{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:16:29Z","timestamp":1790381789313,"version":"4.1.0"},"reference-count":54,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T00:00:00Z","timestamp":1709078400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2024,2,28]],"date-time":"2024-02-28T00:00:00Z","timestamp":1709078400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"HHS | NIH | National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["AI 106269, AI 133797"],"award-info":[{"award-number":["AI 106269, AI 133797"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSphere"],"published-print":{"date-parts":[[2024,2,28]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              <jats:italic>Histoplasma capsulatum<\/jats:italic>\n              is a dimorphic fungal pathogen acquired via inhalation of soil-resident spores. Upon exposure to mammalian body temperatures, these fungal elements transform into yeasts that reside primarily within phagocytes. Macrophages (M\u03a6) provide a permissive environment for fungal replication until T cell-dependent immunity is engaged. M\u03a6 activated by granulocyte macrophage colony stimulating factor (GM-CSF) induces metallothioneins (MTs) that bind zinc (Zn) and deprive yeast cells of labile Zn, thereby disabling fungal growth. Prior work demonstrated that the zinc transporter,\n              <jats:italic>ZRT2<\/jats:italic>\n              , was important for fungal survival\n              <jats:italic>in vivo<\/jats:italic>\n              . Hence, we constructed a yeast cell reporter strain that expresses green fluorescent protein (GFP) under control of the\n              <jats:italic>ZRT2<\/jats:italic>\n              zinc-regulated promoter. This reporter accurately responds to a medium devoid of Zn.\n              <jats:italic>ZRT2<\/jats:italic>\n              expression increased in GM-CSF, but not interferon-\u03b3, stimulated M\u03a6. To examine the\n              <jats:italic>in vivo<\/jats:italic>\n              response, we infected mice with a reporter yeast strain and assessed\n              <jats:italic>ZRT2<\/jats:italic>\n              expression at 0, 3, 7, and 14 days post-infection (dpi).\n              <jats:italic>ZRT2<\/jats:italic>\n              expression minimally increased at 3 dpi and peaked at 7 dpi, corresponding with the onset of adaptive immunity. We discovered that the major M\u03a6 populations that restrict Zn from the fungus are interstitial M\u03a6 and exudate M\u03a6. Neutralizing GM-CSF blunted the control of infection but unexpectedly increased\n              <jats:italic>ZRT2<\/jats:italic>\n              expression. This increase was dependent on another cytokine that activates M\u03a6 to control\n              <jats:italic>H. capsulatum<\/jats:italic>\n              replication, M-CSF. These findings illustrate the reporter\u2019s ability to sense Zn\n              <jats:italic>in vitro<\/jats:italic>\n              and\n              <jats:italic>in vivo<\/jats:italic>\n              and correlate\n              <jats:italic>ZRT2<\/jats:italic>\n              expression with GM-CSF and M-CSF activation of M\u03a6.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                Phagocytes use an arsenal of defenses to control the replication of\n                <jats:italic>Histoplasma<\/jats:italic>\n                yeasts, one of which is the limitation of trace metals. On the other hand,\n                <jats:italic>H. capsulatum<\/jats:italic>\n                combats metal restriction by upregulating metal importers such as the Zn importer\n                <jats:italic>ZRT2<\/jats:italic>\n                . This transporter contributes to\n                <jats:italic>H. capsulatum<\/jats:italic>\n                pathogenesis upon activation of adaptive immunity. We constructed a fluorescent\n                <jats:italic>ZRT2<\/jats:italic>\n                transcriptional reporter to probe\n                <jats:italic>H. capsulatum<\/jats:italic>\n                Zn sensing during infection and exposed the role for M-CSF activation of macrophages when GM-CSF is absent. These data highlight the ways in which fungal pathogens sense metal deprivation\n                <jats:italic>in vivo<\/jats:italic>\n                and reveal the potential of metal-sensing reporters. The work adds a new dimension to study how intracellular pathogens sense and respond to the changing environments of the host.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/msphere.00704-23","type":"journal-article","created":{"date-parts":[[2024,1,23]],"date-time":"2024-01-23T14:01:39Z","timestamp":1706018499000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Utilization of a\n            <i>Histoplasma capsulatum<\/i>\n            zinc reporter reveals the complexities of fungal sensing of metal deprivation"],"prefix":"10.1128","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4662-6554","authenticated-orcid":false,"given":"Logan T.","family":"Blancett","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heather M.","family":"Evans","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"Candor","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA"},{"name":"Immunology Graduate Program, Cincinnati Children\u2019s Hospital Medical Center and University of Cincinnati College of Medicine, Cincinnati, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William R.","family":"Buesing","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio A. 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