern. Seven recurrent CDR3 core motifs were detected in MDR-TB samples; however, because several motifs were short and lacked canonical IMGT-defined full-length CDR3 boundaries, these findings should be interpreted as candidate sequence features rather than validated antigen-specific TCR clonotypes. TCRβ CDR3 repertoires in both groups were mainly composed of 13-, 14-, and 15-amino-acid sequences. This study provides preliminary, hypothesis-generating evidence that peripheral blood TCRβ CDR3 repertoire features may differ between MDR-TB and DS-TB patients. Larger cohorts including healthy controls and independent functional validation are required before disease-specific or antigen-specific conclusions can be drawn.
The World Health Organization’s goal to eradicate hepatitis C by 2030 remains unattainable without an effective vaccine. Consensus generation combined with layering has the potential to introduce vaccines against highly variable viruses like HCV. However, this approach yields numerous candidates that require in vivo testing. Profiling the affinity of these candidates to monoclonal antibodies via in silico methods can help reduce the number of candidates. In this study, we evaluated the differentiating power of in silico affinity profiling on two previously reported genotypic consensus sequences (NOTC1 and NOTC2) and two newly generated pan-genotypic sequences (L1-CON and L2-CON), each derived using distinct consensus generation algorithms. Structural models of these sequences were generated via homology modelling and simulated within a canonical ensemble. The resulting average structures were then used as inputs for antigen–antibody docking studies against all reported antibody structures known to bind to each epitope in the HCV E2 protein. Docking results for epitope II revealed lower binding energies for NOTC1 compared to NOTC2 (p = 0279) and J-2a (p = 0.0132) consistent with their reported in vivo performance. This discrimination was not reproduced in genetic distance from the common ancestor of Genotype 1 and also structural similarity between protein models and crystalographed antigen. Notably, neither sequence nor structural similarity correlated with the docking outcomes, highlighting the predictive value of the proposed affinity profiling pipeline as an initial assessment tool for vaccine candidates.
DLBCL shows clinical heterogeneity, driven by molecular diversity within the tumor along with diverse components in the tumor microenvironment (TME). Immunogenic cell death (ICD) activates adaptive antitumor immunity by inducing immunogenic signals in cancer cells, positioning it as a key mechanism in tumor immunotherapy. However, the role of ICD in DLBCL pathogenesis remains understood. We analyzed integrated single-cell and bulk RNA-seq data from GEO to identify ICD-related genes via AUCell, ssGSEA, and WGCNA. We constructed an ICD-related signature (ICDRS) via the Mime1 package and validated it in four independent cohorts. Prognostic utility was evaluated via Kaplan‒Meier survival analysis, time-dependent receiver operating characteristic (ROC) curves, and multivariate Cox regression. The TME is characterized by ESTIMATE, CIBERSORT, and ImmuCC. Finally, a combination therapy was designed on the basis of risk stratification and validated in DLBCL models. Integration of single-cell and bulk RNA-seq data identified 122 ICD-related genes, enabling the construction of an ICDRS. The ICDRS demonstrated prognostic value in DLBCL patients. The ICDRS score was an independent predictor of OS in all cohorts and was correlated with clinical indicators, including the IPI, ECOG, and Ann Arbor stage. Importantly, the ICDRS characterized an immunosuppressive microenvironment in high-risk patients and predicted immunotherapy and chemotherapy responses. In DLBCL models, radiotherapy reduced the risk score, and fludarabine-radiotherapy-anti-PD-1 inhibited tumor progression by reversing radiotherapy-induced immunosuppression. The ICDRS serves as a prognostic biomarker for DLBCL, and the combination therapy overcomes radioresistance, thereby establishing a translatable strategy.
Severe pneumonia poses a life-threatening risk to children’s physical health. This study aimed to evaluate the diagnostic value of serum miR-331-3p in pediatric severe pneumonia. A total of 97 healthy children, 101 children with non-severe pneumonia and 101 children with severe pneumonia were enrolled. Serum miR‑331‑3p expression was detected by RT‑qPCR. An LPS‑induced MRC‑5 cell model was established to simulate severe pneumonia in vitro. Cell proliferation, apoptosis, and inflammatory cytokine levels were measured. The direct targeting relationship between miR‑331‑3p and B cell lymphoma 2 antagonist/killer 1(BAK1) was verified by dual‑luciferase reporter assay. Serum miR‑331‑3p expression gradually decreased from healthy controls to patients with non-severe pneumonia, and reached the lowest level in children with severe pneumonia. miR-331-3p exhibited good diagnostic efficacy for identifying severe pneumonia from healthy children (AUC = 0.899) and discriminating severe from non-severe pneumonia (AUC = 0.830). Lower miR-331-3p levels were correlated with higher inflammatory indicators in pneumonia patients. In vitro, miR‑331‑3p promoted cell proliferation, inhibited apoptosis, and suppressed the production of TNF‑α and IL‑6 by directly targeting and negatively regulating BAK1. Overexpression of BAK1 abolished the protective effects of miR‑331‑3p. Serum miR‑331‑3p exhibits favorable differential diagnostic potential for identifying severe pediatric pneumonia in this single-center cohort, while multicenter validation with independent cohorts is still needed to confirm its clinical predictive performance.
