Trichinella britovi: muscle larvae, adult worm - LC-MS/MS

ABSTRACT: Of the species of Trichinella spp. that may affect human health, Trichinella britovi is the second most common. As an early diagnosis of trichinellosis is crucial for effective treatment, it is important to identify sensitive, specific and common antigens of adult T. britovi worms and muscle larvae. The present study was undertaken to uncover the stage-specific and common proteins of T. britovi that hold promise for specific diagnostics. To that end, the somatic extracts obtained from two developmental stages, muscle larvae (ML) and adult worms (Ad), were separated using two-dimensional gel electrophoresis (2DE) coupled with immunoblot analysis. The positively-visualized protein spots specific for each stage were identified through liquid chromatography-tandem mass spectrometry (LC-LC/MS). A total of 272 spots were detected in the proteome of T. britovi adult worms (Ad) and 261 in the muscle larvae (ML). The somatic extracts from Ad and ML were specifically recognized by pig T. britovi-infected swine sera at 10 days post infection (dpi) and 60 dpi, with a total of 70 prominent protein spots. According to immunoblotting patterns and LC-MS/MS results, the immunogenic spots recognized by different pig T. britovi-infected sera were divided into three groups for the two developmental stages: adult stage-specific proteins, muscle larvae stage-specific proteins, and common proteins. Forty-five Ad proteins (29 Ad-specific and 16 common) and thirteen ML proteins (nine ML-specific and four common) cross-reacted with sera at 10 dpi. Many of the proteins identified in Ad (myosin-4, myosin light chain kinase, paramyosin, intermediate filament protein B, actin-depolymerizing factor 1, and calreticulin) are involved in structural and motor activity. Among the most abundant proteins identified in ML were 14-3-3 protein zeta, actin-5C, ATP synthase subunit d, deoxyribonuclease-2-alpha, poly-cysteine and histide-tailed protein, enolase, V-type proton ATPase catalytic and serine protease 30. Heat shock protein, intermediate filament protein ifa-1 and intermediate filament protein B were identified in both proteomes. The current study represents the first immunoproteomic identification of the antigenic proteins of adult worm and muscle larvae of T. britovi. These results provide a valuable basis for the development of diagnostic methods. The identification of common components among two developmental stages of T. britovi may be useful in the preparation of parasitic antigens in recombinant forms for diagnostic use.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Trichinella Britovi

TISSUE(S): Whole Body

SUBMITTER: Sylwia Grzelak  

LAB HEAD: Sylwia Grzelak

PROVIDER: PXD011215 | Pride | 2019-11-12

REPOSITORIES: Pride