Peptides are increasingly employed as active ingredients in both therapeutic and cosmeceutical applications due to their high biological specificity, favorable safety profiles, and expanding market relevance. In topical formulations, peptide activity is typically confined to the skin, where widespread dermal proteases may significantly affect their stability and efficacy. Despite the growing use of bioactive peptides in dermatological and cosmeceutical products, robust analytical methodologies for assessing their susceptibility to dermal enzymatic degradation remain limited. In this study, a two-dimensional HPLC-MS/MS (2D-HPLC-MS/MS) method based on ion trap detection was developed and validated for the quantitative evaluation of peptide stability in human skin homogenate (HSH). The analytical setup integrates online cleanup with chromatographic separation on a silica-based pentafluorophenyl (PFP) column, enabling reliable analysis of peptides with different polarity profiles within a single workflow. The system enables direct injection of sample solutions with relatively high organic solvent content, allowing limited sample dilution and preservation of analytical sensitivity. A key feature of the method is the use of isomeric peptide analogues as internal standards, monitored under identical MS/MS conditions as their corresponding analytes. This approach provides effective correction for ionization and fragmentation variability while offering a practical alternative to stable isotope-labelled standards. The ion trap mass analyzer ensured controlled and reproducible fragmentation behavior, supporting robust quantitative performance. The developed method demonstrated satisfactory linearity, sensitivity, precision, accuracy, and minimal matrix effects. Application to stability studies of three peptides with distinct polarity profiles confirmed its suitability for monitoring degradation kinetics in HSH. Overall, this 2D-HPLC-MS/MS strategy provides a versatile analytical platform for dermal peptide stability assessment and supports early-stage screening and preclinical studies of bioactive peptides intended for topical applications.
Peptide-activated G protein-coupled receptors (GPCRs) regulate physiological processes through interaction with neuropeptides and peptide hormones. Identifying endogenous peptide agonists remains challenging, as peptide-GPCR pairings often follow gene-family relationships that offer limited predictive insight for orphan GPCRs without characterized homologs. Using a dataset of experimentally validated peptide-GPCR interactions from Caenorhabditis elegans, we demonstrate that AF-multimer confidence metrics partially discriminate agonist from non-agonist complexes, with improved discrimination using AF-Multistate-derived active-state templates. Feature analysis revealed that AF-multimer's pair representations outperform single representations, with distinct subregions providing complementary signals. Leveraging these insights, we developed DeorphaNN, a graph neural network integrating active-state GPCR-peptide structural predictions, interatomic interactions, and deep learning embeddings to prioritize putative peptide agonists for experimental screening. DeorphaNN generalized across diverse species, as shown by performance on annelid and human retrospective benchmarks. Experimental validation confirmed predicted agonists for two orphan GPCRs, demonstrating its utility for accelerating peptide-GPCR deorphanization.
Numerous natural angiotensin-converting enzyme (ACE) inhibitors (ACEIs) have been extensively studied and applied for the prevention and control of hypertension. This study conducted sequence analysis and computer screening on ACE inhibitory peptides isolated from silkworm pupa, determining their inhibition types through kinetic analysis. By integrating spectral techniques, molecular docking, and molecular dynamics simulations, we investigated the interaction mechanisms between silkworm-derived active peptides and ACE. The results indicated that three novel high-activity ACE inhibitors were identified through computer screening and in vitro validation of DA201-C macroporous resin-purified peptides. MQGFIPE exhibited the strongest inhibitory activity (IC50 = 0.21 ± 0.036 mg/mL), followed by PNDIL (IC50 = 0.569 ± 0.005 mg/mL) and ADNIPIK (IC50 = 0.849 ± 0.001 mg/mL). Lineweaver-Burk plots suggested that MQGFIPE and PNDIL functioned as mixed-type inhibitors, while ADNIPIK acted as a non-competitive inhibitor. Molecular docking simulations revealed that these peptides inhibit ACE through hydrogen bonding formation and hydrophobic interactions with residues. Additionally, spectral studies demonstrated that the peptides MQGFIPE, ADNIPIK and PNDIL effectively quench the intrinsic fluorescence of ACE, alter its conformation and the microenvironment of aromatic amino acid residues, thereby inhibiting its activity. Molecular dynamics simulations confirmed their stable binding to the ACE complex. These findings suggest that silkworm pupa are a significant natural source of ACE-inhibiting peptides, which could serve as functional components in hypertension dietary supplements.
