Search Results (948)

In light of our previous publication, we hypothesized that chronic lymphocytic leukemia (CLL) cells also recruit monocytes to acquire survival advantage. To test this, we treated Buffy coat-driven monocytes with exosomes isolated from the peripheral blood of 45 treatment-naïve patients. The CLL-derived exosomes turned monocytes into IL-6-producing cells as an increase of 13-fold in the IL-6 levels was obtained in the growth medium of the exposed monocytes. Subsequently, we filtered out the monocytes and added CLL cells to this IL-6 enriched medium. As a result, the oncogene STAT3 became phosphorylated, and thus may have provided the cells with a survival advantage. A total of 67 phosphoproteins were upregulated in response to CLL-derived exosomal exposure in the recipient monocytes, with TFIIF being among the top scored proteins in this analysis. Transfection of monocytes with a TFIIF-containing vector increased the levels of IL-6 about 14-fold in the culture medium. Importantly, the CLL-derived exosomes induced the transformation of a portion of the recipient monocytes (45% compared to 30% of the unexposed cells) to become nurse-like fibrocyte cells. Taken together, CLL cells communicate with monocytes through the exosomes that they release. Once they are taken up by monocytes, they turn them into IL-6-producing cells, which provide a survival advantage to the neoplastic cells, creating a vicious circle that promotes disease progression. Full article

Background: Lymphocyte-activation gene 3 (LAG3) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) are immune checkpoint receptors and inhibitory regulators of T-cells. Methods: Here, we analyzed the prevalence and potential impact of the LAG3 gene variant rs870849 and the CTLA4 gene variant rs231775 in AML patients eligible for autologous stem cell transplantation. Results: While CTLA4 rs231775 was prevalent at reduced minor allele frequencies (MAF 0.33), LAG3 rs870849 was prevalent at elevated minor allele frequencies (MAF 0.58) in AML patients, compared to the allele frequencies in the European population (MAF 0.37 and MAF 0.39). The gene risk analysis indicated a dose-dependent risk of AML disease associated with LAG3 rs870849, but no risk associated with CTLA4 rs231775. Baseline blood count profiles differed across LAG3 genotypes, suggesting a link between LAG3 rs870849 and disease-associated levels of anemia, thrombocytopenia and peripheral blast percentage. Conslusions: The germline LAG3 variant rs870849 may be associated with AML disease risk and specific hematological disease features. Full article

Styrene, a constituent of polystyrene food-contact materials, can migrate into hot beverages, but data on short-term consumer exposure and associated biological responses remain limited. In this single-arm longitudinal human biomonitoring pilot study, 40 healthy adults consumed tea or coffee daily in Styrofoam cups for approximately two weeks. Biomarkers were measured at baseline, day 6, and day 11, including urinary mandelic acid (MA) and phenylglyoxylic acid (PGA), salivary malondialdehyde (MDA), comet assay parameters in peripheral blood lymphocytes, and micronucleus (MN) frequency in buccal cells. Measured styrene migration into beverages ranged from 3.3 to 7.1 μg/L, below the World Health Organization guideline value. Urinary metabolites and salivary MDA showed substantial interindividual variability and no consistent temporal pattern. In contrast, generalized estimating equation models showed progressive increases in comet assay indicators over the exposure period. Tail intensity and tail moment increased over time, with stronger changes among participants consuming two cups daily. MN frequency did not change significantly. These findings suggest that repeated short-term consumption of hot beverages in polystyrene cups was associated with modest changes in selected early biomarkers of biological effect under consumer-use conditions. The results should be interpreted cautiously in light of the modest sample size, short follow-up, and absence of more specific mechanistic endpoints, but they support further study of repeated low-level exposure to food-contact materials. Full article

