Characterizing Excretory-Secretory Products Proteome Across Larval Development Stages inAscaris suum

Abstract

IntroductionAscaris lumbricoidesandAscaris suumare parasitic nematodes that primarily infest the small intestines of humans and pigs, respectively. Ascariasis poses a significant threat to human health and swine health. UnderstandingAscarislarval development is crucial for developing novel therapeutic interventions that will prevent ascariasis in both humans and pigs. This study aimed to characterize the excretory-secretory (ES) proteome of differentAscaris suumlarval stages (L3-egg, L3-lung, L3-trachea) to identify potential targets for intervention to preventAscaris-induced global morbidity.MethodsStage-specific larvae were isolated, cultured in vitro and ES-product was collected. Third-stageAscarislarvae (L3) were isolated from embryonated eggs (L3-egg), isolated from the lungs of Balb/c mice infected withAscaris suumeggs at day 8 post infection (L3-lungs) and isolated from the trachea of Balb/c mice infected withAscaris suumeggs at day 12 post infection (L3-trachea). ES products were obtained by culturing larvae. Proteomic analysis was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatic tools including MaxQuant, Perseus, and Andromeda, following a detailed protocol available on GitHub. The analysis encompassed peptide identification, scoring, and quantification against an organism-specific database, with subsequent quality control, correlation assessment, and differential abundance determination using the Amica algorithm.ResultsA total of 58 unique proteins were identified in the ES products. Fourteen proteins were common across all stages, while others were stage-specific. Principal component analysis revealed distinct protein profiles for each stage, suggesting qualitatively different proteomes. Gene ontology analysis indicated stage-specific GO enrichment of specific protein classes, such as nuclear proteins in L3-egg ES products and metabolic enzymes in L3-lung and L3-trachea ES products.DiscussionThis study revealed stage-specific differences in the composition ofAscarisES products. Further investigation into the functional roles of these proteins and their interactions with host cells is crucial for developing novel therapeutic and diagnostic strategies against ascariasis.