{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T02:02:27Z","timestamp":1790647347176,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,24]]},"group-title":"Immunology","reference-count":73,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["K08 AI163407"],"award-info":[{"award-number":["K08 AI163407"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01 AI187662"],"award-info":[{"award-number":["R01 AI187662"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R21 AI167675"],"award-info":[{"award-number":["R21 AI167675"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["75N93019C00071"],"award-info":[{"award-number":["75N93019C00071"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]},{"award":["75N93019C00070"],"award-info":[{"award-number":["75N93019C00070"]}],"id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Control of\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  (Mtb) infection requires CD4\n                  <jats:sup>+<\/jats:sup>\n                  T cell recognition of infected macrophages. However, T cells also colocalize with non-infected macrophages in granulomas. We investigated whether these bystander macrophages present Mtb antigens and shape human CD4\n                  <jats:sup>+<\/jats:sup>\n                  T cell responses. Using\n                  <jats:italic>ex vivo<\/jats:italic>\n                  co-culture systems, non-infected monocyte-derived macrophages (MDMs) were exposed to Mtb-infected MDMs or infection-conditioned supernatants before co-incubation with autologous memory CD4\n                  <jats:sup>+<\/jats:sup>\n                  T cells from individuals with latent Mtb infection (LTBI). Bystander macrophages activated memory CD4\n                  <jats:sup>+<\/jats:sup>\n                  T cells through MHC-II-dependent antigen presentation. Single-cell T cell receptor (TCR) sequencing and TCR-transduced reporter cell lines identified Mtb-specific clonotypes recognizing both infected and bystander macrophages, as well as clonotypes preferentially recognizing infected cells. Strikingly, a subset of TCRs recognized infected but not bystander macrophages. Antigen transfer occurred through soluble Mtb proteins rather than extracellular vesicles. Compared with responses to infected macrophages, bystander macrophages induced attenuated effector responses. These findings reveal antigen-specific recognition of bystander macrophages and suggest that antigens preferentially presented by infected cells may inform TB vaccine design.\n                <\/jats:p>","DOI":"10.64898\/2026.09.19.752467","type":"posted-content","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T07:05:10Z","timestamp":1790319910000},"source":"Crossref","is-referenced-by-count":0,"title":["Human memory CD4\n                  <sup>+<\/sup>\n                  T cells recognize non-infected macrophage bystanders exposed to\n                  <i>Mycobacterium tuberculosis<\/i>\n                  -infected cells"],"prefix":"10.64898","author":[{"given":"Vinicius G.","family":"Suzart","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"},{"name":"Department of Pathology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Schauner","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott M.","family":"Reba","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel P.","family":"Gail","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gwendolyn","family":"Swarbrick","sequence":"additional","affiliation":[{"name":"Portland VA Medical Center, Portland, Oregon, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"Lewinsohn","sequence":"additional","affiliation":[{"name":"Portland VA Medical Center, Portland, Oregon, USA"},{"name":"Pulmonary & Critical Care Medicine, Oregon Health & Science University, Portland, Oregon, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah A.","family":"Lewinsohn","sequence":"additional","affiliation":[{"name":"Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, Oregon, USA"},{"name":"Division of Infectious Diseases, Department of Pediatrics, Oregon Health and Science University, Portland, Oregon, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bianca","family":"Lepe","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"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1716-6712","authenticated-orcid":false,"given":"Bryan D.","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"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3374-6699","authenticated-orcid":false,"given":"Samuel M.","family":"Behar","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Massachusetts Chan Medical School, Worcester, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher M.","family":"Sassetti","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Massachusetts Chan Medical School, Worcester, MA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael L.","family":"Freeman","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"},{"name":"Department of Pathology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. Henry","family":"Boom","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"},{"name":"Department of Pathology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clifford","family":"Harding","sequence":"additional","affiliation":[{"name":"Department of Pathology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9879-327X","authenticated-orcid":false,"given":"Stephen