Background: Leukemic non-nodal variant mantle cell lymphoma (nnMCL) is an uncommon subtype of mantle cell lymphoma that lacks lymphadenopathy and generally follows an indolent clinical course. Adverse genetic alterations such as TP53 inactivation and del(13q) may have prognostic significance. Clinical findings such as splenomegaly may serve as a clue to the diagnosis and should prompt further evaluation. Case Presentation: We describe a 91-year-old woman who presented with a one-month history of anemia (hemoglobin 12.3 g/dL), mild thrombocytopenia (platelets 136 × 10⁹/L), isolated splenomegaly and no palpable lymphadenopathy. Despite a normal total white blood cell count, intermittent relative lymphocytosis with atypical lymphocytes (4%) and smudge cells was detected on the complete blood count. Peripheral blood flow cytometry demonstrated a monoclonal kappa-restricted B-cell population negative for CD5 and CD10, comprising approximately 20% of lymphocytes. Conventional karyotyping and fluorescent in situ hybridization (FISH) analysis identified del(13q), del(17p)/TP53, and IGH-CCND1 rearrangement in 8–19.5% of peripheral blood leukocytes. A month after the initial assessment, the patient presented following a fall. CT imaging of the abdomen revealed marked splenomegaly, a large subcapsular/perisplenic hematoma, and moderate-to-large hemoperitoneum. Emergent laparotomy showed an enlarged spleen (1490 g, 23 × 16 × 7.5 cm) with laceration. Histologic evaluation showed atypical lymphoid cells positive for CD20 and cyclin D1, with strong p53 expression, negative for CD5 and SOX11, and a low Ki-67 index. Similar involvement was identified in the small bowel and appendix. Targeted sequencing of splenic tissue, performed as part of a retrospective molecular characterization, identified a pathogenic TP53 variant (p.His179Gln). Conclusions: This case provides a rare opportunity to evaluate splenic and small intestinal involvement by nnMCL at both the gross and histologic levels. It highlights the importance of integrating clinical findings with flow cytometry, imaging, cytogenetic, and molecular data in establishing the diagnosis. Even when peripheral blood findings suggest a low disease burden, imaging may better define the extent of disease and support appropriate clinical assessment, particularly in elderly patients at risk for complications related to splenomegaly. Full article

Background: Autoimmune encephalitis (AE) is an increasingly well recognized disorder in the past decade both in adults and in children, yet pediatric data are still limited. A full peripheral blood cell count is a routine examination that provides valuable information regarding the immune system. Thus, there are peripheral blood cell count (PBCC)-derived ratios that reflect systemic inflammatory activity and they have been associated with disease severity in adults: the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), monocyte-to-lymphocyte ratio (MLR), systemic immune–inflammation index (SII), systemic inflammation response index (SIRI), and aggregate index of systemic inflammation (AISI). Methods: This study is a retrospective chart review of children under 18 years diagnosed with definite or probable AE and treated in our institution from 1 January 2018 until 1 December 2025. Only patients with available PBCC results at the time of the first hospital admission after neurological/psychiatric symptom onset were included. An age-matched control group was created by selecting the results of PBCC of patients presenting for routine pediatric follow-ups with normal inflammatory and hematologic parameters. The group means were compared using an independent-samples t-test or the Mann–Whitney U test for non-normally distributed data. Analysis of the receiver operating characteristics curve (ROC curve) was conducted, followed by the area under the curve ROC curve (AUC). Results: A total of 45 children with AE and 150 controls were included in the study. Of these, 22 patients (49%) had probable AE and 23 patients (51%) had definite AE. The NLR, PLR, SII, SIRI and AISI values were significantly higher in AE patients compared with the controls, but the AUC values (~0.58–0.66) indicate poor-to-fair discriminative ability. Youden’s index-based cut-off values were associated with high specificity and modest sensitivity. The likelihood ratios in the range of 2–3 (LR+) and 0.6–0.7 (LR−) suggest weak rule-in capacity and limited rule-out utility. Conclusions: Our results suggest that at the time of the initial hospitalization, children with AE already show altered peripheral immune cell profiles compared to their age-matched peers. The high specificity and the low sensitivity of the inflammatory indices make them more suitable for supporting the AE diagnosis in suggestive clinical circumstances, but not for screening. These results represent a foundation for further investigation of the roles that these indices have both as diagnostic and prognostic factors for these children. Full article

