{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,9]],"date-time":"2025-09-09T21:48:26Z","timestamp":1757454506422,"version":"3.41.2"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,8,18]],"date-time":"2022-08-18T00:00:00Z","timestamp":1660780800000},"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. Immunol."],"abstract":"<jats:p>Melioidosis is a fatal infectious disease caused by <jats:italic>Burkholderia pseudomallei<\/jats:italic>. Complications following treatment are usually due to antibiotic resistance and relapse is mainly caused by <jats:italic>B. pseudomallei<\/jats:italic> biofilm. Although the release of neutrophil extracellular traps (NETs) is crucial to capture and eliminate bacterial pathogens, to date response of NETs to <jats:italic>B. pseudomallei<\/jats:italic> biofilm is poorly understood. Here we compare the NETs produced by neutrophils in response to <jats:italic>B. pseudomallei<\/jats:italic> H777 (a biofilm-producing strain containing the <jats:italic>bpsl0618<\/jats:italic> gene), a biofilm-defect strain lacking this gene (<jats:italic>B. pseudomallei<\/jats:italic> M10) and a <jats:italic>bpsl0618<\/jats:italic> biofilm-complemented strain, <jats:italic>B. pseudomallei<\/jats:italic> C17, in which function of <jats:italic>bpsl0618<\/jats:italic> was restored. Co-cultivation of these strains with healthy human neutrophils at MOI 10 with or without cytochalasin D demonstrated that H777 significantly resisted neutrophil-mediated killing and non-phagocytotic mechanisms compared to M10 (<jats:italic>p<\/jats:italic> &amp;lt; 0.0001). Three distinct morphotypes of NETs were seen: \u201caggregated\u201d, \u201cspiky\u201d and \u201ccloudy\u201d. These were induced in different proportions by the different bacterial strains. All types of NETs were shown to confine all <jats:italic>B. pseudomallei<\/jats:italic> strains. Strains H777 and C17 could stimulate production of twice as much extracellular DNA (234.62 ng\/mL and 205.43 ng\/mL, respectively) as did M10 (111.87 ng\/mL). Cells of H777 and C17 were better able to survive in the presence of neutrophil killing mechanisms relative to M10 (<jats:italic>p<\/jats:italic> &amp;lt; 0.0001) and NET formation (<jats:italic>p<\/jats:italic> &amp;lt; 0.0001 and 0.05). These findings suggest that NET stimulation was insufficient to eradicate <jats:italic>B. pseudomallei<\/jats:italic> H777 and C17 despite their possession of <jats:italic>bpsl0618<\/jats:italic>, a sugar-transferase gene associated with biofilm formation ability. Our findings demonstrate that <jats:italic>B. pseudomallei<\/jats:italic> biofilm phenotype may be a key factor in assisting pathogens to escape killing by neutrophils. This work provides a better understanding of how <jats:italic>B. pseudomallei<\/jats:italic> biofilm-associated infections induce and survive NET formation, resulting in bacterial persistence and increased severity of disease.<\/jats:p>","DOI":"10.3389\/fimmu.2022.926788","type":"journal-article","created":{"date-parts":[[2022,8,18]],"date-time":"2022-08-18T07:55:22Z","timestamp":1660809322000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Burkholderia pseudomallei biofilm phenotypes confined but surviving in neutrophil extracellular traps of varying 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