Comparative analysis of excretory-secretory products of muscle larvae of three isolates of Trichinella pseudospiralis by the iTRAQ method.

Trichinella pseudospiralis is a non-encapsulated intracellular parasitic nematode that can possess a strong ability to modulate the host immune response. Here, we compared the differentially expressed proteins of excretory-secretory (ES) products in three isolates of T. pseudospiralis muscle larvae (ML) [from Russia (RUS), United States of America (USA) and Australia (AUS)] using isobaric tags for relative and absolute quantification (iTRAQ)-based technology. A total of 2591 nonredundant proteins were identified, of which 65 (146), 72 (98) and 43 (103) significantly upregulated (downregulated) differentially expressed proteins were detected among pairwise comparisons (T4RUS vs T4USA, T4AUS vs T4USA and T4RUS vs T4AUS). In addition, GO annotation, KEGG and STRING analyses were carried out on the screened differentially altered proteins. The main biological processes involved included carbohydrate metabolic processes, DNA metabolic processes, cellular protein modification processes and homeostatic processes. The majority of KEGG pathways were found to be related to the metabolic pathways, lysosome pathway and protein processing in endoplasmic reticulum. Moreover, all ES protein expression levels involved in the lysosome pathway were significantly higher in the T4USA isolate than in the other two isolates. We also found differences in the expression of some important immunoregulatory proteins, such as protein disulfide-isomerase, thioredoxin protein and deoxyribonuclease-2-alpha, between different isolates of T. pseudospiralis ML. Flow cytometry was used to detect the increase in the CD4+/CD8 + T-cell ratio in pig peripheral blood and to verify the effect of T. pseudospiralis on the Th1/Th2 polarization of the host. Quantitative real-time PCR analysis also confirmed that the changes in the transcriptional level of genes were consistent with those at the proteomic level. These findings reveal the possible role of significantly differentially expressed proteins in ES products of the different isolates of T. pseudospiralis in antagonizing and participating in the regulation of the host immune response and maintaining a stable growth environment.

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