title: Immunoproteomic and bioinformatic approaches to identify secreted Leishmania amazonensis, L. braziliensis, and L. infantum proteins with specific reactivity using canine serum.
abstract: Leishmania spp have a wide range of hosts, and each host can harbor several Leishmania species. Dogs, for example, are frequently infected by Leishmania infantum, where they constitute its main reservoir, but they also serve as hosts for L. braziliensis and L. amazonensis. Serological tests for antibody detection are valuable tools for diagnosis of L. infantum infection due to the high levels of antibodies induced, unlike what is observed in L. amazonensis and L. braziliensis infections. Likewise, serology-based antigen-detection can be useful as an approach to diagnose any Leishmania species infection using different corporal fluid samples. Immunogenic and secreted proteins constitute powerful targets for diagnostic methods in antigen detection. As such, we performed immunoproteomic (2-DE, western blot and mass spectrometry) and bioinformatic screening to search for reactive and secreted proteins from L. amazonensis, L. braziliensis, and L. infantum. Twenty-eight non-redundant proteins were identified, among which, six were reactive only in L. amazonensis extracts, 10 in L. braziliensis extracts, and seven in L. infantum extracts. After bioinformatic analysis, seven proteins were predicted to be secreted, two of which were reactive only in L. amazonensis extracts (52kDa PDI and the glucose-regulated protein 78), one in L. braziliensis extracts (pyruvate dehydrogenase E1 beta subunit) and three in L. infantum extracts (two conserved hypothetical proteins and elongation factor 1-beta). We propose that proteins can be suitable targets for diagnostic methods based on antigen detection.
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, 31279-910 Belo Horizonte, Minas Gerais, Brazil. Electronic address: brunasoaressl@yahoo.com.br.
authors.forename: B S S
authors.initials: BS
authors.lastname: Lima
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, 31279-910 Belo Horizonte, Minas Gerais, Brazil. Electronic address: junior-fialho@hotmail.com.
authors.forename: L C
authors.initials: LC
authors.lastname: Fialho
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, 31279-910 Belo Horizonte, Minas Gerais, Brazil. Electronic address: simonefpires@gmail.com.
authors.forename: S F
authors.initials: SF
authors.lastname: Pires
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Patologia, 31279-910 Belo Horizonte, Minas Gerais, Brazil. Electronic address: wagnertafuri@gmail.com.
authors.forename: W L
authors.initials: WL
authors.lastname: Tafuri
authors.affiliation: Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Departamento de Parasitologia, 31279-910 Belo Horizonte, Minas Gerais, Brazil. Electronic address: helida@icb.ufmg.br.
authors.forename: H M
authors.initials: HM
authors.lastname: Andrade
mesh_terms: D000818:Animals; D019295:Computational Biology / Q000379:methods*; D004283:Dog Diseases / Q000175:diagnosis / Q000469:parasitology*; D004285:Dogs; D007162:Immunoproteins / Q000302:isolation & purification*; D007891:Leishmania / Q000145:classification* / Q000302:isolation & purification; D007896:Leishmaniasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D040901:Proteomics / Q000379:methods*
chemical_list: D007162:Immunoproteins
keywords: Diagnosis; Immunoproteome; Leishmania; Secreted antigens