Article
Analysis of Leishmania chagasi by 2-D Difference Gel Eletrophoresis (2-D DIGE) and Immunoproteomic: Identification of Novel Candidate Antigens for Diagnostic Tests and Vaccine
Registro en:
COSTA, Míriam M. et al. Analysis of Leishmania chagasi by 2-D Difference Gel Eletrophoresis (2-D DIGE) and Immunoproteomic: Identification of Novel Candidate Antigens for Diagnostic Tests and Vaccine. J. Proteome Res., v.10, p.2172–2184, Feb. 2011.
1535-3893
10.1021/pr101286y
1535-3907
Autor
Costa, Míriam M.
Andrade, Hélida M.
Bartholomeu, Daniella C.
Freitas, Leandro M.
Pires, Simone F.
Chapeaurouge, Alexander D.
Perales, Jonas
Ferreira, André T.
Giusta, Mário S.
Melo, Maria N.
Gazzinelli, Ricardo T.
Resumen
Identification of novel antigens is essential for
developing new diagnostic tests and vaccines. We used DIGE to
compare protein expression in amastigote and promastigote
forms of Leishmania chagasi. Nine hundred amastigote and
promastigote spots were visualized. Five amastigote-specific,
25 promastigote-specific, and 10 proteins shared by the two
parasite stages were identified. Furthermore, 41 proteins were
identified in the Western blot employing 2-DE and sera from
infected dogs. From these proteins, 3 and 38 were reactive with
IgM and total IgG, respectively. The proteins recognized by total
IgG presented different patterns in terms of their recognition by
IgG1 and/or IgG2 isotypes. All the proteins selected by Western
blot were mapped for B-cell epitopes. One hundred and eighty
peptides were submitted to SPOT synthesis and immunoassay.
A total of 25 peptides were shown of interest for serodiagnosis to visceral leishmaniasis. In addition, all proteins identified in this
study were mapped for T cell epitopes by using the NetCTL software, and candidates for vaccine development were selected.
Therefore, a large-scale screening of L. chagasi proteome was performed to identify new B and T cell epitopes with potential use for
developing diagnostic tests and vaccines. 2030-01-01