Prediction of CD8(+) Epitopes in Leishmania braziliensis Proteins Using EPIBOT: In Silico Search and In Vivo Validation

BACKGROUND: Leishmaniasis is caused by intracellular Leishmania parasites that induce a T-cell mediated response associated with recognition of CD4(+) and CD8(+) T cell Line 1Lineepitopes. Identification of CD8(+) antigenic determinants is crucial for vaccine and therapy development. Herein, we developed an open-source software dedicated to search and compile data obtained from currently available on line prediction algorithms. METHODOLOGY/PRINCIPAL FINDINGS: We developed a two-phase algorithm and implemented in an open source software called EPIBOT, that consolidates the results obtained with single prediction algorithms, generating a final output in which epitopes are ranked. EPIBOT was initially trained using a set of 831 known epitopes from 397 proteins from IEDB. We then screened 63 Leishmania braziliensis vaccine candidates with the EPIBOT trained tool to search for CD8(+) T cell epitopes. A proof-of-concept experiment was conducted with the top eight CD8(+) epitopes, elected by EPIBOT. To do this, the elected peptides were synthesized and validated for their in vivo cytotoxicity. Among the tested epitopes, three were able to induce lysis of pulsed-target cells. CONCLUSION: Our results show that EPIBOT can successfully search across existing prediction tools, generating a compiled list of candidate CD8(+) epitopes. This software is fast and a simple search engine that can be customized to search over different MHC alleles or HLA haplotypes.