Allergic asthma is a helper T cell-driven chronic airway inflammation. This study evaluated the clinical significance of miR-191-3p in allergic asthma and its role in regulating airway epithelial inflammation via CD247. qRT-PCR was employed to determine miR-191-3p levels in 125 allergic asthma patients and 105 healthy controls. ROC curve assessed diagnostic value. Pearson and logistic regression analyzed clinical correlations and predictive factors. BEAS-2B cells were HDM-stimulated to establish an inflammation model. After miR-191-3p inhibitor transfection, inflammatory and oxidative stress were assessed. The miR-191-3p/CD247 interaction was predicted bioinformatically and confirmed by dual-luciferase assay. Rescue experiments validated the role of this axis in airway inflammation. Serum miR-191-3p was significantly elevated in allergic asthma patients versus controls (AUC = 0.841), was positively correlated with total IgE and eosinophil counts, and was identified as an independent risk factor for asthma. In BEAS-2B cells, HDM stimulation upregulated miR-191-3p expression, increased inflammatory release, and induced oxidative stress imbalance, all of which were reversed by miR-191-3p inhibition. Mechanistically, CD247 was validated as a direct target of miR-191-3p. Rescue experiments showed that CD247 silencing partially abolished the anti-inflammatory effects of miR-191-3p inhibition. miR-191-3p is significantly upregulated both in allergic asthma patients and in house dust mite-stimulated airway epithelial cells. These findings indicate that miR-191-3p may contribute to airway inflammation and oxidative stress through targeted regulation of CD247 expression.
This study aimed to investigate the correlation between serum T helper 1/T helper 2 (Th1/Th2) balance and the risk of osteoporosis (OP) in patients with Helicobacter pylori (Hp) infection. All patients were tested for both Hp and bone mineral density (BMD). A total of 200 patients were categorized into Hp non-infected (n = 100) and Hp infected (n = 100) groups based on 14C urea breath test results. Subsequently, patients were further divided into non-OP (n = 110) and OP (n = 90) groups according to BMD measurements. Serum levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) were measured to calculate the IFN-γ/IL-4 ratio, reflecting the Th1/Th2 balance. Bone metabolism markers, bone alkaline phosphatase (BALP) and bone gal protein (BGP), were also assessed. The Hp infected group exhibited significantly lower BMD (0.97 vs. 1.02 g/cm², P = 0.024), higher BALP (19.28 vs. 18.25 µg/L, P = 0.026), lower BGP (17.35 vs. 18.91 µg/L, P = 0.017), a higher IFN-γ/IL-4 ratio (18.26 vs. 15.87, P < 0.001), and a higher OP prevalence (54
Sepsis-associated acute kidney injury (SA-AKI) is a serious complication for sepsis patients, and microRNA (miRNA) is involved in its occurrence and development. This study investigated the clinical value of miR-379-5p and its potential mechanism on LPS-induced human proximal tubular epithelial cell lines (HK-2). Serum miR-379-5p levels were measured in 85 sepsis patients and 74 SA-AKI patients. The clinical significance of miR-379-5p was explored from the perspectives of SA-AKI diagnosis, kidney injury correlation, and 28-day survival assessment. The LPS-induced HK-2 cell model was constructed, and the regulatory effect of miR-379-5p on LPS-induced HK-2 cell viability and inflammatory response was analyzed. The regulatory mechanism was explored, focusing on the involvement of EDN1. Low expression of serum miR-379-5p could distinguish SA-AKI from sepsis patients and was negatively correlated with serum creatinine (Scr), blood urea nitrogen (BUN), cystatin-C (Cys-C), kidney injury molecule-1 (KIM-1), C-reactive protein (CRP), procalcitonin (PCT), and white blood cell (WBC). Low expression of miR-379-5p was correlated with the 28-day survival rate. LPS could inhibit the expression of miR-379-5p in HK-2 cells, inhibit cell viability, and promote the concentration of inflammatory factors TNF-α, IL-1β, and IL-6. Overexpression of miR-379-5p reversed the effect of LPS on HK-2 cells. EDN1 was a direct target gene of miR-379-5p. Overexpression of EDN1 counteracted the protective effect of miR-379-5p overexpression on HK-2 cells. Downregulated serum miR-379-5p had the potential value in diagnosing SA-AKI and forecasting 28-day patient survival. Overexpression of miR-379-5p protected HK-2 cells from LPS-induced injury by targeting and inhibiting EDN1.
