title: Quantitative proteome profiling informs on phenotypic traits that adapt Leishmania donovani for axenic and intracellular proliferation.
abstract: Protozoan parasites of the genus Leishmania are important human pathogens that differentiate inside host macrophages into an amastigote life cycle stage. Although this stage causes the pathogenesis of leishmaniasis, only few proteins have been implicated in amastigote intracellular survival. Here we compare morphology, infectivity and protein expression of L. donovani LD1S grown in host free (axenic) culture, or exclusively propagated in infected hamsters, with the aim to reveal parasite traits absent in axenic but selected for in hamster-derived amastigotes through leishmanicidal host activities. Axenic and splenic amastigotes showed a striking difference in virulence and the ability to cause experimental hepato-splenomegaly in infected hamsters. 2D-DIGE analysis revealed statistically significant differences in abundance for 152 spots, with 14 spots showing fivefold or higher abundance in splenic amastigotes. Proteins identified by MS analysis include the anti-oxidant enzyme tryparedoxin peroxidase, and enzymes implicated in protein and amino acid metabolism. Analysis of parasite growth in vitro in minimal medium demonstrated increased survival of hamster-derived compared with axenic parasites under conditions that mimic the nutrient poor, cytotoxic phagolysosome. Thus, our comparative proteomics analysis sheds important new light on the biochemistry of bona fide amastigotes and informs on survival factors relevant for intracellular L. donovani infection.
authors.affiliation: Laboratory of Parasite Virulence Trypanosome Cell Biology Unit, Institut Pasteur, CNRS URA 2581/Institut National de la Santé et de la Recherche Médicale AVENIR Program, Paris, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Pescher
authors.forename: Thierry
authors.initials: T
authors.lastname: Blisnick
authors.forename: Philippe
authors.initials: P
authors.lastname: Bastin
authors.forename: Gerald F
authors.initials: GF
authors.lastname: Späth
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005786:Gene Expression Regulation; D007893:Leishmania donovani / Q000737:chemistry* / Q000254:growth & development*; D017737:Macrophages, Peritoneal / Q000469:parasitology; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D020543:Proteome / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis*; D013154:Spleen / Q000469:parasitology; D014774:Virulence
chemical_list: D020543:Proteome; D015800:Protozoan Proteins