Search Results (4,426)

Background: Since the emergence of SARS-CoV-2 in the human population, the virus genome has undergone numerous mutations, enabling it to enhance transmissibility and evade acquired immunity. As a result of these mutations, most monoclonal neutralizing antibodies have lost their efficacy, as they are unable to neutralize new variants. Antibodies that neutralize a broad range of SARS-CoV-2 variants are of significant value in combating both current and potential future variants, making the identification and development of such antibodies an ongoing critical goal. This study discusses the strategy of using heterologous antigens in biopanning rounds. Methods: After four rounds of biopanning, nanobody variants were selected from a phage display library. Immunochemical methods were used to evaluate their specificity to the S protein of various SARS-CoV-2 variants, as well as to determine their competitive ability against ACE2. Viral neutralization activity was analyzed. A three-dimensional model of nanobody interaction with RBD was constructed. Results: Four nanobodies were obtained that specifically bind to the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein and exhibit neutralizing activity against various SARS-CoV-2 strains. Conclusions: The study demonstrates that performing several rounds of biopanning with heterologous antigens allows the selection of nanobodies with a broad reactivity spectrum. However, the fourth round of biopanning does not lead to the identification of nanobodies with improved characteristics. Full article

Exosomes can transport regulatory biomolecules and are mediators of cellular signaling among metabolic tissues through endocrine mechanisms. Understanding the pathways and processes underlying exosome-mediated inter-tissue communication is critical for elucidating the molecular pathophysiology of metabolic diseases such as obesity, type 2 diabetes mellitus (T2DM), and cardiovascular disorders. Consequently, these mechanisms represent novel and promising targets for pharmacological regulation. We examined the current knowledge regarding exosome physiology, the mechanisms of interaction with target tissues, and its role in metabolic tissue communication. We also analyzed the secretory profiles of exosomes in metabolic tissues, emphasizing their regulatory roles in adipose tissue, liver, pancreas, skeletal muscle, and the small intestine, while discussing their association with metabolic diseases. In this sense, we propose the exosomal pentad as a novel framework highlighting exosome-mediated inter-organ communication, where exosomes may regulate a metabolic axis involving these tissues. This model aligns with the ominous octet in type 2 diabetes but emphasizes exosomes as key regulators of metabolic homeostasis and potential therapeutic targets. The role of exosomes for the treatment of metabolic diseases emerges as a critical area of pharmacologic exploration. For instance, therapeutic strategies that prevent target tissue binding or expression of cargo molecules such as miRNAs could be designed, using antagomiRs or nanoparticles. Additionally, integrins like αvβ5 on the exosomal membrane can be blocked with monoclonal antibodies or engineered for targeted delivery of therapeutic molecules. Exosomes, critical mediators of inter-organ communication and metabolic regulation, hold potential to design precise molecular-level therapies while minimizing systemic side effects. Full article

Non-small-cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality worldwide. Immunotherapy targeting the PD-1/PD-L1 axis has revolutionized treatment, providing durable responses in a subset of patients. However, with fewer than 50% of patients achieving significant benefits, there is a critical need to expand therapeutic strategies. This review explores emerging targets in immune checkpoint inhibition beyond PD-1/PD-L1, including CTLA-4, TIGIT, LAG-3, TIM-3, NKG2A, and CD39/CD73. We highlight the biological basis of CD8 T cell exhaustion in shaping the antitumor immune response. Novel therapeutic approaches targeting additional inhibitory receptors (IR) are discussed, with a focus on their distinct mechanisms of action and combinatory potential with existing therapies. Despite significant advancements, challenges remain in overcoming resistance mechanisms and optimizing patient selection. This review underscores the importance of dual checkpoint blockade and innovative bispecific antibody engineering to maximize therapeutic outcomes for NSCLC patients. Full article

