{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T14:09:44Z","timestamp":1790690984520,"version":"4.1.0"},"posted":{"date-parts":[[2026,7,10]]},"group-title":"Immunology","reference-count":89,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T00:00:00Z","timestamp":1783641600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Recognition of infected macrophages by CD4+ T cells is essential to immune protection against\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  (Mtb), the causative agent of tuberculosis (TB). However, not all infected macrophage subsets successfully elicit T cell activation. We recently discovered that M2-like macrophages fail to efficiently activate memory CD4+ T cells when infected with Mtb, yet successfully elicit T cell activation when loaded with peptides, \u03b3-irradiated bacteria, or Mtb whole cell lysate. Since the mechanisms underlying CD4+ T cell evasion by infected M2 but not M1-like macrophages remain underexplored, we sought to determine the genes and pathways unique to Mtb infection of M2-like cells, including alveolar macrophages. RNA sequencing of human macrophages infected with virulent Mtb identified enrichment of IL-10 and type I interferon (IFN) signaling genes, including\n                  <jats:italic>IL10RA<\/jats:italic>\n                  and\n                  <jats:italic>HERC5<\/jats:italic>\n                  , respectively, in infected M2-like monocyte-derived and alveolar macrophages. However, genes involved in MHC-II trafficking, such as\n                  <jats:italic>AP1M2<\/jats:italic>\n                  , were higher in infected M1-like macrophages. In complementary experiments using fluorescence microscopy and flow cytometry, we observed impaired trafficking of newly synthesized MHC-II to the plasma membrane of Mtb-infected M2-like macrophages despite high total surface MHC-II levels. Neutralization of IL-10 or knockdown of\n                  <jats:italic>HERC5<\/jats:italic>\n                  restored MHC-II trafficking to the cell surface among infected M2-like macrophages and significantly enhanced activation of memory CD4+ T cells in an MHC-II-dependent manner. These findings identify coordinated IL-10 and type I IFN signaling as key mechanisms that restrict MHC-II trafficking to the plasma membrane in Mtb-infected M2-like macrophages, thereby limiting antigen presentation and CD4+ T cell activation. We propose that host-directed therapies targeting these pathways in infected alveolar macrophages will facilitate T cell recognition for the prevention or treatment of active TB.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Author Summary<\/jats:title>\n                  <jats:p>\n                    Recognition of infected macrophages by CD4+ T cells is essential to immune protection against\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    (Mtb), the causative agent of tuberculosis (TB). However, not all infected macrophage subsets successfully elicit T cell activation. We recently discovered that M2-like macrophages fail to efficiently activate memory CD4\n                    <jats:sup>+<\/jats:sup>\n                    T cells when infected with Mtb, yet they successfully elicit T cell activation when treated with peptides, \u03b3-irradiated bacteria, or Mtb whole cell lysate. In this study, we identified genes and pathways uniquely upregulated in Mtb-infected M2-like macrophages that are linked to inefficient CD4+ T cell activation, including IL-10 signaling and type I interferon (IFN) pathways. These pathways were linked to reduced MHC-II trafficking to the plasma membrane in Mtb-infected M2-like macrophages. Neutralization of IL-10 or knockdown of\n                    <jats:italic>HERC5<\/jats:italic>\n                    restored MHC-II trafficking and augmented memory CD4+ T cell activation. Our study demonstrates that IL-10 signaling and type I IFN pathways play detrimental roles in macrophages during Mtb infection, impairing MHC-II trafficking and CD4+ T cell activation. Since lung-resident alveolar macrophages express a dominant M2-like phenotype, these findings suggest that targeting IL-10 and type I IFN signaling may offer a strategy to enhance CD4+ T cell-mediated immunity and improve TB outcomes.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.07.736943","type":"posted-content","created":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T11:40:22Z","timestamp":1783770022000},"source":"Crossref","is-referenced-by-count":1,"title":["Restricted MHC-II trafficking in\n                  <i>Mycobacterium tuberculosis<\/i>\n                  -infected M2-like macrophages limits CD4+ T cell activation"],"prefix":"10.64898","author":[{"given":"Avinaash K.","family":"Sandhu","sequence":"first","affiliation":[{"name":"Biomedical Sciences Training Program, Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH, USA"},{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9792-1556","authenticated-orcid":false,"given":"Daniel P.","family":"Gail","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel C.","family":"Simmermon","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"},{"name":"Biomedical Sciences Training Program, Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Demetria","family":"Webb","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Metro Health Research Institute, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Hmiel","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, Metro Health Research Institute, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles","family":"Bark","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1716-6712","authenticated-orcid":false,"given":"Bryan","family":"Bryson","sequence":"additional","affiliation":[{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA"},{"name":"Ragon