Journal of extracellular biology最新文献

{"title":"Investigation Into the Collection and Isolation Workflows of Extracellular Vesicles From Tears","authors":"Nikolay Boychev,&nbsp;Mudigunda V. Sushma,&nbsp;Vincent Yeung,&nbsp;Amy E. Ross,&nbsp;Joseph B. Ciolino","doi":"10.1002/jex2.70158","DOIUrl":"https://doi.org/10.1002/jex2.70158","url":null,"abstract":"<p>Tear extracellular vesicles (tEVs) are a promising source of biomarkers for ocular and systemic diseases. Standardized approaches for their collection and isolation remain underdeveloped. Contact lenses (CLs), routinely used for vision correction and as bandage lenses, offer a practical platform for clinically integrated tear sampling. Using healthy rabbit eyes (<i>n</i> = 4), we compared two tear collection methods, daily soft CLs and Schirmer Strips (SS), and two EV isolation techniques, ExoQuick (EQ) polymer precipitation and ultracentrifugation (UC). tEVs were characterized by particle counts, protein content, zeta potential, mRNA expression of reference genes (GAPDH, SNRPG, TOMM7), and exosomal tetraspanins (CD9, CD63). CL-derived tEVs showed lower protein contamination and reduced inflammatory cytokine expression, whereas SS-derived tEVs contained more cellular debris and higher levels of MMP-9, TNF-α, and Leptin, consistent with tissue stress and remodeling. EQ yielded higher particle recovery and smaller aggregates than UC, but produced vesicles with more positive surface charge, indicating altered surface properties and reduced colloidal stability. GAPDH demonstrated the most stable expression across samples, and CD63 exhibited higher eye-specific RNA scores. These findings provide a methodological framework for tEV profiling, highlighting complementary molecular information captured by CLs and SS and trade-offs between EQ and UC for tEV recovery and stability.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70158","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148386606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating the Effect of Ginger-Derived Nanovesicles on the Growth and Metabolic Activity of Bacteroides thetaiotaomicron: An Isothermal Microcalorimetric Study","authors":"Francesca Loria,&nbsp;Anna Kattel,&nbsp;Marina Junusova,&nbsp;Marco Brucale,&nbsp;Francesco Valle,&nbsp;Paolo Bergese,&nbsp;Eeva-Gerda Kobrin,&nbsp;Antonio Chiesi,&nbsp;Paolo Guazzi,&nbsp;Sirin Korulu,&nbsp;Irina Stulova,&nbsp;Nataša Zarovni,&nbsp;Raivo Vilu","doi":"10.1002/jex2.70163","DOIUrl":"https://doi.org/10.1002/jex2.70163","url":null,"abstract":"<p>Plant-derived extracellular vesicles (EVs) have shown numerous health benefits, including modulation of the human gut microbiota. Herein, we employed isothermal microcalorimetry (IMC) to explore the effects of ginger-derived nanovesicles (G-NVs) on the growth and metabolic activity of <i>Bacteroides thetaiotaomicron (Bt)</i>, a dominant saccharolytic commensal with promising next-generation probiotic potential. <i>Bt</i> was exposed to either G-NVs or the ginger extract separated from G-NVs (G-CTL) in defined media under anaerobic conditions. Both ginger samples enhanced bacterial specific growth rate and maximum metabolic activity, inducing the latter earlier. However, higher biomass production and greater secretion of acetic, succinic and propionic acids occurred only in response to the G-CTL. Complete sugar depletion and unchanged free amino acid levels indicated preferential carbohydrate utilisation by <i>Bt</i>. Overall, these findings revealed that <i>Bt</i>’s metabolic state is shaped by both G-NVs and G-CTL, yet through distinct mechanisms, with G-NVs inducing rapid stimulation without increasing total metabolic output and G-CTL providing a sustained, dose-dependent effect. To our knowledge, this is the first study applying IMC to monitor in real-time the impact of EVs on microbial growth and metabolism, underscoring IMC's utility for mechanistic studies of EV-microbe interactions. Furthermore, this research sets the ground for innovative strategies in nutraceutical and microbiome-targeted therapy development.