{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T01:15:50Z","timestamp":1783732550254,"version":"3.55.0"},"reference-count":70,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004901","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais","doi-asserted-by":"publisher","award":["APQ-02167-21"],"award-info":[{"award-number":["APQ-02167-21"]}],"id":[{"id":"10.13039\/501100004901","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004901","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de Minas Gerais","doi-asserted-by":"publisher","award":["RED-0067-23"],"award-info":[{"award-number":["RED-0067-23"]}],"id":[{"id":"10.13039\/501100004901","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["402417\/2023-2"],"award-info":[{"award-number":["402417\/2023-2"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Brazilian agencies Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Current treatment for visceral leishmaniasis (VL) is associated with toxicity, a high cost, and the emergence of drug-resistant parasite strains. Moreover, no human vaccine is available. In this context, immunotherapeutics combining vaccination and chemotherapy have emerged as a promising alternative. In this study, we combined a recombinant chimeric protein (ChimT) with the adjuvant 3-O-desacyl\u2013monophosphoryl lipid A (MPLA) and the antileishmanial drug miltefosine (Milt) and used it to treat Leishmania infantum-infected mice. Parasitological, immunological, and toxicological assays were performed at 1 and 30 days post treatment. The ChimT\/MPLA\/Milt combination was the most effective therapeutic regimen, inducing robust Th1-type cellular and humoral immune responses, as demonstrated by increased levels of IFN-\u03b3, IL-12, and TNF-\u03b1 cytokines, nitrite, and IgG2a antibodies. These responses were associated with significant reductions in parasite load across various organs. Furthermore, the treatment showed low renal and hepatic toxicity at both evaluation time points. By contrast, ChimT alone, ChimT\/MPLA, and Milt induced lower immunological and parasitological responses when compared with the ChimT\/MPLA\/Milt group. The results observed at 1 and 30 days post treatment demonstrated the potential of ChimT\/MPLA\/Milt combination against VL.<\/jats:p>","DOI":"10.3390\/pathogens14121202","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T10:46:18Z","timestamp":1764067578000},"page":"1202","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Immunotherapeutics Combining a Recombinant Chimeric Protein, Monophosphoryl Lipid A, and Miltefosine Against Visceral Leishmaniasis"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-7343-1660","authenticated-orcid":false,"given":"Marcelo M.","family":"Jesus","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela P.","family":"Lage","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Breno L.","family":"Pimenta","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel J. L.","family":"Moreira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunopatologia, N\u00facleo de Pesquisas em Ci\u00eancias Biol\u00f3gicas\/NUPEB, Departamento de Ci\u00eancias Biol\u00f3gicas, Insituto de Ci\u00eancias Exatas e Biol\u00f3gicas, Universidade Federal de Ouro Preto, Ouro Preto 35402-136, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabela A. G.","family":"Pereira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raquel S. B.","family":"C\u00e2mara","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8193-9551","authenticated-orcid":false,"given":"Ana L.","family":"Silva","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grasiele S. V.","family":"Tavares","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Pesquisa do Programa de P\u00f3s-Gradua\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade, Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Alfredo Balena, 190, Belo Horizonte 30130-100, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5994-2232","authenticated-orcid":false,"given":"Karolina O. M.","family":"Falc\u00e3o","sequence":"additional","affiliation":[{"name":"Departamento de Patologia Cl\u00ednica, COLTEC, Universidade Federal de Minas Gerais, Av. Ant\u00f4nio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saulo S. G.","family":"Dias","sequence":"additional","affiliation":[{"name":"Departamento de Patologia Cl\u00ednica, COLTEC, Universidade Federal de Minas Gerais, Av. 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Available online: http:\/\/www.who.int\/topics\/leishmaniasis\/en\/2025."