Molecular and antigenic characterization of the Leishmania (Viannia) panamensis kinetoplastid membrane protein-11

RESUMO

The kinetoplastid membrane protein 11 (KMP-11) has been recently described in Leishmania (Leishmania) donovani as a major component of the promastigote membrane. Two oligonucleotide primers were synthesized to PCR-amplify the entire coding region of New World Leishmania species. The Leishmania (Viannia) panamensis amplification product was cloned, sequenced and the putative amino acid sequence determined. A remarkably high degree of sequence homology was observed with the corresponding molecule of L. (L) donovani and L. (L) infantum (97 per cent and 96 per cent respectively). Southern blot analysis showed that the KMP-11 locus is conformed by three copies of the gene. The L. (L) panamensis ORF was subsequently cloned in a high expression vector and the recombinant protein was included and purified from Escherichia coli cultures. Immunoblot analysis showed that 80 per cent, 77 per cent and 100 per cent sera from cutaneous, mucocutaneous and visceral leishmaniasis patients, respectively, recognized the recombinant KMP-11 protein. In a similar assay, 86 per cent of a asymptomatic Leishmania-infected individuals showed IgG antibodies against the rKMP-11. We propose that KMP-11 could be used as a serologic marker for infection and disease caused by Leishmania in America.