title: Epitope mapping of the HSP83.1 protein of Leishmania braziliensis discloses novel targets for immunodiagnosis of tegumentary and visceral clinical forms of leishmaniasis.
abstract: Gold standard serological diagnostic methods focus on antigens that elicit a strong humoral immune response that is specific to a certain pathogen. In this study, we used bioinformatics approaches to identify linear B-cell epitopes that are conserved among Leishmania species but are divergent from the host species Homo sapiens and Canis familiaris and from Trypanosoma cruzi, the parasite that causes Chagas disease, to select potential targets for the immunodiagnosis of leishmaniasis. Using these criteria, we selected heat shock protein 83.1 of Leishmania braziliensis for this study. We predicted three linear B-cell epitopes in its sequence. These peptides and the recombinant heat shock protein 83.1 (rHSP83.1) were tested in enzyme-linked immunosorbent assays (ELISAs) against serum samples from patients with tegumentary leishmaniasis (TL) and visceral leishmaniasis (VL) and from dogs infected with Leishmania infantum (canine VL [CVL]). Our data show that rHSP83.1 is a promising target in the diagnosis of TL. We also identified specific epitopes derived from HSP83.1 that can be used in the diagnosis of human TL (peptide 3), both human and canine VL (peptides 1 and 3), and all TL, VL, and CVL clinical manifestations (peptide 3). Receiver operating characteristic (ROC) curves confirmed the superior performance of rHSP83.1 and peptides 1 and 3 compared to that of the soluble L. braziliensis antigen and the reference test kit for the diagnosis of CVL in Brazil (EIE-LVC kit; Bio-Manguinhos, Fiocruz). Our study thus provides proof-of-principle evidence of the feasibility of using bioinformatics to identify novel targets for the immunodiagnosis of parasitic diseases using proteins that are highly conserved throughout evolution.
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Menezes-Souza
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: Tiago Antônio de Oliveira
authors.initials: TA
authors.lastname: Mendes
authors.affiliation: Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Patos de Minas, Brazil.
authors.forename: Matheus de Souza
authors.initials: Mde S
authors.lastname: Gomes
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: João Luís
authors.initials: JL
authors.lastname: Reis-Cunha
authors.affiliation: Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: Ronaldo Alves Pinto
authors.initials: RA
authors.lastname: Nagem
authors.affiliation: Departamento de Análises Clínicas, Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Brazi.
authors.forename: Cláudia Martins
authors.initials: CM
authors.lastname: Carneiro
authors.affiliation: Programa de Pós-Graduação em Ciências da Saúde, Infectologia e Medicina Tropical, Faculdade de Medicina and Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Eduardo Antônio Ferraz
authors.initials: EA
authors.lastname: Coelho
authors.affiliation: Programa de Pós-Graduação em Ciências da Saúde, Universidade Federal do Rio Grande do Norte, Natal, Brazil.
authors.forename: Lúcia Maria da Cunha
authors.initials: LM
authors.lastname: Galvão
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: Ricardo Toshio
authors.initials: RT
authors.lastname: Fujiwara
authors.affiliation: Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil daniella@icb.ufmg.br.
authors.forename: Daniella Castanheira
authors.initials: DC
authors.lastname: Bartholomeu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019295:Computational Biology; D004283:Dog Diseases / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D005260:Female; D006360:Heat-Shock Proteins / Q000276:immunology*; D006801:Humans; D007159:Immunologic Tests; D007892:Leishmania braziliensis / Q000276:immunology*; D018314:Leishmania infantum / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary*; D008297:Male; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000276:immunology*; D016415:Sequence Alignment; D014349:Trypanosoma cruzi / Q000276:immunology
chemical_list: D018985:Epitopes, B-Lymphocyte; D006360:Heat-Shock Proteins; C490310:Hsp83 protein, protozoan; D015800:Protozoan Proteins