Search Results (434)

Glycosylation of surface proteins is a crucial post-translational modification that reflects the activation status of neutrophils, the predominant leukocyte subset in humans and horses. Neutrophils have emerged as active contributors to diseases mediated by the adaptive immune system, such as equine recurrent uveitis (ERU), a sight-threatening disease in horses and a unique model for studying the pathogenesis of autoimmune uveitis in humans. Since changes in surface glycosylation can impact neutrophil function, we were interested in the surface glycosylation landscape on neutrophils from healthy horses and the potential changes in surface glyco-signatures in ERU. Using 35 different plant lectins, we outlined a profile of surface-exposed glycan moieties on equine neutrophils and detected significantly increased O-glycosylation in a diseased state through Jacalin (JAC) binding via flow cytometry. Subsequent molecular weight comparison of JAC pull-down assay data and neutrophil proteomics indicated the surface proteins Integrin beta-2 and CUB domain-containing protein 1 as potential anchors for increased O-glycan levels in ERU. These findings give novel insights into neutrophil surface glycosylation in health and disease and propose O-glycosylation as a possible biomarker for autoimmune uveitis. Full article

Urinary tract infections (UTIs) are among the most common pathologies, with a high incidence in women and hospitalized patients. Their diagnosis is based on the presence of clinical symptoms and signs in addition to the detection of microorganisms in urine trough urine cultures, a time-consuming and resource-intensive test. The goal was to optimize UTI detection through artificial intelligence (machine learning) using non-microbiological laboratory parameters, thereby reducing unnecessary cultures and expediting diagnosis. A total of 4283 urine cultures from patients with suspected UTIs were analyzed in the Microbiology Laboratory of the University Hospital Virgen de las Nieves (Granada, Spain) between 2016 and 2020. Various machine learning algorithms were applied to predict positive urine cultures and the type of isolated microorganism. Random Forest demonstrated the best performance, achieving an accuracy (percentage of correct positive and negative classifications) of 82.2% and an area under the ROC curve of 87.1%. Moreover, the Tree algorithm successfully predicted the presence of Gram-negative bacilli in urine cultures with an accuracy of 79.0%. Among the most relevant predictive variables were the presence of leukocytes and nitrites in the urine dipstick test, along with elevated white cells count, monocyte count, lymphocyte percentage in blood and creatinine levels. The integration of AI algorithms and non-microbiological parameters within the diagnostic and management pathways of UTI holds considerable promise. However, further validation with clinical data is required for integration into hospital practice. Full article

Background: The aim of our study is to assess the risk factors for the development of coronary artery stent thrombosis and restenosis, as well as the main localization of these processes in patients who underwent repeated coronary revascularization during the COVID-19 pandemic. Materials and Methods: Data were retrospectively analyzed from 490 patients who underwent coronary angiography and required repeat revascularization from May 2020 to May 2023. The prevalence and anatomical distribution of coronary stenosis, restenosis, and stent thrombosis were assessed. Results: Coronary artery stenosis was detected in 46.9% of patients. The most affected arteries were the left anterior descending (13.7%), right coronary artery (15.1%), and circumflex branch (9.4%). In-stent restenosis was observed in 19.0% of cases. Coronary thrombosis occurred in 22.8% of patients, while stent thrombosis was found in 11.2%. Multivariate regression revealed that leukocyte count (OR = 1.18, p < 0.05), activated partial thromboplastin time (APTP) (OR = 1.021, p = 0.025), low-density lipoproteins (LDL) (OR = 1.421, p = 0.042), and prior COVID-19 infection (OR = 2.05, p = 0.038) were significant predictors of stent thrombosis. The left ventricular ejection fraction (LVEF) (OR = 0.959, p = 0.017) and hemoglobin levels (OR = 0.975, p = 0.014) have inverse association with risk of stent thrombosis. Conclusion: COVID-19 history is a strong independent risk factor for coronary stent thrombosis, alongside inflammatory and coagulation markers. Full article

