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Background  Trichinellosis is a zoonotic disease in humans caused by  Trichinella  spp. The present study was undertaken to discover excretory-secretory (E-S) proteins from  T. spiralis  and  T. britovi  muscle larvae (ML) that hold promise for species-specific diagnostics. To that end, the purified E-S proteins were analyzed by fluorescent two-dimensional difference gel electrophoresis (2-D DIGE) coupled with protein identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS). To search for immunoreactive proteins that are specifically recognized by host antibodies the E-S proteins were subjected to two-dimensional (2-DE) immunoblotting with antisera derived from pigs experimentally infected with  T. spiralis  or  T. britovi .    Results  According to 2-D DIGE analysis, a total of twenty-two proteins including potentially immunogenic proteins and proteins produced only by one of the two  Trichinella  species were subjected to LC-MS/MS for protein identification. From these proteins seventeen could be identified, of which many were identified in multiple spots, suggesting that they have undergone post-translational modification, possibly involving glycosylation and/or proteolysis. These proteins included 5'-nucleotidase, serine-type protease/proteinase, and p43 glycoprotein (gp43) as well as 49 kDa E-S protein (p49). Our findings also suggest that some of the commonly identified proteins were post-translationally modified to different extents, which in certain cases seemed to result in species-specific modification. Both commonly and specifically recognized immunoreactive proteins were identified by 2-DE immunoblotting; shared antigens were identified as gp43 and different protease variants, whereas those specific to  T. britovi  included multiple isoforms of the 5'-nucleotidase.    Conclusions  Both 2-D DIGE and 2-DE immunoblotting approaches indicate that  T. spiralis  and  T. britovi  produce somewhat distinctive antigen profiles, which contain E-S antigens with potential as species-specific diagnostic markers for  Trichinella . Our results also demonstrate the value of 2-D DIGE as a versatile tool to compare secretomes of different  Trichinella  species for pinpointing factors contributing to the interaction with the host.

proteome science

Comparative analysis of excretory-secretory antigens of Trichinella spiralis and Trichinella britovi muscle larvae by two-dimensional difference gel electrophoresis and immunoblotting