{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T06:34:04Z","timestamp":1773815644599,"version":"3.50.1"},"reference-count":43,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T00:00:00Z","timestamp":1773792000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Chagas disease (CD), caused by the blood borne parasite\n                      <jats:italic>Trypanosoma cruzi<\/jats:italic>\n                      , affects 6\u20138 million people primarily in Latin America. Chronic CD leads to progressive damage in major organs, particularly the heart and gastrointestinal tract, with significant morbidity and mortality. Despite the gut serving as a key parasite reservoir, the immune response within this tissue during chronic infection remains poorly understood. This study investigates the role of double-negative (DN; CD3\n                      <jats:sup>+<\/jats:sup>\n                      \/CD4\n                      <jats:sup>-<\/jats:sup>\n                      \/CD8\n                      <jats:sup>-<\/jats:sup>\n                      ) T cells in gut immunity during\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      infection. Understanding their function could provide insight into mechanisms of parasite persistence and immune regulation in the gastrointestinal tract.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      C57BL\/6 mice were infected with a transgenic strain of\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      Colombiana expressing a luminescence reporter. We collected tissues from infected mice during the acute (~30 days) and chronic phase (~90 days) of the disease and analyze them by bioluminescence imaging. We isolated cells from the gut lamina propria to identify the major immune cell types by multiparameter spectral flow cytometry. Colon tissue was also analyzed by fluorescence microscopy.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We showed that DN T cells were the major lymphocyte population in this tissue by flow cytometry and fluorescence microscopy. The number of DN T cells increased significantly (~1.5 fold) during the acute phase and remained above control levels during the chronic infection. Two major subtypes were identified: (1) DN T cells with inflammatory phenotypes, which peaked in the acute phase and declined in the chronic phase but remained above control levels, and (2) DN T cells with regulatory phenotypes, which were elevated in both phases.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      Our findings highlight the critical role of colonic DN T cells in an experimental\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      infection. Our results support a model in which DN T cells with inflammatory phenotypes may contribute to controlling the parasite burden in the gut during a chronic infection, whereas the sustained presence of DN T cells with regulatory phenotypes may help maintaining gut homeostasis but also create an anti-inflammatory niche that facilitates long-term\n                      <jats:italic>T. cruzi<\/jats:italic>\n                      persistence in this tissue. A better understanding the immune mechanisms occurring in reservoir tissues is important for developing improved therapies for Chagas disease.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1761769","type":"journal-article","created":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T05:38:33Z","timestamp":1773812313000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Double negative T cells (CD4-\/CD8-) are associated with Trypanosoma cruzi persistence in the mouse colon during chronic Chagas disease"],"prefix":"10.3389","volume":"17","author":[{"given":"Jung-Sun","family":"Cho","sequence":"first","affiliation":[{"name":"Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Supriya","family":"Kumar","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erica","family":"Silberstein","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuefei","family":"Ma","sequence":"additional","affiliation":[{"name":"Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic & Allergenic Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander","family":"Zhovmer","sequence":"additional","affiliation":[{"name":"Laboratory of Immunobiochemistry, Division of Bacterial, Parasitic & Allergenic Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alain","family":"Debrabant","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration","place":["Silver Spring, MD, United States"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,3,18]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(10)60061-X","article-title":"Chagas disease","volume":"375","author":"Rassi","year":"2010","journal-title":"Lancet"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(23)01787-7","article-title":"Chagas disease","volume":"403","author":"de Sousa","year":"2024","journal-title":"Lancet"},{"key":"B3","doi-asserted-by":"publisher","first-page":"100832","DOI":"10.1016\/j.lana.2024.100832","article-title":"Clinical features of Chagas disease progression and severity","volume":"37","author":"Nunes","year":"2024","journal-title":"Lancet Reg Health Am"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1590\/S1807-59322009001200013","article-title":"The chronic gastrointestinal manifestations of Chagas disease","volume":"64","author":"Matsuda","year":"2009","journal-title":"Clinics (Sao Paulo)"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.micinf.2010.03.005","article-title":"The innate immune response