Search Results (2,404)

The current knowledge on the histological structure of the gastrointestinal tract (GIT) in cetaceans is based on general descriptions. The aim of this study was to characterize the histology and expression of immune cell markers in samples from the GIT and lymph nodes (LNs) in a harbour porpoise (Phocoena phocoena) bycaught in the Cantabrian Sea. The thickness of the histological layers of the GIT was measured, being greater in the stomach and anal canal, although no significant differences were found among any intestinal segment (p = 0.448). Variation in thickness, morphology of the folds, and the presence of Peyer’s patches allowed the duodenal ampulla and the distal segments to be distinguished from the rest of the intestine. An immunohistochemical technique was performed to identify the following markers: IBA1 for macrophages, CD3 for T lymphocytes, and CD20 for B lymphocytes. The distribution of immune cells varied significantly along the GIT, with higher percentages of all three cell types in the distal intestine and the anal tonsil. Within the LNs, B lymphocytes represented the predominant cell population. This study provides the first description of the histological structure of the GIT and associated lymphoid tissue in a harbour porpoise, which will be useful for future research studies. Full article

Non-muscle invasive bladder cancer (NMIBC) accounts for the majority of bladder cancer diagnoses and remains a clinical challenge due to its high recurrence and progression rates despite intravesical Bacillus Calmette–Guérin (BCG) therapy. In recent years, tumor-infiltrating lymphocytes (TILs) have emerged as promising biomarkers, reflecting the interplay between the tumor and host immune system. However, the evidence regarding their prognostic and predictive role is still conflicting, largely due to methodological heterogeneity, lack of standardized evaluation criteria, and limited prospective validation. This narrative review summarizes the current knowledge on TILs in NMIBC, focusing on their compartmental distribution (stromal, intraepithelial, and tumor–stroma interface), compositional diversity (CD4⁺, CD8⁺, Treg, B cells), and spatial dynamics. Special attention is given to their role in predicting response to BCG immunotherapy, the contribution of tumor-associated macrophages and tertiary lymphoid structures, and the emergence of immune escape pathways, including Programmed Death-Ligand 1 (PD-L1) and the HLA-E/NKG2A axis. Advances in digital pathology, spatial transcriptomics, and integrated immunoscore models provide more accurate metrics compared to simple cell counts, highlighting the importance of functional and spatial signatures. Despite encouraging progress, TILs are not yet ready for routine incorporation into histopathological reporting. Future directions include standardized assessment, integration with molecular biomarkers, and prospective multicenter validation to enable their translation into risk stratification and personalized therapeutic decision-making. Full article

Cancers are diseases described by the irregular spread of cells that have developed invasive features, enabling them to invade adjacent tissues. The specific diagnosis and effective management of oncological treatments depend on the timely detection of circulating tumor cells (CTCs) in a patient’s bloodstream. One of the most promising approaches to CTC separation from blood fractions involves the dielectrophoresis (DEP) technique. This research presents a new DEP-based bionic system designed for MDA-MB-231 breast cancer cell isolation from white blood cell (WBC) subtypes with a viable approach to cell viability. This work leverages the principle that every cell type possesses a unique dielectric fingerprint. This dielectrophoresis microfluidic device is designed to act as a scanner, reading these fingerprints to achieve a continuous, label-free separation of cancer cells from blood components with a high efficiency. In the proposed system that consists of three different stages, the first stage allows for separating B-lymphocytes and Monocytes from Granulocytes and MDA-MB-231 cells. The separation of B-lymphocytes from Monocytes occurs in the second step, while the last step concerns the separation of Granulocytes and MDA-MB-231 cells. In the analysis, x-y graphs of the electric potentials, velocity fields, pressure distributions, and cellular DEP forces applied to the cells, as well as the resulting particle paths, are provided. The model predicts that the system operates with a separation efficiency of nearly 92%. This work focuses on an investigation of the impact of electrode potentials, the velocity of cells, the number of electrodes, the width of the channel, and the output angles on enhancing the separation efficiency of particles. Full article

