{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T02:24:40Z","timestamp":1786501480384,"version":"build-2736575974"},"posted":{"date-parts":[[2025,10,7]]},"group-title":"Immunology","reference-count":50,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T00:00:00Z","timestamp":1759795200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"NIH","award":["AI134245, AI181701, AI128130, AI111914, AI134240, AI138587, AI184581, AI007290, CA20997105, OD01982205, CA24063801A1, AG06827902, CA24663303, CA22952904, CA22430901, AG05791504, AG05628705, AI168439 and AI161943"],"award-info":[{"award-number":["AI134245, AI181701, AI128130, AI111914, AI134240, AI138587, AI184581, AI007290, CA20997105, OD01982205, CA24063801A1, AG06827902, CA24663303, CA22952904, CA22430901, AG05791504, AG05628705, AI168439 and AI161943"]}]},{"name":"NSF","award":["2017242837"],"award-info":[{"award-number":["2017242837"]}]},{"name":"DOD","award":["W81XWH2110143"],"award-info":[{"award-number":["W81XWH2110143"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,10,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Granulomas, the hallmark of tuberculosis (TB) disease, can both restrict\n                  <jats:italic>Mycobacterium tuberculosis (Mtb)<\/jats:italic>\n                  dissemination and impede its clearance. Recent studies indicate that indoleamine 2,3-dioxygenase (IDO1), an immunosuppressive metabolic enzyme, limits infiltration of activated T cells and can contribute to TB disease progression. Treatment with 1-methyl-D-tryptophan (D-1MT), a small molecule inhibitor that restores mTOR signaling, has been shown to improve immune responses\n                  <jats:italic>Mtb<\/jats:italic>\n                  -infected rhesus macaques. Here, we investigated the impact of D-1MT treatment on TB granuloma architecture using 30-plex high-dimensional issue imaging in rhesus macaques. By spatially mapping 13 distinct cell populations, we found D-1MT treatment corresponded with significantly increased infiltration CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells into granulomas compared to untreated controls. Notably, these CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells expressed markers of cell proliferation and cytotoxicity. D-1MT enhanced CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cell infiltration throughout the granuloma, with particularly pronounced effects in the myeloid core, where we observed significantly enhanced spatial interactions between macrophages and CD8\n                  <jats:sup>+<\/jats:sup>\n                  , but not CD4\n                  <jats:sup>+<\/jats:sup>\n                  T cells. Our results demonstrate that: (i) effective intra-granulomatous\n                  <jats:italic>Mtb<\/jats:italic>\n                  control is associated with the close spatial proximity between CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells and macrophages, a feature less abundant in uncontrolled pulmonary TB; (ii) IDO1 induction blocks CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cell infiltration and reduces T cell activation and proliferation; and (iii) therapeutic strategies, including D-1MT, that improve intra-granulomatous killing hold strong translational potential.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Significance statement<\/jats:title>\n                  <jats:p>\n                    Our understanding of immune mechanisms within the TB granuloma has advanced greatly with the advent of high-resolution single cell multiplexed imaging. Using such imaging, we show that TB granulomas in rhesus macaques, a highly translational model of human TB pathology, are characterized by IDO1-mediated immunoregulation. Early pharmacologic restoration of mTOR signaling via D-1MT treatment can reduce IDO1 enzymatic activity and facilitate the recruitment and function of CD8\n                    <jats:sup>+<\/jats:sup>\n                    T cells within the granulomas. These findings reveal specific mechanisms exploited by\n                    <jats:italic>Mtb<\/jats:italic>\n                    to maintain intra-granuloma persistence and underscore immune responses. Future vaccine and therapeutic design should consider these immunoregulatory features to achieve better control of\n                    <jats:italic>Mtb<\/jats:italic>\n                    infection.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.10.02.680083","type":"posted-content","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T07:15:18Z","timestamp":1760080518000},"source":"Crossref","is-referenced-by-count":1,"title":["Therapeutic remodeling of the tuberculosis granuloma with 1-methyl-D-tryptophan enhances CD8\n                  <sup>+<\/sup>\n                  T cell-macrophage interactions"],"prefix":"10.64898","author":[{"given":"Erin F.","family":"McCaffrey","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7959-3165","authenticated-orcid":false,"given":"Alea C.","family":"Delmastro","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bindu","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annu","family":"Devi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadia A.","family":"Golden","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shabaana A.","family":"Khader","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1531-5067","authenticated-orcid":false,"given":"Michael","family":"Angelo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepak","family":"Kaushal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Smriti","family":"Mehra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025101000151318000_2025.10.02.680083v1.1","unstructured":"Global Tuberculosis Report 2024. https:\/\/www.who.int\/teams\/global-programme-on-tuberculosis-and-lung-health\/tb-reports\/global-tuberculosis-report-2024."},{"key":"2025101000151318000_2025.10.02.680083v1.2","doi-asserted-by":"publisher","DOI":"10.1038\/nri3813"},{"key":"2025101000151318000_2025.10.02.680083v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1538"},{"key":"2025101000151318000_2025.10.02.680083v1.4","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-immunol-032712-100022"},{"key":"2025101000151318000_2025.10.02.680083v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-021-01121-x"},{"key":"2025101000151318000_2025.10.02.680083v1.6","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0684.2012.00536.x"},{"key":"2025101000151318000_2025.10.02.680083v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/nri.2017.69"},{"key":"2025101000151318000_2025.10.02.680083v1.8","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0132249"},{"key":"2025101000151318000_2025.10.02.680083v1.9","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jis778"},{"key":"2025101000151318000_2025.10.02.680083v1.10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1711373114"},{"key":"2025101000151318000_2025.10.02.680083v1.11","first-page":"6099","article-title":"Indoleamine 2,3-dioxygenase 1 (IDO1): an up-to-date overview of an eclectic immunoregulatory enzyme","volume":"289","year":"2021","journal-title":"FEBS J"},{"key":"2025101000151318000_2025.10.02.680083v1.12","first-page":"384","article-title":"New Insights into IDO Biology in Bacterial and Viral Infections","volume":"5","year":"2014","journal-title":"Front Immunol"},{"key":"2025101000151318000_2025.10.02.680083v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2016.01.002"},{"key":"2025101000151318000_2025.10.02.680083v1.14","doi-asserted-by":"publisher","DOI":"10.4161\/onci.21716"},{"key":"2025101000151318000_2025.10.02.680083v1.15","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-06-2925"},{"key":"2025101000151318000_2025.10.02.680083v1.16","first-page":"e03189","article-title":"Myeloid-Derived Suppressor Cells Mediate T Cell Dysfunction in Nonhuman Primate TB Granulomas","volume":"12","year":"2021","journal-title":"mBio"},{"key":"2025101000151318000_2025.10.02.680083v1.17","doi-asserted-by":"crossref","unstructured":"Keren, L. et al. 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