M.","family":"Carpenter","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases and HIV Medicine, Department of Medicine, Cleveland, OH, USA"},{"name":"Department of Pathology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026092818351996000_2026.09.19.752467v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jctube.2025.100522"},{"key":"2026092818351996000_2026.09.19.752467v1.2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.55.9.2037-2041.1987"},{"key":"2026092818351996000_2026.09.19.752467v1.3","doi-asserted-by":"publisher","DOI":"10.1084\/jem.193.3.271"},{"key":"2026092818351996000_2026.09.19.752467v1.4","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro.2017.128"},{"key":"2026092818351996000_2026.09.19.752467v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro3230"},{"key":"2026092818351996000_2026.09.19.752467v1.6","doi-asserted-by":"publisher","DOI":"10.1128\/iai.57.2.390-395.1989"},{"key":"2026092818351996000_2026.09.19.752467v1.7","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1000399"},{"key":"2026092818351996000_2026.09.19.752467v1.8","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20071367"},{"key":"2026092818351996000_2026.09.19.752467v1.9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006704"},{"key":"2026092818351996000_2026.09.19.752467v1.10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0801496105"},{"key":"2026092818351996000_2026.09.19.752467v1.11","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.01086"},{"key":"2026092818351996000_2026.09.19.752467v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2014.05.007"},{"key":"2026092818351996000_2026.09.19.752467v1.13","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1301236"},{"key":"2026092818351996000_2026.09.19.752467v1.14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007060"},{"key":"2026092818351996000_2026.09.19.752467v1.15","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002063"},{"key":"2026092818351996000_2026.09.19.752467v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2011.03.022"},{"key":"2026092818351996000_2026.09.19.752467v1.17","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01514-17"},{"key":"2026092818351996000_2026.09.19.752467v1.18","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1010721"},{"key":"2026092818351996000_2026.09.19.752467v1.19","doi-asserted-by":"publisher","DOI":"10.1084\/jem.178.6.2249"},{"key":"2026092818351996000_2026.09.19.752467v1.20","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01089-09"},{"key":"2026092818351996000_2026.09.19.752467v1.21","doi-asserted-by":"crossref","first-page":"43578","DOI":"10.1038\/srep43578","article-title":"Exosomes function in antigen presentation during an in vivo Mycobacterium tuberculosis infection","volume":"7","year":"2017","journal-title":"Sci Rep-uk"},{"key":"2026092818351996000_2026.09.19.752467v1.22","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.19.1.93"},{"key":"2026092818351996000_2026.09.19.752467v1.23","first-page":"365","article-title":"Pathogenesis of post primary tuberculosis: immunity and hypersensitivity in the development of cavities","volume":"44","year":"2014","journal-title":"Ann. Clin. Lab. Sci"},{"key":"2026092818351996000_2026.09.19.752467v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-025-01192-z"},{"key":"2026092818351996000_2026.09.19.752467v1.25","doi-asserted-by":"crossref","first-page":"e20221392","DOI":"10.1084\/jem.20221392","article-title":"Spatial mapping reveals granuloma diversity and histopathological superstructure in human tuberculosis","volume":"220","year":"2023","journal-title":"J. Exp. Med"},{"key":"2026092818351996000_2026.09.19.752467v1.26","doi-asserted-by":"publisher","DOI":"10.1016\/S1473-3099(20)30148-1"},{"key":"2026092818351996000_2026.09.19.752467v1.27","first-page":"462","article-title":"Defective positioning in granulomas but not lung-homing limits CD4 T-cell interactions with Mycobacterium tuberculosis-infected macrophages in rhesus macaques","volume":"11","year":"2017","journal-title":"Mucosal Immunol"},{"key":"2026092818351996000_2026.09.19.752467v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-026-02431-8"},{"key":"2026092818351996000_2026.09.19.752467v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2015.12.003"},{"key":"2026092818351996000_2026.09.19.752467v1.30","doi-asserted-by":"crossref","unstructured":"Gail, D. P. , Suzart, V. G. & Carpenter, S. M . Analyzing human CD4+ T cells activated in response to macrophages infected with Mycobacterium tuberculosis. STAR Protoc. 5, 102939 (2024).","DOI":"10.1016\/j.xpro.2024.102939"},{"key":"2026092818351996000_2026.09.19.752467v1.31","doi-asserted-by":"crossref","unstructured":"Gail, D. P. et al. Mycobacterium tuberculosis impairs human memory CD4+ T cell recognition of M2 but not M1-like macrophages. iScience 26, 107706 (2023).","DOI":"10.1016\/j.isci.2023.107706"},{"key":"2026092818351996000_2026.09.19.752467v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/s41385-019-0217-6"},{"key":"2026092818351996000_2026.09.19.752467v1.33","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1003130"},{"key":"2026092818351996000_2026.09.19.752467v1.34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005760"},{"key":"2026092818351996000_2026.09.19.752467v1.35","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20250460"},{"key":"2026092818351996000_2026.09.19.752467v1.36","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0505-4"},{"key":"2026092818351996000_2026.09.19.752467v1.37","doi-asserted-by":"publisher","DOI":"10.1038\/nature22976"},{"key":"2026092818351996000_2026.09.19.752467v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2012.06.010"},{"key":"2026092818351996000_2026.09.19.752467v1.39","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2300001"},{"key":"2026092818351996000_2026.09.19.752467v1.40","doi-asserted-by":"publisher","DOI":"10.64898\/2026.07.07.736943"},{"key":"2026092818351996000_2026.09.19.752467v1.41","doi-asserted-by":"publisher","DOI":"10.1038\/s41591-022-02110-9"},{"key":"2026092818351996000_2026.09.19.752467v1.42","doi-asserted-by":"publisher","DOI":"10.1078\/0171-9335-00131"},{"key":"2026092818351996000_2026.09.19.752467v1.43","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2007-03-079152"},{"key":"2026092818351996000_2026.09.19.752467v1.44","doi-asserted-by":"publisher","DOI":"10.1172\/JCI44261"},{"key":"2026092818351996000_2026.09.19.752467v1.45","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0002461"},{"key":"2026092818351996000_2026.09.19.752467v1.46","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1402894"},{"key":"2026092818351996000_2026.09.19.752467v1.47","first-page":"16.2.1","article-title":"Presenting Exogenous Antigen to T Cells","volume":"88","year":"2010","journal-title":"Curr. Protoc. Immunol"},{"key":"2026092818351996000_2026.09.19.752467v1.48","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2025.114034"},{"key":"2026092818351996000_2026.09.19.752467v1.49","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2022.04.004"},{"key":"2026092818351996000_2026.09.19.752467v1.50","doi-asserted-by":"publisher","DOI":"10.1038\/s41590-024-01897-8"},{"key":"2026092818351996000_2026.09.19.752467v1.51","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.640725"},{"key":"2026092818351996000_2026.09.19.752467v1.52","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2010.02.013"},{"key":"2026092818351996000_2026.09.19.752467v1.53","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2300107"},{"key":"2026092818351996000_2026.09.19.752467v1.54","doi-asserted-by":"publisher","DOI":"10.1084\/jem.165.2.302"},{"key":"2026092818351996000_2026.09.19.752467v1.55","doi-asserted-by":"publisher","DOI":"10.1038\/nri2131"},{"key":"2026092818351996000_2026.09.19.752467v1.56","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1500710"},{"key":"2026092818351996000_2026.09.19.752467v1.57","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.67.12.6461-6472.1999"},{"key":"2026092818351996000_2026.09.19.752467v1.58","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2021.02.005"},{"key":"2026092818351996000_2026.09.19.752467v1.59","doi-asserted-by":"publisher","DOI":"10.1002\/eji.201040433"},{"key":"2026092818351996000_2026.09.19.752467v1.60","doi-asserted-by":"crossref","first-page":"eadw9184","DOI":"10.1126\/scitranslmed.adw9184","article-title":"Immunopeptidomics can inform the design of mRNA vaccines for the delivery of Mycobacterium tuberculosis MHC class II antigens","volume":"17","year":"2025","journal-title":"Sci. Transl. Med"},{"key":"2026092818351996000_2026.09.19.752467v1.61","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.84070"},{"key":"2026092818351996000_2026.09.19.752467v1.62","doi-asserted-by":"publisher","DOI":"10.1038\/S41587-023-01767-Y"},{"key":"2026092818351996000_2026.09.19.752467v1.63","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.4096"},{"key":"2026092818351996000_2026.09.19.752467v1.64","doi-asserted-by":"publisher","DOI":"10.1101\/2025.03.27.645853"},{"key":"2026092818351996000_2026.09.19.752467v1.65","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1012760"},{"key":"2026092818351996000_2026.09.19.752467v1.66","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-019-1874-1"},{"key":"2026092818351996000_2026.09.19.752467v1.67","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-021-02451-7"},{"key":"2026092818351996000_2026.09.19.752467v1.68","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0619-0"},{"key":"2026092818351996000_2026.09.19.752467v1.69","doi-asserted-by":"publisher","DOI":"10.1186\/s13059-015-0844-5"},{"key":"2026092818351996000_2026.09.19.752467v1.70","doi-asserted-by":"publisher","DOI":"10.1101\/2022.02.28.482303"},{"key":"2026092818351996000_2026.09.19.752467v1.71","doi-asserted-by":"crossref","first-page":"e43","DOI":"10.1002\/imt2.43","article-title":"Complex heatmap visualization","volume":"1","year":"2022","journal-title":"iMeta"},{"key":"2026092818351996000_2026.09.19.752467v1.72","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-15-293"},{"key":"2026092818351996000_2026.09.19.752467v1.73","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1038\/s41541-017-0008-6","article-title":"Comprehensive definition of human immunodominant CD8 antigens in tuberculosis","volume":"2","year":"2017","journal-title":"Npj Vaccines"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.19.752467","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T01:35:32Z","timestamp":1790645732000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.19.752467"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,24]]},"references-count":73,"URL":"https:\/\/doi.org\/10.64898\/2026.09.19.752467","relation":{},"subject":[],"published":{"date-parts":[[2026,9,24]]},"subtype":"preprint"}}