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"750","DOI":"10.12688\/f1000research.11120.1","article-title":"Leishmaniasis: A review","volume":"6","author":"Arenas","year":"2017","journal-title":"F1000Research"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Costa, C.H.N., Chang, K.P., Costa, D.L., and Cunha, F.V.M. (2023). From infection to death: An overview of the pathogenesis of visceral leishmaniasis. Pathogens, 12.","DOI":"10.3390\/pathogens12070969"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ponte-Sucre, A., Gamarro, F., Dujardin, J.C., Barrett, M.P., L\u00f3pez-V\u00e9lez, R., Garc\u00eda-Hern\u00e1ndez, R., Pountain, A.W., Mwenechanya, R., and Papadopoulou, B. (2017). Drug resistance and treatment failure in leishmaniasis: A 21st century challenge. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0006052"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1080\/14728222.2018.1472241","article-title":"Emerging therapeutic targets for treatment of leishmaniasis","volume":"22","author":"Sundar","year":"2018","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1080\/17460441.2022.2002843","article-title":"Assay development in leishmaniasis drug discovery: A comprehensive review","volume":"17","author":"Zulfiqar","year":"2022","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1056\/NEJMoa021556","article-title":"Oral miltefosine for Indian visceral leishmaniasis","volume":"347","author":"Sundar","year":"2002","journal-title":"N. Engl. J. Med."},{"key":"ref_8","first-page":"3634568","article-title":"Miltefosine rescue treatment for visceral leishmaniasis relapse patient","volume":"2019","author":"Bastola","year":"2019","journal-title":"Case Rep. Infect. Dis."},{"key":"ref_9","first-page":"e02507","article-title":"Systematic review of host-mediated activity of miltefosine in leishmaniasis through immunomodulation","volume":"63","author":"Bhairosing","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Melcon-Fernandez, E., Galli, G., Garc\u00eda-Estrada, C., Bala\u00f1a-Fouce, R., Reguera, R.M., and P\u00e9rez-Pertejo, Y. (2023). Miltefosine and Nifuratel combination: A promising therapy for the treatment of Leishmania donovani visceral leishmaniasis. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021635"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Benaim, G., and Paniz-Mondolfi, A. (2024). Unmasking the mechanism behind miltefosine: Revealing the disruption of intracellular Ca2+ homeostasis as a rational therapeutic target in leishmaniasis and Chagas disease. Biomolecules, 14.","DOI":"10.20944\/preprints202402.1176.v1"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1186\/s13071-016-1553-y","article-title":"Possibilities and challenges for developing a successful vaccine for leishmaniasis","volume":"9","author":"Srivastava","year":"2016","journal-title":"Parasites Vectors"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"789","DOI":"10.4269\/ajtmh.18-0949","article-title":"Natural resistance of Leishmania infantum to miltefosine contributes to the low efficacy in the treatment of visceral leishmaniasis in Brazil","volume":"101","author":"Carnielli","year":"2019","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1080\/14760584.2018.1459191","article-title":"Vaccine candidates against leishmania under current research","volume":"17","author":"Iborra","year":"2018","journal-title":"Expert Rev. Vaccines"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Rostamian, M., Bahrami, F., and Niknam, H.M. (2018). Vaccination with whole-cell killed or recombinant leishmanial protein and toll-like receptor agonists against Leishmania tropica in BALB\/c mice. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0204491"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"104272","DOI":"10.1016\/j.cellimm.2020.104272","article-title":"IFN-\u03b3+ CD4+ T cell-driven prophylactic potential of recombinant LDBPK_252400 hypothetical protein of Leishmania donovani against visceral leishmaniasis","volume":"361","author":"Yadav","year":"2021","journal-title":"Cell. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.actatropica.2011.08.007","article-title":"DNA vaccination with KMP11 and Lutzomyia longipalpis salivary protein protects hamsters against visceral leishmaniasis","volume":"120","author":"Silva","year":"2011","journal-title":"Acta Trop."