Excreted Leishmania peruviana and Leishmania amazonensis iron-superoxide dismutase purification: specific antibody detection in Colombian patients with cutaneous leishmaniasis.
Leishmania sp. survival in the vertebrate host depends on the host macrophage immune response as well as on the parasite's defense against free radicals. Iron-superoxide dismutase (Fe-SOD) is a key antioxidant enzyme that contributes to radical superoxide dismutation, preventing the disease from surging and propagating itself. Leishmania sp. has various Fe-SOD isoforms, one of which (Fe-SODe) is excreted into the medium and, being highly immunogenic, can be considered a very good molecular marker. In this work, we purified the Fe-SOD enzymes excreted by L. peruviana and L. amazonensis and studied them as antigens in serodiagnosis. We used ELISA and Western blot techniques to test 51 human cutaneous leishmaniasis sera from Colombia. All 51 patients presented with dermal injuries caused by unknown Leishmania species. The results observed with the purified proteins were compared with those obtained when total soluble lysate and unpurified Fe-SODe were used as the antigen fraction. Thus, we conclude that the purified enzymes are more sensitive and specific than their unpurified counterparts and that there is no cross-reactivity between them.
Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain. Electronic address: longonisilviaste@ugr.es.
Silvia Stefania
SS
Longoni
Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain.
Clotilde
C
Marín
Departamento de Parasitología, Facultad de Ciencias, Universidad de Granada, 18071 Granada, Spain.
Manuel
M
Sánchez-Moreno
D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D003105:Colombia; D005609:Free Radicals / Q000378:metabolism; D006801:Humans; D007891:Leishmania / Q000201:enzymology / Q000276:immunology; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000276:immunology; D013482:Superoxide Dismutase / Q000276:immunology* / Q000302:isolation & purification*
D000906:Antibodies; D005609:Free Radicals; D013482:Superoxide Dismutase
Chromatography; Colombia; ELISA; Free radicals; Iron superoxide dismutase; Leishmania amazonensis; Leishmania peruviana