{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T15:00:49Z","timestamp":1780930849835,"version":"3.54.1"},"posted":{"date-parts":[[2026,6,4]]},"group-title":"Immunology","reference-count":53,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T00:00:00Z","timestamp":1780531200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["5T32HD007233"],"award-info":[{"award-number":["5T32HD007233"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["U19-AI162598"],"award-info":[{"award-number":["U19-AI162598"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["DP2-AI164249"],"award-info":[{"award-number":["DP2-AI164249"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["75N93019C00070"],"award-info":[{"award-number":["75N93019C00070"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100015326","name":"NIH Common Fund","doi-asserted-by":"crossref","award":["5K08AI166072"],"award-info":[{"award-number":["5K08AI166072"]}],"id":[{"id":"10.13039\/100015326","id-type":"DOI","asserted-by":"crossref"}]},{"name":"SEATRAC New2TB","award":["P30-AI168034"],"award-info":[{"award-number":["P30-AI168034"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Although spread of internalized Mtb from the initial infected alveolar macrophages (AMs) is a crucial determinant of infection outcomes, the role of cell death in facilitating this spread, and how it is regulated by Mtb remains poorly understood. Ferroptosis, a lipid peroxidation-mediated form of necrotic cell death, contributes to pathology during later stages of infection. However, the role of ferroptosis in early AM cell death remains inadequately defined, and its induction during infection has been primarily studied with the laboratory strain, H37Rv. Using gene set variation analysis of single cell RNAseq data profiling Mtb-infected murine lungs, we found that the hypervirulent Beijing sublineage clinical strain SA161 is associated with an elevated pro-ferroptotic transcriptional response in AMs compared to H37Rv by 17 days post infection. Consistent with these transcriptional profiles, we found that SA161 induced increased lipid peroxidation in comparison to H37Rv during infection\n                  <jats:italic>in vitro<\/jats:italic>\n                  and\n                  <jats:italic>in vivo<\/jats:italic>\n                  . Administration of the lipid peroxidation inhibitor, ferrostatin-1 (Fer-1), reduces this Mtb-induced lipid peroxidation. Notably, we found that administration of Fer-1 to Mtb-infected mice significantly reduced bacterial burden for SA161 at 14 dpi while having no effect on H37Rv. Microscopic analysis of SA161-infected lung lesions at 14 dpi suggests that inhibition of ferroptosis-driving lipid peroxidation results in a greater proportion of AMs amongst infected cells and decreased neutrophil-associated IFN signaling. Collectively, these findings reveal that ferroptosis plays an important role during early infection with virulent clinical strains of Mtb by influencing bacterial spread and signaling of immune cell responders, potentially informing host-directed intervention strategies.\n                <\/jats:p>","DOI":"10.64898\/2026.05.29.728786","type":"posted-content","created":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T13:20:16Z","timestamp":1780579216000},"source":"Crossref","is-referenced-by-count":0,"title":["Modulation of Ferroptosis During Early\n                  <i>Mycobacterium tuberculosis<\/i>\n                  Infection Contributes to Beijing Lineage Strain SA161 Virulence"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0116-1781","authenticated-orcid":false,"given":"Joshua J","family":"Ivie","sequence":"first","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sylvia","family":"Stull","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ethan","family":"Bustad","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Courtney R","family":"Plumlee","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara B","family":"Cohen","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fergal J","family":"Duffy","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan H","family":"Diercks","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John D","family":"Aitchison","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"},{"name":"Department of Pediatrics, University of Washington Seattle"},{"name":"Department of Global Health, University of Washington Seattle"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kevin B","family":"Urdahl","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"},{"name":"Department of Pediatrics, University of Washington Seattle"},{"name":"Department of Immunology, University of Washington Seattle"},{"name":"Department of Global Health, University of Washington Seattle"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexis","family":"Kaushansky","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"},{"name":"Department of Pediatrics, University of Washington Seattle"},{"name":"Department of Global Health, University of Washington Seattle"},{"name":"Department of Microbiology, University of Washington Seattle"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7484-1193","authenticated-orcid":false,"given":"Alissa C","family":"Rothchild","sequence":"additional","affiliation":[{"name":"Department of Veterinary and Animal Sciences, University of Massachusetts