Targeted protein degradation (TPD) technology, with a particular emphasis on proteolysis-targeting chimeras (PROTAC), has emerged as a pivotal advancement in the field of drug discovery. However, several challenges-including the identification of suitable ligands for traditionally undruggable proteins, issues related to poor solubility and permeability, nonspecific biodistribution, and off-target toxicity-have significantly hindered their clinical translation. Peptides, recognized for their ability to serve as promising ligands for broad molecular recognition, exhibit unique potential to address these limitations in TPD applications. Literature and related information were collected from online resources such as Google Scholar, Web of Science, PubMed, CNKI, Baidu Scholar, and X-mol. Recent advancements in peptide-mediated TPD have shown promise in overcoming these challenges as researchers focus on engineering highly selective peptides that enhance binding affinity for traditionally undruggable proteins while optimizing their solubility and permeability, with next-generation delivery systems also developed to reduce nonspecific biodistribution and off-target toxicity, thereby improving the therapeutic potential of peptide-based TPD approaches. This review summarizes recent advancements in peptide-based PROTAC development, focusing on innovative delivery strategies and methods for enhancing efficiency, while also offering insights into future prospects aimed at optimizing therapeutic precision and efficacy.
Effective cancer immunotherapy is often hindered by V-domain Ig suppressor of T cell activation (VISTA) signaling and myeloid-derived suppressor cells (MDSCs) within tumor microenvironments (TMEs). Here, we report D-peptide-nanodrug conjugates that synergize VISTA blockade and innate activation to overcome these constraints. We developed Reorpn7, a proteolytically stable D-peptide VISTA inhibitor with high binding affinity and in vitro serum stability. This inhibitor was co-loaded with a TLR7/8 agonist (R848) into an MMP-2 responsive PLGA nanoplatform (r7PL-PLGA@R848). Upon enrichment in the TME, MMP-2 responsive cleavage enables the precise release of the VISTA-targeting peptide Reorpn 7 and the TLR7/8 agonist R848, thereby achieving synergistic therapeutic effect of VISTA blockade and innate immune activation. In MC38 tumor models, this nanosystem significantly increased intratumoral CD8+ T cell infiltration and achieved robust antitumor efficacy without obvious systemic toxicity. Mechanistically, the platform reshaped the TME from an inhibitory milieu into an immunosupportive one by coordinating checkpoint blockade, myeloid reprogramming, and T cell recruitment. This stimuli-responsive paradigm offers a translatable strategy to overcome immune checkpoint blockade resistance in challenging malignancies.
Pigskin peptides (PSPs) are bioactive collagen hydrolysates known for their antioxidant, antibacterial, and other biomedical potentials; however, their direct application is limited by poor environmental stability. Herein, we report the use of NU-1000, a robust zirconium-based metal-organic framework (MOF), as a delivery platform. A PSP-loaded NU-1000 composite, denoted as PSPs-1@NU-1000-2, was successfully constructed using PSPs with a molecular weight below 3000 Da, and this composite exhibited superior bioactivity. Structural and morphological characterization by FTIR, SEM, XRD, and BET confirmed the successful incorporation of PSPs into NU-1000. Notably, compared with free PSPs, the PSPs-1@NU-1000-2 composite demonstrated significantly enhanced DPPH radical scavenging activity and stronger antibacterial effects against Escherichia coli and Staphylococcus aureus. This MOF-based immobilization strategy not only preserves but also potentiates the bioactivities of PSPs, offering a promising peptide-nanomaterial composite for applications in the cosmeceutical, pharmaceutical, and food industries.
