Characterisation of novel protein families secreted by muscle stage larvae of Trichinella spiralis

Guiliano, David B.; Oksov, Yelena; Lustigman, Sara; Gounaris, Kleoniki; Selkirk, Murray E.

doi:10.1016/j.ijpara.2008.09.012

Cited by 18 publications

(17 citation statements)

References 41 publications

(41 reference statements)

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“…According to the images showing the morphology ( www. Trichinella .org ) and the results reported by Guiliano [ 28 ], the localisation was in β-stichocytes of the stichosome. We failed to detect secreted Ts-CLP protein in NBL (Figure 4 D), the capsule and muscle tissue (Figure 4 F-H), but we observed Ts-CLP signals in the intestinal epithelium (Figure 4 B).…”

Section: Discussionsupporting

confidence: 63%

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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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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Section: Discussionsupporting

confidence: 63%

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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“…We have previously published the results of an informatics-based screen of the T. spiralis transcriptome that identified three novel proteins secreted by mL1 [6]. Here, we extend this study and describe two novel abundant transcripts encoding low molecular weight secreted proteins of unknown function.…”

Section: Introductionsupporting

confidence: 51%

“…In these colonization processes the excretory-secreted products (ESPs) of the parasites are thought to drive invasion, migration, or the remodeling of host cells and tissues which support parasite development. The identification of these molecules and dissection of their functions continues to be the focus of numerous studies [5][6][7][8], especially with respect to better understanding its unique lifestyle. T. spiralis and other Trichinellids remain important human and agricultural pathogens with millions of people worldwide chronically infected, and are classified as a neglected tropical disease [9,10].…”

Section: Introductionmentioning

confidence: 99%

Vaccination with novel low-molecular weight proteins secreted from Trichinella spiralis inhibits establishment of infection

et al. 2020

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Trichinella spiralis muscle stage larvae (mL1) produce excretory-secreted products (ESPs), a complex mixture of protein, which are believed to be important for establishing or maintaining an infection niche within skeletal muscle and the intestine. Studies of both whole ESPs and individual cloned proteins have shown that some ESPs are potent immunogens capable of eliciting protective immune responses. Here we describe two novel proteins, Secreted from Muscle stage Larvae SML-4 and SML-5 which are 15 kDa and 12 kDa respectively. The genes encoding these proteins are highly conserved within the Trichinellids, are constituents of mL1 ESP and localized in the parasite stichosome. While SML-5 is only expressed in mL1 and early stages of adult nematode development, SML-4 is a tyvosylated glycoprotein also produced by adult nematodes, indicating it may have a function in the enteral phase of the infection. Vaccination with these proteins resulted in an impaired establishment of adult stages and consequently a reduction in the burden of mL1 in BALB/c mice. This suggests that both proteins may be important for establishment of parasite infection of the intestine and are prophylactic vaccine candidates.

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“…Proteins previously identified in T . spiralis SP using alternative methods were also present amongst the results[20–25]). To avoid contamination with proteins released from dead or dying parasites, uncoiled and floating T .…”

Section: Resultsmentioning

confidence: 82%

Ubiquitin-Dependent Modification of Skeletal Muscle by the Parasitic Nematode, Trichinella spiralis

et al. 2016

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Trichinella spiralis is a muscle-specific parasitic worm that is uniquely intracellular. T. spiralis reprograms terminally differentiated skeletal muscle cells causing them to de-differentiate and re-enter the cell cycle, a process that cannot occur naturally in mammalian skeletal muscle cells, but one that holds great therapeutic potential. Although the host ubiquitin pathway is a common target for viruses and bacteria during infection, its role in parasite pathogenesis has been largely overlooked. Here we demonstrate that the secreted proteins of T. spiralis contain E2 Ub-conjugating and E3 Ub-ligase activity. The E2 activity is attributed to TsUBE2L3, a novel and conserved T. spiralis enzyme located in the secretory organ of the parasite during the muscle stages of infection. TsUBE2L3 cannot function with any T.spiralis secreted E3, but specifically binds to a panel of human RING E3 ligases, including the RBR E3 ARIH2 with which it interacts with a higher affinity than the mammalian ortholog UbcH7/UBE2L3. Expression of TsUBE2L3 in skeletal muscle cells causes a global downregulation in protein ubiquitination, most predominantly affecting motor, sarcomeric and extracellular matrix proteins, thus mediating their stabilization with regards to proteasomal degradation. This effect is not observed in the presence of the mammalian ortholog, suggesting functional divergence in the evolution of the parasite protein. These findings demonstrate the first example of host-parasite interactions via a parasite-derived Ub conjugating enzyme; an E2 that demonstrates a novel muscle protein stabilization function.

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Characterisation of novel protein families secreted by muscle stage larvae of Trichinella spiralis

Cited by 18 publications

References 41 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

Vaccination with novel low-molecular weight proteins secreted from Trichinella spiralis inhibits establishment of infection

Ubiquitin-Dependent Modification of Skeletal Muscle by the Parasitic Nematode, Trichinella spiralis

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