Abstract
BackgroundTrichinella spiralis is an important foodborne zoonotic parasite and it is necessary to develop a vaccine in order to interrupt transmission from animals to humans. A 31 kDa protein from T. spiralis (Ts31) is an antigen targeted by protective antibodies, and Ts31 contains a domain of trypsin-like serine protease that might have the function of serine protease. The purpose of this study was to investigate the molecular characteristics of Ts31 and its induced immune protection.MethodsExpression and localization of Ts31 in various T. spiralis phases were investigated using qPCR and immunofluorescent test (IFT). The specific binding between Ts31 and intestinal epithelium cells (IECs) was analyzed by Far-Western blotting, ELISA and IFT, and the cellular localization of binding sites was examined on confocal microscopy. The mice were subcutaneously vaccinated with recombinant Ts31 protein (rTs31), serum specific IgG was determined by ELISA, and immune protection induced by immunization with rTs31 was evaluated. Inhibition of anti-rTs31 IgG on IL1 invasion of IECs and ADCC-mediated killing of newborn larvae (NBL) was also determined.ResultsTs31 was expressed at different life-cycle stages and located principally at the stichosome and cuticle of this parasite. rTs31 was capable to specially bond to IECs, and binding site was located in the cytoplasm of IECs. Immunization of mice with rTs31 elicited a significant humoral response and protection, as demonstrated by a 56.93% reduction of adult worms at 6 days post-infection (dpi) and a 53.50% reduction of muscle larvae at 42 dpi after larval challenge. Anti-rTs31 antibodies impeded T. spiralis penetration of enterocytes in a dose-dependent pattern, and participated in the destruction of NBL by an ADCC-mediated manner.ConclusionsTs31 facilitated the T. spiralis penetration of intestinal epithelium, which could make it a vaccine candidate target molecule against Trichinella infection.
Highlights
Trichinella spiralis is an important foodborne zoonotic parasite and it is necessary to develop a vaccine in order to interrupt transmission from animals to humans
Transcription level of the Trichinella spiralis 31 kDa protein (Ts31) gene at different stage worms quantitative PCR (qPCR) analysis revealed that the Ts31 gene was transcribed at different life-cycle stages for T. spiralis worms (ML, intestinal L1 larvae (IL1), 3 and 6 dpi Adult worms (AW), and newborn larvae (NBL)) (Fig. 1)
Expression and localization of Ts31 at different stage worms The immunofluorescent test (IFT) results indicated that the immunofluorescent staining was observed at different stages (ML, IL1, female and male at 3 dpi) by using anti-recombinant Ts31 protein (rTs31) serum
Summary
Trichinella spiralis is an important foodborne zoonotic parasite and it is necessary to develop a vaccine in order to interrupt transmission from animals to humans. Humans acquire this infection mainly by ingesting raw or poorly processed animal meat contaminated by muscle larvae (ML) of Trichinella [2, 3]. If the IL1 invasion of IECs, larval development and newborn larvae (NBL) deposition are interrupted, or IL1, AW and NBL are expelled from the intestine by vaccination, Trichinella infection will be prevented or eliminated in the early stage of enteral infection [14, 15]. Development of an anti-Trichinella vaccine is necessary for the control and elimination of porcine Trichinella infection to ensure pork safety [16,17,18]
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