Generation of Leishmania donovani axenic amastigotes: their growth and biological characteristics

In this report, we describe an in vitro culture system for the generation and propagation of axenic amastigotes from the well characterised 1S-CL2D line of Leishmania donovani. Fine structure analyses of these in vitro-grown amastigotes demonstrated that they possessed morphological features charact...

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Bibliographic Details
Published in:International journal for parasitology 2004-02, Vol.34 (2), p.205-217
Main Authors: Debrabant, Alain, Joshi, Manju B, Pimenta, Paulo F.P, Dwyer, Dennis M
Format: Article
Language:English
Subjects:
Acid Phosphatase - secretion
Animals
Biological and medical sciences
Cricetinae
Culture system
Fundamental and applied biological sciences. Psychology
Host-Parasite Interactions
Human protozoal diseases
Humans
Infectious diseases
Infectivity
Leishmania donovani
Leishmania donovani - physiology
Leishmania donovani - ultrastructure
Leishmaniasis
Leshmaniasis
Life cycle. Host-agent relationship. Pathogenesis
Macrophages - parasitology
Medical sciences
Microscopy, Electron, Scanning
Parasite
Parasitic diseases
Parasitology - methods
Protozoa
Protozoal diseases
Trypanosomatid
Virulence
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Summary:In this report, we describe an in vitro culture system for the generation and propagation of axenic amastigotes from the well characterised 1S-CL2D line of Leishmania donovani. Fine structure analyses of these in vitro-grown amastigotes demonstrated that they possessed morphological features characteristic of L. donovani tissue-derived amastigotes. Further, these axenic amastigotes ( LdAxAm) were shown to synthesise and release a secretory acid phosphatase isoform similar to that produced by intracellular amastigotes. Such LdAxAm also expressed surface membrane 3′-nucleotidase enzyme activity similar to that of tissue-derived amastigotes. Moreover, LdAxAm, in contrast to promastigotes, expressed significant levels of the amastigote-specific A2 proteins. In addition, LdAxAm, derived from long term cultures of Ld 1S-CL2D promastigotes, had significant infectivity for both human macrophages in vitro and for hamsters in vivo. Thus, the in vitro culture system described herein provides a useful tool for the generation of large quantities of uniform populations of axenic amastigotes of the L. donovani 1S-CL2D line. The availability of such material should greatly facilitate studies concerning the cell and molecular biology of this parasite developmental stage.
ISSN:0020-7519
1879-0135
DOI:10.1016/j.ijpara.2003.10.011