Contents lists available at ScienceDirect Cytokine journal homepage: www.elsevier.com/locate/cytokine Leishmania infantum amastin protein incorporated in distinct adjuvant systems induces protection against visceral leishmaniasis Patrícia A.F. Ribeiro a , Danniele L. Vale a , Daniel S. Dias a , Daniela P. Lage a , Débora V.C. Mendonça a , Fernanda F. Ramos a , Lívia M. Carvalho b , Ana Maria R.S. Carvalho a , Bethina T. Steiner c , Marjorie C. Roque d , João A. Oliveira-da-Silva a , Jamil S. Oliveira e , Grasiele S.V. Tavares a , Nathália C. Galvani a , Vívian T. Martins a , Miguel A. Chávez-Fumagalli a , Bruno M. Roatt b , Ricardo L.F. Moreira f , Daniel Menezes-Souza a,g , Mônica C. Oliveira d , Ricardo A. Machado-de-Ávila c , Antônio L. Teixeira a,h , Eduardo A.F. Coelho a,g, ⁎ a Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil b Laboratório de Imunopatologia, Núcleo de Pesquisas em Ciências Biológicas/NUPEB, Departamento de Ciências Biológicas, Insituto de Ciências Exatas e Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil c Programa de Pós-Graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma 88806-000, Santa Catarina, Brazil d Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil e Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil f Fundação Hospitalar do Estado de Minas Gerais, Hospital Eduardo de Menezes, Belo Horizonte 30622-020, Minas Gerais, Brazil g Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, 31270-901 Belo Horizonte, Minas Gerais, Brazil h Neuropsychiatry Program, Department of Psychiatry and Behavioral Sciences, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX, USA ARTICLE INFO Keywords: Visceral leishmaniasis Amastin Vaccine Immunogenicity Adjuvants ABSTRACT The control measures against visceral leishmaniasis (VL) include a precise diagnosis of disease, the treatment of human cases, and reservoir and vector controls. However, these are insufficient to avoid the spread of the disease in specific countries worldwide. As a consequence, prophylactic vaccination could be interesting, although no effective candidate against human disease is available. In the present study, the Leishmania infantum amastin protein was evaluated regarding its immunogenicity and protective efficacy against experimental VL. BALB/c mice immunized with subcutaneous injections of the recombinant protein with or without liposome/saponin (Lip/Sap) as an adjuvant. After immunization, half of the animals per group were euthanized and immunological evaluations were performed, while the others were challenged with L. infantum promastigotes. Forty-five days after infection, the animals were euthanized and parasitological and immunological evaluations were performed. Results showed the development of a Th1-type immune response in rAmastin-Lip and rAmastin-Sap/vaccinated mice, before and after infection, which was based on the production of protein and parasite-specific IFN-γ, IL-12, GM-CSF, and nitrite, as well as the IgG2a isotype antibody. CD4 + T cells were mainly responsible for IFN-γ production in vaccinated mice, which also presented significant reductions in parasitism in their liver, spleen, draining lymph nodes, and bone marrow. In addition, PBMC cultures of treated VL patients and healthy subjects stimulated with rAmastin showed lymphoproliferation and higher IFN-γ production. In conclusion, the present study shows the first case of an L. infantum amastin protein associated with distinct delivery systems inducing protection against L. infantum infection and demonstrates an immunogenic effect of this protein in human cells. https://doi.org/10.1016/j.cyto.2020.155031 Received 11 October 2019; Received in revised form 21 January 2020; Accepted 6 February 2020 ⁎ Corresponding author at: Laboratório de Pesquisa do Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Prof. Alfredo Balena, 190, 30.130-100, Belo Horizonte, Minas Gerais, Brazil. E-mail address: eduardoferrazcoelho@yahoo.com.br (E.A.F. Coelho). Cytokine 129 (2020) 155031 1043-4666/ © 2020 Elsevier Ltd. All rights reserved. T