{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T06:26:54Z","timestamp":1788848814838,"version":"build-2803163510"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T00:00:00Z","timestamp":1701043200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>\n                    We used cultured human conjunctival goblet cells to determine (i) whether the toxigenic\n                    <jats:italic>S. aureus-<\/jats:italic>\n                    induced activation of the epithelial goblet cells requires two signals to activate the NLRP3 inflammasome, (ii) if one signal is mediated by TLR1, TLR2, or TLR6, and (iii) if the\n                    <jats:italic>S. aureus<\/jats:italic>\n                    toxin \u03b1 toxin is another signal for the activation of the inflammasome and secretion of mature IL-1\u03b2. Cultured cells were incubated with siRNA to knock down the different TLRs. After stimulation with toxigenic\n                    <jats:italic>S. aureus<\/jats:italic>\n                    RN6390, pro-IL-1\u03b2 synthesis, caspase-1 activity, and mature IL-1\u03b2 secretion were measured. In a separate set of experiments, the cells were incubated with toxigenic\n                    <jats:italic>S. aureus<\/jats:italic>\n                    RN6390 or mutant\n                    <jats:italic>S. aureus<\/jats:italic>\n                    ALC837 that does not express \u03b1 toxin with or without exogenous \u03b1 toxin. A gentamicin protection assay was used to determine if intracellular bacteria were active. We conclude that \u03b1 toxin from toxigenic\n                    <jats:italic>S. aureus<\/jats:italic>\n                    triggers two separate mechanisms required for the activation of the NLRP3 inflammasome and secretion of mature IL-1\u03b2. In the first mechanism, \u03b1 toxin secreted from internalized\n                    <jats:italic>S. aureus<\/jats:italic>\n                    produces a pore, allowing the internalized bacteria and associated pathogen-associated molecular patterns to interact with intracellular TLR2 and, to a lesser extent, TLR1. In the second mechanism, \u03b1 toxin forms a pore in the plasma membrane, leading to an efflux of cytosolic K\n                    <jats:sup>+<\/jats:sup>\n                    and influx of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    . We conclude that \u03b1 toxin by these two different mechanisms triggers the synthesis of pro-IL-1\u03b2 and NLRP3 components, activation of capase-1, and secretion of mature IL-1\u03b2 to defend against bacterial infection.\n                  <\/jats:p>","DOI":"10.3389\/fcimb.2023.1265471","type":"journal-article","created":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T01:02:25Z","timestamp":1701046945000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Staphylococcus aureus activates NRLP3-dependent IL-1\u03b2 secretion from human conjunctival goblet cells using \u03b1 toxin and toll-like receptors 2 and 1"],"prefix":"10.3389","volume":"13","author":[{"given":"Dayu","family":"Li","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robin","family":"Hodges","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"AukrustNaqvi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey","family":"Bair","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulo J. M.","family":"Bispo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael S.","family":"Gilmore","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meredith","family":"Gregory-Ksander","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darlene A.","family":"Dartt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,11,27]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"24035","DOI":"10.1074\/jbc.M109.014266","article-title":"Biglycan, a danger signal that activates the NLRP3 inflammasome via toll-like and P2X receptors","volume":"284","author":"Babelova","year":"2009","journal-title":"J. Biol. Chem."},{"key":"B2","doi-asserted-by":"publisher","first-page":"787","DOI":"10.4049\/jimmunol.0901363","article-title":"Cutting edge: NF-kappaB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression","volume":"183","author":"Bauernfeind","year":"2009","journal-title":"J. Immunol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1140","DOI":"10.3390\/toxins5061140","article-title":"Staphylococcus aureus alpha-toxin: nearly a century of intrigue","volume":"5","author":"Berube","year":"2013","journal-title":"Toxins."},{"key":"B4","doi-asserted-by":"publisher","first-page":"e80743","DOI":"10.1371\/journal.pone.0080743","article-title":"TLR2 ligands induce NF-kappaB activation from endosomal compartments of human monocytes","volume":"8","author":"Brandt","year":"2013","journal-title":"PloS One"},{"key":"B5","doi-asserted-by":"publisher","first-page":"37169","DOI":"10.1074\/jbc.274.52.37169","article-title":"SarA, a global regulator of virulence determinants in Staphylococcus aureus, binds to a conserved motif essential for sar-dependent gene regulation","volume":"274","author":"Chien","year":"1999","journal-title":"J. Biol. Chem."},{"key":"B6","doi-asserted-by":"publisher","first-page":"e1003047","DOI":"10.1371\/journal.ppat.1003047","article-title":"Neutrophil-derived IL-1beta is sufficient for abscess formation in immunity against Staphylococcus aureus in mice","volume":"8","author":"Cho","year":"2012","journal-title":"PloS pathogens."