{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:17:06Z","timestamp":1785403026626,"version":"3.56.0"},"reference-count":67,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T00:00:00Z","timestamp":1770768000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T00:00:00Z","timestamp":1770768000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2026,2,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Listeria monocytogenes<\/jats:italic>\n                      is a facultative intracellular bacterial pathogen that is a potent inducer of cell-mediated immunity, which has led to the development of attenuated,\n                      <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                      -based cancer vaccines.\n                      <jats:italic toggle=\"yes\">L. monocytogenes<\/jats:italic>\n                      strains, such as live-attenuated double-deleted\n                      <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                      (LADD), lacking two key virulence factors, \u0394\n                      <jats:italic toggle=\"yes\">actA<\/jats:italic>\n                      and \u0394\n                      <jats:italic toggle=\"yes\">inlB<\/jats:italic>\n                      , have been used safely in clinical trials and showed promising anti-tumor activity. Despite early clinical success, improving potency and safety by preventing extracellular bacterial growth is paramount for the development of further clinical applications. We describe a quadruple attenuated intracellular\n                      <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                      (QUAIL) strain that, in addition to \u0394\n                      <jats:italic toggle=\"yes\">actA<\/jats:italic>\n                      \u0394\n                      <jats:italic toggle=\"yes\">inlB,<\/jats:italic>\n                      lacks\n                      <jats:italic toggle=\"yes\">ribC<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">ribF<\/jats:italic>\n                      , which encode enzymes required for generating the essential flavin cofactors flavin mononucleotide (FMN) and flavin adenine nucleotide (FAD). QUAIL imported FMN and FAD during intracellular growth but was unable to grow extracellularly in blood or on vascular catheters in mice, which reduced its lethality. Despite its lack of extracellular growth, QUAIL maintained its immunoprotective properties, which were comparable to LADD. Furthermore, we showed that QUAIL can be engineered to synthesize riboflavin, leading to expansion and activation of mucosal-associated invariant T cells. Together, our data support the use of QUAIL as a promising therapeutic platform with an improved safety profile that is amenable to further modifications to expand its immune-activating potential.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                        -based live-attenuated cancer vaccines represent a promising therapy in many different pre-clinical tumor models and in clinical trials. Enhancing its anti-cancer immunity and increasing its safety profile will advance the clinical applications of\n                        <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                        vaccines. By manipulating\n                        <jats:italic toggle=\"yes\">Listeria monocytogenes<\/jats:italic>\n                        flavin metabolism, we engineered a quadruple attenuated intracellular\n                        <jats:italic toggle=\"yes\">Listeria<\/jats:italic>\n                        (QUAIL) vaccine candidate strain that has limited toxicity associated with extracellular growth in major extracellular niches\n                        <jats:italic toggle=\"yes\">in vivo,<\/jats:italic>\n                        including blood and implanted catheter ports. Furthermore, we showed that QUAIL can be effectively programmed to engage innate-like T cells known as mucosal-associated invariant T cells, which could be harnessed for future cancer immunotherapies. The results presented here lay the foundation for further analysis of QUAIL as a safer, yet immunopotent\n                        <jats:italic toggle=\"yes\">L. monocytogenes<\/jats:italic>\n                        vaccine or therapeutic vector.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.03652-25","type":"journal-article","created":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T14:24:03Z","timestamp":1767191043000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Reprogramming\n                    <i>Listeria monocytogenes<\/i>\n                    flavin metabolism to improve its therapeutic safety profile and broaden innate T-cell 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