Unexplained seizures represent a substantial diagnostic and therapeutic challenge in clinical neurology. Immune-mediated mechanisms coupled with prior infections are increasingly recognized as pivotal etiological contributors, yet large-scale epidemiological data from western China remain scarce. This retrospective cohort study enrolled 471 patients with unexplained seizures from eight provinces in western China between January 2017 and June 2022. Serum and cerebrospinal fluid samples were tested for neural-specific autoantibodies, thyroid peroxidase antibody (TPO-Ab), thyroglobulin antibody (TgAb), antinuclear antibody (ANA), and immunoglobulin M/G antibodies against Toxoplasma gondii, rubella virus, cytomegalovirus, and herpes simplex virus. Patients were stratified into definite autoimmune-associated seizures (AAS), probable AAS, and non-AAS subgroups. This trial was registered with the Chinese Clinical Trial Registry (ChiCTR1900027074) on October 30, 2019, and was retrospectively registered as the first participant was enrolled in January 2017. Overall, 14.65
To systematically evaluate the key modeling parameters of an allergic rhinitis (AR) model induced by ovalbumin (OVA) combined with aluminum hydroxide [Al(OH)₃] adjuvant in BALB/c mice, and to compare the effects of different sensitization and challenge protocols on model induction, thereby providing a reference for model standardization. A systematic search was conducted in PubMed, Embase, Web of Science and the Cochrane Library to identify studies using OVA combined with Al(OH)₃ to establish mouse models of AR. Data were extracted on animal strain, modeling protocols, and behavioral, immunological, and histopathological outcome measures. The model induction effects were evaluated using the standardized mean difference (SMD) and 95
To investigate the prevalence and distribution characteristics of unexpected antibodies through a retrospective analysis of 115 positive cases and explore transfusion strategies for patients with unexpected antibodies. Clinical data of 33,013 patients who underwent unexpected antibody testing in a hospital from May 2023 to October 2025 were collected, and 115 positive cases were retrospectively analyzed. The overall positivity rate of unexpected antibodies and their distribution across different sexes and ABO blood types were statistically analyzed. The effects of transfusion history, pregnancy history, disease types, and treatment history on the production of unexpected antibodies were examined. Among 33,013 tested individuals, 115 were positive for unexpected antibodies, yielding an overall positivity rate of 0.35
Allergic rhinitis (AR) is a prevalent inflammatory condition with limited long-term treatment options. Probiotics have been investigated as immunomodulatory agents, yet evidence remains inconsistent. This umbrella review with updated meta-analysis evaluated probiotic efficacy on immunological and clinical outcomes in children and adults with AR. We systematically searched PubMed, Scopus, Web of Science, Cochrane Library, and Embase through May 2026. Randomized controlled trials assessing probiotics effects on IgE, antigen-specific IgE, eosinophil count, Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ), and Total Rhinitis Symptom Score (RTSS) were included. Weighted mean differences (WMD) with 95
MicroRNAs (miRNAs) are key regulators and potential biomarkers for early diagnosis and prognosis of sepsis. To investigate the diagnostic and prognostic value, and molecular mechanism of miR-1290 in sepsis. Enrollment comprised 127 healthy controls and 133 patients with sepsis. RT‑qPCR measured serum miR-1290 and TAO Kinase 1 (TAOK1) levels. ROC curve and Logistic regression assessed diagnostic efficacy and risk prediction of miR-1290 for sepsis. Kaplan-Meier and Cox regression evaluated its short-term prognostic value. An LPS-induced macrophage (THP-1) inflammation model was established to examine the effects of miR-1290 on cell viability (CCK-8), inflammatory cytokines TNF-α, IL-6, and IL-1β (ELISA), and macrophage polarization markers (RT-qPCR). Bioinformatic prediction and dual-luciferase reporter verified the interaction between miR-1290 and TAOK1, and rescue experiments were conducted to determine that TAOK1 reverses the biological functions of miR-1290. miR-1290 was downregulated in patients with sepsis and showed high diagnostic value. Its expression level was negatively correlated with both SOFA and APACHE II scores, and demonstrated good predictive performance for short-term mortality. In the LPS-induced macrophage model, overexpression of miR-1290 suppressed TAOK1 expression, elevated cell viability, suppressed the secretion of TNF-α, IL-6, and IL-1β, inhibited M1 polarization (downregulation of iNOS and CD86), and enhanced M2 polarization (upregulation of CD206 and Arg-1). Moreover, TAOK1 overexpression reversed the anti-inflammatory and protective effects of miR-1290. Downregulated miR-1290 shows potential as both a diagnostic and prognostic indicator in sepsis. It may exert a protective effect on macrophages by targeting TAOK1, thereby inhibiting disease progression.