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5494","DOI":"10.1016\/j.vaccine.2022.08.005","article-title":"Immunoprophylaxis using polypeptide chimera vaccines plus adjuvant system promote Th1 response controlling the spleen parasitism in hamster model of visceral leishmaniasis","volume":"40","author":"Ostolin","year":"2022","journal-title":"Vaccine"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Campos, M.P., Figueiredo, F.B., Morgado, F.N., Renzetti, A.R.D.S., Souza, S.M.M., Pereira, S.A., Rodrigues-da-Silva, R.N., Lima-Junior, J.D.C., and Luca, P.M. (2018). Leishmania infantum virulence factor A2 protein: Linear B-cell epitope mapping and identification of three main linear B-cell epitopes in vaccinated and naturally infected dogs. Front. Immunol., 9.","DOI":"10.3389\/fimmu.2018.01690"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Alonso, A., Alcolea, P.J., Larraga, J., Peris, M.P., Esteban, A., Cort\u00e9s, A., Ruiz-Garc\u00eda, S., Castillo, J.A., and Larraga, V. (2023). A non-replicative antibiotic resistance-free DNA vaccine delivered by the intranasal route protects against canine leishmaniasis. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1213193"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1585","DOI":"10.1016\/j.vaccine.2008.01.026","article-title":"Evaluation of an immunochemotherapeutic protocol constituted of N-methyl meglumine antimoniate (Glucantime) and the recombinant Leish-110f + MPL-SE vaccine to treat canine visceral leishmaniasis","volume":"26","author":"Miret","year":"2008","journal-title":"Vaccine"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1017\/S0031182021001906","article-title":"Immunochemotherapy for visceral leishmaniasis: Combinatorial action of miltefosine plus LBSapMPL vaccine improves adaptative Th1 immune response with control of splenic parasitism in experimental hamster model","volume":"149","author":"Carvalho","year":"2022","journal-title":"Parasitology"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Yadagiri, G., Singh, A., Arora, K., and Mudavath, S.L. (2023). Immunotherapy and immunochemotherapy in combating visceral leishmaniasis. Front. Med., 10.","DOI":"10.3389\/fmed.2023.1096458"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2755","DOI":"10.1016\/j.vaccine.2021.04.004","article-title":"A chimeric vaccine combined with adjuvant system induces immunogenicity and protection against visceral leishmaniasis in BALB\/c mice","volume":"39","author":"Ostolin","year":"2021","journal-title":"Vaccine"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Oliveira, D.S., Zald\u00edvar, M.F., Gon\u00e7alves, A.A.M., Resende, L.A., Mariano, R.M.D.S., Pereira, D.F.S., Conrado, I.D.S.S., Costa, M.A.F., Lair, D.F., and Vilas-Boas, D.F. (2024). New Approaches to the Prevention of Visceral Leishmaniasis: A Review of Recent Patents of Potential Candidates for a Chimeric Protein Vaccine. Vaccines, 12.","DOI":"10.3390\/vaccines12030271"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1016\/j.parint.2016.07.005","article-title":"Treatment of murine visceral leishmaniasis using an 8-hydroxyquinoline-containing polymeric micelle system","volume":"65","author":"Duarte","year":"2016","journal-title":"Parasitol. Int."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Lage, D.P., Ribeiro, P.A.F., Dias, D.S., Mendon\u00e7a, D.V.C., Ramos, F.F., Carvalho, L.M., Steiner, B.T., Tavares, G.S.V., Martins, V.T., and Machado, A.S. (2020). Liposomal Formulation of ChimeraT, a Multiple T-Cell Epitope-Containing Recombinant Protein, Is a Candidate Vaccine for Human Visceral Leishmaniasis. Vaccines, 8.","DOI":"10.3390\/vaccines8020289"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Coelho, V.T., Oliveira, J.S., Valadares, D.G., Ch\u00e1vez-Fumagalli, M.A., Duarte, M.C., Lage, P.S., Soto, M., Santoro, M.M., Tavares, C.A., and Fernandes, A.P. (2012). Identification of proteins in promastigote and amastigote-like Leishmania using an immunoproteomic approach. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001430"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"106412","DOI":"10.1016\/j.actatropica.2022.106412","article-title":"A recombinant Leishmania amastigote-specific protein, rLiHyG, with adjuvants, protects against infection with Leishmania infantum","volume":"230","author":"Machado","year":"2022","journal-title":"Acta Trop."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.cellimm.2017.11.001","article-title":"Recombinant prohibitin protein of Leishmania infantum acts as a vaccine candidate and diagnostic marker against visceral leishmaniasis","volume":"323","author":"Dias","year":"2018","journal-title":"Cell. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1038\/s41541-020-00224-0","article-title":"A candidate vaccine for human visceral leishmaniasis based on a specific T cell epitope-containing chimeric protein protects mice against Leishmania infantum infection","volume":"5","author":"Lage","year":"2020","journal-title":"NPJ Vaccines"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1590\/0037-8682-0138-2015","article-title":"New delivery systems for amphotericin B applied to the improvement of leishmaniasis treatment","volume":"48","author":"Ribeiro","year":"2015","journal-title":"Rev. Soc. Bras. Med. Trop."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1017\/S0031182017002116","article-title":"Chemotherapeutics of visceral leishmaniasis: Present and future developments","volume":"145","author":"Sundar","year":"2018","journal-title":"Parasitology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.idc.2018.10.005","article-title":"Visceral Leishmaniasis: Recent Advances in Diagnostics and Treatment Regimens","volume":"33","author":"Diro","year":"2019","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lage, D.P., Vale, D.L., Linhares, F.P., Freitas, C.S., Machado, A.S., Cardoso, J.M.O., Oliveira, D., Galvani, N.C., Oliveira, M.P., and Oliveira-da-Silva, J.A. (2022). A recombinant chimeric protein-based vaccine containing T-cell epitopes from amastigote proteins and combined with distinct adjuvants, induces immunogenicity and protection against Leishmania infantum infection. Vaccines, 10.","DOI":"10.3390\/vaccines10071146"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Vale, D.L., Freitas, C.S., Martins, V.T., Moreira, G.J.L., Machado, A.S., Ramos, F.F., Pereira, I.A.G., Bandeira, R.S., Jesus, M.M., and Tavares, G.S.V. (2023). Efficacy of an immunotherapy combining immunogenic chimeric protein plus adjuvant and amphotericin B against murine visceral leishmaniasis. Biology, 12.","DOI":"10.3390\/biology12060851"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1051\/parasite\/2020027","article-title":"A clioquinol-containing Pluronic F127 polymeric micelle system is effective in the treatment of visceral leishmaniasis in a murine model","volume":"27","author":"Tavares","year":"2020","journal-title":"Parasite"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"104884","DOI":"10.1016\/j.micinf.2021.104884","article-title":"Intranasal immunization with chitosan microparticles enhances LACK-DNA vaccine protection and induces specific long-lasting immunity against visceral leishmaniasis","volume":"24","author":"Gomes","year":"2022","journal-title":"Microbes Infect."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1006\/meth.2001.1261","article-title":"An overview of real-time quantitative PCR: Applications to quantify cytokine gene expression","volume":"25","author":"Giulietti","year":"2001","journal-title":"Methods"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Moreira, N.D.D., Vitoriano-Souza, J., Roatt, B.M., Vieira, P.M.D.A., Ker, H.G., de Oliveira Cardoso, J.M., Giunchetti, R.C., Carneiro, C.M., de Lana, M., and Reis, A.B. (2012). Parasite burden in hamsters infected with two different strains of leishmania (Leishmania) infantum: \u201cLeishman Donovan units\u201d versus real-time PCR. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0047907"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"100317","DOI":"10.1016\/j.lansea.2023.100317","article-title":"Estimating the proportion of relapse following treatment of visceral leishmaniasis: Meta-analysis using Infectious Diseases Data Observatory (IDDO) systematic review","volume":"22","author":"Chhajed","year":"2023","journal-title":"Lancet Reg. Health-Southeast Asia"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Roatt, B.M., Aguiar-Soares, R.D., Coura-Vital, W., Ker, H.G., Moreira, N., Vitoriano-Souza, J., Giunchetti, R.C., Carneiro, C.M., and Reis, A.B. (2014). Immunotherapy and immunochemotherapy in visceral leishmaniasis: Promising treatments for this neglected disease. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00272"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.crimmu.2021.10.003","article-title":"Heterologous vaccine therapy associated with half course of Miltefosine promote activation of the proinflammatory response with control of splenic parasitism in a hamster model of visceral leishmaniasis","volume":"2","author":"Carvalho","year":"2021","journal-title":"Curr. Res. Immunol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/j.vaccine.2009.09.071","article-title":"Immunotherapy with the saponin enriched-Leishmune vaccine versus immunochemotherapy in dogs with natural canine visceral leishmaniasis","volume":"28","author":"Santos","year":"2010","journal-title":"Vaccine"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Dayakar, A., Chandrasekaran, S., Kuchipudi, S.V., and Kalangi, S.K. (2019). Cytokines: Key determinants of resistance or disease progression in visceral leishmaniasis: Opportunities for novel diagnostics and immunotherapy. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.00670"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Agallou, M., Margaroni, M., Kotsakis, S.D., and Karagouni, E. (2020). A Canine-Directed Chimeric Multi-Epitope Vaccine Induced Protective Immune Responses in BALB\/c Mice Infected with Leishmania infantum. Vaccines, 8.","DOI":"10.3390\/vaccines8030350"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Ayari-Riabi, S., Ben Khalaf, N., Bouhaouala-Zahar, B., Verrier, B., Trimaille, T., Benlasfar, Z., Chenik, M., and Elayeb, M. (2022). Polylactide nanoparticles as a biodegradable vaccine adjuvant: A study on safety, protective immunity and efficacy against human leishmaniasis caused by Leishmania major. Molecules, 27.","DOI":"10.3390\/molecules27248677"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Fern\u00e1ndez, L., Solana, J.C., S\u00e1nchez, C., Jim\u00e9nez, M.A., Requena, J.M., Coler, R., Reed, S.G., Valenzuela, J.G., Kamhawi, S., and Oliveira, F. (2021). Protective efficacy in a hamster model of a multivalent vaccine for human visceral leishmaniasis (Mulevaclin) consisting of the KMP11, LEISH-F3+, and LJL143 antigens in virosomes, plus GLA-SE adjuvant. Microorganisms, 9.","DOI":"10.3390\/microorganisms9112253"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1186\/s13071-025-06756-5","article-title":"Screening helper T lymphocyte epitopes based on IFN-gamma\/IL-10 ratio for developing a novel multi-epitope vaccine candidate using Wolbachia surface protein as an adjuvant against visceral leishmaniasis","volume":"18","author":"Zhang","year":"2025","journal-title":"Parasite Vectors"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1080\/14760584.2016.1213632","article-title":"Adjuvant system AS01: Helping to overcome the challenges of modern vaccines","volume":"16","author":"Didierlaurent","year":"2017","journal-title":"Expert Rev. Vaccines"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1080\/14760584.2024.2382725","article-title":"Adjuvant system AS01: From mode of action to effective vaccines","volume":"23","author":"Roman","year":"2024","journal-title":"Expert Rev. Vaccines"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Oliveira-Maciel, D., Dos-Santos, J.S., Oliveira-Silva, G., Mello, M.F., da Fonseca-Martins, A.M., Carneiro, M.P.D., Ramos, T.D., Firmino-Cruz, L., Gomes, D.C.O., and Rossi-Bergmann, B. (2021). MPLA and AddaVax\u00ae Adjuvants Fail to Promote Intramuscular LaAg Vaccine Protectiveness against Experimental Cutaneous Leishmaniasis. Microorganisms, 9.","DOI":"10.3390\/microorganisms9061272"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"105820","DOI":"10.1016\/j.rvsc.2025.105820","article-title":"Potential immunoprophylactic use of a multi-epitope protein of CD4+ and CD8+ T cells against visceral leishmaniasis in Balb\/c mice","volume":"194","author":"Costa","year":"2025","journal-title":"Res. Veter-Sci."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Ismail, N., Karmakar, S., Bhattacharya, P., Sepahpour, T., Takeda, K., Hamano, S., Matlashewski, G., Satoskar, A.R., Gannavaram, S., and Dey, R. (2022). Leishmania Major Centrin Gene-Deleted Parasites Generate Skin Resident Memory T-Cell Immune Response Analogous to Leishmanization. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.864031"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Soto, M., Corvo, L., Garde, E., Ram\u00edrez, L., Iniesta, V., Bonay, P., G\u00f3mez-Nieto, C., Gonz\u00e1lez, V.M., Mart\u00edn, M.E., and Alonso, C. (2015). Coadministration of the three antigenic Leishmania infantum poly (a) binding proteins as a DNA vaccine induces protection against Leishmania major infection in BALB\/c mice. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0003751"},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Alves-Silva, M.V., Nico, D., Luca, P.M., and Palatnik-de-Sousa, C.B. (2019). The F1F3 recombinant chimera of Leishmania donovani-nucleoside hydrolase (NH36) and its epitopes induce cross-protection against Leishmania (V.) braziliensis infection in mice. Front. Immunol., 10.","DOI":"10.3389\/fimmu.2019.00724"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Devender, M., Sebastian, P., Maurya, V.K., Kumar, K., Anand, A., Namdeo, M., and Maurya, R. (2024). Immunogenicity and protective efficacy of tuzin protein as a vaccine candidate in Leishmania donovani-infected BALB\/c mice. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1294397"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1111\/j.1365-2249.2007.03501.x","article-title":"The immunodominant T helper 2 (Th2) response elicited in BALB\/c mice by the Leishmania LiP2a and LiP2b acidic ribosomal proteins cannot be reverted by strong Th1 inducers","volume":"150","author":"Iborra","year":"2007","journal-title":"Clin. Exp. Immunol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1177\/039463200902200119","article-title":"Vaccination with Trichinella spirallis antigens increases CD8+ peripheral T cells and enhances the Th2 immune response in Leishmania infantum challenged mice","volume":"22","author":"Daoudaki","year":"2009","journal-title":"Int. J. Immunopathol. Pharmacol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.actatropica.2015.11.006","article-title":"A new Leishmania-specific hypothetical protein, LiHyT, used as a vaccine antigen against visceral leishmaniasis","volume":"154","author":"Martins","year":"2016","journal-title":"Acta Trop."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Bhowmick, S., Ravindran, R., and Ali, N. (2014). IL-4 contributes to failure, and colludes with IL-10 to exacerbate Leishmania donovani infection following administration of a subcutaneous leishmanial antigen vaccine. BMC Microbiol., 14.","DOI":"10.1186\/1471-2180-14-8"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Tedla, M.G., Nahar, M.F., Every, A.L., and Scheerlinck, J.Y. (2024). The Immune Memory Response of In Vitro-Polarised Th1, Th2, and Th17 Cells in the Face of Ovalbumin-Transgenic Leishmania major in a Mouse Model. Int. J. Mol. Sci., 25.","DOI":"10.1101\/2024.07.10.601391"},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Nateghi-Rostami, M., and Sohrabi, Y. (2024). Memory T cells: Promising biomarkers for evaluating protection and vaccine efficacy against leishmaniasis. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1304696"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"2849","DOI":"10.1007\/s00436-022-07628-y","article-title":"A new immunochemotherapy schedule for visceral leishmaniasis in a hamster model","volume":"121","author":"Santana","year":"2022","journal-title":"Parasitol. Res."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"3461","DOI":"10.1128\/AAC.00137-11","article-title":"CpG oligodeoxynucleotide and miltefosine, a potential combination for treatment of experimental visceral leishmaniasis","volume":"55","author":"Gupta","year":"2011","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Van Henten, S., Tesfaye, A.B., Abdela, S.G., Tilahun, F., Fikre, H., Buyze, J., Kassa, M., Cnops, L., Pareyn, M., and Mohammed, R. (2021). Miltefosine for the treatment of cutaneous leishmaniasis\u2014A pilot study from Ethiopia. PLoS Negl. Trop. Dis., 15.","DOI":"10.1371\/journal.pntd.0009460"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.ebiom.2018.09.029","article-title":"A Leishmania infantum genetic marker associated with miltefosine treatment failure for visceral leishmaniasis","volume":"36","author":"Carnielli","year":"2018","journal-title":"EBioMedicine."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"3794","DOI":"10.1128\/AAC.01787-15","article-title":"Preclinical studies evaluating subacute toxicity and therapeutic efficacy of LQB-118 in experimental visceral leishmaniasis","volume":"60","author":"Martins","year":"2016","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1051\/parasite\/2025009","article-title":"Leishmaniases: Strategies in treatment development","volume":"32","author":"Majoor","year":"2025","journal-title":"Parasite"},{"key":"ref_70","first-page":"147","article-title":"Renal involvement in leishmaniasis: A review of the literature","volume":"4","author":"Clementi","year":"2011","journal-title":"Nephrol. Dial. Transplant. Plus"}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1202\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T11:14:33Z","timestamp":1764069273000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/14\/12\/1202"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,25]]},"references-count":70,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["pathogens14121202"],"URL":"https:\/\/doi.org\/10.3390\/pathogens14121202","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,25]]}}}