This study examined decade-long trends and differences in sodium-glucose cotransporter 2 inhibitor (SGLT2i) and glucagon-like peptide-1 receptor agonist (GLP-1 RA) prescriptions among adults with type 2 diabetes using real-world data from Epic Cosmos. We analyzed electronic health records of 1 517 594 adults with type 2 diabetes without end-stage renal disease from 2014 to 2024. Annual prescribing trends were evaluated by patient race and insurance type using negative binomial regression. Medication exposure was defined using active prescriptions/orders recorded in Epic Cosmos during the calendar year. In pooled descriptive analyses, we also characterized patients prescribed these medications by clinical characteristics, neighborhood-level social vulnerability, and prescriber specialty. From 2014 to 2024, SGLT2i use rose from 0.5% to 12.1% and GLP-1 RA use increased from 0.9% to 15.9%. Black patients had consistently lower prescription rates than White patients across insurance groups. Primary care physicians prescribed about one-third of these medications. In pooled descriptive analyses, endocrinology was associated with higher observed prescribing rates than primary care or cardiology. Patients from neighborhoods with lower social vulnerability were more likely to receive these therapies. Use of SGLT2i and GLP-1 RA increased substantially over the past decade, significant racial, socioeconomic, and insurance-related differences persist in prescribing these therapies.
Headache is the most common symptom of idiopathic intracranial hypertension (IIH) and may persist despite therapy, with a significant impact on quality of life. Since calcitonin gene-related peptide (CGRP) plays a crucial role in the pathophysiology and management of primary headaches such as migraine, this raises the question whether CGRP also contributes to headache in IIH. Therefore, we compared tear fluid CGRP levels between IIH patients with headache and healthy controls, and in IIH patients before and after CSF pressure normalization by therapeutic lumbar puncture. IIH patients with headache attributed to IIH and healthy controls were included. To avoid confounding with chronic migraine, IIH patients with a chronic migraine phenotype were excluded. Tear fluid was collected from IIH patients and controls. In IIH patients, an additional measurement was performed approximately 3 h after therapeutic lumbar puncture. CGRP levels were analyzed using a commercially available ELISA. Twenty-three IIH patients (all female; age: 34.0 ± 8.8 years) and 20 healthy controls (all female, age: 25.7 ± 5.5 years) were included. IIH patients had 16.4 ± 12.3 headache days per month and headache was mostly bilateral and pressing. Baseline tear fluid CGRP levels were significantly lower in IIH patients compared to healthy controls (2.4 ± 1.2 ng/ml vs. 4.9 ± 4.2 ng/ml, p < 0.001). There was no significant change in CGRP levels in IIH patients before vs. after therapeutic lumbar puncture (2.4 ± 1.2 vs. 2.4 ± 1.7 ng/ml, p = 0.236). Similarly, in the subgroup with immediate headache improvement, CGRP levels remained unchanged (before: 2.2 ± 0.9 ng/ml, after: 2.8 ± 2.4 ng/ml; p = 0.674). Tear fluid CGRP levels were lower in IIH patients with headache but without a chronic migraine phenotype compared to healthy controls. In addition, CSF pressure normalization was not associated with changes in CGRP levels after 3 h. These results do not support a major role of CGRP in IIH-associated headache without a chronic migraine phenotype. The study was previously registered at the German Clinical Trial Register (DRKS www.drks.de) (DRKS00025278), Trial registration date 25.06.2021.