Background: Monoclonal antibodies (mAbs) are potent treatment options for infectious diseases. The rapid isolation and in vivo validation of therapeutic mAb candidates, including mAb cocktails, are essential to combat novel or rapidly mutating pathogens. The rapid selection and production of mAb candidates in sufficient amount and quality for preclinical studies are a major limiting step in the mAb development pipeline. Methods: Here, we developed a method to facilitate the screening of therapeutic mAbs in mouse models. Four conventional mAbs were transformed into single-chain variable fragments fused to the fragment crystallizable (Fc) region of a human IgG1 (scFv-IgG). These scFv-IgG were expressed individually or as a cocktail in vitro and in mice following transfection or hydrodynamic delivery of the corresponding plasmids. Results: This method induced high expression of all scFv-IgG and provided protection in two murine infection models. Conclusions: This study highlights the benefits of this approach for the rapid, low-cost screening of therapeutic mAb candidates. Full article

Superoxide dismutase (SOD), a key antioxidant enzyme, plays a crucial role in neutralizing reactive oxygen species (ROS) and maintaining redox balance. However, SOD is highly susceptible to glycation, a non-enzymatic modification induced by reducing sugars and reactive carbonyl species such as methylglyoxal. This review aims to provide a comprehensive analysis of SOD glycation, examining its biochemical mechanisms, its impact on enzymatic function, and its role in the progression of oxidative stress-related diseases. Additionally, it explores potential therapeutic strategies to prevent SOD glycation and restore its activity, highlighting translational applications for disease management. The review examines research on SOD glycation and its pathological consequences in diabetes complications, neurodegenerative disorders, and cardiovascular diseases. Key therapeutic interventions, including advanced glycation end-product (AGE) inhibitors (aminoguanidine, pyridoxamine), antioxidants (N-acetylcysteine, alpha-lipoic acid), SOD mimetics (MnTBAP, Tempol), enzyme stabilizers (thymoquinone, alliin), and receptor for advanced glycation end-products (RAGE) blockade, are analyzed for their efficacy in mitigating oxidative stress. SOD glycation reduces enzymatic activity, leading to elevated ROS levels and inflammation. Glycated SOD interacts with RAGE, increasing oxidative stress biomarkers. AGE inhibitors reduce carbonyl stress, whereas antioxidants lower ROS levels. SOD mimetics restore up to 85% of enzymatic activity, and enzyme stabilizers protect SOD from structural degradation. Additionally, monoclonal antibodies targeting RAGE have been shown to reduce inflammatory cytokines and improve mitochondrial function. SOD glycation is a major contributor to oxidative stress-related diseases. Preventing glycation and restoring SOD function through a multifaceted therapeutic approach is crucial for mitigating disease progression. By elucidating the role of SOD in disease pathogenesis, this review contributes to the advancement of targeted therapies for oxidative stress-related conditions, including diabetes, neurodegeneration, and cardiovascular diseases. Full article

The interleukin (IL)-17 family encompasses six structurally related pro-inflammatory cystine knot proteins, designated as IL-17A to IL-17F. Over the last decades, evidence has pointed to its role as a critical player in the development of inflammatory diseases such as psoriasis (PsO), axial spondyloarthritis (axSpA), and psoriatic arthritis (PsA). More specifically, IL-17A and IL-17F are overexpressed in the skin and synovial tissues of patients with these diseases, and recent studies suggest their involvement in promoting inflammation and tissue damage in axSpA and PsA. Bimekizumab is a monoclonal antibody targeting both IL-17A and IL-17F, playing an important role in the treatment of these diseases. This review details the implications of bimekizumab in the therapeutic armamentarium of axSpA and PsA. Full article

Ebola virus disease (EVD) is an acute illness with a high-case fatality rate (CFR) caused by an RNA virus belonging to the Filoviridae family. Over the past 50 years, regular EVD outbreaks have been reported. The West African EVD outbreak of 2013–2016 proved to be significantly more widespread and complex than previous ones, resulting in approximately 11,000 deaths. A coordinated international effort was required to bring the outbreak under control. One of the main challenges faced by clinicians and researchers combating EVD was the absence of vaccines and preventive treatments. Only recently have efforts led to the development of effective therapeutic options. Among these, monoclonal antibody-based drugs have emerged as the most promising agents for the urgent treatment of EVD. This article aims to review the key milestones in the development of antibody-based therapies for EVD, tracing the journey from the use of convalescent serum to the creation of effective monoclonal antibody-based drugs and their combinations. Full article

