Fast high yield of pure Leishmania (Leishmania) infantum axenic amastigotes and their infectivity to mouse macrophages |
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Authors: | Juliana Dias Costa Renata Soares Léa Cysne Finkelstein Suzana C?rte-Real Maria de Nazareth Meirelles and Renato Porrozzi |
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Institution: | (1) Laborat?rio de Pesquisas em Leishmaniose, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil 4365, Rio de Janeiro, RJ, 21045-900, Brazil;(2) Laborat?rio de Ultra-Estrutura Celular, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil 4365, Rio de Janeiro, RJ, 21045-900, Brazil;(3) Laborat?rio de Imunoparasitologia, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil 4365, Rio de Janeiro, RJ, 21045-900, Brazil;(4) Laborat?rio de Biologia Estrutural, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil 4365, Rio de Janeiro, RJ, 21045-900, Brazil;(5) Centro Universit?rio de Volta Redonda—UniFOA, Avenida Paulo Erlei Alves Abrantes 1325, Tr?s Po?os, Volta Redonda, RJ, 27240-560, Brazil; |
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Abstract: | Leishmania (L.) infantum (syn. Leishmania chagasi) is a dimorphic protozoan parasite that lives in promastigote and amastigote form in its sandfly vector and mammalian hosts,
respectively. Here, we describe an in vitro culture system for the generation of a pure population of L. infantum axenic amastigotes after only 4 days incubation in culture medium supplemented with fetal calf serum, human urine, l-glutamine, and HEPES at 37oC (pH 5.5). Ultrastrutural analysis and infection assays in two macrophage populations (Kupffer
cells (KUP) and peritoneal macrophages (PM)) infected with axenic amastigotes demonstrated that they maintained morphological
and biochemical (A2 expression) features and a similar infection pattern to tissue-derived L. infantum amastigotes. The susceptibility of the macrophage lines to axenic or tissue-derived amastigotes and promastigotes was investigated.
We found a completely different susceptibility profile for KUP and PM. Liver macrophages, both KUP and immigrant macrophages,
are intimately involved in the response to L. infantum infection; this difference in susceptibility is probably related to their capacity to eliminate these parasites. Our in vitro
system was thus able to generate axenic amastigotes that resemble tissue-derived amastigotes both in morphology and infectivity
pattern; this will help in further investigation of the biological characteristics of the host–parasite relationship as well
as the process of pathogenesis. |
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