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However, the biological basis of incomplete parasite clearance after treatment remains poorly understood. Here, we investigated the tissue distribution of an intrinsically drug-resistant T. cruzi strain in a murine model of acute infection and evaluated the effects of short-term treatment with benznidazole or E1224 on parasite burden and host immune responses. Parasite loads were quantified by qPCR in multiple tissues before and after cyclophosphamide-induced immunosuppression, and plasma cytokines were measured to assess systemic immune responses. The heart, skeletal muscle, and white adipose tissue harbored the highest parasite burdens. Although both drugs significantly reduced parasite loads, incomplete clearance and tissue-specific recrudescence were observed following immunosuppression. Benznidazole-treated mice showed parasite rebound in the heart, adipose tissue, and large intestine, whereas E1224-treated animals exhibited recrudescence in the heart, skeletal muscle, and liver. Acute infection induced a robust pro-inflammatory response, and parasite reduction was accompanied by decreased Th1 cytokine levels. These findings demonstrate parasite persistence within defined tissue reservoirs despite chemotherapy, suggesting tissue-specific survival as a potential biological basis for therapeutic failure.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00228","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T20:02:10Z","timestamp":1787083330000},"page":"3068-3079","source":"Crossref","is-referenced-by-count":0,"title":["Tissue-Specific\nParasite Persistence Following Chemotherapy\nin a Drug-Resistant\n                    <i>Trypanosoma cruzi<\/i>\n                    Preclinical\nModel"],"prefix":"10.1021","volume":"12","author":[{"given":"Valqui\u0301ria Angelis","family":"Fernandes","sequence":"first","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]},{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda Late\u0301rcia Tranches","family":"Dias","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Se\u0301rgio","family":"Caldas","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Ezequiel Dias , , , ,","place":["Belo Horizonte, Minas Gerais, Brazil, 30510-010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2591-8654","authenticated-orcid":true,"given":"Gilson","family":"Faria","sequence":"additional","affiliation":[{"name":"Fundac\u0327a\u0303o Ezequiel Dias , , , ,","place":["Belo Horizonte, Minas Gerais, Brazil, 30510-010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Luiza Leite","family":"Pereira","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriella Martiniano","family":"Pereira","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo Dian","family":"de Oliveira Aguiar Soares","sequence":"additional","affiliation":[{"name":"Universidade Federal de Ouro Preto , , , ,","place":["Ouro Preto, Minas Gerais, Brazil, 35402-133"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thais Lopes Valentim Di Paschoale","family":"Ostolin","sequence":"additional","affiliation":[{"name":"Universidade Federal de Ouro Preto , , , ,","place":["Ouro Preto, Minas Gerais, Brazil, 35402-133"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivo Santana","family":"Caldas","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3186-5328","authenticated-orcid":true,"given":"Ro\u0302mulo Dias","family":"Novaes","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9398-6478","authenticated-orcid":true,"given":"Li\u0301via","family":"de Figueiredo Diniz","sequence":"additional","affiliation":[{"name":"Universidade Federal de Alfenas , , , ,","place":["Alfenas, Minas Gerais, Brazil, 37130-001"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"2026091109073650200_cit1","unstructured":"World Health Organization.\n          . Chagas Disease (American Trypanosomiasis). https:\/\/www.who.int\/health-topics\/chagas-disease#tab=tab_1 (accessed\n2025)."