{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T18:56:23Z","timestamp":1779216983090,"version":"3.51.4"},"posted":{"date-parts":[[2026,1,12]]},"group-title":"Immunology","reference-count":64,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T00:00:00Z","timestamp":1768176000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"crossref","award":["439662427"],"award-info":[{"award-number":["439662427"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Institute of Health","award":["R01AI166313"],"award-info":[{"award-number":["R01AI166313"]}]},{"name":"National Institute of Health","award":["R35GM142900"],"award-info":[{"award-number":["R35GM142900"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,1,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Rationale<\/jats:title>\n                  <jats:p>\n                    Tuberculosis vaccines aim to elicit protective immunity, with IFN\u03b3-producing CD4\u207a T cells considered central mediators of host defense. In murine models, IFN\u03b3 activates macrophages to induce inducible nitric oxide synthase (iNOS), resulting in nitric oxide\u2013dependent control of\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    (\n                    <jats:italic>Mtb<\/jats:italic>\n                    ).\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Objective<\/jats:title>\n                  <jats:p>To define the relevance of IFN\u03b3-induced iNOS activity in humans and other natural host of virulent mycobacterial species.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    We systematically compared IFN\u03b3-induced effector responses in\n                    <jats:italic>Mtb<\/jats:italic>\n                    -infected myeloid cells across species. Using bulk RNA sequencing, functional infection assays, and nitric oxide measurements, we assessed IFN\u03b3 responsiveness in human and mouse macrophages. These analyses were extended to monocytes from seven mammalian species and complemented by reanalysis of publicly available single-cell RNA-sequencing datasets and spatial proteomic imaging of tuberculous granulomas.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Measurements and Main Results<\/jats:title>\n                  <jats:p>\n                    Single-cell transcriptomic reanalysis revealed minimal\n                    <jats:italic>NOS2<\/jats:italic>\n                    expression in myeloid cells from human and non-human primate granulomas.\n                    <jats:italic>In vitro<\/jats:italic>\n                    , IFN\u03b3 pretreatment failed to induce\n                    <jats:italic>NOS2<\/jats:italic>\n                    transcription, iNOS activity, or\n                    <jats:italic>Mtb<\/jats:italic>\n                    growth restriction in human macrophages, in stark contrast to murine cells. Across species, iNOS activity was largely restricted to mice, with limited induction in cattle monocytes. Instead, human respiratory epithelial cells consistently expressed\n                    <jats:italic>NOS2<\/jats:italic>\n                    , and multiplexed ion beam imaging localized iNOS protein to epithelial compartments adjacent to granulomatous lesions.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>IFN\u03b3 signaling is uncoupled from iNOS induction in primate myeloid cells and epithelial compartments represent dominant sources of iNOS in human tuberculosis. These findings, challenge murine macrophage\u2013centric paradigms and IFN\u03b3-based correlates used in TB vaccine development and central to pathophysiology of tuberculosis and other pneumonias.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.01.11.697682","type":"posted-content","created":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T04:15:13Z","timestamp":1768277713000},"source":"Crossref","is-referenced-by-count":1,"title":["Species-specific iNOS expression distinguishes epithelial and myeloid IFN\u03b3 responses in tuberculosis"],"prefix":"10.64898","author":[{"given":"Fabian","family":"Stei","sequence":"first","affiliation":[{"name":"Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald - Isle of Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erin F.","family":"McCaffrey","sequence":"additional","affiliation":[{"name":"Spatial Immunology Unit, T-Lymphocyte Biology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bj\u00f6rn","family":"Zessin","sequence":"additional","affiliation":[{"name":"Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald - Isle of Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonathan","family":"Pioch","sequence":"additional","affiliation":[{"name":"Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald - Isle of Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefan","family":"Simm","sequence":"additional","affiliation":[{"name":"Institute of Bioanalytics, University of Applied Sciences Coburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beate","family":"Stubbe","sequence":"additional","affiliation":[{"name":"Internal Medicine B, Pneumology, University Hospital Greifswald, Greifswald, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ralf","family":"Ewert","sequence":"additional","affiliation":[{"name":"Internal Medicine B, Pneumology, University Hospital Greifswald, Greifswald, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Verreck","sequence":"additional","affiliation":[{"name":"Biomedical Primate Research Centre, Rijswijk, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bianca E","family":"Schneider","sequence":"additional","affiliation":[{"name":"Priority Area Infections, Research Center Borstel-Leibniz Lung Center, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anca","family":"Dorhoi","sequence":"additional","affiliation":[{"name":"Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald - Isle of Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1716-6712","authenticated-orcid":false,"given":"Bryan D","family":"Bryson","sequence":"additional","affiliation":[{"name":"Ragon Institute of MGH, Harvard, and MIT, Cambridge, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bj\u00f6rn","family":"Corleis","sequence":"additional","affiliation":[{"name":"Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald - Isle of Riems, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026020421094655000_2026.01.11.697682v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-019-0174-z"},{"key":"2026020421094655000_2026.01.11.697682v1.2","doi-asserted-by":"crossref","unstructured":"Lefrancais E , Hudrisier D , Neyrolles O , Behar SM , Ernst JD . 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