Background: Pig islets have the potential to address the limited supply of human islets available for transplantation. However, the knowledge of the biology of pig islets is currently limited. Thus, this study evaluated the molecules involved in cell-to-cell adhesion and insulin secretion pathways during the in vitro development of neonatal pig islets to understand the tissue we hope to use as a possible solution to the shortage of human islets for transplantation. Methods: Through RT-qPCR, immunoassays, and assessments of islet function, we explored the expression of E-cadherin and its correlation with the molecules involved in the insulin secretion pathway including GTPase, RAC1, and the membrane fusion protein SNAP25 during neonatal pig islet development. Results: Despite no significant difference observed in gross morphology and viability, as well as variable expression of RAC1, insulin, and SNAP25 in islets from 1-, 3-, and 7-day-old neonatal pigs, there was an apparent trend towards improved function in islets obtained from 3- and 7-day-old pigs compared with 1-day-old pigs. In the presence of 30 mM KCl, the amount of insulin secreted by islets from 3- and 7-day-old pigs but not from 1-day-old pigs was increased. Disruption of E-cadherin interactions with monoclonal antibodies resulted in decreased insulin secretion capacity of islets from 3-day old pigs. Conclusions: Our results show that blocking E-cadherin interactions with monoclonal antibodies resulted in disrupted peri-islet capsule and impaired islet insulin secretion under high glucose conditions. Thus, E-cadherin is important in the in vitro postnatal development and function of pig islets. Full article

Background: Macrophages play an important role in eliminating diseased and damaged cells through programmed cell death. Signal regulatory protein alpha (SIRPα) is a crucial immune checkpoint primarily expressed on myeloid cells and macrophages. It initiates a ‘do not eat me’ signal when engaged with CD47, which is typically expressed at elevated levels on multiple solid tumors. The phospholipase A2 Group 7 (PLA2G7), which is mainly secreted by macrophages, interacts with oxidized low-density lipoprotein (oxLDL) and associates with several vascular diseases and cancers. Methods: To identify potent fully human monoclonal antibodies (mAbs) against human SIRPα and PLA2G7, we conducted bio-panning of phage antibody libraries. Results: We isolated one human Fab (1B3) and VH (1A3) for SIRPα, as well as one human Fab (1H8) and one VH (1A9) for PLA2G7; the 1B3 Fab and 1A3 VH are competitively bound to SIRPα, interfering with CD47 binding. The 1B3 IgG and 1A3 VH-Fc augmented macrophage-mediated phagocytic activity when combined with the anti-EGFR antibody, cetuximab. The anti-PLA2G7 antibodies exhibited high specificity for the PLA2G7 antigen and effectively blocked the PLA2G7 enzymatic activity with half-maximal inhibitory concentrations (IC₅₀) in the single-digit nanomolar range. Additionally, 1H8 IgG and its derivative bispecific antibody exhibited the ability to block PLA2G7-mediated tumor cell migration. Conclusions: Our anti-SIRPα mAbs are expected to serve as potent and fully human immune checkpoint inhibitors of SIRPα, enhancing the antitumor responses of SIRPα-positive immune cells. Moreover, our anti-PLA2G7 mAbs represent promising fully human PLA2G7 enzymatic blockade antibodies with the potential to enhance both anti-tumor and anti-aging responses. Anti-SIRPα and PLA2G7 mAbs can modulate macrophage phagocytic activity and inflammatory responses against tumors. Full article

