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Leishmania (Viannia) panamensis-specific IgE and IgA antibodies were quantified in patients with parasitologically confirmed American tegumentary leishmaniasis using a radioallergosorbent test (RAST) and an enzyme-linked immunosorbent assay (ELISA), respectively. The RAST values, presented as the mean ± SEM percentage of total isotope added, were significantly elevated in patients having disease evolution greater than 12 months (3.14 ± 0.91), as compared with those with an evolution time of 12 or fewer months (1.66 ± 0.15) (P = 0.017). A separate group of patients, those with eosinophils in the biopsy specimen of their lesion, also had elevated mean RAST values (2.55 ± 0.58) when compared with patients who did not demonstrate these cells in their biopsy specimens (1.32 ± 0.24) (P = 0.038). Leishmania-specific IgA levels, presented as the mean ± SEM optical density, were significantly higher for patients with mucocutaneous disease (0.40 ± 0.03) than for patients with cutaneous disease (0.28 ± 0.023) (P = 0.0063). Inhibition testing with homologous and heterologous antigens confirmed the specificity of these assays, and were used to assess cross-reactivity among L. (Viannia) subspecies and other kinetoplastid hemoparasites. Results demonstrate that patients with more severe forms of American tegumentary leishmaniasis, defined either as increased duration of disease or invasion of the mucosa, have elevated levels of Leishmania-specific IgE and IgA antibodies, respectively.