Lymphocyte activation gene-3 (LAG-3) is a pivotal immune checkpoint receptor that exerts a negative regulatory effect on T-cell function. Although LAG-3-blocking antibodies have shown promising clinical potential, the inherent limitations of conventional monoclonal antibodies necessitate the development of novel antibody formats with enhanced biological and pharmacological properties. In this study, a panel of single-domain antibodies (sdAbs) targeting human LAG-3 was generated via phage display technology. Among these candidates, 2H-G7 was identified as a high-affinity sdAb that binds to LAG-3 with an equilibrium dissociation constant (K_D) in the nanomolar range. Notably, 2H-G7 potently blocks the interactions of LAG-3 with both of its key ligands, fibrinogen-like protein 1 (FGL1) and major histocompatibility complex class II (MHC-II). Its capacity to restore impaired T-cell function was validated by quantifying interleukin-2 (IL-2) secretion and CD69 expression in stimulated primary human peripheral blood mononuclear cells (PBMCs). Epitope mapping studies localized the binding site of 2H-G7 to the D1D2 extracellular domains of LAG-3, distinct from relatlimab, a clinically approved LAG-3-blocking antibody serving as the benchmark. In a xenogeneic mouse model of non-small-cell lung cancer (NSCLC), 2H-G7-Fc exhibited superior tumor growth inhibition efficacy compared with relatlimab. These findings demonstrate that 2H-G7 is a promising lead candidate for the development of next-generation LAG-3-targeted tumor immunotherapies. Full article

This study investigated cytogenetic damage in peripheral blood lymphocytes of 10 differentiated thyroid cancer patients who received multiple ¹³¹I radioiodine (RAI) treatments following total thyroidectomy. Blood samples were collected before the RAI therapy course and 2–3 days after the course for a few selected courses (from 1 to 3) for each patient. The cumulative average number of chromosome aberrations (CAs) per cell and its increment due to a selected RAI course were evaluated using the multiplex fluorescence in situ hybridization method (mFISH). An increase in the number of CAs was observed with the accumulation of RAI activity. The yield of these CAs per unit of accumulated RAI activity was, however, approximately three-fold lower than the respective yield for the incremented number of CAs in a selected course, thus demonstrating the elimination of CAs and/or aberrant cells with time. Biological dosimetry was performed based on the number of all types of CAs and in vitro mFISH calibration curves. With an average administered RAI activity of 3.40 ± 0.39 GBq, the average absorbed blood dose was 0.61 Gy (0.31–0.89:95% CI). Our results demonstrate that one-time administration of such activities of RAI was safe, since the commonly accepted threshold of 2 Gy for the blood dose was not exceeded. Full article

Background and Objectives: Age-related macular degeneration (AMD) is a multifactorial retinal disease in which inflammation and blood-retinal barrier dysfunction may contribute to disease pathogenesis. Claudin-5 is a key tight-junction protein involved in endothelial barrier integrity. Hemogram-derived indices such as the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), monocyte-to-lymphocyte ratio (MLR), and pan-immune-inflammation value (PIV) reflect systemic inflammatory status. This study aimed to evaluate circulating claudin-5 levels and systemic inflammatory indices in patients with wet and dry AMD and to investigate their associations with visual function. Materials and Methods: This prospective case–control study included 90 participants: 30 patients with wet AMD, 30 patients with dry AMD, and 30 healthy controls. All participants underwent detailed ophthalmologic examination, including best-corrected visual acuity (BCVA) assessment and optical coherence tomography. Serum claudin-5 levels were analyzed by enzyme-linked immunosorbent assay, and NLR, PLR, MLR, and PIV were calculated from complete blood count parameters. Group comparisons, correlation analyses, and age-adjusted analyses were performed using appropriate statistical methods. Results: Age differed significantly among the groups (p = 0.032), with the highest median age in the dry AMD group. BCVA (logMAR) also differed significantly (p < 0.001), and both AMD groups had worse visual acuity than controls. Median serum claudin-5 levels were 2.42 in controls, 3.28 in the wet AMD group, and 3.10 in the dry AMD group, with no significant between-group difference (p = 0.280). NLR, MLR, and PIV were also comparable among the groups (p = 0.310, p = 0.410, and p = 0.752, respectively). PLR differed among the groups (p = 0.019), and post hoc analysis showed higher PLR values in the dry AMD group than in the wet AMD group (p = 0.013). However, this difference was no longer statistically significant after adjustment for age (adjusted p = 0.098). Serum claudin-5 was not significantly correlated with age, BCVA, NLR, PLR, MLR, or PIV. Conclusions: Circulating claudin-5 did not differ significantly across AMD phenotypes and was not associated with age, visual function, or systemic inflammatory indices. Although PLR differed between wet and dry AMD before adjustment for age, the overall findings suggest that single-point peripheral serum measurements of claudin-5 may have limited utility in reflecting local retinal barrier-related changes in AMD. Larger longitudinal studies are needed to clarify its potential biomarker role. Full article