The prevalence of cerebral amyloid angiopathy (CAA) has been shown to increase with age, with rates reported to be around 50–60% in individuals over 80 years old who have cognitive impairment. The disease often presents as spontaneous lobar intracerebral hemorrhage (ICH), which carries a high risk of recurrence, along with transient focal neurologic episodes (TFNE) and progressive cognitive decline, potentially leading to Alzheimer’s disease (AD). In addition to ICH, neuroradiologic findings of CAA include cortical and subcortical microbleeds (MB), cortical subarachnoid hemorrhage (cSAH) and cortical superficial siderosis (cSS). Non-hemorrhagic pathologies include dilated perivascular spaces in the centrum semiovale and multiple hyperintense lesions on T2-weighted magnetic resonance imaging (MRI). A definitive diagnosis of CAA still requires histological confirmation. The Boston criteria allow for the diagnosis of a probable or possible CAA by considering specific neurological and MRI findings. The recent version, 2.0, which includes additional non-hemorrhagic MRI findings, increases sensitivity while maintaining the same specificity. The characteristic MRI findings of autoantibody-related CAA-related inflammation (CAA-ri) are similar to the so-called “amyloid related imaging abnormalities” (ARIA) observed with amyloid antibody therapies, presenting in two variants: (a) vasogenic edema and leptomeningeal effusions (ARIA-E) and (b) hemorrhagic lesions (ARIA-H). Clinical and MRI findings enable the diagnosis of a probable or possible CAA-ri, with biopsy remaining the gold standard for confirmation. In contrast to spontaneous CAA-ri, only about 20% of patients treated with monoclonal antibodies who show proven ARIA on MRI also experience clinical symptoms, including headache, confusion, other psychopathological abnormalities, visual disturbances, nausea and vomiting. Recent findings indicate that treatment should be continued in cases of mild ARIA, with ongoing MRI and clinical monitoring. This review offers a concise update on CAA and its associated consequences. Full article

Objectives: This study aimed to assess the effectiveness of gold nanoparticles conjugated with anti-EGFR monoclonal antibodies (GNPs-EGFR) in distinguishing between benign and malignant salivary gland tumors. Methods: A total of 49 oral salivary gland tissue samples were analyzed, including 22 malignant salivary gland tumors (MSGTs), 15 benign salivary gland tumors (BSGTs), and 12 control samples. For each sample, three 5 μm consecutive tissue sections were prepared. The first section was stained with hematoxylin and eosin (H&E) to confirm the diagnosis, the second was immunohistochemically stained for anti-EGFR, and the third was treated with GNPs-EGFR followed by hyperspectral microscopy to analyze the reflectance spectrum. Results: Reflectance intensity was significantly higher (p < 0.001) in MSGTs compared to BSGTs and controls, with intensity levels increasing alongside tumor grade. The average hyperspectral reflectance values were strongly correlated with the GNPs-EGFR immunohistochemical score and varied significantly between subgroups (p < 0.001). Conclusions: GNPs-EGFR reflection measurements effectively differentiate MSGTs from BSGTs with high sensitivity. This diffusion–reflection technique holds potential as a valuable tool for tumor detection, surgical margin assessment, and intraoperative identification of residual disease in salivary gland tumors. Full article

Objectives: Identifying immune-protective biomarkers is crucial for the effective management and mitigation of current and future COVID-19 outbreaks, particularly in preventing or counteracting the immune evasion exhibited by the Omicron variants. The emergence of SARS-CoV-2 variants, especially those within the Omicron lineage, has highlighted their capacity to evade neutralizing antibodies, emphasizing the need to understand the role of antibody-dependent cell-mediated cytotoxicity (ADCC) in combating these infections. Methods: This study, conducted in Qichun City, Hubei province, from December 2021 to March 2023, involved 50 healthy Chinese adults who had received two doses of inactivated vaccines and had subsequently experienced mild infections with the Omicron BA.5 variant. Blood samples from these 50 healthy Chinese adults were collected at six distinct time points: at baseline and at the 1st, 3rd, 6th, and 9th months following the third dose of the inactivated vaccine, as well as 3 months post-breakthrough infection. Their sera were analyzed to assess ADCC and neutralization effects. Results: The results indicated that the antibodies elicited by the inactivated SARS-CoV-2 vaccine targeted the spike protein, exhibiting both pre-existing neutralizing and ADCC activities against Omicron variants BA.5 and XBB.1.5. Notably, the ADCC activity demonstrated greater stability compared to that of the neutralizing effects, persisting for at least 15 months post-vaccination, and could be augmented by additional vaccine doses and breakthrough infections. The ADCC effect associated with hybrid immunity effectively targets a spectrum of prospective Omicron variants, including BA.2.86, CH.1.1, EG.5.1, and JN.1. Conclusions: In light of its stability and broad-spectrum efficacy, we recommend the use of the ADCC effect as a biomarker for assessing protective immunity and guiding the development of vaccines and monoclonal antibodies. Full article

