Antigenicity, immunogenicity and protective efficacy of three proteins expressed in the promastigote and amastigote stages of leishmania infantum against visceral leishmaniasis
AuthorMartins, Vivian Tamietti; Chávez-Fumagalli, Miguel Angel; Lage, Daniela P.; Duarte, Mariana Costa; Garde, Esther; Costa, Lourena Emanuele; Silva, Viviane Gomes da; Oliveira, Jamil Silvano; Magalhães-Soares, Danielle Ferreira de; Teixeira, Santuza Maria Ribeiro; Fernandes, Ana Paula Salles Moura; Soto Álvarez, Manuel; Tavares, Carlos Alberto Pereira; Coelho, Eduardo Antonio Ferraz
EntityUAM. Departamento de Biología Molecular
PublisherPublic Library of Science
10.1371/journal.pone.0137683PLoS ONE 10.9 (2015): e0137683
Funded byThis work was supported by grants from Pró-Reitoria de Pesquisa from UFMG (Edital 01/ 2014), Instituto Nacional de Ciência e Tecnologia em Nanobiofarmacêutica (INCT-Nanobiofar), FAPEMIG (CBB-APQ-00819-12), and CNPq (APQ-472090/ 2011-9, APQ-482976/2012-8, APQ-488237/2013-0). MACF is a grant recipient of FAPEMIG/CAPES. EAFC, APF and SMRT are grant recipient of CNPq. This study was also, in part, supported in Spain by grants from Ministerio de Ciencia e Innovación (FIS PI14/00366). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript
ProjectGobierno de España. FIS PI14/00366
SubjectsAmastigote; Dog; Female; Nonhuman; Biología y Biomedicina / Biología
Rights© 2015 Martins et al.
Esta obra está bajo una Licencia Creative Commons Atribución 4.0 Internacional.
In the present study, two Leishmania infantum hypothetical proteins present in the amastigote stage, LiHyp1 and LiHyp6, were combined with a promastigote protein, IgE-dependent histamine-releasing factor (HRF); to compose a polyproteins vaccine to be evaluated against L. infantum infection. Also, the antigenicity of the three proteins was analyzed, and their use for the serodiagnosis of canine visceral leishmaniasis (CVL) was evaluated. The LiHyp1, LiHyp6, and HRF DNA coding sequences were cloned in prokaryotic expression vectors and the recombinant proteins were purified. When employed in ELISA assays, all proteins were recognized by sera from visceral leishmaniasis (VL) dogs, and presented no cross-reactivity with either sera from dogs vaccinated with a Brazilian commercial vaccine, or sera of Trypanosoma cruzi-infected or Ehrlichia canis-infected animals. In addition, the antigens were not recognized by antibodies from non-infected animals living in endemic or non-endemic areas for leishmaniasis. The immunogenicity and protective efficacy of the three proteins administered in the presence of saponin, individually or in combination (composing a polyproteins vaccine), were evaluated in a VL murine model: BALB/c mice infected with L. infantum. Spleen cells from mice inoculated with the individual proteins or with the polyproteins vaccine plus saponin showed a protein-specific production of IFN-γ, IL-12, and GM-CSF after an in vitro stimulation, which was maintained after infection. These animals presented significant reductions in the parasite burden in different evaluated organs, when compared to mice inoculated with saline or saponin. The decrease in parasite burden was associated with an IL-12-dependent production of IFN-γ against parasite total extracts (produced mainly by CD4+ T cells), correlated to the induction of parasite proteins-driven NO production. Mice inoculated with the recombinant protein-based vaccines showed also high levels of parasite-specific IgG2a antibodies. The polyproteins vaccine administration induced a more pronounced Th1 response before and after challenge infection than individual vaccines, which was correlated to a higher control of parasite dissemination to internal organs
Google Scholar:Martins, Vivian Tamietti - Chávez-Fumagalli, Miguel Angel - Lage, Daniela P. - Duarte, Mariana Costa - Garde, Esther - Costa, Lourena Emanuele - Silva, Viviane Gomes da - Oliveira, Jamil Silvano - Magalhães-Soares, Danielle Ferreira de - Teixeira, Santuza Maria Ribeiro - Fernandes, Ana Paula Salles Moura - Soto Álvarez, Manuel - Tavares, Carlos Alberto Pereira - Coelho, Eduardo Antonio Ferraz
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Subtractive phage display selection from canine visceral leishmaniasis identifies novel epitopes that mimic leishmania infantum antigens with potential serodiagnosis applications Costa, Lourena Emanuele; Lima, Mayara Ingrid Sousa; Chávez-Fumagalli, Miguel Angel; Menezes-Souza, Daniel; Martins, Vivian Tamietti; Duarte, Mariana Costa; Lage, Paula Sousa; Lopes, Eliane Gonçalves Paiva; Lage, Daniela P.; Ribeiro, Tatiana Gomes; Andrade, Pedro H R; De Magalhães Soares, Danielle Ferreira; Soto Álvarez, Manuel; Tavares, Carlos Alberto Pereira; Goulart, Luiz Ricardo; Coelho, Eduardo Anton̂io Ferraz
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