A proteomic road to acquire an accurate serological diagnosis for human tegumentary leishmaniasis.
Diagnostic tools are important for clinical management and epidemiological evaluation of Tegumentary (TL) and Visceral (VL) Leishmaniasis. Serology is not frequently used for the diagnosis of the TL form because low antibody titers and cross-reaction with VL. Therefore, it is crucial to identify specific and immunogenic antigens from species associated with the TL form. Here we employed a proteomic approach coupled to an in silico analysis and identified the most abundant and immunogenic proteins from Leishmania amazonensis, Leishmania braziliensis and Leishmania infantum. Of 16 species specific proteins, nine were from the species causative of the TL form (L. amazonensis and L. braziliensis). In silico analysis revealed 18 B-cell epitopes with 0% similarity to Trypanosoma cruzi orthologs and, therefore, less likely to crossreact with sera of patients with Chagas disease. Two proteins reacted exclusively with serum from TL patients and presented several B-cell epitopes without similarity to T. cruzi orthologs: the hypothetical protein GI 134063939 and the metallo-peptidase Clan MA(E)-Family M3. The immunoassay revealed nine peptides with strong reactivity to sera from TL patients. These proteins and peptides may be good candidates to improve the specificity and sensibility of serological tests aiming to diagnose the TL of this neglected human disease.



As no gold-standard test for tegumentary leishmaniasis (TL) exists, a combination of different diagnostic techniques is often necessary to obtain precise results. Thus, the identification of species-specific, highly immunogenic and abundant proteins that stimulate the humoral immune response in the host should help in the development of serological tests for human TL. Herein we searched for these potential antigens in Leishmania species related to American Leishmaniasis (L. amazonensis, L. braziliensis and L. infantum). To this end, we employed an immunoproteomic approach using proteins from these Leishmania species and sera from TL and Visceral Leishmaniasis (VL) patients. Our study unveils specific proteins and peptides that may represent antigens that will help the efforts to improve the accuracy of serological tests to diagnose the TL form.
Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil.
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BS
Lima
Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil.
S F
SF
Pires
Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil.
L C
LC
Fialho
Laboratório de Pesquisas Clínicas, Centro de Pesquisas René Rachou, CP: 1742 - CEP: 30190-002 Belo Horizonte, Minas Gerais, Brazil.
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EJ
Oliveira
Unidade Acadêmica de Ciências da Saúde, Universidade do Extremo Sul Catarinense, CEP: 88.806-000 Criciúma, Santa Catarina, Brazil.
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RA
Machado-de-Avila
Departamento Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil.
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Chávez-Olórtegui
Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fiocruz, CEP: 21040-360, Rio de Janeiro, Brazil.
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Chapeaurouge
Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fiocruz, CEP: 21040-360, Rio de Janeiro, Brazil.
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Perales
Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. Electronic address: helidandrade@gmail.com.
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HM
Andrade
D000953:Antigens, Protozoan / Q000032:analysis*; D003429:Cross Reactions; D003937:Diagnosis, Differential; D006801:Humans; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000276:immunology; D058069:Neglected Diseases / Q000175:diagnosis; D040901:Proteomics / Q000379:methods; D015800:Protozoan Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods* / Q000592:standards; D013045:Species Specificity
D000953:Antigens, Protozoan; D015800:Protozoan Proteins
Diagnostic; Epitopes; Immunoassays; Immunoproteomic; Mass spectrometry; Spot synthesis; Tegumentary leishmaniasis