Rheumatoid arthritis (RA) is a chronic autoimmune disease that may lead to progressive joint damage and disability. Although methotrexate is a first-line treatment, some patients have inadequate responses or adverse reactions. This study aimed to evaluate the efficacy and safety of methotrexate combined with iguratimod versus methotrexate monotherapy in patients with RA. Based on existing medical records, the clinical data of 80 patients with rheumatoid arthritis treated at our hospital between October 2021 and October 2023 were retrospectively analyzed. This was a single-center, retrospective, non-randomized controlled study, and the investigators were not involved in treatment allocation. According to the actual treatment regimens documented in the medical records, patients were divided into a conventional treatment group, which received methotrexate, and a combination treatment group, which received methotrexate plus iguratimod. Treatment efficacy, bone metabolism, CRP, RF, ESR, immune function, and adverse reactions were compared between the two groups. As the treatment regimens were not randomly assigned, the findings were mainly used to evaluate the associations between different treatment regimens and clinical outcomes. The combination therapy group demonstrated significantly higher response rates for ACR20, ACR50, ACR70 and considerably higher levels of N-MID and T-PINP than the conventional treatment group following treatment. This group demonstrated significantly decreased levels of β-CTX, CRP, RF, and ESR compared to the conventional treatment group (p < 0.05). After treatment, except for the similar changes in CD4 + and CD8 + levels, the levels of IgG, IgA, and IgM in the combination treatment group were significantly lower than the conventional treatment group (p < 0.05). The combination therapy group showed a more significant drop in DAS28 values than conventional treatment group (p < 0.05). However, there was no significant difference in adverse effects (p > 0.05). In this single-center retrospective study, compared with methotrexate monotherapy, methotrexate combined with iguratimod was associated with higher ACR20, ACR50, and ACR70 response rates, as well as greater improvements in inflammatory markers, bone metabolism indicators, and DAS28 scores, without a significant increase in the incidence of adverse reactions. However, because this study used a non-randomized design and did not include propensity score matching, the findings should be further validated in prospective randomized controlled trials.
Systemic lupus erythematosus (SLE) is an autoimmune disease in which genetic susceptibility contributes to immune dysregulation and increased disease risk. However, the specific mechanisms by which these SNPs influence gene and protein expression and ultimately affect disease pathology remain unclear. Thus, this study integrated genotype and protein expression results to thoroughly investigate the influence of genetic variation on the immune mechanisms underlying systemic lupus erythematosus (SLE). A total of 38 SLE patients and 34 healthy controls were included in this study. The cell percentage and expression level of CD28, CTLA-4, and PD-1 on the CD3+CD4+ cells were assessed by flow cytometry, in which phytohemagglutinin, interleukin-7, and interleukin-2 were used to stimulate cells to detect CTLA-4 and PD-1. Associations between SNP genotypes and protein expression or cell proportions were subsequently evaluated using the Mann-Whitney U test, with significance at p = 0.017. Finally, the data were integrated with our previous functional analyses of SNPs to elucidate the role of co-stimulatory gene variants in SLE. The GT genotype of the CD28 rs1879877 was associated with increased CD3⁺CD28⁺ expression (p = 0.017) exclusively in patients with SLE. Additionally, the TT genotype of the CTLA4 rs733618 was associated with increased cell percentage (p = 0.007 in SLE group and 0.011 in combined group), and the TT genotype of rs16840252 were associated with increased CTLA-4 expression (p = 0.004) in the resting status. Furthermore, the CC genotype of rs36084323 and the GG genotype of rs2227982 were associated with increased PD-1 expression in the SLE group (p = 0.014) and combined group (p = 0.002 and 0.001). Notably, rs36084323 and rs2227982 consistently correlated strongly with increased PD-1 expression after stimulation (p = 0.002 and 0.001). These findings highlight that SNPs in the CTLA4 and PDCD1 genes act as general regulatory variants, while only CD28 rs1879877 appears to have an enhanced effect on CD28 expression specifically under the SLE disease condition. Additionally, the rs733618 and rs36084323 SNPs were associated with SLE and exhibited significant differences in transcriptional activity.