},{"key":"B7","doi-asserted-by":"publisher","first-page":"e7446","DOI":"10.1371\/journal.pone.0007446","article-title":"Staphylococcus aureus alpha-hemolysin activates the NLRP3-inflammasome in human and mouse monocytic cells","volume":"4","author":"Craven","year":"2009","journal-title":"PloS One"},{"key":"B8","doi-asserted-by":"publisher","first-page":"4455","DOI":"10.4049\/jimmunol.1000833","article-title":"Conjunctival goblet cell secretion stimulated by leukotrienes is reduced by resolvins D1 and E1 to promote resolution of inflammation","volume":"186","author":"Dartt","year":"2011","journal-title":"J. Immunol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1111\/imr.12286","article-title":"Initiation and perpetuation of NLRP3 inflammasome activation and assembly","volume":"265","author":"Elliott","year":"2015","journal-title":"Immunol. Rev."},{"key":"B10","doi-asserted-by":"publisher","first-page":"241","DOI":"10.1038\/ni.1703","article-title":"The inflammasome: a caspase-1-activation platform that regulates immune responses and disease pathogenesis","volume":"10","author":"Franchi","year":"2009","journal-title":"Nat. Immunol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1111\/j.1463-5224.2010.00844.x","article-title":"Expression of Toll-like receptors 2, 3, 4, 6, 9, and MD-2 in the normal equine cornea, limbus, and conjunctiva","volume":"14","author":"Gornik","year":"2011","journal-title":"Vet. Ophthalmol."},{"key":"B12","doi-asserted-by":"publisher","first-page":"5823","DOI":"10.4049\/jimmunol.0900444","article-title":"Activation of the Nlrp3 inflammasome by Streptococcus pyogenes requires streptolysin O and NF-kappa B activation but proceeds independently of TLR signaling and P2X7 receptor","volume":"183","author":"Harder","year":"2009","journal-title":"J. Immunol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/S0022-4804(03)00314-7","article-title":"Intracellular survival of Staphylococcus aureus within cultured enterocytes","volume":"114","author":"Hess","year":"2003","journal-title":"J. Surg. Res."},{"key":"B14","doi-asserted-by":"publisher","first-page":"517","DOI":"10.1165\/rcmb.2013-0047OC","article-title":"The nucleotide-binding domain and leucine-rich repeat protein-3 inflammasome is not activated in airway smooth muscle upon toll-like receptor-2 ligation","volume":"49","author":"Hirota","year":"2013","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"12873","DOI":"10.1073\/pnas.0904958106","article-title":"NOD2 contributes to cutaneous defense against Staphylococcus aureus through alpha-toxin-dependent innate immune activation","volume":"106","author":"Hruz","year":"2009","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B16","doi-asserted-by":"publisher","first-page":"16","DOI":"10.1016\/j.molmed.2013.10.002","article-title":"License for destruction: tumor-specific cytokine targeting","volume":"20","author":"Johansson","year":"2014","journal-title":"Trends Mol. Med."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.3390\/ijms20133328","article-title":"The NLRP3 inflammasome: an overview of mechanisms of activation and regulation","volume":"20","author":"Kelley","year":"2019","journal-title":"Int. J. Mol. Sci."},{"key":"B18","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1369\/jhc.6A7101.2006","article-title":"Inflammasome components NALP 1 and 3 show distinct but separate expression profiles in human tissues suggesting a site-specific role in the inflammatory response","volume":"55","author":"Kummer","year":"2007","journal-title":"J. Histochem Cytochem."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1013","DOI":"10.1016\/j.cell.2014.04.007","article-title":"Mechanisms and functions of inflammasomes","volume":"157","author":"Lamkanfi","year":"2014","journal-title":"Cell."},{"key":"B20","doi-asserted-by":"publisher","first-page":"6928","DOI":"10.1167\/iovs.12-10163","article-title":"Effect of histamine on Ca(2+)-dependent signaling pathways in rat conjunctival goblet cells","volume":"53","author":"Li","year":"2012","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B21","doi-asserted-by":"publisher","first-page":"e000101","DOI":"10.1136\/bmjophth-2017-000101","article-title":"Neither non-toxigenic Staphylococcus aureus nor commensal S. epidermidi activates NLRP3 inflammasomes in human conjunctival gobelt cells","volume":"2","author":"Li","year":"2016","journal-title":"BMJ Open Ophthalmology."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1119","DOI":"10.1038\/mi.2013.7","article-title":"Resolvin D1 and aspirin-triggered resolvin D1 regulate histamine-stimulated conjunctival goblet cell secretion","volume":"6","author":"Li","year":"2013","journal-title":"Mucosal Immunol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"5661","DOI":"10.1167\/iovs.13-12473","article-title":"Role of PKCalpha activation of Src, PI-3K\/AKT, and ERK in EGF-stimulated proliferation of rat and human conjunctival goblet cells","volume":"54","author":"Li","year":"2013","journal-title":"Invest. Ophthalmol. Visual science."