Glucagon-like peptide-1 receptor agonists (GLP-1RAs) reduce stroke incidence, but their impact on the severity of acute ischemic stroke (AIS) when events occur remains unclear. The aim of this study is to evaluate whether pre-stroke GLP-1RA use is associated with reduced ischemic stroke severity and improved post-stroke outcomes. We conducted a retrospective cohort study using the 2016-2024 data from the TriNetX US Collaborative Network. Patients were categorized based on documented use of GLP-1RAs for at least six months prior to the index stroke event. After, propensity score matching, primary outcome was measured by the National Institutes of Health Stroke Scale (NIHSS), with secondary outcomes including rates of severe stroke (NIHSS ≥10), intensive care unit admission, neurological symptoms, in-hospital complications, and 30-day mortality. A total of 209,180 first-time AIS patients were identified, of whom 4773 (2.3%) were GLP-1RA users. After PSM, 4769 patients remained in each group. GLP-1RA users had lower median NIHSS (3 vs 4, p < 0.001). GLP-1RA users also had lower rates of severe stroke (18.4% vs 25.5%, p < 0.001), dysphagia/dysarthria (43.5% vs 48.0%, p < 0.001), aphasia (25.2% vs 28.7%, p < 0.001), hemiplegia/hemiparesis (37.6% vs 43.3%, p < 0.001), intracranial hemorrhage (13.2% vs 14.7%, p = 0.031), and 30-day mortality (6.3% vs 7.6%, p = 0.016). Among adult patients who suffered a first-time AIS, pre-stroke use of GLP-1RA was associated with reduced stroke severity, lower rates of symptoms and complications, and lower mortality. These findings suggest that the benefits GLP-1RA may extend beyond preventing the occurrence of cerebrovascular events.
The locus coeruleus (LC), a cluster of noradrenergic neurons in the dorsal pons, is the brain's main source of norepinephrine (NE), crucial for memory, cognition, and stress response. NE dysregulation has been linked to mood disorders, chronic stress, and neurodegenerative diseases such as Alzheimer's disease (AD). Amyloid-β42 (Aβ42) is a key protein in AD pathology, and forms plaques that trigger neurodegeneration. Studies show elevated Aβ42 levels are associated with anxiety symptoms, even in cognitively normal individuals. Previous research suggests a positive correlation between NE and Aβ42, with Aβ42 present in NE-producing LC neurons. NE may affect Aβ42 levels through adrenergic receptors on neurons or microglia. However, the exact role of NE in modulating Aβ42 remains unclear. Using the NE depletion models, N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP-4) lesions and dopamine β-hydroxylase (DBH) knockout (KO) mice, researchers found reduced Aβ42 levels without changes in amyloid precursor protein processing. A newer model, DBH internal ribosome entry site-Cre × floxed tyrosine hydroxylase (DBH-IRES-Cre × floxed-TH) preserved dopamine (DA) but eliminated NE in adrenergic neurons, further supporting the specific influence of NE on Aβ42. Findings indicate that decreased Aβ42 in NE-depleted mice stems not from altered DA but from NE loss. This highlights the NE-mediated regulation of endogenous Aβ42 and the role of NE in AD-related pathology.
Medullary thymic epithelial cells (mTECs) are central to immune self-tolerance owing to their capacity to express and present a diverse repertoire of self-derived peptides to developing thymocytes. This diversity arises from both promiscuous gene expression and specialised differentiation programmes, including thymic mimetic-cell populations, and is largely shaped by AIRE/Aire and FEZF2/Fezf2. Although the roles of Aire and Fezf2 in regulating peripheral tissue antigen transcription are well established, a critical conceptual gap remains regarding how transcriptional promiscuity is translated into the repertoire of peptides presented by major histocompatibility complex class II (MHC-II) molecules. In this review and perspective, we synthesize knowledge on mTEC biology, Aire- and Fezf2-dependent transcriptional programmes, and advances in immunopeptidomics. We discuss regulatory layers that decouple mRNA abundance from peptide presentation and highlight how single-cell transcriptomics and mass spectrometry-based immunopeptidomics provide complementary insights into mTEC heterogeneity and antigenic output. We argue that integrating these approaches is essential to understand central tolerance mechanistically and to explain selective defects in thymic self-antigen presentation. This framework offers a refined basis for interpreting autoimmune disease origins and guiding antigen-focused therapeutic strategies.