Egg yolk IgY antibody has significant application potential in aquaculture as a form of passive immunotherapy against various bacterial infections owing to its capacity for large-scale and cost-effective production. In this research, laying hens were immunized with live or inactivated Aeromonas hydrophila to produce IgY antibodies. Following this, experiments were carried out to assess the passive immune protection rates of the two types of IgY antibodies when used to immunize goldfish (Carassius auratus), which were then infected with A. hydrophila or Aeromonas veronii. ELISA experiments were conducted to demonstrate the interaction between the IgY antibodies and the bacteria. The kidneys of C. auratus were coated on a Luria–Bertani (LB) medium to evaluate bacterial content. The leukocyte phagocytosis was detected by a cell phagocytosis assay. The serum of C. auratus was used to assess the expression of antioxidant factors, and a qRT-PCR was conducted to evaluate the mRNA expression of inflammatory factors in visceral tissue. Furthermore, histopathology and immunofluorescence analysis were performed to evaluate the structural integrity, apoptosis, and DNA damage of visceral tissues. The results indicated that the live or inactivated A. hydrophila IgY antibodies exhibited passive immune protection rates against A. hydrophila and A. veronii and could recognize these two bacteria in vitro. Additionally, these two IgY improved the phagocytic ability of leukocytes, diminished renal bacterial concentration, and decreased the levels of antioxidant factors and mRNA expression of inflammatory factors. Meanwhile, the two IgY antibodies did not cause any pathology of the kidney, spleen, and intestine, and decreased the levels of DNA damage factor (γH2A.X) and cell apoptosis factor (p53) in renal tissue. Therefore, live and inactivated A. hydrophila IgY antibodies can resist bacterial infections, with live bacteria IgY providing greater protection than inactivated bacteria IgY. Further, A. hydrophila is an aquatic pathogen that causes minimal damage to laying hens, and the immunity of live A. hydrophila conforms to animal welfare. Altogether, live A. hydrophila IgY antibody can serve as a polyvalent passive immune vaccine candidate in aquaculture. Full article

Background/Objectives: The maintenance of peri-implant soft tissue health is critical for the long-term success of implant therapy, particularly in edentulous patients rehabilitated with mandibular overdentures. Leukocyte- and platelet-rich fibrin (L-PRF) has been proposed as an autologous biomaterial to enhance peri-implant tissue quality. This randomized controlled clinical trial evaluated the effect of L-PRF on peri-implant mucosal thickness in edentulous patients treated with mandibular implant-retained overdentures. Methods: Edentulous patients received two interforaminal implants to retain a mandibular overdenture and were randomly assigned to a test group (L-PRF applied during surgery) or a control group (standard protocol without L-PRF). Clinical measurements of keratinized mucosal thickness and width were recorded at baseline, 12 weeks, and 24 weeks. Volumetric analyses of soft and hard tissue changes were performed using digital superimposition of STL models. The trial was conducted in accordance with the Declaration of Helsinki and approved by the Scientific Ethics Committee of the Aconcagua Health Service. All participants provided written informed consent. Results: A significant increase in keratinized mucosal thickness was observed in the L-PRF group at 12 and 24 weeks compared with baseline (p < 0.01). No significant differences were detected between the groups in soft tissue volume (p = 0.12) or bone volume (p = 0.45). Mucosal width remained stable in both groups throughout follow-up. Conclusions: The application of L-PRF at implant placement resulted in a significant gain in peri-implant mucosal thickness, suggesting a soft tissue modulating effect. Enhancing keratinized mucosal thickness during implant surgery may improve peri-implant tissue quality and support long-term stability of mandibular overdentures. Full article

Mesenchymal stromal cells (MSCs) exert their immunoregulatory properties after licensing by inflammatory signaling cues, e.g., interferon (IFN)-γ. However, MSCs licensing by IFN-γ may result in increased expression of human leukocyte antigen (HLA) class II, which is related to rapid cell elimination, impairment of their immunosuppressive properties, and patient sensitization. The aim of this study was to evaluate the impact of IFN-γ on mediated immunoregulation and HLA class II expression. In this study, Wharton’s jelly (WJ) MSCs were isolated from human umbilical cords. Well-defined WJ-MSCs were submitted to IFN-γ exposure, and after 96 h, evaluation of biomolecule secretion and HLA class II expression was performed. Typing of HLA alleles using a next-generation sequencing (NGS) platform was performed. IFN-γ-primed WJ-MSCs secreted a high amount of immunoregulatory biomolecules, while elevated expression of HLA-DRB1 was observed. Analyses the NGS results showed the possibility of WJ-MSCs cluster formation based on their frequency of detected HLA alleles and immunoregulatory potential. Taking into consideration that IFN-γ-primed WJ-MSCs express HLA class II alleles, it is suggested that the HLA histocompatibility between allogeneic donor and recipient should be strongly considered to acquire the most beneficial outcome for the MSCs therapeutic strategy. Full article