},{"key":"B24","doi-asserted-by":"publisher","first-page":"e74010","DOI":"10.1371\/journal.pone.0074010","article-title":"Staphylococcus aureus activates the NLRP3 inflammasome in human and rat conjunctival goblet cells","volume":"8","author":"McGilligan","year":"2013","journal-title":"PloS One"},{"key":"B25","doi-asserted-by":"publisher","first-page":"3942","DOI":"10.4049\/jimmunol.0900729","article-title":"A critical role for hemolysins and bacterial lipoproteins in Staphylococcus aureus-induced activation of the Nlrp3 inflammasome","volume":"183","author":"Munoz-Planillo","year":"2009","journal-title":"J. Immunol."},{"key":"B26","doi-asserted-by":"publisher","first-page":"2324","DOI":"10.1182\/blood-2008-03-146720","article-title":"Differential requirement for the activation of the inflammasome for processing and release of IL-1beta in monocytes and macrophages","volume":"113","author":"Netea","year":"2009","journal-title":"Blood."},{"key":"B27","doi-asserted-by":"publisher","first-page":"1867","DOI":"10.1517\/14712590802494212","article-title":"The role of NLRs and TLRs in the activation of the inflammasome","volume":"8","author":"Netea","year":"2008","journal-title":"Expert Opin. Biol. Ther."},{"key":"B28","doi-asserted-by":"publisher","first-page":"280","DOI":"10.1189\/jlb.0907656","article-title":"Cellular trafficking of lipoteichoic acid and Toll-like receptor 2 in relation to signaling: role of CD14 and CD36","volume":"84","author":"Nilsen","year":"2008","journal-title":"J. Leukoc. Biol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"4365","DOI":"10.1128\/jb.170.9.4365-4372.1988","article-title":"Cloning, characterization, and sequencing of an accessory gene regulator (agr) in Staphylococcus aureus","volume":"170","author":"Peng","year":"1988","journal-title":"J. Bacteriol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"799","DOI":"10.5301\/ijao.5000030","article-title":"Toll-like receptors (TLRs) in innate immune defense against Staphylococcus aureus","volume":"34","author":"Pietrocola","year":"2011","journal-title":"Int. J. Artif. Organs."},{"key":"B31","doi-asserted-by":"publisher","first-page":"442","DOI":"10.1016\/j.it.2015.06.002","article-title":"Inflammasomes of the intestinal epithelium","volume":"36","author":"Sellin","year":"2015","journal-title":"Trends Immunol."},{"key":"B32","doi-asserted-by":"publisher","first-page":"617","DOI":"10.1083\/jcb.201602089","article-title":"The cell biology of inflammasomes: Mechanisms of inflammasome activation and regulation","volume":"213","author":"Sharma","year":"2016","journal-title":"J. Cell Biol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"2477","DOI":"10.1167\/iovs.02-0550","article-title":"Isolation and characterization of cultured human conjunctival goblet cells","volume":"44","author":"Shatos","year":"2003","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"31002","DOI":"10.1074\/jbc.M602794200","article-title":"Membrane sorting of toll-like receptor (TLR)-2\/6 and TLR2\/1 heterodimers at the cell surface determines heterotypic associations with CD36 and intracellular targeting","volume":"281","author":"Triantafilou","year":"2006","journal-title":"J. Biol. Chem."},{"key":"B35","doi-asserted-by":"publisher","first-page":"110","DOI":"10.1167\/iovs.12-10655","article-title":"NLRP3 inflammasome activation in retinal pigment epithelial cells by lysosomal destabilization: implications for age-related macular degeneration","volume":"54","author":"Tseng","year":"2013","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"B36","doi-asserted-by":"publisher","first-page":"3337","DOI":"10.4049\/jimmunol.173.5.3337","article-title":"Intracellularly expressed TLR2s and TLR4s contribution to an immunosilent environment at the ocular mucosal epithelium","volume":"173","author":"Ueta","year":"2004","journal-title":"J. Immunol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"3174","DOI":"10.1073\/pnas.1915829117","article-title":"Orchestration of human macrophage NLRP3 inflammasome activation by Staphylococcus aureus extracellular vesicles","volume":"117","author":"Wang","year":"2020","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B38","doi-asserted-by":"publisher","first-page":"7125","DOI":"10.4049\/jimmunol.165.12.7125","article-title":"Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses","volume":"165","author":"Wyllie","year":"2000","journal-title":"J. Immunol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"1","DOI":"10.4049\/jimmunol.163.1.1","article-title":"Cutting edge: recognition of Gram-positive bacterial cell wall components by the innate immune system occurs via Toll-like receptor 2","volume":"163","author":"Yoshimura","year":"1999","journal-title":"J. Immunol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"344","DOI":"10.1111\/cei.12458","article-title":"Serum amyloid A induces interleukin-1beta secretion from keratinocytes via the NACHT, LRR and PYD domains-containing protein 3 inflammasome","volume":"179","author":"Yu","year":"2015","journal-title":"Clin. Exp. Immunol."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1265471\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T01:02:45Z","timestamp":1701046965000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1265471\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,27]]},"references-count":40,"alternative-id":["10.3389\/fcimb.2023.1265471"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1265471","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,11,27]]},"article-number":"1265471"}}