{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,20]],"date-time":"2026-05-20T03:16:02Z","timestamp":1779246962318,"version":"3.51.4"},"reference-count":45,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,5,8]],"date-time":"2025-05-08T00:00:00Z","timestamp":1746662400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p><jats:italic>Klebsiella variicola<\/jats:italic> is a member of <jats:italic>Klebsiella pneumoniae<\/jats:italic> complex and an emerging zoonotic pathogen. As part of the lymphatic system, the spleen plays a pivotal role in destroying invading pathogens. Various microRNAs (miRNAs) are involved in host resistance to pathogens. However, specific miRNAs that act against <jats:italic>K. variicola<\/jats:italic> remain unknown. Therefore, RNA sequencing (RNA-Seq) of the miRNA profile of the chicken spleen was conducted to further clarify the host immune response to <jats:italic>K. variicola<\/jats:italic> infection. Challenge of 7-day-old chicks with <jats:italic>K. variicola<\/jats:italic> strain AHKV-S01 caused severe damage and enlargement of the spleen. In total, 22 differentially expressed (DE) miRNAs (fold change&amp;gt;2, <jats:italic>q<\/jats:italic>&amp;lt; 0.05) were identified. Functional annotation analysis of the target genes of DE miRNAs found that signaling pathways related to innate immunity, inflammation, and metabolism were significantly enriched. Notably, expression of gga-miR-2954 was significantly upregulated in the infection group as compared to the control group. <jats:italic>In vitro<\/jats:italic>, gga-miR-2954 directly repressed luciferase reporter gene activity by binding to the 3\u2032 untranslated regions of <jats:italic>STAB1<\/jats:italic>. Overexpression of gga-miR-2954 in HD11 macrophages significantly inhibited expression of <jats:italic>STAB1<\/jats:italic>, which is involved in activation of several proinflammatory cytokines. <jats:italic>K. variicola<\/jats:italic> induced damage to the spleen by over activation of inflammatory and innate immune responses. The observed changes to the miRNA expression profile of the chicken spleen elucidate host immune responses to <jats:italic>K. variicola<\/jats:italic> infection, providing critical insights for developing novel therapeutic strategies to enhance chicken resistance against this pathogen.<\/jats:p>","DOI":"10.3389\/fcimb.2025.1544506","type":"journal-article","created":{"date-parts":[[2025,5,8]],"date-time":"2025-05-08T06:04:42Z","timestamp":1746684282000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["MicroRNA warfare: how chickens combat Klebsiella variicola with gga-miR-2954"],"prefix":"10.3389","volume":"15","author":[{"given":"Lei","family":"Yin","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuehuai","family":"Shen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dongdong","family":"Yin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hongyan","family":"Hou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jieru","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruihong","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kezong","family":"Qi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yin","family":"Dai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaocheng","family":"Pan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,5,8]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1152","DOI":"10.1016\/j.immuni.2023.04.004","article-title":"The spleen in local and systemic regulation of immunity","volume":"56","author":"Bronte","year":"2023","journal-title":"Immunity"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vetpar.2024.110200","article-title":"MicroRNA expression profile of chicken liver at different times after Histomonas meleagridis infection","volume":"329","author":"Chen","year":"2024","journal-title":"Vet Parasitol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"705","DOI":"10.1038\/s41591-020-0825-4","article-title":"Adaptive evolution of virulence and persistence in carbapenem-resistant Klebsiella pneumoniae","volume":"26","author":"Ernst","year":"2020","journal-title":"Nat. Med."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.01170-23","article-title":"The role of integration host factor in biofilm and virulence of high-alcohol-producing Klebsiella pneumoniae","volume":"11","author":"Fan","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1767","DOI":"10.2147\/idr.s404551","article-title":"Emergence of OXA-484-producing klebsiella variicola in China","volume":"16","author":"Ge","year":"2023","journal-title":"Infect. Drug Resist."},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00044-22","article-title":"Multidrug-resistant klebsiella variicola isolated in the urine of healthy bovine heifers, a potential risk as an emerging human pathogen","volume":"88","author":"Giannattasio-Ferraz","year":"2022","journal-title":"Appl. Environ. Microbiol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1425","DOI":"10.1083\/jcb.113.6.1425","article-title":"Identification of a novel high molecular weight protein preferentially expressed by sinusoidal endothelial cells in normal human tissues","volume":"113","author":"Goerdt","year":"1991","journal-title":"J. Cell Biol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"D154","DOI":"10.1093\/nar\/gkm952","article-title":"miRBase: tools for microRNA genomics","volume":"36","author":"Griffiths-Jones","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.psj.2022.102296","article-title":"The targeted anti-Salmonella bacteriophage attenuated the inflammatory response of laying hens challenged with Salmonella Gallinarum","volume":"102","author":"Hosseindoust","year":"2023","journal-title":"Poult. Sci."