},{"key":"2026091109073650200_cit2","doi-asserted-by":"publisher","first-page":"1262","DOI":"10.1016\/j.amjmed.2020.05.022","article-title":"Chagas\nDisease: Epidemiology and Barriers to Treatment","volume":"133","author":"Mills","year":"2020","journal-title":"Am. J. Med."},{"key":"2026091109073650200_cit3","doi-asserted-by":"publisher","first-page":"100881","DOI":"10.1016\/j.lana.2024.100881","article-title":"The Epidemiology\nof Chagas Disease in the Americas","volume":"37","author":"Cucunuba\u0301","year":"2024","journal-title":"Lancet Reg.\nHealth Am."},{"key":"2026091109073650200_cit4","doi-asserted-by":"publisher","first-page":"105161","DOI":"10.1016\/j.actatropica.2019.105161","article-title":"Mapping Benznidazole\nResistance in Trypanosomatids and Exploring Evolutionary Histories\nof Nitroreductases and ABCG Transporter Protein Sequences","volume":"200","author":"Petravicius","year":"2019","journal-title":"Acta Trop."},{"key":"2026091109073650200_cit5","first-page":"7","article-title":"V. II Consenso\nBrasileiro em Doenc\u0327a de Chagas, 2015","volume":"25","author":"Dias","year":"2016","journal-title":"Epidemiol. Serv. Saude"},{"key":"2026091109073650200_cit6","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1590\/S0036-46652002000100006","article-title":"Long Term\nEvaluation of Etiological Treatment of Chagas Disease with Benznidazole","volume":"44","author":"Canc\u0327ado","year":"2002","journal-title":"Rev. Inst. Med. Trop. Sa\u0303o Paulo."},{"key":"2026091109073650200_cit7","doi-asserted-by":"publisher","first-page":"609","DOI":"10.1586\/eri.11.31","article-title":"Current Status of\nChagas Disease Chemotherapy","volume":"9","author":"Guedes","year":"2011","journal-title":"Expert Rev. Anti\nInfect. Ther."},{"key":"2026091109073650200_cit8","doi-asserted-by":"publisher","first-page":"939","DOI":"10.1016\/j.jacc.2016.12.023","article-title":"Benznidazole and Posaconazole\nin Eliminating Parasites\nin Asymptomatic T. cruzi Carriers: The STOP-CHAGAS Trial","volume":"69","author":"Morillo","year":"2017","journal-title":"J. Am. Coll. Cardiol."},{"key":"2026091109073650200_cit9","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1016\/S0140-6736(17)31612-4","article-title":"Chagas Disease","volume":"391","author":"Pe\u0301rez-Molina","year":"2018","journal-title":"Lancet"},{"key":"2026091109073650200_cit10","doi-asserted-by":"publisher","first-page":"524","DOI":"10.1136\/hrt.2008.159624","article-title":"Challenges and Opportunities\nfor Primary, Secondary, and Tertiary Prevention of Chagas\u2019\nDisease","volume":"95","author":"Rassi","year":"2009","journal-title":"Heart"},{"key":"2026091109073650200_cit11","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1128\/AAC.44.1.150-155.2000","article-title":"Activities of the Triazole Derivative SCH 56592 (Posaconazole)\nagainst Drug-Resistant Strains of the Protozoan Parasite Trypanosoma\n(Schizotrypanum) cruzi in Immunocompetent and Immunosuppressed Murine\nHosts","volume":"44","author":"Molina","year":"2000","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026091109073650200_cit12","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/S0924-8579(02)00273-X","article-title":"In Vitro\nand In Vivo Activities of Ravuconazole on Trypanosoma cruzi, the Causative Agent of Chagas Disease","volume":"21","author":"Urbina","year":"2003","journal-title":"Int.\nJ. Antimicrob. Agents"},{"key":"2026091109073650200_cit13","doi-asserted-by":"publisher","first-page":"e00401-18","DOI":"10.1128\/AAC.00401-18","article-title":"Outcome\nof E1224-Benznidazole Combination Treatment for Infection with a Multidrug-Resistant Trypanosoma cruzi Strain in Mice","volume":"62","author":"Diniz","year":"2018","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026091109073650200_cit14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0001870","article-title":"Fexinidazole: A Potential New Drug\nCandidate for Chagas Disease","volume":"6","author":"Bahia","year":"2012","journal-title":"PLoS Negl. Trop.