to Trypanosoma cruzi infection","volume":"12","author":"Kayama","year":"2010","journal-title":"Microbes Infect"},{"key":"B6","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1016\/0169-4758(95)80095-6","article-title":"The role of T cells in Trypanosoma cruzi infections","volume":"11","author":"Tarleton","year":"1995","journal-title":"Parasitol Today"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1007\/s00281-015-0481-9","article-title":"CD8+ T cells in Trypanosoma cruzi infection","volume":"37","author":"Tarleton","year":"2015","journal-title":"Semin Immunopathol"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1038\/nature01441","article-title":"CD4+ T cells are required for secondary expansion and memory in CD8+ T lymphocytes","volume":"421","author":"Janssen","year":"2003","journal-title":"Nature"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2009.07.006","article-title":"CD8+ T cells in Trypanosoma cruzi infection","volume":"21","author":"Padilla","year":"2009","journal-title":"Curr Opin Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1093\/intimm\/dxh387","article-title":"Trypanosoma cruzi modulates the profile of memory CD8+ T cells in chronic Chagas\u2019 disease patients","volume":"18","author":"Albareda","year":"2006","journal-title":"Int Immunol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0900852","article-title":"Chronic human infection with Trypanosoma cruzi drives CD4+ T cells to immune senescence","volume":"183","author":"Albareda","year":"2009","journal-title":"J Immunol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01245-06","article-title":"Limited role for CD4+ T-cell help in the initial priming of Trypanosoma cruzi-specific CD8+ T cells","volume":"75","author":"Padilla","year":"2007","journal-title":"Infect Immun"},{"key":"B13","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1016\/j.pt.2016.08.009","article-title":"Putting infection dynamics at the heart of chagas disease","volume":"32","author":"Lewis","year":"2016","journal-title":"Trends Parasitol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01242-20","article-title":"In Vivo Analysis of Trypanosoma cruzi Persistence Foci at Single-Cell Resolution","volume":"11","author":"Ward","year":"2020","journal-title":"mBio"},{"key":"B15","doi-asserted-by":"publisher","first-page":"e0195879","DOI":"10.1371\/journal.pone.0195879","article-title":"A novel nanoluciferase-based system to monitor Trypanosoma cruzi infection in mice by bioluminescence imaging","volume":"13","author":"Silberstein","year":"2018","journal-title":"PloS One"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1016\/j.humpath.2007.01.020","article-title":"Megacolon in Chagas disease: a study of inflammatory cells, enteric nerves, and glial cells","volume":"38","author":"da Silveira","year":"2007","journal-title":"Hum Pathol"},{"key":"B17","doi-asserted-by":"publisher","first-page":"e0038221","DOI":"10.1128\/iai.00382-21","article-title":"Incomplete recruitment of protective T cells is associated with trypanosoma cruzi persistence in the mouse colon","volume":"90","author":"Ward","year":"2022","journal-title":"Infect Immun"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.bcp.2011.05.019","article-title":"CD3+CD4-CD8- (double negative) T cells: saviours or villains of the immune response","volume":"82","author":"D\u2019Acquisto","year":"2011","journal-title":"Biochem Pharmacol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1111\/imm.13441","article-title":"Double-negative T cells: Setting the stage for disease control or progression","volume":"165","author":"Velikkakam","year":"2022","journal-title":"Immunology"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiw266","article-title":"Blocking of CD1d decreases trypanosoma cruzi-induced activation of CD4-CD8- T cells and modulates the inflammatory response in patients with chagas heart disease","volume":"214","author":"Passos","year":"2016","journal-title":"J Infect Dis"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12992","article-title":"Specific activation of CD4(-) CD8(-) double-negative T cells by Trypanosoma cruzi-derived glycolipids induces a proinflammatory profile associated with cardiomyopathy in Chagas patients","volume":"190","author":"Passos","year":"2017","journal-title":"Clin Exp Immunol"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-0849-4_13","article-title":"Introduction to multiparametric flow cytometry and analysis of high-dimensional data","volume":"2194","author":"Sun","year":"2021","journal-title":"Methods Mol Biol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"768113","DOI":"10.3389\/fimmu.2021.768113","article-title":"How to prepare spectral flow cytometry datasets for high dimensional data analysis: A practical workflow","volume":"12","author":"den Braanker","year":"2021","journal-title":"Front Immunol"},{"key":"B24","doi-asserted-by":"publisher","first-page":"235","DOI":"10.1038\/s41392-023-01471-y","article-title":"T cells in health and disease","volume":"8","author":"Sun","year":"2023","journal-title":"Signal Transduct Target Ther"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.021908.132723","article-title":"Regulatory T cells in the control of host-microorganism interactions (*)","volume":"27","author":"Belkaid","year":"2009","journal-title":"Annu Rev Immunol"},{"key":"B26","doi-asserted-by":"publisher","first-page":"659","DOI":"10.3389\/fimmu.2015.00659","article-title":"Evasion of the Immune Response by Trypanosoma cruzi during Acute