Background and Objectives: The incidence of colorectal cancer (CRC) in developed Western countries is constantly growing. CRC represents the third most common cancer and the second leading cancer-related cause of death worldwide. Innate and adaptive immunity play a pivotal role in the tumor response, but many of these interactions are still not well understood. Granulysin (GNLY) is an effector, cytolytic molecule, present in human cytotoxic granules of different lymphocyte subpopulations, mainly in cytotoxic T cells and NK cells. Pore-forming proteins GNLY, perforin and granzymes play a key role in cell-mediated immune responses against tumors and infections. Materials and Methods: We aimed to analyze perforin and GNLY-mediated cytotoxicity in the peripheral blood of patients with CRC by flow cytometry. Simultaneously, the cells were labeled with monoclonal antibodies against perforin, GNLY and different surface antigens (CD3, CD4, CD8 and CD56). Phenotypes of lymphocyte subpopulation and expression of perforin and GNLY were analyzed using intracellular and surface immunofluorescence. Results: Total perforin and GNLY expressions in peripheral blood mononuclear cells (PBMC) were significantly lower than in the control group. Statistically significant differences were observed in the distribution of perforin and GNLY expression in different stages of tumors classified according to Dukes’, indicating that the percentage of total perforin and GNLY was significantly diminished in accordance with tumor progression. Perforin and GNLY expression were significantly reduced in NK and NKT cells, accompanied by reduced cytolytic potential in patients with CRC and a consequent reduction in their ability to eliminate tumors and infected cells. Conclusions: The determination of cytotoxic potential may provide a valuable assessment of a patient’s immune status and represent a novel therapeutic target. Patients with CRC exhibit markedly impaired perforin- and GNLY-mediated cytotoxicity that correlates with disease progression. Assessment and restoration of cytolytic potential may therefore serve as indicators of immune competence and promising therapeutic strategies to improve perioperative and oncologic outcomes. Full article

Thalassemia is a hereditary hemoglobinopathy characterized by ineffective erythropoiesis, chronic hemolysis, and transfusion-related iron overload, which collectively contribute to oxidative stress and organ dysfunction. The present study aimed to investigate the relationships between iron metabolism, oxidative stress biomarkers, and immune cell function across different clinical conditions. Peripheral blood samples were obtained from healthy individuals and patients with iron deficiency anemia, obesity, thalassemia trait (TT), β-thalassemia HbE (BTE), and β-thalassemia major (BTM). Hematological parameters were measured using automated hematology analyzers, while biochemical indicators, including liver enzymes and bilirubin, were determined using clinical chemistry assays. Iron overload was evaluated using serum iron parameters and T2*-weighted magnetic resonance imaging. Oxidative stress biomarkers, including reduced glutathione, thiobarbituric acid-reactive substances, and total antioxidant capacity, were assessed spectrophotometrically. Flow cytometric analysis was used to measure reactive oxygen species, redox-active iron, and lipid peroxide levels in granulocytes and lymphocytes. Thalassemia patients exhibited severe anemia, elevated liver enzymes, increased bilirubin levels, and significant alterations in iron metabolism compared with healthy controls. Hepatic iron accumulation was more common than cardiac iron deposition, particularly in BTE patients. Granulocyte oxidative burst activity was significantly reduced in thalassemia patients, whereas lymphocyte responses remained relatively preserved. Increased variability in glutathione levels suggested activation of intracellular antioxidant defense mechanisms in response to chronic oxidative stress. These findings highlight the complex interplay between iron overload, oxidative stress, and the immune cell dysfunction associated with thalassemia, thereby providing insights into improved monitoring and therapeutic strategies. Full article