\nDis."},{"key":"2026091109073650200_cit15","doi-asserted-by":"publisher","first-page":"1899","DOI":"10.1056\/NEJMoa1313122","article-title":"Randomized\nTrial of Posaconazole and Benznidazole for Chronic Chagas\u2019\nDisease","volume":"370","author":"Molina","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"2026091109073650200_cit16","doi-asserted-by":"publisher","first-page":"e1186","DOI":"10.1093\/cid\/ciac579","article-title":"A Phase 2, Randomized, Multicenter,\nPlacebo-Controlled, Proof-of-Concept Trial of Oral Fexinidazole in\nAdults with Chronic Indeterminate Chagas Disease","volume":"76","author":"Torrico","year":"2023","journal-title":"Clin. Infect. Dis."},{"key":"2026091109073650200_cit17","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1016\/S1473-3099(17)30538-8","article-title":"Treatment of\nAdult Chronic Indeterminate Chagas Disease with Benznidazole and Three\nE1224 Dosing Regimens: A Proof-of-Concept, Randomised, Placebo-Controlled\nTrial","volume":"18","author":"Torrico","year":"2018","journal-title":"Lancet Infect. Dis."},{"key":"2026091109073650200_cit18","doi-asserted-by":"publisher","first-page":"395","DOI":"10.1016\/S1473-3099(23)00651-5","article-title":"Efficacy and Safety\nof Fexinidazole for Treatment of Chronic Indeterminate Chagas Disease\n(FEXI-12): A Multicentre, Randomised, Double-Blind, Phase 2 Trial","volume":"24","author":"Pinazo","year":"2024","journal-title":"Lancet Infect. 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Acta Trop."},{"key":"2026091109073650200_cit21","doi-asserted-by":"publisher","first-page":"828","DOI":"10.1590\/0074-0276140156","article-title":"Drug Discovery\nfor Chagas Disease\nShould Consider Trypanosoma cruzi Strain Diversity","volume":"109","author":"Zingales","year":"2014","journal-title":"Mem. Inst. Oswaldo Cruz"},{"key":"2026091109073650200_cit22","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.34039","article-title":"Spontaneous\nDormancy Protects Trypanosoma cruzi during Extended\nDrug Exposure","volume":"7","author":"Sa\u0301nchez-Valde\u0301z","year":"2018","journal-title":"eLife"},{"key":"2026091109073650200_cit23","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1080\/14787210.2021.1834849","article-title":"Benznidazole for\nthe Treatment of Chagas Disease","volume":"19","author":"Losada Galva\u0301n","year":"2021","journal-title":"Expert Rev.\nAnti Infect. Ther."},{"key":"2026091109073650200_cit24","doi-asserted-by":"publisher","first-page":"150","DOI":"10.1093\/infdis\/jit420","article-title":"New, Combined,\nand Reduced Dosing Treatment Protocols Cure Trypanosoma cruzi Infection in Mice","volume":"209","author":"Bustamante","year":"2014","journal-title":"J. Infect. Dis."},{"key":"2026091109073650200_cit25","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.bcp.2018.01.005","article-title":"Time- and\nDose-Dependence Evaluation of Nitroheterocyclic Drugs for Improving\nEfficacy Following Trypanosoma cruzi Infection: A\nPreclinical Study","volume":"148","author":"Mazzeti","year":"2018","journal-title":"Biochem. Pharmacol."},{"key":"2026091109073650200_cit26","doi-asserted-by":"publisher","first-page":"21648","DOI":"10.1021\/acsomega.5c00996","article-title":"Amiodarone and Ravuconazole\nCombination Potentiates\nChemotherapy against Drug-Resistant Trypanosoma cruzi Strains","volume":"10","author":"Raimundo","year":"2025","journal-title":"ACS Omega"},{"key":"2026091109073650200_cit27","doi-asserted-by":"publisher","first-page":"e00284","DOI":"10.1128\/aac.00284-22","article-title":"Drug Repurposing in Chagas Disease: Chloroquine Potentiates\nBenznidazole Activity against Trypanosoma cruzi In\nVitro and In Vivo","volume":"66","author":"Pandey","year":"2022","journal-title":"Antimicrob. Agents Chemother."