Amherst"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0142-5970","authenticated-orcid":false,"given":"Benjamin H","family":"Gern","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"},{"name":"Department of Pediatrics, University of Washington Seattle"},{"name":"Department of Immunology, University of Washington Seattle"},{"name":"Department of Global Health, University of Washington Seattle"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuyi","family":"Ma","sequence":"additional","affiliation":[{"name":"Center for Global Infectious Disease Research, Seattle Children\u2019s Research Institute"},{"name":"Department of Global Health, University of Washington Seattle"},{"name":"Department of Microbiology, University of Washington Seattle"},{"name":"Department of Chemical Engineering, University of Washington Seattle"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026060807203030000_2026.05.29.728786v1.1","unstructured":"World Health Organization. Global Tuberculosis Report 2025. https:\/\/www.who.int\/teams\/global-tuberculosis-programme\/tb-reports\/global-tuberculosis-report-2025. Updated November 12, 2025. Accessed March 12, 2026."},{"key":"2026060807203030000_2026.05.29.728786v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/ng.2744"},{"key":"2026060807203030000_2026.05.29.728786v1.3","doi-asserted-by":"publisher","DOI":"10.1111\/imr.12264"},{"key":"2026060807203030000_2026.05.29.728786v1.4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-pathol-011811-132458"},{"key":"2026060807203030000_2026.05.29.728786v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2018.08.001"},{"key":"2026060807203030000_2026.05.29.728786v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-28506-2"},{"key":"2026060807203030000_2026.05.29.728786v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.11.044"},{"key":"2026060807203030000_2026.05.29.728786v1.8","doi-asserted-by":"crossref","first-page":"1330461","DOI":"10.3389\/fimmu.2024.1330461","article-title":"The dance of macrophage death: the interplay between the inevitable and the microenvironment","volume":"15","year":"2024","journal-title":"Front Immunol"},{"key":"2026060807203030000_2026.05.29.728786v1.9","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00246.2022"},{"key":"2026060807203030000_2026.05.29.728786v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2017.09.003"},{"key":"2026060807203030000_2026.05.29.728786v1.11","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20181776"},{"key":"2026060807203030000_2026.05.29.728786v1.12","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20080767"},{"key":"2026060807203030000_2026.05.29.728786v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2008.03.003"},{"key":"2026060807203030000_2026.05.29.728786v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-37148-x"},{"key":"2026060807203030000_2026.05.29.728786v1.15","doi-asserted-by":"crossref","unstructured":"Rothchild AC , Olson GS , Nemeth J , Amon LM , Mai D , Gold ES , Diercks AH , Aderem A. Alveolar macrophages generate a noncanonical NRF2-driven transcriptional response to Mycobacterium tuberculosis in vivo. Sci Immunol. 2019;4(37).","DOI":"10.1126\/sciimmunol.aaw6693"},{"key":"2026060807203030000_2026.05.29.728786v1.16","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20172020"},{"key":"2026060807203030000_2026.05.29.728786v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.12.033"},{"key":"2026060807203030000_2026.05.29.728786v1.18","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20210615"},{"key":"2026060807203030000_2026.05.29.728786v1.19","doi-asserted-by":"crossref","unstructured":"Chen X , Kang R , Kroemer G , Tang D. Ferroptosis in infection, inflammation, and immunity. J Exp Med.2021;218(6).","DOI":"10.1084\/jem.20210518"},{"key":"2026060807203030000_2026.05.29.728786v1.20","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.03.042"},{"key":"2026060807203030000_2026.05.29.728786v1.21","doi-asserted-by":"crossref","unstructured":"Amaral EP , Foreman TW , Namasivayam S , Hilligan KL , Kauffman KD , Barbosa Bomfim CC , Costa DL , Barreto-Duarte B , Gurgel-Rocha C , Santana MF , Cordeiro-Santos M , Du Bruyn E , Riou C , Aberman K , Wilkinson RJ , Barber DL , Mayer-Barber KD , Andrade BB , Sher A. GPX4 regulates cellular necrosis and host resistance in Mycobacterium tuberculosis infection. J Exp Med. 2022;219(11).","DOI":"10.1084\/jem.20220504"},{"issue":"1","key":"2026060807203030000_2026.05.29.728786v1.22","first-page":"120","article-title":"BACH1 promotes tissue necrosis and Mycobacterium tuberculosis susceptibility","volume":"9","year":"2024","journal-title":"Nat Microbiol"},{"key":"2026060807203030000_2026.05.29.728786v1.23","doi-asserted-by":"crossref","first-page":"RP106814","DOI":"10.7554\/eLife.106814.2","article-title":"Lipid peroxidation and type I interferon coupling fuels pathogenic macrophage activation causing tuberculosis susceptibility","volume":"14","year":"2025","journal-title":"Elife"},{"key":"2026060807203030000_2026.05.29.728786v1.24","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.091096998"},{"key":"2026060807203030000_2026.05.29.728786v1.25","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20250466"},{"issue":"10","key":"2026060807203030000_2026.05.29.728786v1.26","doi-asserted-by":"crossref","first-page":"e20250161","DOI":"10.1084\/jem.20250161","article-title":"Early and opposing neutrophil and CD4 T cell responses shape