Background: Exercise-induced bronchoconstriction (EIB) is common in athletes, being more frequent in outdoor endurance-based/long-distance sports. We followed a World-Class marathon paddler’s season with recurrent episodes of EIB, which intensified during cold exposure workouts. This unique immunophenotype profile during the season and its variations were reflected in acute and chronic inflammatory markers. Methods: A longitudinal case study was conducted with blood sampling obtained from a single paddler after overnight fasting at three timepoints: T1 (beginning of season, after 15-day rest period), T2 (post-Winter National Championship), and T3 (post-Summer National Championship). Complete blood counts and lymphocyte immunophenotyping were performed using automated hematology analysis and multiparametric flow cytometry. Results: The total numbers of leukocytes (T1: 6.3; T2: 5.0; T3: 5.5 × 10⁹/L), neutrophils (3.1; 2.5; 2.8 × 10⁹/L), and lymphocytes (2.4; 1.8; 2.2 × 10⁹/L) declined between T1 and T2, followed by a partial recovery at T3. In contrast, monocyte counts exhibited the reverse pattern (0.41; 0.62; 0.31 × 10⁹/L). The two T cell subsets (αβ and γδ) remained relatively stable, showing only minor seasonal fluctuations. CD19+ B cells, initially at very low levels, increased steadily as the season progressed (0.05; 0.07; 0.16 × 10⁹/L). During T2, the proportion of memory lymphocytes (CD45RO+) rose, while naive cells (CD45RA+) declined; this trend was subsequently inverted at M3. Although the CD4+/CD8+ ratio varied over time, it consistently stayed below the normal reference range established for healthy controls (0.50; 0.83; 0.60 for T1, T2, and T3, respectively). Conclusions: The immune assessment of the World-Class marathon paddler revealed transient immunosuppression early in the season, marked by reduced neutrophils, a low CD4+/CD8+ ratio, and diminished CD19+ lymphocytes. Over time, immune parameters showed signs of recovery, indicating a temporary imbalance that did not impair the athlete’s physical performance. Conclusions: This case study of an elite marathon kayaker revealed transient immune fluctuations across a competitive season, including early immunosuppression (low neutrophils, CD4+/CD8+ ratio 0.50, and minimal CD19+ B cells) followed by partial recovery mid- and late-season. Despite persistently inverted CD4+/CD8+ ratios suggesting chronic immune dysregulation, the athlete maintained competitive performance, highlighting the temporary nature of these changes and emphasizing that regular immune monitoring can help optimize health and performance in elite athletes. Full article

Background/Objectives: NOTCH1 is frequently mutated in chronic lymphocytic leukemia (CLL) and is a marker of poor prognosis. In addition to NOTCH1, mutations in the NOTCH1 regulatory pathway including SPEN have been described in a limited number of CLL cases and others have suggested that these mutations are also associated with adverse patient outcomes Methods: In this study, 1617 CLL cases were assessed using targeted sequencing and a 29-gene panel and the results were correlated with prognosis. Results: SPEN mutations were detected in 48 (2.9%) CLL patients: 92.4% were deleterious (frameshift or truncating nonsense mutations) and the remaining (7.6%) were missense. Compared with SPEN wild type CLL patients, SPEN mutated patients had a statistically higher frequency of IGHV unmutated status (79.5% vs. 57.8%, p = 0.004), CD38 positivity (73.3% vs. 52.4%, p = 0.01), ZAP70 positivity (77.3% vs. 58.3%, p = 0.01) and trisomy 12 (43.5% vs. 13.7%, p < 0.001). The most common gene mutations co-occurring with SPEN mutations were as follows: NOTCH1 (43.7%), TP53 (22.9%), BIRC3 (12.5%), SF3B1 (10.4%), XPO1 (8.3%), MUC2 (6.2%), ATM (4.2%), FBXW7 (4.2%), and BTK (4.2%). Patients with SPEN mutated CLL had a significantly shorter time-to-first treatment compared to CLL patients with wild type SPEN (2.5 vs. 4.07 years, p = 0.01). The finding of shorter time-to-first treatment in SPEN mutated CLL patients was not maintained in a multivariable analysis. IGHV unmutated status, TP53 disruption, and trisomy 12 remained independently predictive of a shorter time-to-first treatment in a multivariable analysis. Conclusions: These data show that SPEN mutations in CLL are associated with adverse prognostic impact and should be included in sequencing assays performed for the prognostic workup of CLL patients. Full article

