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Immunoproteomics approach for the discovery of antigens applied to the diagnosis of canine visceral leishmaniasis

2023; Elsevier BV; Volume: 241; Linguagem: Inglês

10.1016/j.actatropica.2023.106865

1873-6254

Scarleth Silva Costa, Lucas Magno Oliveira Santos, Larissa Chaves Freire, Ana Luiza Filizzola Tedeschi, Naianda Rezende Ribeiro, Mariana Helena Rodrigues Queiroz, Emídio Beraldo-Neto, Daniel C. Pimenta, Nathália Coral Galvani, Gabriel Paulino Luiz, Maria Eduarda de Oliveira, Ricardo Andrez Machado‐de‐Ávila, Ana Maria Ravena Severino Carvalho, Bryan Victor Serafim Brigido, Alexandre Barbosa Reis, Ana Paula Fernandes, Eduardo Antônio Ferraz Coelho, Bruno Mendes Roatt, Daniel Menezes‐Souza, Mariana C. Duarte,

Toxin Mechanisms and Immunotoxins

In the present study, an immunoproteomic approach using Leishmania infantum parasites isolated from naturally infected dogs from an endemic region of the disease, was carried out to identify new antigens to be used in the diagnosis of canine visceral leishmaniasis (CVL). Protein extracts, obtained from parasites isolated from asymptomatic (CanLA) and symptomatic (CanLS) dogs, were used to perform the two-dimensional gels. Western Blotting assays were carried out by employing a pool of sera from dogs with visceral leishmaniasis (CanLA or CanLS), healthy dogs from an endemic area, or dogs with similar diseases associated with cross-reactions (babesiosis and ehrlichiosis). With these results, it was possible to exclude the spots that showed a cross-reactivity of the sera from groups of healthy dogs, and those with babesiosis or ehrlichiosis. Taken together, 20 proteins were identified, 15 of which have already been described in the literature and 5 of which are hypothetical. An immunogenomic screen strategy was applied to identify conserved linear B-cell epitopes in the identified hypothetical proteins. Two peptides were synthesized and tested in ELISA experiments as a proof of concept for the validation of our immunoproteomics findings. The results demonstrated that the antigens presented sensitivity and specificity values ranging from 81.93% to 97.59% and 78.14 to 85.12%, respectively. As a comparative antigen, a preparation of a Leishmania extract showed sensitivity and specificity values of 75.90% and 74.88%, respectively. The present study was able to identify proteins capable of being used for the serodiagnosis of canine visceral leishmaniasis.