Infection","volume":"6","author":"Cardoso","year":"2015","journal-title":"Front Immunol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12297","article-title":"Bioluminescence imaging of chronic Trypanosoma cruzi infections reveals tissue-specific parasite dynamics and heart disease in the absence of locally persistent infection","volume":"16","author":"Lewis","year":"2014","journal-title":"Cell Microbiol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1111\/cmi.12584","article-title":"Host and parasite genetics shape a link between Trypanosoma cruzi infection dynamics and chronic cardiomyopathy","volume":"18","author":"Lewis","year":"2016","journal-title":"Cell Microbiol"},{"key":"B29","doi-asserted-by":"publisher","first-page":"e0244710","DOI":"10.1371\/journal.pone.0244710","article-title":"Memory impairment in chronic experimental Chagas disease: Benznidazole therapy reversed cognitive deficit in association with reduction of parasite load and oxidative stress in the nervous tissue","volume":"16","author":"Vilar-Pereira","year":"2021","journal-title":"PloS One"},{"key":"B30","doi-asserted-by":"publisher","first-page":"975106","DOI":"10.3389\/fimmu.2022.975106","article-title":"Central nervous system commitment in Chagas disease","volume":"13","author":"Useche","year":"2022","journal-title":"Front Immunol"},{"key":"B31","doi-asserted-by":"publisher","first-page":"168","DOI":"10.3390\/v11020168","article-title":"Myeloid cells during viral infections and inflammation","volume":"11","author":"Stegelmeier","year":"2019","journal-title":"Viruses"},{"key":"B32","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.coi.2021.09.004","article-title":"Regulation of myeloid-cell activation","volume":"73","author":"Greene","year":"2021","journal-title":"Curr Opin Immunol"},{"key":"B33","doi-asserted-by":"publisher","first-page":"2794","DOI":"10.3389\/fimmu.2018.02794","article-title":"Immunosuppression in experimental chagas disease is mediated by an alteration of bone marrow stromal cell function during the acute phase of infection","volume":"9","author":"Muller","year":"2018","journal-title":"Front Immunol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1016\/j.pt.2019.08.006","article-title":"Understanding CD8(+) T cell immunity to trypanosoma cruzi and how to improve it","volume":"35","author":"Acosta Rodriguez","year":"2019","journal-title":"Trends Parasitol"},{"key":"B35","doi-asserted-by":"publisher","first-page":"e1002645","DOI":"10.1371\/journal.ppat.1002645","article-title":"CD8+ T-cells expressing interferon gamma or perforin play antagonistic roles in heart injury in experimental Trypanosoma cruzi-elicited cardiomyopathy","volume":"8","author":"Silverio","year":"2012","journal-title":"PloS Pathog"},{"key":"B36","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1186\/s12967-022-03761-5","article-title":"Chagasic cardiomyopathy is marked by a unique signature of activated CD4(+) T cells","volume":"20","author":"Almeida","year":"2022","journal-title":"J Transl Med"},{"key":"B37","doi-asserted-by":"publisher","first-page":"816005","DOI":"10.3389\/fimmu.2022.816005","article-title":"CD3(+)CD4(-)CD8(-) (Double-negative) T cells in inflammation, immune disorders and cancer","volume":"13","author":"Wu","year":"2022","journal-title":"Front Immunol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00179-10","article-title":"Trypanosoma cruzi-induced activation of functionally distinct alphabeta and gammadelta CD4- CD8- T cells in individuals with polar forms of Chagas\u2019 disease","volume":"78","author":"Villani","year":"2010","journal-title":"Infect Immun"},{"key":"B39","doi-asserted-by":"publisher","first-page":"761795","DOI":"10.3389\/fimmu.2021.761795","article-title":"Distinct CD4(-)CD8(-) (Double-negative) memory T-cell subpopulations are associated with indeterminate and cardiac clinical forms of chagas disease","volume":"12","author":"Passos","year":"2021","journal-title":"Front Immunol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0901533","article-title":"Human TCR-alpha beta+ CD4- CD8- T cells can derive from CD8+ T cells and display an inflammatory effector phenotype","volume":"183","author":"Crispin","year":"2009","journal-title":"J Immunol"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2006-10-050625","article-title":"New differentiation pathway for double-negative regulatory T cells that regulates the magnitude of immune responses","volume":"109","author":"Zhang","year":"2007","journal-title":"Blood"},{"key":"B42","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1038\/sj.jid.5700003","article-title":"Immunosurveillance and immunoregulation by gammadelta T cells","volume":"126","author":"Girardi","year":"2006","journal-title":"J Invest Dermatol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02762009000700021","article-title":"A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI","volume":"104","author":"Zingales","year":"2009","journal-title":"Mem Inst Oswaldo Cruz"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1761769\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,18]],"date-time":"2026-03-18T05:38:34Z","timestamp":1773812314000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2026.1761769\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,18]]},"references-count":43,"alternative-id":["10.3389\/fimmu.2026.1761769"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2026.1761769","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,18]]},"article-number":"1761769"}}