Expressed Sequence Tags of Trichinella spiralis Muscle Stage Larvae

BackgroundThe intestinal phase is the early invasion stage of Trichinella spiralis (T. spiralis), in which muscle larvae invade intestine epithelial cells and then develop into adult worms to breed newborn larvae. Thus, intestinal infective larvae are first exposed to the immune system of the host, and antigens from the worms may be the earliest marker in the diagnosis of trichinellosis and may contribute to vaccine development to prevent Trichinella infections in pigs.MethodsA cDNA library of intestinal infective larvae of T. spiralis at 6 hours post infection (p.i.) was constructed and immunoscreened using serum collected from pigs that were infected with T. spiralis at 26 days p.i. T. spiralis cystatin-like protein (Ts-CLP) gene encoding a 45.9 kDa protein was cloned and expressed in Escherichia coli. The rabbit antisera were generated and used to determine the location of Ts-CLP in the parasite. Transcription levels of Ts-CLP in different developmental stages of T. spiralis were observed by RT-PCR. The potential application of recombinant Ts-CLP in diagnosis against T. spiralis infection was tested by ELISA. The immune protection of recombinant Ts-CLP protein against T. spiralis infection was evaluated in mice.ResultsThirty-three positive clones were selected from cDNA library, among which 20 clones encoded the same novel cystatin-like protein (Ts-CLP). Immunolocalisation and real-time quantitative PCR revealed that native Ts-CLP was localised primarily to β-stichocytes and that the Ts-clp gene was transcribed and expressed in all developmental stages of T. spiralis. The recombinant protein rTs-CLP was recognised by pig antiserum as early as 15 days p.i., and could induce protective immunity in mice, with a 61.21% reduction in the number of muscle larvae.ConclusionsThese data preliminarily suggested that Ts-CLP may play an important role in the early infection of T. spiralis and that recombinant Ts-CLP protein is a candidate antigen for diagnosis and vaccine development in Trichinella infections.

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“…[ 26 ] and Park et al . [ 27 ]) revealed 73, 57 and 0 ESTs of this gene in the ML (5 020 ESTs), Ad (2 981 ESTs) and NBL (3 234 ESTs) databases.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of a high-frequency gene encoding a strongly antigenic cystatin-like protein from Trichinella spiralis at its early invasion stage

et al. 2015

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“…Recently Kouzaki et al [ 29 ] reported that proteases induce production of TSLP through PAR2, and TSLP triggers dendritic cell-mediated Th2 type inflammation. Trichinella also excreted and/or secreted various kinds of proteases for invasion and digestion of host tissues [ 11 - 13 ], and these proteases might elicit TSLP via PAR2.…”

Section: Discussionmentioning

confidence: 99%

“…T. spiralis larvae have been shown to be able to establish chronic infections in the skeletal muscles of immunocompetent hosts [ 10 ]. They also expressed a greater variety of proteases, particularly serine protease and aspartic protease, in larval stages as compared to the adult worm stage, and these proteases might be necessary to infect new hosts [ 11 - 13 ]. T. spiralis could evoke a host Th2 immune response, and the host also elevates its Th2 immune responses in order to eliminate the worms [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Protease-Activated Receptor 2 Is Involved in Th2 Responses againstTrichinella spiralisInfection

et al. 2011

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In order to get a better understanding of the role of protease-activated receptor 2 (PAR2) in type 2 helper T (Th2) cell responses against Trichinella spiralis infection, we analyzed Th2 responses in T. spiralis-infected PAR2 knockout (KO) mice. The levels of the Th2 cell-secreted cytokines, IL-4, IL-5, and IL-13 were markedly reduced in the PAR2 KO mice as compared to the wild type mice following infection with T. spiralis. The serum levels of parasite-specific IgE increased significantly in the wild type mice as the result of T. spiralis infection, but this level was not significantly increased in PAR2 KO mice. The expression level of thymic stromal lymphopoietin, IL-25, and eotaxin gene (the genes were recently known as Th2 response initiators) of mouse intestinal epithelial cells were increased as the result of treatment with T. spiralis excretory-secretory proteins. However, the expression of these chemokine genes was inhibited by protease inhibitor treatments. In conclusion, PAR2 might involve in Th2 responses against T. spiralis infection.

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Identification of host immune regulation candidate genes of Toxascaris leonina by expression sequenced tags (ESTs) analysis

Cho

Lee

Kang

et al. 2009

Veterinary Parasitology

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Expressed Sequence Tags of Trichinella spiralis Muscle Stage Larvae

Cited by 4 publications

References 35 publications

Characterisation of a high-frequency gene encoding a strongly antigenic cystatin-like protein from Trichinella spiralis at its early invasion stage

Characterisation of a high-frequency gene encoding a strongly antigenic cystatin-like protein from Trichinella spiralis at its early invasion stage

Protease-Activated Receptor 2 Is Involved in Th2 Responses againstTrichinella spiralisInfection

Identification of host immune regulation candidate genes of Toxascaris leonina by expression sequenced tags (ESTs) analysis

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