Hirudin is a potent thrombin inhibitor limited by short circulation half-life and bleeding complications. We engineered an FXa-activatable hirudin thrombus-targeted prodrug, yet the N-terminal IEGR peptide introduced unwanted basal activity. After further screening, we found this residual function was fully silenced by a P-selectin-binding peptide upstream of IEGR, with no compromise to FXa-mediated cleavage. Our final construct PXHV2 incorporates both peptides, C-terminal human serum albumin to extend half-life, and an albumin-embedded cyclic RGD sequence for constitutive platelet recruitment. PXHV2 remains catalytically inert until FXa proteolysis restores thrombin-suppressive capacity. In murine electrical and laser-induced thrombosis models, PXHV2 delayed arterial and microvascular occlusion and maintained 120 min pre-injury antithrombotic protection, unlike rapidly ineffective free hirudin. PXHV2 elicited no increase in tail bleeding relative to saline controls. Our data identify PXHV2 as a long-lived, thrombus-selective prodrug with durable efficacy and minimal bleeding risk.
Tirzepatide, a once-weekly glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptor agonist, has demonstrated its efficacy in clinical trials; however, real-world evidence on treatment satisfaction and experiences among people with obesity or overweight (PwO) and their prescribing physicians remains limited. This study aimed to assess the real-world clinical characteristics, treatment patterns, and satisfaction with tirzepatide among PwO without type 2 diabetes (T2D) and their prescribing physicians in the United States. This secondary analysis utilized data from a cross-sectional survey of physicians and PwO from October 2023 to April 2024, and from October 2024 to January 2025. Physicians provided data for PwO meeting selection criteria, at data capture/previous body mass index (BMI) ≥30 or ≥27 and < 30 kg/m2 with at least one obesity-related complication (ORC), without T2D, enrolled in a weight management program, and were receiving tirzepatide at the time of data collection. Sociodemographic and clinical characteristics, treatment history, and satisfaction measures were analyzed. Data from 151 physicians and 199 PwO without T2D using tirzepatide were included. Most PwO (84.4%) had ≥1 ORC, with hypertension and dyslipidemia being the most prevalent. Over 90% (n = 45/48) of PwO reported likelihood to recommend tirzepatide reflecting high satisfaction and willingness to recommend tirzepatide to others. Physicians reported long-term safety, ease of dosing, and reduced appetite as key reasons for prescribing tirzepatide. Physicians reported that tirzepatide treatment was successful in the majority of people (78.5%) being treated for obesity. The data in these surveys reflected that PwO without T2D prescribed tirzepatide had high motivation, adherence, and satisfaction. These findings provide real-world insights into treatment and satisfaction with tirzepatide.
Nesfatin-1 is an 82-amino acid polypeptide derived from the precursor protein nucleobindin 2 (NUCB2), which is a conserved multifunctional peptide in vertebrates that plays a key role in metabolic regulation, inflammatory response and ferroptosis. Although Nesfatin-1 has been characterized in various vertebrates, from mammals to teleosts, its structural and functional characteristics in primitive vertebrates remain unclear. In this study, Nesfatin-1 orthologs from the most primitive jawless vertebrate lamprey (Lethenteron camtschaticum) were cloned and identified, designated as Lc-Nesfatin-1. Bioinformatic analysis revealed that Nesfatin-1 of lamprey is relatively conserved in both sequence similarity and three-dimensional (3D) structure compared to that of higher vertebrates. Lc-Nesfatin-1 was significantly upregulated under lipopolysaccharide (LPS) stimulation, which also markedly induced the expression of pro-inflammatory cytokines, the key signaling molecule NF-κB, and anti-inflammatory cytokines. Notably, Flow cytometry analysis showed that Lc-Nesfatin-1 did not significantly inhibit LPS-induced intracellular ROS production. Furthermore, Nesfatin-1 serves as an effective negative regulator that significantly inhibits ferroptosis. These results reveal that Lc-Nesfatin-1 plays a critical role in modulating both LPS-induced inflammatory responses and ferroptosis. This study has successfully elucidated the characteristics and functions of Nesfatin-1 in primitive vertebrates and provided valuable insights into the investigation of inflammatory responses and ferroptosis in jawless vertebrates.