Reactive oxygen species (ROS) are among the most effective tools of the innate immune response against pathogenic microbes. The respiratory burst (RB) of polymorphonuclear leukocytes (PMNs) generates an electron current that reduces molecular oxygen to superoxide. Superoxide reacts to form hydrogen peroxide as a precursor to the highly bactericidal hypochlorous acid. Here, we investigated whether alterations in extracellular potassium concentration impact H₂O₂ production. Such changes may occur, for example, during massive cell death due to necrosis or due to trauma injuries when potassium diffuses out of the cells. We recorded H₂O₂ release over a 2 h period of RB under varying potassium concentrations. Except for 100 mM potassium chloride, which increased the time delay before detectable H₂O₂ production, none of the potassium concentrations had a substantial effect on RB. We further examined whether this effect depended on the specific monovalent ion species. When sodium or methanesulfonate was used instead of potassium or chloride, respectively, no changes in H₂O₂ production were observed. Cell volume measurements under different potassium concentrations showed that only 100 mM potassium chloride significantly shrank the cells. We propose that hypertonic stress is crucial for delaying RB in human granulocytes, whereas the RB itself is independent of the tested ionic species. Additionally, the conducted hypertonic stress experiments revealed an unexpected time-dependence during the course of the RB, showing that the first 6 min were almost inert to hyperosmotic stress. Full article

Background: Bovine alphaherpesvirus 1 (BoAHV-1) is a major respiratory and reproductive pathogen in cattle worldwide. Both innate and adaptive immune responses contribute to protection against this virus; however, virus-host interactions remain partly undefined. In this study, the impact of BoAHV-1 infection on calves’ immune responses was investigated in detail. Methods: Six calves were intranasally infected with wild-type BoAHV-1, and blood samples were collected longitudinally. Leukocyte subset dynamics were assessed by complete haematological assay and flow cytometry, while multiplex ELISA was used to quantify serum levels of ten cytokines. For each parameter, post-infection values (days 2, 4, 8, 10, and 14) were compared with pre-infection baseline values (day 0). Results: Infection induced an initial phase of immunosuppression, reflected by decreased circulating αβ and γδ-T cells. However, infected animals rapidly developed a protective immune response, characterised by increased circulating classical and intermediate monocytes and elevated levels of the related chemokine MIP-1β. Early post-infection, rises in serum IFN-γ and IL-10 were also detected. Conclusions: Our data suggest that monocyte recruitment and increased serum levels of IFN-γ and IL-10 are positively associated with the ability to overcome infection. A better understanding of the immunopathogenic mechanisms underlying BoAHV-1 infection will support the development of more effective vaccines against this virus. Full article

Donor-specific anti-HLA antibodies (DSAs) bind to donor vascular endothelial cells and mediate allograft rejection (AMR), but a clinical challenge for which targeted therapeutic options remain limited. We used a multiplexed single-antigen bead (SAB) assay to detect anti-human leukocyte antigen (HLA) antibodies. Based on the antigens which patient’s antibodies aganist to, we developed bivalent HLA-Fc fusion proteins composed of HLA-derived antigenic domains and human IgG1-Fc effector regions (rA24-Fc and rB13-Fc). Specific binding and functional activity of the HLA-Fc proteins were further validated by flow cytometry, ELISA, complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) assays. Our findings demonstrate that the fusion proteins rA24-Fc and rB13-Fc significantly reduced HLA-specific antibody reactivity in vitro. Notably, rA24-Fc and rB13-Fc selectively bound to B-cell hybridomas (e.g., mouse W6/32 cells) expressing membrane immunoglobulins (BCR) which bound to the most HLA class I antigens. Importantly, rA24-Fc and rB13-Fc elicited antigen-specific, Fc-dependent elimination of the specific B-cell hybridomas. This study highlights HLA-Fc fusion proteins as a promising therapeutic strategy for the antigen-specific suppression of depletion of alloreactive B cells through dual cytotoxic mechanisms. This precision targeted to BCR of B cells approach is used to apply to the treatment of antibody-mediated rejection. Full article

