{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T01:15:15Z","timestamp":1790903715952,"version":"4.1.0"},"reference-count":81,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T00:00:00Z","timestamp":1767744000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T00:00:00Z","timestamp":1767744000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100013281","name":"Joint Programming Initiative on Antimicrobial Resistance","doi-asserted-by":"publisher","award":["Co-Action"],"award-info":[{"award-number":["Co-Action"]}],"id":[{"id":"10.13039\/100013281","id-type":"DOI","asserted-by":"publisher"}]},{"name":"EU Horizon Europe DIGIBIO project","award":["101060066"],"award-info":[{"award-number":["101060066"]}]},{"DOI":"10.13039\/501100004543","name":"China Scholarship Council","doi-asserted-by":"publisher","award":["none"],"award-info":[{"award-number":["none"]}],"id":[{"id":"10.13039\/501100004543","id-type":"DOI","asserted-by":"publisher"}]},{"name":"UKRI Future Leaders Fellowship","award":["MR\/V027204\/1"],"award-info":[{"award-number":["MR\/V027204\/1"]}]},{"name":"EOS (Excellence of Science) program, Belgian FNRS-FRS","award":["O.0011.22"],"award-info":[{"award-number":["O.0011.22"]}]},{"name":"Belgian Fonds de la Recherche Scientifique","award":["T.0189.16"],"award-info":[{"award-number":["T.0189.16"]}]},{"name":"Belgian Fonds de la Recherche Scientifique","award":["T.0205.22"],"award-info":[{"award-number":["T.0205.22"]}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2026,1,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      Antibiotic-resistant bacteria, particularly the ESKAPE (\n                      <jats:italic toggle=\"yes\">Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa,<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">Enterobacter<\/jats:italic>\n                      spp.) pathogens, pose a major public health threat. Their ability to reside inside cells contributes to their persistence and resistance. Combining rifampicin with polymyxins is a much-discussed approach against multidrug-resistant Gram-negative bacterial infections. We therefore evaluated a combination of polymyxin B and rifampicin against Gram-negative clinical isolates in extracellular and intracellular\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      models of infection. The combination was synergistic against intra- and extra-cellular forms of\n                      <jats:italic toggle=\"yes\">P. aeruginosa, A. baumannii, E. coli<\/jats:italic>\n                      , and\n                      <jats:italic toggle=\"yes\">K. pneumoniae<\/jats:italic>\n                      . This synergy was enhanced in an acidic environment resembling the host vacuole where intracellular bacteria reside. The combination remained synergistic against rifampicin and polymyxin B-resistant\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      . To reveal the molecular underpinnings of the synergy, we used reverse genetics to identify and describe\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      mutants more susceptible to the combination. They show altered membrane properties and more pronounced oxidative damage when exposed to the combination. This work sheds a new light on the mechanisms of the synergy between rifampicin and polymyxins, demonstrates its applicability to Gram-negative ESKAPE pathogens, including when residing intracellularly. Overall, the data suggest that repurposing rifampicin with polymyxin B can effectively target hard-to-eradicate intracellular bacteria.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aac.01319-25","type":"journal-article","created":{"date-parts":[[2025,11,28]],"date-time":"2025-11-28T12:49:32Z","timestamp":1764334172000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Synergistic activity of rifampicin and polymyxin B against intracellular Gram-negative ESKAPE pathogens involves bacterial membrane alterations and enhanced oxidative damages"],"prefix":"10.1128","volume":"70","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6900-6986","authenticated-orcid":true,"given":"Vallo","family":"Varik","sequence":"first","affiliation":[{"name":"Pharmacologie cellulaire et mol\u00e9culaire, Louvain Drug Research Institute, Universit\u00e9 catholique de Louvain","place":["Brussels, Belgium"]},{"id":[{"id":"https:\/\/ror.org\/010jaxs89","id-type":"ROR","asserted-by":"publisher"}],"name":"European Molecular Biology Laboratory","place":["Heidelberg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-0228-0120","authenticated-orcid":false,"given":"Gang","family":"Wang","sequence":"additional","affiliation":[{"name":"Pharmacologie cellulaire et mol\u00e9culaire, Louvain Drug Research Institute, Universit\u00e9 catholique de Louvain","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1434-5176","authenticated-orcid":true,"given":"George","family":"Kritikos","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/010jaxs89","id-type":"ROR","asserted-by":"publisher"}],"name":"European Molecular Biology Laboratory","place":["Heidelberg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4682-1037","authenticated-orcid":true,"given":"Manuel","family":"Banzhaf","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00eae9z71","id-type":"ROR","asserted-by":"publisher"}],"name":"Newcastle University Biosciences Institute, Faculty of Medical Sciences, Newcastle University","place":["Newcastle upon Tyne, United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilien","family":"Drouot","sequence":"additional","affiliation":[{"name":"Pharmacologie cellulaire et mol\u00e9culaire, Louvain Drug Research Institute, Universit\u00e9 catholique de Louvain","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8368-4193","authenticated-orcid":true,"given":"Alexandra","family":"Koumoutsi","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/010jaxs89","id-type":"ROR","asserted-by":"publisher"}],"name":"European Molecular Biology Laboratory","place":["Heidelberg, Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0052-7991","authenticated-orcid":true,"given":"Fran\u00e7oise","family":"Van Bambeke","sequence":"additional","affiliation":[{"name":"Pharmacologie cellulaire et mol\u00e9culaire, Louvain Drug Research Institute, Universit\u00e9 catholique de Louvain","place":["Brussels, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","first-page":"318","volume-title":"Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics","author":"Tacconelli E","year":"2017","unstructured":"Tacconelli E, Magrini N, Carmeli Y, Harbarth S, Kahlmeter G, Kluytmans J, Mendelson M, Pulcini C, Singh N, Theuretzbacher U. 2017. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics, p 318\u2013327. Vol. 27. World Health Organization."