Different cyanobacterial species have been shown to be a valuable source of biologically active compounds with immunomodulatory activity. To date, little is known about members of the genus Tolypothrix (Cyanophyceae). Therefore, the present study focuses on five Tolypothrix strains (T. tenuis PACC 5497, T. tenuis PACC 8648, T. distorta SAG 1482-2, T. distorta CCALA 194, Tolypothrix sp. PACC 5501) that were not previously evaluated for specific morphological characteristics and immunomodulatory potential toward human immune cells. Cyanobacterial cultures were studied by light and transmission electron microscopy (TEM). Peripheral blood leukocytes were isolated from patients with inflammatory conditions and treated ex vivo with Tolypothrix non-polar extract fractions. Following treatment, the cells were analyzed by flow cytometry, and cytokine concentrations in culture supernatants were quantified by ELISA. Light microscopy observations showed that the cultures established from four of the strains have morphological features that correspond to T. tenuis Kützing (1843) ex Bornet et Flahault 1887 and T. distorta Kützing (1843) ex Bornet et Flahault. TEM analyses indicated parietal arrangement of cellular thylakoids in all strains, but T. distorta CCALA194 and Tolypothrix sp. PACC 5501 also displayed fascicular thylakoid arrangement. Immunophenotypic analyses revealed significantly increased proportions of T, NK, and B lymphocytes in leukocyte cultures treated with Tolypothrix extracts compared to the untreated controls. The concentrations of proinflammatory cytokines were lower in the culture medium of treated cells, while levels of the anti-inflammatory cytokine interleukin-10 remained stable, except in cultures treated with T. distorta SAG 1482-2 extract. The present study provides the detailed morphological characteristics of five strains of the genus Tolypothrix and indicate that non-polar extract fractions derived from the strains exert immunomodulatory effects on human leukocytes. Full article

Immune dysregulation is a critical driver of various pathological processes. Fish maw (FM) serves as a traditional immunomodulatory food. However, the immunomodulatory properties and mechanisms of fish maw hydrolysate (FMH) remain unclear. Here, low-molecular-weight FMH was prepared from Perca fluviatilis, exhibiting a major molecular weight distribution of 73–580 Da (83.89%), enriched in charged and hydrophobic amino acids (28.61% and 67.33%, respectively). Moreover, high-resolution mass spectrometry (HRMS) analysis identified 5 small peptides, including Asp-Leu and Gly-Pro-Ala, alongside 7 collagen-derived polypeptides with characteristic Gly-X-Y repetitive motifs. In cyclophosphamide (CTX)-induced immunosuppressed C57BL/6J mice, FMH significantly ameliorated alterations in peripheral blood cell parameters, regulated cytokine homeostasis, attenuated splenic histopathological lesions, and enhanced splenic lymphocyte proliferation. Mechanistically, thymic transcriptomic profiling identified 2237 DEGs in the CTX vs. CON comparison and 212 DEGs in the CTX+FMH vs. CTX comparison, with the NF-κB signaling pathway significantly enriched. Furthermore, qRT-PCR validated the expression of key NF-κB-related genes, including IκBα, P50, P65, CHUK, IL-1β, and IL-6, while immunohistochemical analysis confirmed reduced PI3K and P65 expression, thereby partly restoring immune homeostasis. These findings support FMH as a potential dietary immunomodulator. Full article

Chronic lymphocytic leukemia (CLL) exhibits marked clinical heterogeneity that is closely associated with genomic instability. Although cytogenetic abnormalities are widely used for risk stratification, they do not fully capture the biological complexity of the disease. Telomere dysfunction and alterations in DNA damage response pathways have been implicated in disease progression, but their relationship with cytogenetic risk in CLL remains incompletely characterized. In this study, peripheral blood mononuclear cells (PBMCs) from 48 CLL patients were analyzed. The analyzed PBMC fractions were enriched in leukemic B cells, with an estimated median tumor content above 85–90%. Cytogenetic profiles were obtained by conventional karyotyping following in vitro immunostimulation with DSP30 and interleukin-2 and classified according to ERIC and Döhner criteria. Telomere length was assessed by quantitative PCR, and CHEK1 and CHEK2 expression levels were quantified by RT–qPCR. Molecular parameters were compared across cytogenetic risk groups. Distinct molecular profiles were observed across cytogenetic categories. Favorable-risk CLL cases showed preserved telomere length, low CHEK1 expression, and maintained CHEK2 levels. Intermediate-risk cases, predominantly characterized by trisomy 12, exhibited moderate telomere shortening accompanied by increased CHEK1 expression and partial reduction of CHEK2. High-risk CLL cases, defined by del(11q), del(17p), or complex karyotypes, displayed pronounced telomere shortening, marked CHEK1 upregulation, and strong suppression of CHEK2. Telomere length was inversely correlated with cytogenetic risk (Spearman’s ρ = −0.68, p < 0.0001), and the CHEK1/CHEK2 expression ratio increased progressively with genomic complexity. These findings indicate that telomere length and CHEK1/CHEK2 expression patterns are closely associated with cytogenetic risk in CLL and may provide complementary biological information for risk stratification. Full article