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70163","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148386605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineered Red Blood Cell-Derived Extracellular Vesicles With Klotho Peptide Protect the Kidney From Fibrosis","authors":"Tunahan Ergunay,&nbsp;Alessia Brossa,&nbsp;Michela Arena,&nbsp;Stefania Bruno,&nbsp;Alessandro Gori,&nbsp;Anne Metje van Genderen,&nbsp;Rosalinde Masereeuw,&nbsp;Benedetta Bussolati","doi":"10.1002/jex2.70159","DOIUrl":"https://doi.org/10.1002/jex2.70159","url":null,"abstract":"<p>Chronic kidney disease is characterized by progressive tubular injury and fibrosis, ultimately leading to irreversible loss of renal function. Extracellular vesicles (EVs) have emerged as promising therapeutic platforms due to their biocompatibility and engineering potential. In this study, we engineered red blood cell–derived EVs to deliver an active peptide (RBC-EVKP1) from the anti-aging hormone klotho, as a targeted antifibrotic strategy. EVs were isolated from healthy donors and surface-functionalized with the peptide. Fibrosis was evaluated in proximal tubular epithelial cells exposed to TGFβ and in a 3D proximal tubule-on-chip model. RBC-EVKP1 treatment significantly attenuated TGFβ-induced fibrotic and mesenchymal gene expression, reduced extracellular matrix accumulation, and suppressed SMAD signaling. In addition, RBC-EVKP1 reduced cell migration and preserved epithelial organization. In the tubule-on-chip system, engineered EVs maintained cytoskeletal integrity and reduced injury-associated marker expression under fibrotic conditions. Collectively, these findings demonstrate that klotho-engineered EVs effectively inhibit TGFβ-driven fibrosis and preserve tubular integrity, highlighting a promising therapeutic strategy for targeting kidney fibrosis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70159","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148386603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterisation and Comparative Evaluation of miRNA Contained in Small Extracellular Vesicles Isolated From Bovine Milk and Whey","authors":"Caterina Trevisan,&nbsp;Giulia Polacchini,&nbsp;Bruno Stefanon,&nbsp;Monica Colitti","doi":"10.1002/jex2.70165","DOIUrl":"10.1002/jex2.70165","url":null,"abstract":"<p>Small extracellular vesicles (EVs) from bovine milk and whey are emerging as biologically active carriers of microRNA (miRNA), yet their composition and variability among different dairy sources remain incompletely understood. We conducted an integrated characterisation of EV-associated miRNAs isolated from milk and whey collected from three dairy farms. EVs were purified and validated according to MISEV2023 criteria, and small RNA sequencing was performed on milk and whey samples collected on three different days at each farm (18 samples). After quality filtering and adaptor trimming, mapped reads were normalised to counts per million (CPM). To ensure robust detection, only miRNAs present in at least 67% of samples were retained, resulting in a high-confidence set of 329 miRNAs for downstream analyses. Milk-derived EVs showed a higher overall miRNA read count than whey-derived EVs (9.45 ± 5.3 million vs. 5.03 ± 2.5 million reads; <i>p</i> &lt; 0.10), with Farm Z displaying the greatest overall abundance. Stability assessment using <i>Z</i>-scored coefficients of variation demonstrated that miRNA reproducibility varied considerably between farms and collections, indicating that farm- and collection-specific factors, rather than matrix origin alone, were major contributors to expression variability. Differential expression analysis (DESeq2) identified 32 miRNAs significantly modulated between milk and whey EVs, with most (<i>n</i> = 29) enriched in whey. Functional enrichment of predicted targets indicated involvement in immune and inflammation-related pathways, including NF-κB signalling, cytokine–receptor interaction and Ras signalling. Pairwise comparison of farms revealed only a small number of differentially expressed miRNAs in both matrices, and KEGG analyses did not identify significant pathways after multiple-testing correction. This study presents a comparative overview of bovine milk and whey EV-associated miRNAs, highlighting higher miRNA abundance in milk, significant farm-dependent variability, and enrichment of immune-related pathways that distinguish the two matrices.