This study investigates the immunomodulatory effects of a well-characterized cannabidiol (CBD)-rich cannabis extract, CAN296, on T lymphocytes (T cells), particularly Cluster of Differentiation 4 (CD4⁺) helper and Cluster of Differentiation 8 (CD8⁺) cytotoxic subsets, by examining T-cell activation, cytokine secretion, and cytotoxic molecule expression in comparison with the conventional treatments dexamethasone (DEX) and tacrolimus (TAC). It addresses key processes involved in the formation of premalignant immune-mediated lesions, such as those seen in oral lichen planus (OLP) and oral manifestations of graft-versus-host disease (oGVHD). CD4⁺ and CD8⁺ T cells were isolated from healthy donors and assessed in vitro for T cell activation via CD69 expression, secreted tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) levels according to enzyme-linked immunosorbent assay (ELISA), and cytotoxic molecule expression Granzyme B, Perforin, Fas Ligand (Fas-L) quantified by flow cytometry. Cells were treated with different doses of CAN296 (2, 4, 8 µg/mL), DEX (0.4, 4, 40 µg/mL), or TAC (0.1, 1, 10 ng/mL), and all parameters were compared to untreated controls. CAN296 significantly inhibited T cell activation, reducing CD69 expression in CD4⁺ T cells to 2–11% and in CD8⁺ T cells to 5–17%. It also markedly suppressed TNF-α secretion in CD4⁺ T cells at all concentrations (p < 0.0001). In CD8⁺ T cells, CAN296 led to a near-complete reduction in TNF-α and IFN-γ, leaving both cytokines barely detectable at all tested doses (p < 0.0001). The effect of cell inhibition was significantly more pronounced than that observed with DEX or TAC, displaying dose-dependent reductions. TAC inconsistently lowered TNF-α while paradoxically increasing IFN-γ at lower concentrations. Additionally, CAN296 consistently suppressed cytotoxic molecule expression, reducing Granzyme B by 81–82%, Perforin by 40–53%, and Fas-L by 40–44%. DEX showed variable effects on cytotoxic molecule expression. At the same time, TAC demonstrated inconsistent modulation of Perforin and Granzyme B. Overall, CAN296 outperformed DEX and TAC, demonstrating more potent and consistent immunomodulatory effects. CBD-rich cannabis extract, CAN296, exhibits potent immunomodulatory properties by effectively inhibiting T cell activation, lowering pro-inflammatory cytokines, and suppressing cytotoxic molecule expression. Its efficacy surpasses conventional therapies like DEX and TAC, offering a promising novel treatment modality for T cell-mediated disorders, including OLP and oGVHD. These findings support further development of CAN296 formulations to optimize dosing and delivery, followed by clinical trials to validate its therapeutic potential. Full article

The reconstruction of the foreskin using autografts is a complex procedure. A novel decellularisation method for epithelial tissue has been developed, producing an extracellular matrix scaffold from human donor foreskin. This study evaluated the immune response and integration of this scaffold after implantation in rats, focusing on inflammatory infiltrate, neovascularization, recellularization, and foreign body reaction. Twenty-six rats underwent a 1 cm infrascapular incision with scaffold implantation in the hypodermal layer. Group A (13 rats) was subject to a 30-day follow-up period, while Group B (13 rats) was subject to a 5-day follow-up period. Inflammation at the implantation site was scored from 0 (none) to 3 (severe). Tissue explants were. After 5 days (Group B) and 30 days (Group A), a tissue explant was performed and examined histologically and immunohistochemically. The clinical evaluation revealed slight signs of inflammation during the initial five days following the implantation procedure. Neutrophil (0.87 ± 0.35; 1 ± 0.53) and eosinophil (0.61 ± 0.51; 0.75 ± 0.46) presence was slight, with no significant differences between groups. Lymphocyte infiltration was moderate (1.87 ± 0.35; 1.75 ± 0.46), exceeding macrophage presence (1.25 ± 0.46; 1.12 ± 0.35). Neovascularization and cellular colonization were significantly greater at 30 days (2 ± 0.53; 2.42 ± 0.53) compared to 5 days (0.57 ± 0.21; 0.62 ± 0.32). Encapsulation remained mild in all cases, with no intergroup differences (0.87 ± 0.35). These findings indicate that the decellularized extracellular matrix derived from human foreskin elicits minimal immune response while promoting neovascularization and cellular repopulation. This supports its potential use as a biocompatible scaffold in reconstructive procedures. Full article

