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Metacyclogenesis of Leishmania (Viannia) guyanensis: a comprehensive study of the main transformation features in axenic culture and purification of metacyclic promastigotes by negative selection with Bauhinia purpurea lectin

Published online by Cambridge University Press:  27 December 2018

Bárbara Pinheiro Mendes
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Ildefonso Alves da Silva Jr.
Affiliation:
Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goias, 74605-050, Goiânia, GO, Brazil
Jarina Pena DaMata
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Thiago Castro-Gomes
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Leda Quercia Vieira
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
Fátima Ribeiro-Dias
Affiliation:
Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goias, 74605-050, Goiânia, GO, Brazil
Maria Fátima Horta*
Affiliation:
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31.270-901, Belo Horizonte, MG, Brazil
*
Author for correspondence: M. Fátima Horta, E-mail: [email protected]

Abstract

Leishmania (Viannia) guyanensis is one species that causes cutaneous leishmaniasis in the New World. The incidence of infections with this parasite is probably underestimated and few studies exist on this species, despite its epidemiological importance. In particular, there are no studies concerning L. guyanensis metacyclogenesis and no technique for obtaining metacyclic promastigotes for this species is presently available. Here, we have studied L. guyanensis metacyclogenesis in axenic culture, describing the main changes that occur during this process, namely, in morphology and size, sensitivity to complement-mediated lysis, surface carbohydrates and infectivity to macrophages. We have shown that metacyclogenesis in L. guyanensis promastigotes is basically complete on the 4th day of culture, as determined by decreased body size, increased flagellum length, resistance to complement-mediated lysis and infectivity. We have also found that only a fraction of the parasites is agglutinated by Bauhinia purpurea lectin. The non-agglutinated parasites, which also peaked on the 4th day of culture, had all morphological traits typical of the metacyclic stage. This is the first report describing metacyclogenesis in L. guyanensis axenic promastigotes and a simple and efficient method for the purification of metacyclic forms. Furthermore, a model of human macrophage infection with L. guyanensis was established.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2018 

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