Antigenicity, immunogenicity and protective efficacy of three proteins expressed in the promastigote and amastigote stages of leishmania infantum against visceral leishmaniasis

Vivian Tamietti Martins, Miguel Angel Chávez-Fumagalli, Daniela Pagliara Lage, Mariana Costa Duarte, Esther Garde, Lourena Emanuele Costa, Viviane Gomes Da Silva, Jamil Silvano Oliveira, Danielle Ferreira De Magalhães-Soares, Santuza Maria Ribeiro Teixeira, Ana Paula Fernandes, Manuel Soto, Carlos Alberto Pereira Tavares, Eduardo Antonio Ferraz Coelho

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27 Citas (Scopus)


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.

Idioma originalInglés
Número de artículoe0137683
PublicaciónPLoS ONE
EstadoPublicada - 14 set. 2015
Publicado de forma externa


Profundice en los temas de investigación de 'Antigenicity, immunogenicity and protective efficacy of three proteins expressed in the promastigote and amastigote stages of leishmania infantum against visceral leishmaniasis'. En conjunto forman una huella única.

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