Background: The main type of treatment of unresectable NSCLC is chemoradiotherapy, which has a relatively high toxicity. One option allowing to reduce the toxicity of this approach may be immunocorrective therapy. The appointment of this type of treatment should be warranted in terms of patient’s immune system response. This confirms the importance of verifying systemic immune disorders in primary patients with NSCLC. Goal: To assess the features of the population of immune cells in peripheral blood in patients with stage IIIB, IIIC primary NSCLC and to identify any signs of secondary immunodeficiency in this cohort. Methods: We analyzed the frequencies of circulating T cells (CD3+, CD4+, CD8+), B-cells (CD19+), NK-cells (CD3-CD16+CD56+ cell), and NKT-cells (CD3+CD56+ cells) within CD45+ cells (lymphocytes) in 80 patients with stage IIIB-IIIC NSCLC, and in 40 healthy controls using eight-color flow cytometry. Results: In patients with stages IIIB-IIIC primary NSCLC, changes within immunocompetent blood cells were found. Moreover, it was unveiled that quantitative changes affected all major immunocompetent cells. A decrease in the proportion of CD4+ T cells and B lymphocytes and an increase in the number of NK and NKT cells were found. Also, an increase in the number of double-positive CD4+CD8+ T cells was revealed, as well as a significant increase in the proportion of B1a (CD5+CD19+) cells among B lymphocytes (qualitative disorders). Conclusion: The revealed multidirectional changes among immunocompetent peripheral blood cells in patients with locally advanced NSCLC (stages IIIB-IIIC) can be beyond doubt considered as signs of systemic immune disorders in this cohort (secondary immunodeficiency). Full article

Introduction: Vitamin D is involved in numerous processes and is obtained both exogenously and endogenously. Its active form is 1,25-dihydroxycholecalciferol, which exerts its biological effects via the vitamin D receptor (VDR). The main factors influencing VDR density are polymorphisms of the VDR gene, which may affect, e.g., gene mRNA stability and also VDR gene expression. There are four main polymorphic sites within the gene, BsmI, ApaI, FokI and TaqI, and two polymorphisms related to the gene promoter: GATA and Cdx2. One of the functions of vitamin D is to modulate the immune system. It affects T lymphocytes, B lymphocytes and dendritic cells. Currently, vitamin D deficiency is a common global problem that is associated with an increased risk of autoimmune diseases, including Hashimoto’s thyroiditis. Numerous studies have demonstrated an association between low vitamin D levels and elevated thyroid-stimulating hormone (TSH) levels, and have also proven the existence of a negative correlation between vitamin D levels andanti-thyroid peroxidase (anti-TPO) and anti-thyroglobulin (anti-Tg) antibody titers. Review objectives and a concise summary of the methodology: The review aims to analyze studies examining the relationship between specific VDR polymorphisms, vitamin D levels, and the development of various diseases, with a particular emphasis on Hashimoto’s thyroiditis. This review is based on original and review articles written in English published between March 2018–November 2024 searched primarily in the PubMed, and additionally in Google Scholar databases. A narrative review of the literature was conducted. Conclusions: The presence of specific VDR polymorphisms influences the effectiveness of vitamin D supplementation, but the role of supplementation in the prevention of autoimmune diseases has not been definitively confirmed. To date, studies have primarily involved relatively small groups of patients with significant population heterogeneity, with case–control investigations being the most common. Therefore, further research on larger, more homogeneous groups is recommended to achieve more standardized results. Additionally, the influence of epigenetic factors modulating VDR activity and its interactions with the environmental factors is also important. Full article