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13317565/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148364304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exosome Secretion Drives Chemo-Resistance of Temozolomide in Glioblastoma","authors":"Wenpei Li,&nbsp;Shaoping Jiang,&nbsp;Kun Liu,&nbsp;Zhi Li,&nbsp;Jun Yang,&nbsp;Weimin Liu,&nbsp;Yuanyu Huang,&nbsp;Minghui Yang,&nbsp;Shuai Guo,&nbsp;Shaobo Ruan","doi":"10.1002/jex2.70161","DOIUrl":"10.1002/jex2.70161","url":null,"abstract":"<p>Temozolomide (TMZ) is a first-line chemotherapeutic agent for the treatment of glioblastoma (GBM), while a majority of patients do not effectively respond to TMZ owing to the multiple resistance mechanisms. In this study, we found that TMZ treatment substantially increased the exosome secretion from GBM cell line. The secreted exosomes from TMZ-treated C6 cells maintained typical physicochemical properties, with only slight fluctuation in expression profile of surface markers compared to untreated cell-derived exosomes. Further investigation revealed that increased expression of p53 might be the predominant reason to promote exosome secretion, potentially through the upregulation of six-transmembrane epithelial antigen of prostate 3 (STEAP3) expression under stress condition. Moreover, we identified that pre-treatment with TMZ-induced exosomes could reduce the sensitivity of C6 cells to TMZ and considered that exosome secretion may serve as a potent TMZ resistance mechanism via enhancing anti-apoptotic ability of GBM cells towards TMZ exposure. Proteomics analysis revealed a strategic mechanism to resist TMZ-induced apoptosis by upregulating exosomal proteins associated with biogenesis, chemoresistance, and therapeutic adaption. This finding revealed a considerable TMZ resistance mechanism and provided a new inspiration for preventing exosome biogenesis to resensitise TMZ cytotoxicity towards GBM.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13317785/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148364253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Salivary Extracellular Vesicles: Paradigm Shift in Liquid Biopsy Diagnostics","authors":"Kwanele Xulu,&nbsp;Usri H. Ibrahim,&nbsp;Irene Mackraj,&nbsp;Carola Niesler","doi":"10.1002/jex2.70166","DOIUrl":"10.1002/jex2.70166","url":null,"abstract":"<p>Extracellular vesicles (EVs), lipid bilayer nanoparticles released by virtually all cells, serve as essential messengers for intercellular communication. Due to their involvement in several pathophysiological processes, EVs have recently gained considerable attention as potentially diagnostic and prognostic biomarkers for various illnesses. The widespread distribution of EVs across all biofluids positions them as ideal, minimally invasive biomarkers for disease progression using a liquid biopsy approach. Among biofluids, saliva is uniquely accessible and has a low soluble protein content, making its EV population a highly promising source of diagnostic biomarkers. Salivary EVs have been investigated for their promising potential for diagnosing local and systemic diseases, including cancers, autoimmune diseases and neuropsychiatric disorders. In this review, we present a synopsis of the current landscape related to salivary EVs, highlighting the unique characteristics and potential of these vesicles, the technical challenges related to their application, and their future prospects for clinical translation as powerful diagnostic tools.