University of Khartoum

Generation of leishmania axenic forms from leishmania donovani and leishmania major parasites.

Generation of leishmania axenic forms from leishmania donovani and leishmania major parasites.

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Title: Generation of leishmania axenic forms from leishmania donovani and leishmania major parasites.
Author: Awad, Nazik; Mukhtar, Moawia M.
Abstract: Leishmania parasite is an obligatory intracellular parasite which cycles between the mid gut of the sandflies (extracellular promastigote) and the phagolysosomes of mammalian macrophages (intracellular amastigote). Promastigotes have been readily cultured, whereas axenic cultures of amastigotes have only recently been developed. This work described a simple method to yield large amount of leishmania amastigote like forms in axenic cultures using promastigotes as the starting population and two types of culture media RPMI 1640 and Dullbeco's modified Egal's medium. Promastigote are exposed to environmental changes that mimic the in vivo process by subjecting it to 37? C , 5_5.5 acidic pH and 7%_10% CO2. The method described induced extracellular amastigote transformation of L. donovani, L. major, L. infantum, and L.archibaldi. The rounded parasites obtained in axenic cultures were morphologically similar to intracellular amastigote. Moreover, the axenic amastigote remained viable as measured by their ability to revert back to promastigotes by transforming them to promastigote growth conditions (RPMI1640 medium at pH 7.2 and 26? C), and their ability to infect J774 and THP1 macrophage cell line in the presence and absence of serum component. The proteins were extracted from starting promastigote and axenic form during the time of transformation. Proteins were fractionated using 12% SDS-PAGE. Shared protein bands ranging from 26- 116 kda were detected by SDS-PAGE in both stages with variation in protein size and intensities between the starting population promastigote and axenic form (some band started to faint and other band became abundant with increasing the time of exposure.
URI: http://hdl.handle.net/123456789/6303
Date: 2008-09


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