Recent studies propose that nanomaterials, either independently or coupled with biomolecular conjugates, have the ability to influence immune activity directly, creating new opportunities for advancing immunotherapies targeting infections and cancer. This review highlights current findings on how functionalized carbon nanotubes (f-CNTs), graphene, and carbon nanohorns interact with immune cells. Among these, f-CNTs have been the most thoroughly explored, though research interest in graphene has been rising steadily. Analysis of published work shows that macrophages are the most frequently studied immune cells (56%), followed by lymphocytes (30%), particularly T cells (22%). Investigations into monocytes and dendritic cells represent 7%, mixed populations such as peripheral blood mononuclear cells make up 6%, and studies on B cells and natural killer (NK) cells remain minimal (1%). Much of the available research has focused on assessing cytotoxicity and compatibility rather than uncovering precise mechanisms of immune modulation. Nonetheless, recent large-scale gene expression profiling has revealed novel immunomodulatory properties of f-CNTs, including stimulation of certain inflammatory signaling pathways. Research on graphene’s immune interactions is still developing. Overall, this review consolidates evidence on the immunological potential of biocompatible f-CNTs and graphene, offering groundwork for their future application in immunology and medicine. Full article

Background/Objectives: Allergen-specific immunotherapy remains the only disease-modifying treatment for allergic diseases, and the use of recombinant hypoallergenic derivatives is a promising therapeutic approach. Among these, BTH2 has previously shown efficacy in an acute murine model of allergy induced by Blomia tropicalis. The present study aimed to evaluate both the efficacy and safety of BTH2 in a chronic asthma model induced by B. tropicalis. Methods: A/J male mice (n = 6) were sensitized and chronically challenged with B. tropicalis extract over four months. One group repeatedly received subcutaneous doses of BTH2 (25 µg) for three months (65 doses). Parameters of allergic airway inflammation, antibody profiles, cytokine levels, and markers of AIT success were evaluated in bronchoalveolar lavage fluid, lung tissue, serum, and splenocyte cultures. Results: Repeated BTH2 administration was well tolerated, with no signs of systemic toxicity. BTH2 significantly reduced neutrophilic and eosinophilic airway inflammation, while increasing lymphocytes and regulatory cytokines in the lungs. It suppressed IgE against B. tropicalis allergens, while inducing mucosal IgA responses and systemic IgG, which may be linked to the observed blocking antibody activity in BTH2-treated mice. The treatment also led to downregulation of Th2 cytokines and enhanced expression of regulatory and Th1-associated cytokines, especially IL-10, TGF-β and IFN-γ. Correlation matrix analyses indicated that regulatory cytokines were correlated with beneficial antibody responses and reduced inflammation. Conclusions: BTH2 shows strong therapeutic and immunomodulatory effects in a chronic asthma model induced by B. tropicalis, with a favorable safety profile. These findings support its potential for future clinical trials, including those involving patients with allergic asthma. Full article

Background: Mammalian orthoreovirus is a ubiquitous double-stranded RNA virus that causes mild respiratory and enteric infections, primarily in infants and young children. Its significant environmental stability and association with conditions like celiac disease highlight an unmet medical need, as no licensed vaccine or antiviral treatment currently exist. Methods: An immunoinformatics-driven approach was employed to design a multi-epitope vaccine. The highly antigenic inner capsid protein Sigma-2 was used to predict cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and linear B cell epitopes using NetCTL, NetMHCpan, NetMHCIIpan, and IEDB tools. Selected epitopes were fused with appropriate linkers. The construct’s antigenicity, allergenicity, and physicochemical properties were evaluated. The tertiary structure was predicted with AlphaFold2, refined, and validated. Molecular docking with TLR2 and TLR4 was performed using HDOCK, and immune response simulation was conducted with C-ImmSim. Finally, the sequence was codon-optimized for E. coli expression using JCat. Results: The final vaccine construct comprises one CTL, four HTLs, and one B cell epitope. It is antigenic (VaxiJen score: 0.5026), non-allergenic, and non-toxic and possesses favorable physicochemical properties, including stability (instability index: 32.28). Molecular docking revealed exceptionally strong binding to key immune receptors, particularly TLR2 (docking score: −324.37 kcal/mol). Immune simulations predicted robust antibody production (elevated IgM, IgG1, and IgG2) and lasting memory cell formation. Codon optimization yielded an ideal CAI value of 0.952 and a GC content of 57.15%, confirming high potential for recombinant expression. Conclusions: This study presents a novel multi-epitope vaccine candidate against reovirus, designed to elicit broad cellular and humoral immunity. Comprehensive in silico analyses confirm its structural stability, potent interaction with innate immune receptors, and high potential for expression. These findings provide a strong rationale for further wet-lab studies to validate its efficacy and advance it as a promising prophylactic candidate. Full article