Glucagon-like peptide-1 receptor (GLP-1R) agonists have transformed obesity pharmacotherapy, producing clinically meaningful weight loss and robust improvements in glycemic control. Yet their efficacy remains constrained by dose-limiting gastrointestinal adverse effects, including nausea and vomiting, that reduce adherence and limit escalation to maximally effective doses. Thus, the next-generation of obesity therapeutics must not only enhance weight loss but also expand the therapeutic window by dissociating metabolic efficacy from aversive side effects. The recent clinical success of dual GLP-1R/glucose-dependent insulinotropic polypeptide (GIP) receptor agonists has catalyzed renewed interest in GIP biology. However, this resurgence has exposed a fundamental paradox: both pharmacological activation and blockade of GIP receptor (GIPR) signaling reduce body weight and enhance the efficacy of GLP-1-based therapies. These seemingly opposing pharmacological strategies therefore converge on a shared therapeutic outcome through distinct biological mechanisms. GIPR agonism appears to promote weight loss in part through recruitment of anorectic neural circuits, increased thermogenesis, and attenuation of GLP-1-induced aversive effects, whereas GIPR antagonism may enhance GLP-1R signaling and counteract the lipogenic actions of endogenous GIP. In this review, we examine the mechanistic basis underlying the GIPR agonism-antagonism paradox and discuss how GIPR signaling may regulate the balance between metabolic efficacy and treatment tolerability. Understanding these mechanisms may guide the rational design of next-generation incretin-based therapies that maximize weight loss while minimizing adverse effects.
Cardiovascular-kidney-metabolic (CKM) syndrome, formally defined by the American Heart Association in 2023, affects approximately 90% of US adults, who meet criteria for stage 1 or higher. The rapid convergence of multiple drug classes on CKM pathways-SGLT2 inhibitors, finerenone, GLP-1 receptor agonists, ARNI, and interleukin-directed therapies-has created an urgent need for pharmacologically grounded frameworks that guide drug selection, interpret biomarker responses, and monitor target engagement across interconnected organ systems. This review proposes a three-dimensional biomarker-guided approach to precision pharmacotherapy in CKM syndrome. In the organ-specific dimension, we map key biomarkers to their corresponding drug targets and elucidate the molecular mechanisms underlying drug-biomarker interactions: SGLT2 inhibitors attenuate myocardial injury through metabolic substrate shifting toward ketone body utilization and, based on preclinical evidence, NHE1 inhibition; neprilysin selectivity of sacubitril/valsartan explains the differential natriuretic peptide response; and tubuloglomerular feedback mediates the renoprotective hemodynamic effects of SGLT2 inhibitors. In the pathway-specific dimension, we identify cross-system biomarkers-hs-CRP, IL-6, galectin-3, GDF-15, and FGF21-that reveal shared druggable targets spanning the IL-1β/NLRP3 inflammasome axis (canakinumab, colchicine), IL-6 trans-signaling (ziltivekimab), and FGF21/β-klotho metabolic signaling. In the temporal dimension, we demonstrate how serial biomarker trajectories serve as pharmacodynamic readouts that distinguish therapeutic drug effects from disease progression, including the initial eGFR dip with SGLT2 inhibitors and natriuretic peptide changes during combination therapy. Central to this framework is the concept of "pharmacological phenotyping"-using multi-biomarker panels to define drug-responsive pathophysiological states that directly inform therapeutic selection, analogous to companion diagnostics in oncology. We further present a comprehensive drug-biomarker interaction matrix with pharmacological rationale and analyze the emerging drug development pipeline, including RNA-based Lp(a) therapeutics, FGF21 analogues, galectin-3 inhibitors, and in vivo CAR-T anti-fibrotic approaches. This framework provides a practical roadmap for biomarker-guided precision pharmacotherapy in CKM syndrome.
