A simple and reproducible method to obtain large numbers of axenic amastigotes of different Leishmania species.
This work describes a simple method to yield large amounts of Leishmania amastigote-like forms in axenic cultures using promastigotes as the starting population. The method described induced extracellular amastigote transformation of Leishmania amazonensis (97%), Leishmania braziliensis (98%) and Leishmania chagasi (90%). The rounded parasites obtained in axenic cultures were morphologically similar, even at the ultrastructural level, to intracellular amastigotes. Moreover, the axenic amastigotes remained viable as measured by their ability to revert back to promastigotes and to infect BALB/c mice. L. amazonensis and L. braziliensis promastigotes and axenic amastigotes differed in terms of their Western blot profiles. A 46 kDa protein was recognized by specific antibodies only in axenic and lesion-derived L. amazonensis amastigotes and not in promastigotes.
Teixeira
Márcia Cristina Aquino
MC
Laboratório Central de Saúde Pública Professor Gonçalo Moniz, Av Waldemar Falcão 123, 40295-001, Salvador, Brazil.
de Jesus Santos
Regilene
R
Sampaio
Romina Barreto
RB
Pontes-de-Carvalho
Lain
L
dos-Santos
Washington L C
WL
D000818:Animals; D015153:Blotting, Western; D006224:Cricetinae; D003470:Culture Media; D006801:Humans; D007891:Leishmania / Q000254:growth & development* / Q000472:pathogenicity / Q000648:ultrastructure; D007896:Leishmaniasis / Q000469:parasitology / Q000503:physiopathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D010274:Parasitology / Q000379:methods*; D015203:Reproducibility of Results
D003470:Culture Media