pulmonary tuberculosis pathology","volume":"222","year":"2025","journal-title":"J Exp Med"},{"issue":"3","key":"2026060807203030000_2026.05.29.728786v1.27","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.tube.2009.01.005","article-title":"Clinical strains of Mycobacterium tuberculosis display a wide range of virulence in guinea pigs","volume":"89","year":"2009","journal-title":"Tuberculosis (Edinb)"},{"key":"2026060807203030000_2026.05.29.728786v1.28","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1427846"},{"key":"2026060807203030000_2026.05.29.728786v1.29","doi-asserted-by":"publisher","DOI":"10.3389\/FIMMU.2024.1427510"},{"issue":"4","key":"2026060807203030000_2026.05.29.728786v1.30","doi-asserted-by":"crossref","first-page":"e1013635","DOI":"10.1371\/journal.ppat.1013635","article-title":"Single-cell profiling reveals that dynamic lung immune responses distinguish protection from susceptibility to tuberculosis","volume":"22","year":"2026","journal-title":"PLoS Pathog"},{"key":"2026060807203030000_2026.05.29.728786v1.31","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-14-7"},{"key":"2026060807203030000_2026.05.29.728786v1.32","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkac935"},{"issue":"1","key":"2026060807203030000_2026.05.29.728786v1.33","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1186\/s40001-023-01371-5","article-title":"Bioinformatics-led discovery of ferroptosis-associated diagnostic biomarkers and molecule subtypes for tuberculosis patients","volume":"28","year":"2023","journal-title":"Eur J Med Res"},{"key":"2026060807203030000_2026.05.29.728786v1.34","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-04579-w"},{"key":"2026060807203030000_2026.05.29.728786v1.35","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-024-01348-9"},{"issue":"1","key":"2026060807203030000_2026.05.29.728786v1.36","doi-asserted-by":"crossref","first-page":"10611","DOI":"10.1038\/s41598-022-14964-7","article-title":"Predicting prognosis and clinical features of the tumor microenvironment based on ferroptosis score in patients with breast cancer","volume":"12","year":"2022","journal-title":"Sci Rep"},{"key":"2026060807203030000_2026.05.29.728786v1.37","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005809"},{"key":"2026060807203030000_2026.05.29.728786v1.38","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.jare.2024.12.028","article-title":"The role of cGAS-STING signaling pathway in ferroptosis","volume":"76","year":"2025","journal-title":"J Adv Res"},{"key":"2026060807203030000_2026.05.29.728786v1.39","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.166.7.4604"},{"key":"2026060807203030000_2026.05.29.728786v1.40","doi-asserted-by":"publisher","DOI":"10.1126\/science.1224026"},{"key":"2026060807203030000_2026.05.29.728786v1.41","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-0270-8"},{"issue":"1","key":"2026060807203030000_2026.05.29.728786v1.42","doi-asserted-by":"crossref","first-page":"e0252622","DOI":"10.1128\/spectrum.02526-22","article-title":"Mycobacterium tuberculosis Rv1324 Protein Contributes to Mycobacterial Persistence and Causes Pathological Lung Injury in Mice by Inducing Ferroptosis","volume":"11","year":"2023","journal-title":"Microbiol Spectr"},{"issue":"Pt 4","key":"2026060807203030000_2026.05.29.728786v1.43","first-page":"146294","article-title":"Mycobacterium tuberculosis Rv2521 promotes ferroptosis-dependent pathogenicity by inhibiting NF-kappaB activation","volume":"321","year":"2025","journal-title":"Int J Biol Macromol"},{"key":"2026060807203030000_2026.05.29.728786v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/S41587-023-01767-Y"},{"key":"2026060807203030000_2026.05.29.728786v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.11.026"},{"key":"2026060807203030000_2026.05.29.728786v1.46","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0550-8"},{"issue":"6","key":"2026060807203030000_2026.05.29.728786v1.47","doi-asserted-by":"crossref","first-page":"1607","DOI":"10.1038\/s41564-024-01697-8","article-title":"Propionate prevents loss of the PDIM virulence lipid in Mycobacterium tuberculosis","volume":"9","year":"2024","journal-title":"Nat Microbiol"},{"key":"2026060807203030000_2026.05.29.728786v1.48","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-6581-6_22"},{"key":"2026060807203030000_2026.05.29.728786v1.49","doi-asserted-by":"publisher","DOI":"10.1136\/jcp.56.8.619"},{"key":"2026060807203030000_2026.05.29.728786v1.50","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.170.4.1987"},{"key":"2026060807203030000_2026.05.29.728786v1.51","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2012.07.011"},{"key":"2026060807203030000_2026.05.29.728786v1.52","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.107523"},{"key":"2026060807203030000_2026.05.29.728786v1.53","unstructured":"R Core Team (2024). R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria. https:\/\/www.R-project.org\/"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.05.29.728786","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T14:21:14Z","timestamp":1780928474000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.05.29.728786"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,4]]},"references-count":53,"URL":"https:\/\/doi.org\/10.64898\/2026.05.29.728786","relation":{},"subject":[],"published":{"date-parts":[[2026,6,4]]},"subtype":"preprint"}}