},{"key":"2026091109073650200_cit28","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1016\/S0020-7519(01)00158-8","article-title":"Parasite\nPersistence in the Aetiology of Chagas Disease","volume":"31","author":"Tarleton","year":"2001","journal-title":"Int. J. Parasitol."},{"key":"2026091109073650200_cit29","doi-asserted-by":"publisher","first-page":"102615","DOI":"10.1016\/j.mib.2025.102615","article-title":"The Importance of\nPersistence and\nDormancy in Trypanosoma cruzi Infection and Chagas\nDisease","volume":"86","author":"Bunkofske","year":"2025","journal-title":"Curr. Opin. Microbiol."},{"key":"2026091109073650200_cit30","first-page":"154","article-title":"Comportamento de\numa Cepa do Trypanosoma cruzi em Hospedeiros com\nBaixa Resiste\u0302ncia","volume":"14","author":"Andrade","year":"1972","journal-title":"Rev. Inst. Med. Trop.\nSa\u0303o Paulo."},{"key":"2026091109073650200_cit31","article-title":"Drug Screening and Development Cascade for Chagas Disease:\nAn Update of In Vitro and In Vivo Experimental Models","volume":"119","author":"Soeiro","year":"2024","journal-title":"Mem. Inst. Oswaldo Cruz"},{"key":"2026091109073650200_cit32","first-page":"32","article-title":"Caracterizac\u0327a\u0303o Morfo-Biolo\u0301gica\ne Histopatolo\u0301gica de Diferentes Cepas do Trypanosoma cruzi","volume":"70","author":"Andrade","year":"1970","journal-title":"Gaz. Me\u0301d. Bahia"},{"key":"2026091109073650200_cit33","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1111\/j.1365-2613.2006.00476.x","article-title":"Trypanosoma cruzi: Clones Isolated from the Colombian Strain Reproduce\nthe Parental Strain Characteristics, with Ubiquitous Histotropism","volume":"87","author":"Camandaroba","year":"2006","journal-title":"Int. J. Exp. Pathol."},{"key":"2026091109073650200_cit34","doi-asserted-by":"publisher","first-page":"1285","DOI":"10.1111\/cmi.12297","article-title":"Bioluminescence\nImaging of Chronic Trypanosoma cruzi Infections Reveals\nTissue-Specific Parasite Dynamics and Heart Disease in the Absence\nof Locally Persistent Infection","volume":"16","author":"Lewis","year":"2014","journal-title":"Cell. Microbiol."},{"key":"2026091109073650200_cit35","doi-asserted-by":"publisher","first-page":"106581","DOI":"10.1016\/j.actatropica.2022.106581","article-title":"The Entrance\nRoute: Oral, Mucous, Cutaneous, or Systemic\nHas a Marked Influence on the Outcome of Trypanosoma cruzi Experimental Infection","volume":"234","author":"Gonc\u0327alves","year":"2022","journal-title":"Acta Trop."},{"key":"2026091109073650200_cit36","first-page":"69","article-title":"Trypanosoma cruzi: Paninfectivity of CL Strain during Murine Acute Infection","volume":"82","author":"Lenzi","year":"1996","journal-title":"Parasitol. Res."},{"key":"2026091109073650200_cit37","doi-asserted-by":"publisher","first-page":"105736","DOI":"10.1016\/j.actatropica.2020.105736","article-title":"The Tissue-Specific Tropism in Trypanosoma cruzi: Is It True?","volume":"213","author":"An\u0303ez","year":"2021","journal-title":"Acta Trop."},{"key":"2026091109073650200_cit38","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1016\/S0166-6851(99)90035-X","article-title":"Differential\nTissue Distribution of Diverse Clones of Trypanosoma cruzi in Infected Mice","volume":"100","author":"Andrade","year":"1999","journal-title":"Mol. Biochem. Parasitol."},{"key":"2026091109073650200_cit39","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1177\/1087057114552623","article-title":"A New Experimental\nModel for Assessing Drug Efficacy against Trypanosoma cruzi Infection Based on Highly Sensitive In Vivo Imaging","volume":"20","author":"Lewis","year":"2015","journal-title":"J. Biomol. Screen."