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13317764/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148364276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bovine Milk–Derived Extracellular Vesicles as Emerging Drug Delivery Platforms: Current Advances, Applications and Challenges","authors":"Martina Brattini,&nbsp;Michelle D'Urso,&nbsp;Sofia Mariotto,&nbsp;Elena Butturini","doi":"10.1002/jex2.70160","DOIUrl":"https://doi.org/10.1002/jex2.70160","url":null,"abstract":"<p>Extracellular vesicles (EVs) are nanosized lipid bilayer particles naturally secreted by cells, mediating intercellular communication and transporting diverse bioactive molecules. Among alternative EV sources, bovine milk-derived EVs (BMEVs) have emerged as promising platforms for drug delivery due to their accessibility, scalability, stability and promising biocompatibility, although their long-term safety profile, particularly for engineered or cargo-loaded formulations, still requires further investigation. BMEVs can encapsulate small molecules, natural polyphenols and nucleic acids, enhancing bioavailability, protecting cargo from degradation and facilitating functional delivery. Recent studies demonstrate their potential for oral administration and their intrinsic therapeutic properties, including antioxidant, anti-inflammatory, and immunomodulatory activities. Moreover, the increasing number of patents highlights their translational and commercial relevance in therapeutic, cosmetic and nutraceutical applications. Despite these promising features, challenges remain in standardizing isolation protocols, optimizing cargo loading, ensuring batch reproducibility and evaluating long-term safety. Addressing these gaps is essential to enable clinical translation and establish BMEVs as versatile drug delivery platforms.</p><p><b>Trial Registration</b>: ClinicalTrials.gov identifier: NCT07402083</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 7","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70160","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148324888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stage-Specific Molecular Cargo of Schwann Cell–Derived Extracellular Vesicles is Associated With Peripheral Nerve Repair","authors":"Manju Sharma,&nbsp;Supasek Kongsomros,&nbsp;Maulee Sheth,&nbsp;Somchai Chutipongtanate,&nbsp;Leyla Esfandiari","doi":"10.1002/jex2.70157","DOIUrl":"https://doi.org/10.1002/jex2.70157","url":null,"abstract":"<p>Schwann cells (SCs) play a critical role in peripheral nerve regeneration, undergoing dynamic phenotype transitions from myelinating to repair stages following injury. While SC-derived extracellular vesicles (SC-EVs) have emerged as key mediators of intercellular communication during nerve repair, their stage-specific molecular cargo and functional roles remained incompletely understood. Here, we delineate protein, microRNA and long non-coding RNA (lncRNA) landscapes of SC-EVs across distinct differentiation stages, including immature, myelinating, and repair phenotypes, using an in vitro model of primary rat SCs. We show that repair SC-EVs carry distinct microRNAs predicted to modulate genes involved in myelin ensheathment, neuronal differentiation, and neurogenesis. Treatment of immature Schwann cells with repair SC-EVs increased SOX2 expression, suggesting activation of repair-associated Schwann cell responses. Furthermore, modulation of miR-330-5p, miRNA identified in repair SC-EVs, altered SOX2 expression, Schwann cell proliferation, and migration. Repair SC-EVs also contain lncRNAs that may bind and sequester miRNAs, potentially relieving repression of pro-regenerative genes. These findings suggest possible mechanisms by which SC-EVs may regulate Schwann cell repair functions and provide a molecular framework for mechanistic studies.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 6","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70157","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148324028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cell Envelope Remodeling and Lipid Metabolism Coordinate Extracellular Vesicle Output in Mycobacterium Tuberculosis","authors":"José L. Serrano-Mestre,&nbsp;Laura Lerma,&nbsp;Lucía Vázquez-Iniesta,&nbsp;Vivian C. Salgueiro-Toledo,&nbsp;Ainhoa Palacios,&nbsp;Sogol Alebouyeh,&nbsp;Andrea Doddi,&nbsp;Felix Elortza,&nbsp;Mikel Azkargorta,&nbsp;Leticia Sampedro,&nbsp;Estibaliz Atondo,&nbsp;Miguel Ángel Pacual-Itoiz,&nbsp;Oren Mayer,&nbsp;Torin Westwood,&nbsp;William R. Jacobs Jr,&nbsp;Rafael