Background/Objectives: Insect-derived lipids are emerging as circular-economy feed ingredients, but their implementation in ruminant diets requires robust safety assessment beyond productive endpoints. This study evaluated genome integrity in 26 lactating Valdostana Red Pied cows fed concentrates containing either hydrogenated palm fat (HPF; n = 13) or black soldier fly oil (Hermetia illucens oil, HIO; n = 13) for 50 days. Methods: Peripheral blood lymphocytes were analyzed using Sister Chromatid Exchanges (SCE), reflecting replication-associated chromosomal instability, and the alkaline Comet assay, quantifying primary DNA damage at the single-cell level (Tail DNA and Olive tail moment) at T0 (the day before the start of the two experimental diets), T1 (30 d) and T2 (50 d). Results: Baseline SCE frequencies were comparable between groups. Over time, SCE values decreased in both groups, but a significant reduction occurred only in HIO at day 50, with lower SCE frequency than HPF (5.73 ± 0.11 vs. 6.29 ± 0.13; p = 0.002). Comet tail DNA showed a significant time effect (T0 vs. T1: mean difference = 179,846.6; p < 0.001; T0 vs. T2: mean difference = 138,395.2; p = 0.012), with diet-dependent modulation. In fact, in HIO, tail DNA decreased from 387,886 ± 94,606 (T0) to 147,006 ± 30,592 (T1; p < 0.001), remained lower at day 50 (155,723 ± 29,357; p = 0.024), and was lower than HPF at both T1 (p = 0.006) and T2 (p = 0.009). Olive tail moment also decreased over time (T0 vs. T1: mean difference = 1.925 × 10¹⁵; p = 0.008; T0 vs. T2: mean difference = 1.676 × 10¹⁵; p = 0.025), and it differed between diets at day 50 in favor of HIO (5.99 × 10¹⁵ ± 5.45 × 10¹⁴ vs. 7.26 × 10¹⁵ ± 5.98 × 10¹⁴; p = 0.017). Conclusions: Overall, no evidence of genotoxicity was observed in cows fed HIO; conversely, the results support compatibility with genome stability and suggest a modest time-dependent improvement detectable mainly after prolonged supplementation. Full article

Treatment-associated regression of tumors outside the irradiated field has occasionally been reported, but the underlying mechanisms remain unclear, particularly in the context of central nervous system (CNS)–directed therapy. Glioblastoma (GBM) is commonly treated with radiotherapy and temozolomide, both of which may influence tumor biology and the systemic environment. We report a patient with synchronous primary GBM and early-stage lung adenocarcinoma who underwent craniotomy followed by intensity-modulated radiotherapy with concurrent temozolomide for GBM. During GBM-directed chemoradiotherapy, the untreated pulmonary lesion demonstrated progressive regression without any lung-specific therapy, temporally coinciding with CNS-targeted treatment. Although comprehensive immunophenotyping was not feasible, longitudinal changes in the proportion of peripheral blood lymphocytes were observed during therapy. These findings represent a clinical observation characterized by a temporal association between CNS-directed treatment and regression of a distant, non-irradiated tumor. However, the underlying mechanism remains uncertain, and a contribution from systemic temozolomide exposure cannot be excluded. While treatment-related systemic effects may be considered, no specific causal mechanism can be established based on this single case. This case highlights an unusual clinical observation that may warrant further investigation. Further studies are needed to clarify the relationship between CNS-directed therapies and systemic tumor behavior. Full article