Leishmania donovani: In vitro culture and [1H] NMR characterization of amastigote-like forms

Abstract

WhenLeishmania donovani promastigote forms, were cultured in TC-199 medium at 28°C and subsequently incubated at 38°C, they turned into aflagellate (amastogote-like) forms. A return of the incubation-culture temperature to 28°C these amastigote-like forms to revert to promastigotes. The amastigotes obtained by heat-shock, were viable and retained antigenic capacity being recognized by the sera of naturally infected patients. These forms, remained also capable of multiplying inside the J-774A. 1 macrophages. When the amastigote-like forms are kept in culture at 38°C retained their rounded appearance and their biological characteristics for more than 3 months subculturing every 6 days. These amastigote-like forms, when used for subcultures at 28°C, transformed into promastigotes capable of multiplying, as flagellate forms. The amastigotelike forms obtainedin vitro can be used in biochemical studies related to chemotherapy and immunology studies, as part of an effort to combat this parasite. The end-products of of glycolysis were studied in both the amastigote-like and promastigote forms ofL. donovani, by proton magnetic resonance analysis of the culture media. Alanine, succinate, and acetate, were predominant, and to a lesser extent pyruvate, glycine and D-lactate. Our results suggest that both forms ofLeishmania use different biochemical strategies to obtain their energy.