Despite ongoing debate about the "amyloid hypothesis", the imbalance between the production and clearance of β-amyloid (Aβ) peptides in the brain remains one of the most compelling explanations for the progression of Alzheimer's disease. Current strategies therefore focus on discovering clinically relevant therapeutic agents that target Aβ peptides and amyloid structures. Because of their unique and attractive properties - biocompatibility, non-immunogenicity, non-toxicity, and ease of functionalization and production - the use of glycopolymers as amyloid inhibitors has generated interest in therapeutic research for Alzheimer's disease. This review provides a comprehensive and critical overview of the literature on glycopolymers in the treatment of Alzheimer's disease. It begins with a description of the disease's neuropathological mechanisms and the formulations approved by the FDA or currently in clinical trials. The second part discusses the use of glycopolymers as amyloid inhibitors, which prevent the formation of neurotoxic soluble oligomers and subsequent plaques observed in Alzheimer's disease. This is achieved by binding to monomers, blocking self-aggregation, and interrupting toxic interactions, offering a therapeutic strategy to halt disease progression. Finally, the main conclusions and perspectives on the use of glycopolymers as amyloid inhibitors are presented.
Frequent antigenic drift in influenza viruses necessitates broadly protective vaccines. This study evaluated an intranasal adenoviral-vector vaccine expressing influenza A nucleoprotein (NP) fused to the autophagy-inducing peptide C5. A heterologous prime-boost regimen using chimpanzee and bovine adenoviral platforms induced strong NP-specific humoral and cellular immune responses in mice. High serum and lung IgG/IgA level were detected, accompanied by enhanced antibody-dependent cellular cytotoxicity. Cellular analyses revealed potent NP-specific cytokine responses and expansion of effector memory (TEM) and tissue-resident memory (TRM) T cells, particularly CD8+ TRM, in the lungs. Experiments using immune-deficient mice showed that B cells and CD8+ T cells independently contributed to early viral clearance. Adoptive transfer studies demonstrated that lung-derived T cells conferred the strongest heterosubtypic viral restriction, whereas systemic T cells and antibodies provided partial protection. Together, these findings highlight the critical role of lung-resident T cells in cross-protective viral control, supporting intranasal NP-based adenoviral vaccines as promising universal influenza vaccine candidates.
Allergen immunotherapy (AIT) remains the only disease-modifying treatment for immunoglobulin (Ig) E-mediated allergic diseases, including allergic rhinitis (AR), asthma, food allergy, and atopic dermatitis (AD). This review provides a comprehensive update on recent advances in AIT, focusing on immunological mechanisms, novel vaccines, biomarkers, clinical outcomes across different diseases, and emerging combination strategies with biologics. We discussed the pivotal roles of regulatory T and B cells, innate lymphoid cells (ILCs), and blocking antibodies (immunoglobulin G4 [IgG4], immunoglobulin A [IgA]) in establishing long-term tolerance. Novel allergen vaccines-including recombinant and hypoallergenic derivatives, virus-like particle (VLP) conjugates, and transgenic peptide vaccines-aim to improve safety and efficacy. We critically evaluate current biomarkers for predicting and monitoring AIT response, such as the allergen-specific IgE (sIgE)/total immunoglobulin E (tIgE) ratio, basophil activation test, and cellular markers. Clinical evidence supports AIT efficacy in asthma, AR, and AD, with emerging applications in food allergies. Furthermore, combining AIT with biologics (e.g., anti-IgE and anti-interleukin [IL]-4/IL-4 receptor [IL-4R]) shows promise in enhancing safety and efficacy, particularly in severe asthma. Despite these advances, challenges remain in patient selection, treatment duration, and extract standardization. Future research should prioritize predictive biomarkers, short-course regimens, and synergistic biologic-AIT approaches.