Leishmania donovani: in vitro culture and [1H] NMR characterization of amastigote-like forms.
When Leishmania donovani promastigote forms, were cultured in TC-199 medium at 28 degrees C and subsequently incubated at 38 degrees C, they turned into aflagellate (amastigote-like) forms. A return of the incubation-culture temperature to 28 degrees 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 degrees 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 degrees C, transformed into promastigotes capable of multiplying as flagellate forms. The amastigote-like forms obtained in 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 of L. 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 of Leishmania use different biochemical strategies to obtain their energy.
Castilla
J J
JJ
Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, Spain.
Sanchez-Moreno
M
M
Mesa
C
C
Osuna
A
A
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology; D002460:Cell Line; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism; D005407:Flagella / Q000648:ultrastructure; D006019:Glycolysis; D006863:Hydrogen-Ion Concentration; D007893:Leishmania donovani / Q000254:growth & development* / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D008264:Macrophages / Q000469:parasitology; D009682:Magnetic Resonance Spectroscopy; D013696:Temperature
D000953:Antigens, Protozoan; D003470:Culture Media