Flumequine is an antibiotic that is used to treat bacterial diseases in aquaculture. Fish express drug-metabolizing genes in response to antibiotic exposure. However, studies on the effects of high flumequine concentrations on drug metabolism genes and histopathology of the olive flounder are limited. To investigate the response of olive flounder to flumequine, we administered it at different concentrations. We analyzed the expression of drug metabolism genes (CYP) in the liver and histopathological lesions in the liver, spleen, and kidneys. The gene expression levels of CYP were higher at the highest flumequine concentration tested (4×) than at the lowest flumequine concentration (1×). The highest CYP gene expression level was observed for CYP2B4 (46.6-fold) at 4× flumequine compared to that in the control group. Hepatic atrophy, lymphocytic infiltration, and hematopoiesis were observed in the liver, spleen, and kidney at 4× flumequine between 3 and 24 h compared to 1× flumequine, respectively. These results contribute to a better understanding of drug metabolism and the general toxicity of pharmaceutical exposure in olive flounder. Full article

Background/Objectives: The solitary fibrous tumor is an uncommon benign mesenchymal neoplasm with relatively indolent and rarely metastasizing behavior. This retrospective study includes 26 cases of head and neck solitary fibrous tumors diagnosed between 2017 and 2024. Methods: The morphological examination with Hematoxylin–Eosin staining was completed via immunohistochemical reactions with specific antibodies. Results: The Ki-67 proliferation index had a median of 11.2%, with an interquartile range of 5% to 15%. CD20-positive B-cells had a score of 0 in 50% of cases (n = 26), while CD3 and CD5 T-cells had a score of 3 in 81% of cases (n = 21). CD4-positive T-cells had a majority score of 1 (81%, n = 21). CD8-positive T-cells had a broader distribution: 65% (n = 17) of cases presented a score of 1, 27% (n = 7) a score of 2, and 8% (n = 8) a score of 0. Antigen-presenting dendritic cells and mast cells presented a majority score of 0 in the entire cohort, being undetectable in 85% (n = 22) and 88% (n = 23) of cases, respectively. CD20-positive B-lymphocytes demonstrated moderately strong correlations with the Ki-67 cell proliferation index (r = 0.77). The time to recurrence was most strongly associated with the Ki-67 mitotic index (r = 0.81), CD4-positive (r = 0.85), and CD5-positive T-lymphocytes (r = 0.55), and CD20-positive B-lymphocyte expression (r = 0.68). Conclusions: This research illustrates our experience with head and neck solitary fibrous tumors, the surgical decisions, and the morphological and immunohistochemical features, while reviewing the cases published in English in the specialized literature. Full article

The impact of protein bound uremic toxins (PBUTs) and lymphocyte alterations in morbidity and mortality in patients on hemodialysis (HD) is of great concern. The aim of this study was the assessment of association between PBUTs, immunosenescent lymphocytes’ phenotype and clinical events [cardiovascular, severe infections (hospitalization due to infection, respiratory infection), all-cause mortality] during 2-year follow-up. In this prospective observational study, lymphocytes’ phenotype of 54 patients on HD and 31 age-matched controls was analyzed by flow cytometry, and simultaneously, PBUT serum levels [hippuric acid (HA), indoxyl sulfate (IxS), p-cresyl sulfate (pCS), p-cresyl glycuronide (pCG), in-dole-3-acetic acid (IAA), and 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF)] were quantified by ultra-performance liquid chromatography. Patients with increased levels of free IxS and total and free HA had higher mortality within a 2-year follow-up period (p = 0.049, p = 0.01, p = 0.01, respectively). In patients who experienced cardiovascular events, higher concentrations of CMPF (p = 0.015) were observed. Higher total and free HA levels associate with increased all-cause mortality in patients on HD, independently of age, dialysis vintage, and decreased count of CD4+CD45RA+CD31+ and naïve B cells (CD19+IgD+CD27−). In patients on HD, increased levels of total and free HA associate with an increased risk of death. Full article