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A simple and reproducible method to obtain large numbers of axenic amastigotes of different Leishmania species.

By Márcia Cristina Aquino Teixeira, Regilene de Jesus Santos, Romina Barreto Sampaio, Lain Carlos Pontes-de-Carvalho and Washington Luis Conrado dosSantos

Abstract

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

Topics: Leishmania/crescimento & desenvolvimento, Parasitologia/métodos, Animais, Western Blotting, Cricetinae, Meios de Cultura, Humanos, Leishmania/patogenicidade, Leishmania/ultraestrutura, Leishmaniose/parasitologia, Leishmaniose/fisiopatologia, Camundongos Endogâmicos BALB C, Microscopia Eletrônica, Reprodutibilidade dos Testes
Year: 2002
OAI identifier: oai:agregador.ibict.br.RI_FIOCRUZ:oai:localhost:icict/4212
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