Prados-Rosales","doi":"10.1002/jex2.70156","DOIUrl":"https://doi.org/10.1002/jex2.70156","url":null,"abstract":"<p>Tuberculosis (TB), caused by <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>), remains a leading global cause of death due to the pathogen's highly adaptive physiology and its ability to manipulate host immunity. Despite extensive study, our understanding of <i>Mtb</i> biology during infection remains incomplete, limiting the development of new diagnostics, therapeutic and vaccine strategies. The <i>Mtb</i> cell envelope and its associated secretion systems are central to virulence and immune evasion. <i>Mtb</i> releases extracellular vesicles (EVs) that carry immunomodulatory molecules, including lipoproteins, which influence immune responses. Although individual genes and environmental cues have been shown to alter vesiculogenesis, a genome-wide dissection of EV biogenesis has not been attempted. Here, we establish a high-throughput screening system to identify <i>Mtb</i> mutants with altered EV production using a 96-well filtration platform combined with a lipophilic dye or an <i>Mtb</i>-EV (MEV)-specific antibody. Our genome-wide screen revealed genes in cell envelope remodeling and fatty acid metabolism are major regulators of vesiculogenesis. These findings provide a robust platform for defining the molecular determinants of MEV production and lay the groundwork for mechanistic characterization of vesicle biogenesis in <i>Mtb</i>.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 6","pages":""},"PeriodicalIF":4.8,"publicationDate":"2026-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148282135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular Vesicles of Giardia duodenalis: Unravelling Their Virulence Factors and Potential to Induce Protection Against Experimental Giardiasis","authors":"Corral-Ruiz Gerardo,&nbsp;Argüello-García Raúl,&nbsp;Ortiz-Lozano Dalia,&nbsp;Salazar-Villatoro Lizbeth,&nbsp;Pech-Santiago Edar Onam,&nbsp;Espinosa-Cantellano Martha,&nbsp;Hernández-Cueto Daniel,&nbsp;Silva-Olivares D. Angélica,&nbsp;Betanzos Abigail,&nbsp;Ortega-Pierres M. Guadalupe","doi":"10.1002/jex2.70155","DOIUrl":"https://doi.org/10.1002/jex2.70155","url":null,"abstract":"<p>Extracellular vesicles (EVs) are key mediators of intercellular communication, enabling trans-kingdom exchange of proteins, nucleic acids, lipids, and metabolites. In <i>Giardia duodenalis</i>, EVs contribute to parasite survival, host–pathogen interactions, and immune modulation. In this study, EVs were isolated from trophozoites grown in two serum-free media of distinct nutritional complexity (TYI-33 and DMEM) and characterized to define their molecular composition, pathogenic potential, and immunogenic properties. Nanoparticle tracking analysis showed vesicles predominantly 51–150 nm in diameter. Proteomic profiling identified 227 proteins in TYI-33 EVs and 97 in DMEM EVs, including molecules involved in metabolism, antioxidant defence, and virulence. Among these, giardipains 1–3, enolase, fructose-bisphosphate aldolase, ornithine carbamoyltransferase, and arginine deiminase were detected. TYI-33 EVs induced disruption of epithelial junctions and apoptotic-like changes in IEC-6 monolayers, consistent with contact-independent cytopathic activity. To assess their immunoprotective capacity, gerbils were intranasally immunized with TYI-33 EVs and challenged with <i>G. duodenalis</i>. Immunized animals showed significant reductions in intestinal trophozoite burden and faecal cyst shedding at 7, 14, and 21 days post-infection, accompanied by increased serum IL-17F and IL-22. Collectively, these findings indicate that <i>Giardia</i> EVs, while carrying virulence-associated factors, also possess strong immunogenic properties, supporting their potential as a novel mucosal vaccine platform against giardiasis.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"5 6","pages":""},"PeriodicalIF":0.0,"publicationDate":"2026-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.70155","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148237790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}