{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T19:49:35Z","timestamp":1791229775421,"version":"4.1.0"},"reference-count":55,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T00:00:00Z","timestamp":1788912000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T00:00:00Z","timestamp":1788912000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["FDN143248"],"award-info":[{"award-number":["FDN143248"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["FDN154327"],"award-info":[{"award-number":["FDN154327"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2026,9,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Staphylococcus aureus<\/jats:italic>\n                      forms biofilms on biotic and abiotic surfaces, enabling persistent infections that evade immune clearance. Although many\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      strains can produce the biofilm-associated exopolysaccharide, poly-\u03b2-1,6-N-acetyl-D-glucosamine (PNAG), they often form protein-dominated biofilm matrices\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      , leaving a role for PNAG unclear. Using intravital imaging in a foreign-body infection model, we found that PNAG in biofilms hinders neutrophil access and delays bacterial clearance. Neutrophil elastase was crucial for eventual biofilm clearance.\n                      <jats:italic toggle=\"yes\">In vivo<\/jats:italic>\n                      PNAG labeling revealed that the exopolysaccharide forms a physical barrier that prevents neutrophils from reaching bacterial clusters. In contrast, PNAG-deficient strains permitted greater neutrophil infiltration and were cleared more rapidly than wild-type bacteria. Enzymatic degradation of PNAG with the glycoside hydrolases PgaB or dispersin B (DspB) disrupted the biofilm, restored neutrophil access, and enhanced bacterial clearance. Together, these findings identify PNAG as a key structural barrier protecting\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      from innate immunity and suggest that targeting PNAG with glycoside hydrolases may offer a promising therapeutic strategy for biofilm-associated\n                      <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                      infections.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        The biofilm-associated exopolysaccharide PNAG is frequently expressed in\n                        <jats:italic toggle=\"yes\">Staphylococcus aureus<\/jats:italic>\n                        clinical isolates but is often reduced during laboratory passage, with expression highly dependent on growth conditions. While\n                        <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                        analyses have revealed that PNAG is not a dominant matrix component, our intravital imaging of community-acquired methicillin-resistant\n                        <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                        (CA-MRSA) skin infections demonstrates that PNAG is robustly produced\n                        <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                        and plays a central role in immune evasion. These findings highlight how PNAG function in tissue environments may be non-obvious\n                        <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                        and underscore the need for\n                        <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                        models to understand biofilm pathogenesis. By revealing PNAG as a key barrier to neutrophil-mediated clearance, this work positions PNAG and PNAG-targeting glycoside hydrolases as compelling therapeutic candidates for treating antibiotic-resistant\n                        <jats:italic toggle=\"yes\">S. aureus<\/jats:italic>\n                        biofilm infections, a major cause of morbidity in both healthcare and community settings.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.00522-26","type":"journal-article","created":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T13:00:44Z","timestamp":1785502844000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Imaging\n                    <i>Staphylococcus aureus<\/i>\n                    biofilms\n                    <i>in vivo<\/i>\n                    reveals neutrophil exclusion by PNAG exopolysaccharide"],"prefix":"10.1128","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3927-2508","authenticated-orcid":true,"given":"Rachel M.","family":"Kratofil","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03yjb2x39","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Physiology and Pharmacology, University of Calgary","place":["Calgary, Canada"]},{"id":[{"id":"https:\/\/ror.org\/03yjb2x39","id-type":"ROR","asserted-by":"publisher"}],"name":"Calvin, Phoebe, and Joan Snyder Institute for Chronic Diseases, University of Calgary","place":["Calgary, Canada"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rehnuma T.","family":"Sejuty","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05dte6685","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of Calgary","place":["Calgary, 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