},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00181-19"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/bph.13664"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.2020.2717"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2011.00065"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02609-12"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0019970"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.59.3.822-828.1991"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.62.8.3485-3493.1994"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.73.12.8298-8305.2005"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.7.4398-4406.2001"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-010-1036-y"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1164\/rccm.202308-1451OC"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aad3292"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0278-9"},{"key":"e_1_3_4_17_2","unstructured":"Co-ACTION. 2015. Developing combinations of CO-ACTIVE antimicrobials and non-antimicrobials - JPIAMR. Available from: https:\/\/www.jpiamr.eu\/projects\/co-action. Retrieved 28 Oct 2025."},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmi.2020.03.007"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01065-21"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02111-19"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkm509"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jinf.2005.11.019"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1469-0691.2005.01198.x"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.15585\/mmwr.rr6901a1"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a027011"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature22308"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.56.3.395-411.1992"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00064-16"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF01952257"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkg301"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-2854-5_13"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0511100103"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00147-19"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.90.3.665"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2013.01.008"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01479-12"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2036282100"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0029113"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2019.00819"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1558-5646.2011.01302.x"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics2020206"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmicrobiol.2017.14"},{"key":"e_1_3_4_43_2","volume-title":"Microplate selection and recommended practices in high-throughput screening and quantitative biology","author":"Auld DSP","year":"2004","unstructured":"Auld DSP, Coassin PB, Coussens NPP, Hensley P, Klumpp-Thomas C, Michael S, Sittampalam GSP, Trask OB, Wagner BKP, Weidner JRP, Wildey MJP, Dahlin Jl Md P, et al.. 2004. Microplate selection and recommended practices in high-throughput screening and quantitative biology. In Markossian S, Grossman A, Arkin M, Auld D, Austin C, Baell J, Brimacombe K, Chung TDY, Coussens NP, Dahlin JL (ed),. Assay Guidance Manual, Bethesda (MD)."},{"key":"e_1_3_4_44_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2003.03673.x"},{"key":"e_1_3_4_45_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.05829-11"},{"key":"e_1_3_4_46_2","doi-asserted-by":"publisher","DOI":"10.2174\/157489106777452692"},{"key":"e_1_3_4_47_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.21.2.299"},{"key":"e_1_3_4_48_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.24.1.107"},{"key":"e_1_3_4_49_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.accounts.3c00572"},{"key":"e_1_3_4_50_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms1938"},{"key":"e_1_3_4_51_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens10091187"},{"key":"e_1_3_4_52_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0891-5849(99)00107-0"},{"key":"e_1_3_4_53_2","doi-asserted-by":"publisher","DOI":"10.1080\/1071576031000148736"},{"key":"e_1_3_4_54_2","doi-asserted-by":"publisher","DOI":"10.1517\/14656560903334185"},{"key":"e_1_3_4_55_2","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01641-22"},{"key":"e_1_3_4_56_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-02875-z"},{"key":"e_1_3_4_57_2","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00176-16"},{"key":"e_1_3_4_58_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-micro-090817-062444"},{"key":"e_1_3_4_59_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3448"},{"key":"e_1_3_4_60_2","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.9b00204"},{"key":"e_1_3_4_61_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.177.1.114-122.1995"},{"key":"e_1_3_4_62_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-10988-6"},{"key":"e_1_3_4_63_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2008.06585.x"},{"key":"e_1_3_4_64_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.14531"},{"key":"e_1_3_4_65_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12929-022-00874-3"},{"key":"e_1_3_4_66_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2020.114400"},{"key":"e_1_3_4_67_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.06.049"},{"key":"e_1_3_4_68_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbi.2023.110479"},{"key":"e_1_3_4_69_2","doi-asserted-by":"publisher","DOI":"10.1111\/liv.15459"},{"key":"e_1_3_4_70_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-8-196"},{"key":"e_1_3_4_71_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.676534"},{"key":"e_1_3_4_72_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02028-20"},{"key":"e_1_3_4_73_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1744-7348.1939.tb06990.x"},{"key":"e_1_3_4_74_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-022-03836-5"},{"key":"e_1_3_4_75_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0154490"},{"key":"e_1_3_4_76_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.665364"},{"key":"e_1_3_4_77_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2019.05.020"},{"key":"e_1_3_4_78_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.05230-11"},{"key":"e_1_3_4_79_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-18068-5"},{"key":"e_1_3_4_80_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv1227"},{"key":"e_1_3_4_81_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaa1074"},{"key":"e_1_3_4_82_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1241"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.01319-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/aac.01319-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,7]],"date-time":"2026-01-07T14:01:41Z","timestamp":1767794501000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/aac.01319-25"}},"subtitle":[],"editor":[{"given":"James E.","family":"Leggett","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,1,7]]},"references-count":81,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1,7]]}},"alternative-id":["10.1128\/aac.01319-25"],"URL":"https:\/\/doi.org\/10.1128\/aac.01319-25","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"value":"0066-4804","type":"print"},{"value":"1098-6596","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,7]]},"assertion":[{"value":"2025-08-29","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-05","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01319-25"}}