Institute of Mass General, Harvard, and MIT, Cambridge, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard F.","family":"Silver","sequence":"additional","affiliation":[{"name":"Division of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, OH, USA"},{"name":"Department of Medicine, The Louis Stokes Cleveland V.A. Medical Center, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9879-327X","authenticated-orcid":false,"given":"Stephen","family":"Carpenter","sequence":"additional","affiliation":[{"name":"Biomedical Sciences Training Program, Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH, USA"},{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026071406350667000_2026.07.07.736943v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jctube.2025.100522"},{"key":"2026071406350667000_2026.07.07.736943v1.2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.193.3.271"},{"key":"2026071406350667000_2026.07.07.736943v1.3","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1301236"},{"key":"2026071406350667000_2026.07.07.736943v1.4","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006704"},{"key":"2026071406350667000_2026.07.07.736943v1.5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007060"},{"key":"2026071406350667000_2026.07.07.736943v1.6","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-024-01124-3"},{"key":"2026071406350667000_2026.07.07.736943v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-023-01452-1"},{"key":"2026071406350667000_2026.07.07.736943v1.8","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0400983101"},{"key":"2026071406350667000_2026.07.07.736943v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2023.107706"},{"key":"2026071406350667000_2026.07.07.736943v1.10","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20250466"},{"key":"2026071406350667000_2026.07.07.736943v1.11","doi-asserted-by":"publisher","DOI":"10.1126\/sciimmunol.aaw6693"},{"key":"2026071406350667000_2026.07.07.736943v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2018.08.001"},{"key":"2026071406350667000_2026.07.07.736943v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.11.002"},{"key":"2026071406350667000_2026.07.07.736943v1.14","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20071367"},{"key":"2026071406350667000_2026.07.07.736943v1.15","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0801496105"},{"key":"2026071406350667000_2026.07.07.736943v1.16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005380"},{"key":"2026071406350667000_2026.07.07.736943v1.17","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20050265"},{"key":"2026071406350667000_2026.07.07.736943v1.18","doi-asserted-by":"publisher","DOI":"10.1183\/23120541.00915-2024"},{"key":"2026071406350667000_2026.07.07.736943v1.19","doi-asserted-by":"publisher","DOI":"10.1183\/09031936.00030610"},{"key":"2026071406350667000_2026.07.07.736943v1.20","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.171.1.175"},{"key":"2026071406350667000_2026.07.07.736943v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.02.006"},{"key":"2026071406350667000_2026.07.07.736943v1.22","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2015-0146oc"},{"key":"2026071406350667000_2026.07.07.736943v1.23","doi-asserted-by":"publisher","DOI":"10.21203\/rs.3.rs-2986649\/v1"},{"key":"2026071406350667000_2026.07.07.736943v1.24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0040348"},{"key":"2026071406350667000_2026.07.07.736943v1.25","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2300885"},{"key":"2026071406350667000_2026.07.07.736943v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2024.102495"},{"key":"2026071406350667000_2026.07.07.736943v1.27","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2025.04.004"},{"key":"2026071406350667000_2026.07.07.736943v1.28","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.181.8.5545"},{"key":"2026071406350667000_2026.07.07.736943v1.29","doi-asserted-by":"publisher","DOI":"10.1038\/nature09247"},{"key":"2026071406350667000_2026.07.07.736943v1.30","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1601340"},{"key":"2026071406350667000_2026.07.07.736943v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/nri3787"},{"key":"2026071406350667000_2026.07.07.736943v1.32","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20171704"},{"key":"2026071406350667000_2026.07.07.736943v1.33","doi-asserted-by":"publisher","DOI":"10.1165\/rcmb.2010-0319OC"},{"key":"2026071406350667000_2026.07.07.736943v1.34","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201040433"},{"key":"2026071406350667000_2026.07.07.736943v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80404-5"},{"key":"2026071406350667000_2026.07.07.736943v1.36","doi-asserted-by":"publisher","DOI":"10.1111\/imm.12010"},{"key":"2026071406350667000_2026.07.07.736943v1.37","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1401088"},{"key":"2026071406350667000_2026.07.07.736943v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2023.02.026"},{"key":"2026071406350667000_2026.07.07.736943v1.39","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1701120"},{"key":"2026071406350667000_2026.07.07.736943v1.40","doi-asserted-by":"publisher","DOI":"10.1016\/s0140-6736(15)01316-1"},{"key":"2026071406350667000_2026.07.07.736943v1.41","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006687"},{"key":"2026071406350667000_2026.07.07.736943v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80468-9"},{"key":"2026071406350667000_2026.07.07.736943v1.