Background/Objectives: Cryofibrinogenemia, characterized by plasma cryoprecipitation of fibrinogen and related proteins, is a rare and often under-recognized entity that can present with significant renal involvement. Methods: we describe a 66-year-old woman with progressive renal failure due to membranoproliferative glomerulonephritis driven by cryofibrinogen deposits. Her clinical course was marked by relapsing–remitting disease with limited response to high-dose corticosteroids but significant improvement following plasma exchange. Over seven years, she underwent three kidney biopsies, revealing progressive histopathological changes, including glomerular cryofibrinogen deposits and evolving chronicity. A detailed review of the literature identified 50 cases of cryofibrinogenemia, highlighting its association with monoclonal gammopathies, malignancies, and autoimmune diseases. Results: our case uniquely underscores the pathogenic interplay between cryofibrinogenemia and a monoclonal IgG-kappa paraprotein, which was found to directly stabilize fibrinogen and drive cryoprecipitation. This novel observation aligns cryofibrinogenemia with monoclonal gammopathy of renal significance, expanding the diagnostic and therapeutic landscape for this entity. Conclusions: this report also highlights the pivotal role of kidney biopsy with electron microscopy in diagnosing cryofibrinogen-associated renal disease, particularly when conventional biomarkers are insufficient. Moreover, our findings emphasize the therapeutic utility of plasmapheresis and the potential need for therapies aimed at eliminating the pathogenetic monoclonal antibody in managing refractory cases. Enhanced awareness and further research into this rare entity are essential for advancing patient care and outcomes. Full article

Hendra virus (HeV) is a bat-borne zoonotic agent which can cause a severe and highly fatal disease and can be transferred from animals to humans. It has caused over 100 deaths in horses since it was discovered in 1994. Four out of seven infected humans have died. Since the release of the HeV vaccine (Equivac^® HeV Hendra Virus Vaccine for Horses, Zoetis Australia Pty Ltd., Rhodes, NSW 2138) in Australia, there has been an urgent requirement for a serological test for differentiating infected from vaccinated animals (DIVA). All first-line diagnostic serological assays at the Australian Centre for Disease Preparedness (ACDP) incorporate recombinant HeV soluble G glycoprotein (sG) as the antigen, which is also the only immunogen present in the Equivac^® HeV vaccine. Problems therefore arose in that antibody testing results were unable to distinguish between prior vaccination or infection with HeV. This study describes the development of a HeV DIVA ELISA strategy using recombinant sG and HeV nucleoprotein (N), paired with specific monoclonal antibodies in a competition ELISA format. The validation of this assay strategy was performed using a positive cohort of 19 serum samples representing post-infection sera, a negative cohort of 1138 serum samples representing horse sera collected pre-vaccine release and a vaccination cohort of 502 serum samples from horses previously vaccinated with Equivac^® HeV vaccine. For the sG glycoprotein, the diagnostic sensitivity (DSe) was 100.0% (95% CI: 99.3–100.0%) and diagnostic specificity (DSp) 99.91% (95% CI: 99.5–100.0%), using a percentage inhibition cut-off value of >36, whereas for the N protein, DSe was 100.0% (95% CI: 82.4–100.0%) and DSp 100.0% (95% CI: 99.7–100.0%), using a percentage inhibition cut-off value of >49. Taken together, these results demonstrate that the HeV DIVA ELISA strategy developed here is now an essential and critical component of the testing algorithm for HeV serology testing in Australia. Full article