Background and Objectives: Dyslipidemia, a modifiable cardiovascular risk factor, is associated with chronic low-grade inflammation. While leukocyte-derived indices have been investigated in this context, eosinophil-related inflammatory markers remain underexplored. This study examined patterns of eosinophil-to-lymphocyte ratio (ELR) and eosinophil-adjusted systemic inflammation response index (EA-SIRI) across dyslipidemia phenotypes. Materials and Methods: In this retrospective study, adult subjects were classified into six dyslipidemia phenotypes. Leukocyte-derived indices were evaluated across groups, and analyses included comparisons of medians, prevalence rates, tertile distributions, odds ratios, and risk estimates. Results: Both ELR and EA-SIRI were significantly higher in individuals with atherogenic dyslipidemia (ELR: 0.18; EA-SIRI: 1.53) and combined dyslipidemia (ELR: 0.17; EA-SIRI: 1.49) compared to the normolipidemic group (ELR: 0.11; EA-SIRI: 0.92). Notably, these patterns were more pronounced in males aged <40 years and younger females (<40), suggesting sex- and age-related variations in eosinophil-related inflammatory responses to dyslipidemia. Moreover, the highest tertiles of both ELR and EA-SIRI exhibited higher triglycerides and lower HDL-C compared to the lowest tertiles (p < 0.001). The odds of atherogenic dyslipidemia were more than doubled in individuals with elevated ELR (OR = 2.02; p < 0.001) and EA-SIRI (OR = 2.19; p < 0.001). ROC curve analysis indicated modest discriminative power for identifying atherogenic dyslipidemia, with ELR and EA-SIRI yielding AUC of 0.60 (p < 0.001) and 0.62 (p < 0.001), respectively. Conclusions: Our findings suggest eosinophil-related inflammation contributes to immunometabolic dysregulation underlying dyslipidemia. ELR and EA-SIRI may offer insights into inflammation-driven lipid disturbances and help detect subclinical inflammatory activity associated with atherogenic lipid profiles. Full article

Periprosthetic joint infection (PJI) is a devastating complication of joint arthroplasty surgery that is difficult to both diagnose and treat. Misdiagnosing a prosthetic infection has terrible consequences for both the patient and healthcare system. No currently used diagnostic test fulfills the requirements to be considered a gold standard. This shortcoming has been overcome through the implementation of multi-criteria diagnostic protocols elaborated by societies including the Infectious Diseases Society of America, International Consensus Meeting and European Bone and Joint Infection Society, using a combination of clinical, paraclinical and molecular findings in order to achieve the best accuracy in diagnosing PJI. This review aims to survey the current state of the techniques and technologies used for the diagnosis of PJI, investigating the accuracies of serum biomarkers (e.g., C-reactive protein, Interleukin-6, procalcitonin, D-dimers, Serum Intercellular Adhesion Molecule-1), synovial biomarkers (e.g., Antimicrobial peptides, lipocalin-2, leukocyte esterase, calprotectin), tissue biomarkers (e.g., Toll-like receptors, CD15) and advanced molecular techniques (e.g., Polymerase chain reaction, Metagenomic next-generation sequencing), as well as describing their ongoing limitations. In the search for an accurate, inexpensive and fast diagnostic test for PJI, we conclude that the accuracies of the currently studied biomarkers could be further enhanced through the development of novel detection technologies. Full article

Introduction: X-linked agammaglobulinemia (XLA) is a prototypical inborn error of immunity (IEI) caused by mutations in the BTK gene, leading to a profound deficiency of mature B cells and severe pan-hypogammaglobulinemia. The Epstein-Barr virus (EBV), which primarily infects B lymphocytes, is believed to be unable to establish persistence in these patients due to the lack of its natural reservoir. Indeed, current evidence supports that EBV infection is typically refractory in individuals with XLA. Methods: We describe the clinical and molecular characterization of a 10-year-old male patient with genetically confirmed XLA who developed EBV viremia, hemophagocytic lymphohistiocytosis (HLH), and EBV-positive cutaneous T cell lymphoma. Diagnosis was supported by flow cytometry, serology, quantitative PCR, EBER in situ hybridization, histopathology, and whole-exome sequencing. Results: Despite the complete absence of peripheral B cells, EBV was detected in leukocytes and multiple tissues, indicating active infection. The patient developed HLH and a T cell lymphoma with EBER-positive infiltrates. Genetic analysis revealed a nonsense mutation in BTK (1558C>T, R520*), confirming XLA. The clinical course included multiple episodes of neutropenia, viral and bacterial infections, and severe systemic inflammation. Conclusions: This is the first documented case of an XLA patient with confirmed BTK mutation presenting with clinical features more consistent with chronic active EBV infection. These findings challenge the prevailing paradigm that XLA confers protection against EBV-related diseases and further support the possibility of EBV noncanonical reservoirs leading to immune dysregulation. EBV should also be considered in the differential diagnosis of XLA patients presenting with systemic inflammation or lymphoproliferative disease. Full article