},{"key":"2026091109073650200_cit40","doi-asserted-by":"publisher","first-page":"e01242","DOI":"10.1128\/mBio.01242-20","article-title":"In Vivo\nAnalysis\nof Trypanosoma cruzi Persistence Foci at Single-Cell\nResolution","volume":"11","author":"Ward","year":"2020","journal-title":"mBio"},{"key":"2026091109073650200_cit41","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0195879","article-title":"A Novel Nanoluciferase-Based System\nto Monitor Trypanosoma cruzi Infection in Mice by\nBioluminescence Imaging","volume":"13","author":"Silberstein","year":"2018","journal-title":"PLoS One"},{"key":"2026091109073650200_cit42","doi-asserted-by":"publisher","first-page":"4653","DOI":"10.1128\/AAC.00520-15","article-title":"Limited Ability\nof Posaconazole to Cure Both Acute\nand Chronic Trypanosoma cruzi Infections Revealed\nby Highly Sensitive In Vivo Imaging","volume":"59","author":"Francisco","year":"2015","journal-title":"Antimicrob.\nAgents Chemother."},{"key":"2026091109073650200_cit43","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.abb7656","article-title":"A Modified Drug Regimen Clears Active and Dormant Trypanosomes\nin Mouse Models of Chagas Disease","volume":"12","author":"Bustamante","year":"2020","journal-title":"Sci. Transl.\nMed."},{"key":"2026091109073650200_cit44","doi-asserted-by":"publisher","first-page":"1539","DOI":"10.1016\/j.cmi.2025.03.018","article-title":"Long-Term Follow-up of Individuals\nwith Chagas Disease Treated with Posaconazole and Benznidazole in\na Non-Endemic Region: The CHAGASAZOL Cohort","volume":"31","author":"Roig-Sanchis","year":"2025","journal-title":"Clin. Microbiol. Infect."},{"key":"2026091109073650200_cit45","doi-asserted-by":"publisher","first-page":"24085","DOI":"10.1074\/jbc.M412802200","article-title":"The Adipocyte as\nan Important Target Cell for Trypanosoma cruzi Infection","volume":"280","author":"Combs","year":"2005","journal-title":"J. Biol. Chem."},{"key":"2026091109073650200_cit46","doi-asserted-by":"publisher","first-page":"1002","DOI":"10.1016\/j.micinf.2011.06.002","article-title":"Evidence for Trypanosoma cruzi in Adipose Tissue in Human Chronic Chagas\nDisease","volume":"13","author":"Matos Ferreira","year":"2011","journal-title":"Microbes Infect."},{"key":"2026091109073650200_cit47","doi-asserted-by":"publisher","first-page":"276","DOI":"10.1016\/j.pt.2016.11.008","article-title":"Adipose\nTissue: A Safe Haven for Parasites?","volume":"33","author":"Tanowitz","year":"2017","journal-title":"Trends\nParasitol."},{"key":"2026091109073650200_cit48","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1111\/j.1462-5822.2012.01764.x","article-title":"Mechanisms of Trypanosoma cruzi Persistence in Chagas Disease","volume":"14","author":"Nagajyothi","year":"2012","journal-title":"Cell. Microbiol."},{"key":"2026091109073650200_cit49","doi-asserted-by":"publisher","first-page":"830","DOI":"10.1093\/infdis\/jir840","article-title":"Response\nof Adipose Tissue to Early Infection with Trypanosoma cruzi (Brazil Strain)","volume":"205","author":"Nagajyothi","year":"2012","journal-title":"J. Infect. Dis."},{"key":"2026091109073650200_cit50","doi-asserted-by":"publisher","DOI":"10.1111\/jeu.70027","article-title":"Host Organelle\nInteractions Facilitate Cholesterol Acquisition by Trypanosoma cruzi Amastigotes","volume":"72","author":"Alcantara","year":"2025","journal-title":"J. Eukaryot. Microbiol."},{"key":"2026091109073650200_cit51","doi-asserted-by":"publisher","first-page":"339","DOI":"10.3390\/pathogens14040339","article-title":"Adipose Tissue\nin Chagas Disease: A Neglected Component of Pathogenesis","volume":"14","author":"Pessoa","year":"2025","journal-title":"Pathogens"},{"key":"2026091109073650200_cit52","doi-asserted-by":"publisher","DOI":"10.1093\/femspd\/ftv082","article-title":"Immunity and Immune\nModulation in Trypanosoma cruzi Infection","volume":"73","author":"Cardillo","year":"2015","journal-title":"Pathog. Dis."},{"key":"2026091109073650200_cit53","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0011474","article-title":"Differential Expression Profile of Genes Involved in\nthe Immune Response Associated to Progression of Chronic Chagas Disease","volume":"17","author":"Go\u0301mez","year":"2023","journal-title":"PLoS Negl. Trop. 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