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A new Leishmania-specific hypothetical protein and its non-described specific B cell conformational epitope applied in the serodiagnosis of canine visceral leishmaniasis

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Abstract

The serodiagnosis of canine visceral leishmaniasis (CVL) presents problems related to its sensitivity and/or specificity. In the present study, a new Leishmania-specific hypothetical protein, LiHyD, was produced as a recombinant protein (rLiHyD) and evaluated in ELISA experiments for the CVL serodiagnosis. LiHyD was characterized as antigenic in a recent immunoproteomic search performed with Leishmania infantum proteins and the sera of dogs developing visceral leishmaniasis (VL). Aiming to compare the efficacy between whole proteins and synthetic peptides, two linear and one conformational B cell epitopes of LiHyD were synthesized and also evaluated as diagnostic markers. The four antigens were recognized by the sera of dogs suffering VL. On the contrary, low reactivity was observed when they were assayed with sera from non-infected healthy dogs living in endemic or non-endemic areas of leishmaniasis. In addition, no reactivity was found against them using sera from dogs experimentally infected by Trypanosoma cruzi, Babesia canis, or Ehrlichia canis, or sera from animals vaccinated with the Leish-Tec® vaccine, a prophylactic preparation commercially available for CVL prevention in Brazil. As comparative diagnostic tools, a recombinant version of the amastigote-specific A2 protein and a soluble crude Leishmania extract were studied. Both antigens presented lower sensitivity and/or specificity values than the LiHyD-based products. The rLiHyD presented better results for the CVL serodiagnosis than its linear epitopes, although the peptide recreating the conformational epitope resulted also appropriate as a diagnostic marker of CVL. To the best of our knowledge, this is the first study showing the use of a conformational epitope derived from a Leishmania protein for serodiagnosis of CVL.

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Acknowledgments

This work was supported by grants from Instituto Nacional de Ciência e Tecnologia em Nano-biofarmacêutica (INCT-NanoBiofar), FAPEMIG (CBB-APQ-00496-11, CBB-APQ-00819-12, and CBB-APQ-01778-2014), and CNPq (APQ-472090/2011-9 and APQ-488237/2013-0). In addition, this study was partially funded in Madrid by the Spanish grant from Ministerio de Economía y Competitividad-FEDER (FISPI14/00366 from the Instituto de Salud Carlos III), and from Universidade do Extremo Sul Catarinense (UNESC). MACF is a grant recipient of FAPEMIG/CAPES. EAFC is a grant recipient of CNPq.

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Authors and Affiliations

  1. 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, Belo Horizonte, Minas Gerais, 30130-100, Brazil

    Daniela P. Lage, Lourena E. Costa, Miguel A. Chávez-Fumagalli & Eduardo A. F. Coelho

  2. Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, 31270-901, Brazil

    Vívian T. Martins & Carlos A. P. Tavares

  3. Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, 31270-901, Brazil

    Mariana C. Duarte, Daniel Menezes-Souza & Bruno M. Roatt

  4. Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Departamento de Biología Molecular, Universidad Autónoma de Madrid, 28049, Madrid, Spain

    Esther Garde & Manuel Soto

  5. Programa de Pós-Graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, Santa Catarina, 88806-000, Brazil

    Laura M. Dimer, Amanda C. S. Kursancew & Ricardo A. Machado-de-Ávila

  6. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, 31270-901, Brazil

    Danielle F. de Magalhães-Soares

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  1. Daniela P. Lage
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Lage, D.P., Martins, V.T., Duarte, M.C. et al. A new Leishmania-specific hypothetical protein and its non-described specific B cell conformational epitope applied in the serodiagnosis of canine visceral leishmaniasis. Parasitol Res 115, 1649–1658 (2016). https://doi.org/10.1007/s00436-016-4904-x

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