title: The Schistosoma mansoni soluble proteome: a comparison across four life-cycle stages.
abstract: Differential analysis of immune responses to schistosomes has routinely been performed using complex mixtures of soluble proteins from various life-cycle stages, on the assumption that these differed significantly in composition. Proteomic techniques now allow us to characterise and compare such mixtures. The soluble proteins from cercariae, lung-schistosomula, adult worms and eggs of Schistosoma mansoni were separated by high-resolution two-dimensional electrophoresis and the resulting images analysed using appropriate software. A high degree of quantitative and qualitative similarity in spot pattern was revealed across the life-cycle, greatest between adjacent stages. To initiate mapping of these soluble proteomes, the 40 most abundant spots in each preparation, accounting for 21-46% of the total protein, were subjected to peptide fingerprinting by mass spectrometry. On average 55% of the spots were identified, but overall, these comprised only 32 different protein species. With one exception all proteins originated in the cytosol and 24 of the 32 had previously been pinpointed by virtue of their immunoreactivity, including four of the WHO priority vaccine candidates. The similarity in composition between the four preparations means that they are unlikely to discriminate adequately between immune responses to different life-cycle stages and argues strongly for the need to identify true stage-specific marker proteins. Equally, it is difficult to reconcile the abundance and immunogenicity of such cytosolic proteins with their status as vaccine candidates, as it is unlikely they will be accessible to the immune system in an intact parasite.
authors.affiliation: Department of Biology, University of York, Heslington, York YO10 5DD, UK. rsc105@york.ac.uk
authors.forename: Rachel S
authors.initials: RS
authors.lastname: Curwen
authors.forename: Peter D
authors.initials: PD
authors.lastname: Ashton
authors.forename: David A
authors.initials: DA
authors.lastname: Johnston
authors.forename: R Alan
authors.initials: RA
authors.lastname: Wilson
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D015801:Helminth Proteins / Q000378:metabolism*; D006801:Humans; D008018:Life Cycle Stages; D013058:Mass Spectrometry; D051379:Mice; D010449:Peptide Mapping; D020543:Proteome; D012550:Schistosoma mansoni / Q000254:growth & development* / Q000378:metabolism; D012995:Solubility
chemical_list: D000947:Antigens, Helminth; D015801:Helminth Proteins; D020543:Proteome