Human leukocyte antigen class I (HLA-I) molecules present intracellular peptides on the cell surface to enable CD8+ T cells to effectively control viral infections. Many viruses disrupt this antigen presentation pathway to evade immune detection. In this study, we demonstrate that SARS-CoV-2 Nsp1 impairs both the constitutive and interferon-γ (IFN-γ)-induced upregulation of HLA-I. Moreover, Nsp1 also blocks IFN-γ-induced expression of HLA-II. We found that, contrary to previously published work, the early SARS-CoV-2 B 1.1.7 Alpha variant lacking the accessory protein ORF8 retained full capacity to downregulate HLA-I, comparable to an ORF8-expressing wild-type isolate. While ectopic overexpression of ORF8 could reduce HLA-I surface levels, this effect was only observed at high expression levels. In contrast, moderate expression of the viral protein Nsp1 was sufficient to potently suppress both basal and IFN-γ-induced HLA-I, as well as HLA-II expression. To probe the underlying mechanism, we analyzed HLA-I-associated genes in previously published RNA-sequencing datasets and confirmed that Nsp1 reduces expression of components required for HLA-I biosynthesis and antigen processing. These findings identify Nsp1 as a key factor that impairs antigen presentation pathways, potentially contributing to the ability of SARS-CoV-2 to modulate immune recognition. Full article

Type 1 diabetes mellitus (T1DM) is a chronic autoimmune disorder characterized by the destruction of insulin-producing pancreatic beta cells, resulting in lifelong insulin dependence. While genetic susceptibility—particularly human leukocyte antigen (HLA) class II alleles—is a major risk factor, accumulating evidence implicates viral infections as potential environmental triggers in disease onset and progression. This narrative review synthesizes current findings on the role of viral pathogens in T1DM pathogenesis. Enteroviruses, especially Coxsackie B strains, are the most extensively studied and show strong epidemiological and mechanistic associations with beta-cell autoimmunity. Large prospective studies—including Diabetes Virus Detection (DiViD), The environmental determinans of diabetes in the young (TEDDY), Miljøfaktorer i utvikling av type 1 diabetes (MIDIA), and Diabetes Autoimmunity Study in the Young (DAISY)—consistently demonstrate correlations between enteroviral presence and the initiation or acceleration of islet autoimmunity. Other viruses—such as mumps, rubella, rotavirus, influenza A (H1N1), and SARS-CoV-2—have been investigated for their potential involvement through direct cytotoxic effects, immune activation, or molecular mimicry. Interestingly, certain viruses like varicella-zoster virus (VZV) and cytomegalovirus (CMV) may exert modulatory or even protective influences on disease progression. Proposed mechanisms include direct beta-cell infection, molecular mimicry, bystander immune activation, and dysregulation of innate and adaptive immunity. Although definitive causality remains unconfirmed, the complex interplay between genetic predisposition, immune responses, and viral exposure underscores the need for further mechanistic research. Elucidating these pathways may inform future strategies for targeted prevention, early detection, and vaccine or antiviral development in at-risk populations. Full article

The white button mushroom (Agaricus bisporus) is a widely cultivated edible and medicinal mushroom, which contains various active substances, and has application value against pathogenic bacteria in aquaculture. Firstly, A. bisporus water extract (AB-WE) was prepared. Through the detection kits, it was found that the polysaccharide, protein, and polyphenol components of AB-WE were 9.11%, 3.3%, and 1.5%, respectively. The 246 compounds were identified in AB-WE, and the major small-molecule components included L-Isoleucine, L-Tyrosine, L-Valine, and Linoleic acid by HPLC-Q Exactive-Orbitrap-MS. Secondly, the AB-WE was evaluated for its immunological activities through dietary administration and pathogen challenge (Aeromonas hydrophila and Vibrio fluvialis) in goldfish (Carassius auratus). The results showed that the levels of immune factors of acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) increased (p < 0.05) in goldfish, and the relative percentage survival of AB-WE against A. hydrophila and V. fluvialis were 80.00% (p < 0.05) and 81.82% (p < 0.05), respectively. The AB-WE reduced the bacterial content in renal tissue, enhanced the phagocytic activity of leukocytes, and exhibited antioxidant and anti-inflammatory effects by reducing the expression of antioxidant-related factors and inflammatory factors. Through histopathological and immunofluorescence techniques, it was found that AB-WE maintained the integrity of visceral tissues and reduced renal tissue apoptosis and DNA damage. Therefore, AB-WE exhibits immunoprotective activity against A. hydrophila and V. fluvialis infections in fish, and holds promise as an immunotherapeutic agent against major pathogenic bacteria in aquaculture. Full article