Epitope mapping of a single repetitive unit of the B13 Trypanosoma cruzi antigen as fusions to Escherichia coli LamB protein

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Pereira, Catia Mendes [UNIFESP]
Franco da Silveira, José [UNIFESP]
Duranti, M.
Cunha-Neto, E.
Zingales, B.
Castilho, Beatriz Amaral de [UNIFESP]
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B13, one of the immunodominant antigens of Trypanosoma cruzi, is composed of repeats of a 12-amino-acid motif. Using synthetic peptides, the sequence FGQAAAGDK was previously shown to contain the B13 immunodominant epitope recognized by chagasic patients sera. To investigate the effects of neighboring sequences in the immunodominance, we tested serum recognition of two B13 sequences fused to LamB. GDKPSPFGQAAA-LamB and FGQAAAGDKPSP-LamB were recognized, respectively, by 15% and 80% of 80 sera reactive to B13 antigen. Recognition of FGQAAAGDKPSP-LamB was inhibited by AAAGDK-containing synthetic peptides. FGQAAAGDKPSP-LamB competed with a B13 recombinant protein containing 16.6 repeats for binding to chagasic antibodies. These results strengthen previous conclusions on the immunodominant epitope of B13 and provide a comparison of two methods for epitope mapping. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Microbiological Societies.
Fems Microbiology Letters. Amsterdam: Elsevier B.V., v. 235, n. 2, p. 237-242, 2004.