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2024.102939"},{"key":"2026071406350667000_2026.07.07.736943v1.44","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20250460"},{"key":"2026071406350667000_2026.07.07.736943v1.45","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0600397103"},{"key":"2026071406350667000_2026.07.07.736943v1.46","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2024.113870"},{"key":"2026071406350667000_2026.07.07.736943v1.47","doi-asserted-by":"publisher","DOI":"10.7554\/elife.106814"},{"key":"2026071406350667000_2026.07.07.736943v1.48","doi-asserted-by":"publisher","DOI":"10.1007\/s00011-022-01638-3"},{"key":"2026071406350667000_2026.07.07.736943v1.49","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1103426"},{"key":"2026071406350667000_2026.07.07.736943v1.50","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2017.11.024"},{"key":"2026071406350667000_2026.07.07.736943v1.51","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae1113"},{"key":"2026071406350667000_2026.07.07.736943v1.52","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.00090-15"},{"key":"2026071406350667000_2026.07.07.736943v1.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2015.02.002"},{"key":"2026071406350667000_2026.07.07.736943v1.54","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2007.12.002"},{"key":"2026071406350667000_2026.07.07.736943v1.55","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.01624-21"},{"key":"2026071406350667000_2026.07.07.736943v1.56","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0913671107"},{"key":"2026071406350667000_2026.07.07.736943v1.57","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0854.2009.00948.x"},{"key":"2026071406350667000_2026.07.07.736943v1.58","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-05345-8"},{"key":"2026071406350667000_2026.07.07.736943v1.59","doi-asserted-by":"publisher","DOI":"10.1038\/nmicrobiol.2016.232"},{"key":"2026071406350667000_2026.07.07.736943v1.60","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2011.7"},{"key":"2026071406350667000_2026.07.07.736943v1.61","doi-asserted-by":"publisher","DOI":"10.2174\/156652408785160907"},{"key":"2026071406350667000_2026.07.07.736943v1.62","doi-asserted-by":"publisher","DOI":"10.1016\/s1473-3099(23)00818-6"},{"key":"2026071406350667000_2026.07.07.736943v1.63","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2012.03.007"},{"key":"2026071406350667000_2026.07.07.736943v1.64","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.06.040"},{"key":"2026071406350667000_2026.07.07.736943v1.65","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-023-01354-6"},{"key":"2026071406350667000_2026.07.07.736943v1.66","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201646548"},{"key":"2026071406350667000_2026.07.07.736943v1.67","doi-asserted-by":"publisher","DOI":"10.1038\/35055027"},{"key":"2026071406350667000_2026.07.07.736943v1.68","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00774-22"},{"key":"2026071406350667000_2026.07.07.736943v1.69","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1249731"},{"key":"2026071406350667000_2026.07.07.736943v1.70","doi-asserted-by":"publisher","DOI":"10.3390\/v13061102"},{"key":"2026071406350667000_2026.07.07.736943v1.71","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adr6166"},{"key":"2026071406350667000_2026.07.07.736943v1.72","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-023-42039-2"},{"key":"2026071406350667000_2026.07.07.736943v1.73","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0707206105"},{"key":"2026071406350667000_2026.07.07.736943v1.74","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2022.111205"},{"key":"2026071406350667000_2026.07.07.736943v1.75","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2024.108820"},{"key":"2026071406350667000_2026.07.07.736943v1.76","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.05.003"},{"key":"2026071406350667000_2026.07.07.736943v1.77","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2016.06.006"},{"key":"2026071406350667000_2026.07.07.736943v1.78","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.109195"},{"key":"2026071406350667000_2026.07.07.736943v1.79","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-023-01459-4"},{"key":"2026071406350667000_2026.07.07.736943v1.80","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2023.01.011"},{"key":"2026071406350667000_2026.07.07.736943v1.81","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2018.02.009"},{"key":"2026071406350667000_2026.07.07.736943v1.82","doi-asserted-by":"publisher","DOI":"10.1016\/j.jinf.2026.106755"},{"key":"2026071406350667000_2026.07.07.736943v1.83","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009943"},{"key":"2026071406350667000_2026.07.07.736943v1.84","doi-asserted-by":"publisher","DOI":"10.1126\/science.1224026"},{"key":"2026071406350667000_2026.07.07.736943v1.85","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1900230"},{"key":"2026071406350667000_2026.07.07.736943v1.86","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-014-0550-8"},{"key":"2026071406350667000_2026.07.07.736943v1.87","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-15-293"},{"key":"2026071406350667000_2026.07.07.736943v1.88","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkad393"},{"key":"2026071406350667000_2026.07.07.736943v1.89","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkw419"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.07.07.736943","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T13:35:14Z","timestamp":1784036114000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.07.07.736943"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,10]]},"references-count":89,"URL":"https:\/\/doi.org\/10.64898\/2026.07.07.736943","relation":{},"subject":[],"published":{"date-parts":[[2026,7,10]]},"subtype":"preprint"}}