},{"key":"B10","doi-asserted-by":"publisher","first-page":"305","DOI":"10.3390\/nu15020305","article-title":"The pivotal role of aryl hydrocarbon receptor-regulated tight junction proteins and innate immunity on the synergistic effects of postbiotic butyrate and active vitamin D3 to defense against microbial invasion in salmonella colitis","volume":"15","author":"Huang","year":"2023","journal-title":"Nutrients"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1128\/iai.00688-16","article-title":"Novel microRNA involved in host response to avian pathogenic escherichia coli identified by deep sequencing and integration analysis","volume":"85","author":"Jia","year":"2017","journal-title":"Infect. Immun."},{"key":"B12","doi-asserted-by":"publisher","first-page":"8869510","DOI":"10.1155\/2024\/8869510","article-title":"A protective role of canonical wnt\/\u03b2-catenin pathway in pathogenic bacteria-induced inflammatory responses","volume":"27","author":"Jiang","year":"2024","journal-title":"Mediators Inflammation"},{"key":"B13","doi-asserted-by":"publisher","first-page":"1577","DOI":"10.1111\/j.1476-5381.2010.00872.x","article-title":"Animal research: reporting in vivo experiments: the ARRIVE guidelines","volume":"160","author":"Kilkenny","year":"2010","journal-title":"Br. J. Pharmacol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1007\/s00011-022-01542-w","article-title":"Wogonin inhibits tight junction disruption via suppression of inflammatory response and phosphorylation of AKT\/NF-\u03baB and ERK1\/2 in rhinovirus-infected human nasal epithelial cells","volume":"71","author":"Kim","year":"2022","journal-title":"Inflammation Res."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1128\/cmr.00015-23","article-title":"MicroRNAs in infectious diseases: potential diagnostic biomarkers and therapeutic targets","volume":"36","author":"Kimura","year":"2023","journal-title":"Clin. Microbiol. Rev."},{"key":"B16","doi-asserted-by":"publisher","first-page":"1637","DOI":"10.1093\/infdis\/jiad597","article-title":"Chlamydia trachomatis T3SS effector CT622 induces proinflammatory cytokines through TLR2\/TLR4-mediated MAPK\/NF-\u03baB pathways in THP-1 cells","volume":"229","author":"Lei","year":"2024","journal-title":"J. Infect. Dis."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1893","DOI":"10.2147\/idr.s460569","article-title":"Global Emergence and Genomic Epidemiology of bla(NDM)-Carrying Klebsiella variicola","volume":"17","author":"Li","year":"2024","journal-title":"Infect. Drug Resist."},{"key":"B18","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"B19","doi-asserted-by":"publisher","first-page":"699","DOI":"10.1093\/bib\/bby128","article-title":"miR+Pathway: the integration and visualization of miRNA and KEGG pathways","volume":"21","author":"Pian","year":"2020","journal-title":"Brief Bioinform."},{"key":"B20","doi-asserted-by":"publisher","first-page":"2088","DOI":"10.1080\/21505594.2021.1958606","article-title":"Stabilin-1 plays a protective role against Listeria monocytogenes infection through the regulation of cytokine and chemokine production and immune cell recruitment","volume":"12","author":"Pombinho","year":"2021","journal-title":"Virulence"},{"key":"B21","doi-asserted-by":"publisher","first-page":"3684","DOI":"10.1093\/bioinformatics\/btab319","article-title":"IntAct App: a Cytoscape application for molecular interaction network visualization and analysis","volume":"37","author":"Ragueneau","year":"2021","journal-title":"Bioinformatics"},{"key":"B22","doi-asserted-by":"publisher","first-page":"9298","DOI":"10.1073\/pnas.1604780113","article-title":"Stabilin-1 expression defines a subset of macrophages that mediate tissue homeostasis and prevent fibrosis in chronic liver injury","volume":"113","author":"Rantakari","year":"2016","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2022.111575","article-title":"Cholinergic receptor-Wnt pathway controls immune activation by sensing intestinal dysfunction","volume":"41","author":"Ren","year":"2022","journal-title":"Cell Rep."},{"key":"B24","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1080\/22221751.2019.1634981","article-title":"Klebsiella variicola: an emerging pathogen in humans","volume":"8","author":"Rodr\u00edguez-Medina","year":"2019","journal-title":"Emerg. Micro. Infect."},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.04393-22","article-title":"Role of Toll-Like Receptor 4 in Mycobacterium avium subsp. paratuberculosis Infection of Bovine Mammary Epithelial (MAC-T) Cells In Vitro","volume":"11","author":"Shandilya","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"B26","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1038\/ni.1828","article-title":"Negative regulation of TLR4 via targeting of the proinflammatory tumor suppressor PDCD4 by the microRNA miR-21","volume":"11","author":"Sheedy","year":"2010","journal-title":"Nat. Immunol."},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1016\/j.meegid.2021.104768","article-title":"Comprehensive genomic analysis and characterization of a new ST 174 type Klebsiella variicola strain isolated from chicken embryos","volume":"90","author":"Shen","year":"2021","journal-title":"Infect. Genet. Evol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1007\/s00442-003-1409-y","article-title":"On the use of spleen mass as a measure of avian immune system strength","volume":"138","author":"Smith","year":"2004","journal-title":"Oecologia"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2400446121","article-title":"Adaptive evolution of carbapenem-resistant hypervirulent Klebsiella pneumoniae in the urinary tract of a single patient","volume":"121","author":"Song","year":"2024","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.03429-23","article-title":"The Hippo kinases control inflammatory Hippo signaling and restrict bacterial infection in phagocytes","volume":"15","author":"St Louis","year":"2024","journal-title":"mBio"},{"key":"B31","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1186\/s40001-023-01484-x","article-title":"The role of non-canonical Hippo pathway in regulating immune homeostasis","volume":"28","author":"Tang","year":"2023","journal-title":"Eur. J. Med. Res."},{"key":"B32","first-page":"255","article-title":"Role of the spleen in immunity. Immunologic consequences of splenectomy","volume":"103","author":"Tiron","year":"2008","journal-title":"Chirurgia (Bucur)"},{"key":"B33","doi-asserted-by":"publisher","first-page":"110","DOI":"10.1186\/s13567-023-01240-3","article-title":"Chicken miR-148a-3p regulates immune responses against AIV by targeting the MAPK signalling pathway and IFN-\u03b3","volume":"54","author":"Vu","year":"2023","journal-title":"Vet Res."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1769","DOI":"10.3390\/cells12131769","article-title":"TLR4 overexpression aggravates bacterial lipopolysaccharide-induced apoptosis via excessive autophagy and NF-\u03baB\/MAPK signaling in transgenic mammal models","volume":"12","author":"Wang","year":"2023","journal-title":"Cells"},{"key":"B35","doi-asserted-by":"publisher","first-page":"322","DOI":"10.1016\/j.antiviral.2017.07.004","article-title":"Adenoviral-expressed recombinant granulocyte monocyte colony-stimulating factor (GM-CSF) enhances protective immunity induced by inactivated Newcastle Disease Virus (NDV) vaccine","volume":"144","author":"Wang","year":"2017","journal-title":"Antiviral Res."},{"key":"B36","doi-asserted-by":"publisher","first-page":"3397","DOI":"10.1007\/s00705-017-3502-2","article-title":"MicroRNA transcriptome analysis in chicken kidneys in response to differing virulent infectious bronchitis virus infections","volume":"162","author":"Yang","year":"2017","journal-title":"Arch. Virol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1080\/03079457.2023.2181146","article-title":"Identification and characterization of microRNAs, especially gga-miR-181b-5p, in chicken macrophages associated with avian pathogenic E. coli infection","volume":"52","author":"Yang","year":"2023","journal-title":"Avian Pathol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"W71","DOI":"10.1093\/nar\/gky400","article-title":"WEGO 2.0: a web tool for analyzing and plotting GO annotations 2018 update","volume":"46","author":"Ye","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2023.105029","article-title":"Integrated analysis of noncoding RNAs and mRNAs reveals their potential roles in chicken spleen response to Klebsiella variicola infection","volume":"164","author":"Yin","year":"2023","journal-title":"Res. Vet Sci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"2767","DOI":"10.1007\/s00253-016-8060-0","article-title":"Analysis of microRNA expression profile in specific pathogen-free chickens in response to reticuloendotheliosis virus infection","volume":"101","author":"Yu","year":"2017","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1016\/j.rvsc.2023.03.027","article-title":"Transcriptomics analysis reveals key lncRNAs and genes related to the infection of feline kidney cell line by panleukopenia virus","volume":"158","author":"Zhang","year":"2023","journal-title":"Res. Vet Sci."},{"key":"B42","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/s13567-024-01285-y","article-title":"Pasteurella multocida activates Rassf1-Hippo-Yap pathway to induce pulmonary epithelial apoptosis","volume":"55","author":"Zhao","year":"2024","journal-title":"Vet Res."},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1016\/j.mito.2024.101921","article-title":"Escherichia coli infection induces ferroptosis in bovine mammary epithelial cells by activating the Wnt\/\u03b2-catenin pathway-mediated mitophagy","volume":"78","author":"Zhuang","year":"2024","journal-title":"Mitochondrion"},{"key":"B44","doi-asserted-by":"publisher","first-page":"345","DOI":"10.24425\/pjvs.2019.129226","article-title":"Effects of porcine epidemic diarrhea virus infection on tight junction protein gene expression and morphology of the intestinal mucosa in pigs","volume":"22","author":"Zong","year":"2019","journal-title":"Pol. J. Vet Sci."},{"key":"B45","doi-asserted-by":"publisher","first-page":"2126","DOI":"10.3390\/microorganisms11082126","article-title":"Baicalin weakens the porcine exPEC-induced inflammatory response in 3D4\/21 cells by inhibiting the expression of NF-\u03baB\/MAPK signaling pathways and reducing NLRP3 inflammasome activation","volume":"11","author":"Zong","year":"2023","journal-title":"Microorganisms"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1544506\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,8]],"date-time":"2025-05-08T06:04:43Z","timestamp":1746684283000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2025.1544506\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,8]]},"references-count":45,"alternative-id":["10.3389\/fcimb.2025.1544506"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2025.1544506","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,8]]},"article-number":"1544506"}}