Stage-dependent transcriptional changes and characterization of paramyosin of the bovine lungworm Dictyocaulus viviparus

Strübe, Christina; Buschbaum, Sandra; Samson‐Himmelstjerna, Georg von; Schnieder, Thomas

doi:10.1016/j.parint.2009.07.003

Cited by 10 publications

(15 citation statements)

References 33 publications

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“…Upon deep transcriptomic sequencing of B. malayi life-cycle stages, paramyosin and ELF1α were identified across the life-cycle of the parasite [59]. Similarly, in Dictyocaulus viviparus (cattle lungworm) and Clonorchis sinensis (liver fluke) paramyosin is expressed throughout the life-cycle [60, 61]. Undoubtedly, next generation sequencing of the O. lupi transcriptome could be useful in determining the number of transcripts and confirming the expression patterns of paramyosin across the parasite life-cycle.…”

Section: Discussionmentioning

confidence: 99%

Paramyosin of canine Onchocerca lupi: usefulness for the diagnosis of a neglected zoonotic disease

et al. 2016

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BackgroundOf increasing importance to the medical and veterinary communities is the zoonotic filarioid nematode Onchocerca lupi. Onchocercosis, thus far found in wolves, dogs, cats and humans, is diagnosed via skin snips to detect microfilariae and surgical removal of adults from the eye of the host. These methods are time-consuming, laborious and invasive, highlighting the need for new tools for the diagnosis of O. lupi in susceptible hosts. Symptoms related to the presence of the adults in the eye can range from none apparent to severe, including blindness. No reliable chemotherapeutic protocols are available, as yet, to eliminate the infection. Paramyosin, an invertebrate-specific protein, has been well-studied as an allergen, diagnostic marker and vaccine candidate. The aim of this study, therefore, was to isolate and characterise paramyosin from O. lupi to assess its suitability for the development of a serological diagnostic assay.MethodsThe adult and microfilarial stages of O. lupi were isolated from the eyes and skin of a 3-year-old male dog. Total RNA was extracted and reverse transcribed into single stranded cDNA. Reverse-transcription PCR was used to isolate a full-length paramyosin cDNA from adult worms and to investigate the temporal expression patterns of this gene. All amplicons were sequenced using dideoxy chain termination sequencing. Bioinformatics was used to predict the amino acid sequence of the gene, to compare the DNA and protein sequences with those available in public databases and to investigate the phylogenetic relationship of all molecules. Antibody binding sites were predicted using bioinformatics and mapped along with published antigenic epitopes against the O. lupi paramyosin protein. The native protein, and three smaller recombinantly expressed peptides, were subjected to western blot using serum from dogs both positive and negative for O. lupi.ResultsParamyosin of O. lupi was herein molecularly characterized, encoded by a transcript of 2,643 bp and producing a protein of 881 amino acids (101.24 kDa). The paramyosin transcript was detected, by reverse transcription PCR, in adults and microfilariae, but not in eggs. Phylogenetic analysis indicates that this molecule clusters with paramyosins from other filarioids to the exclusion of those from other taxa. A total of 621 unique antibody binding epitopes were predicted for this protein and another 28 were conserved in other organisms. This information was used to design three peptides, for recombinant expression, to identify the antibody binding epitope(s) and reduce potential cross-reactivity with serum from dogs infected with other filarioid nematodes. Native paramyosin, purified from microfilariae and adults, was detected by antibodies present in serum from dogs with known O. lupi infections.ConclusionsData provided herein may assist in the development of a serological diagnostic test, based on antibodies to O. lupi paramyosin, for the diagnosis of this infection, in order to gain more information on the real distribution of this l...

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Section: Discussionmentioning

confidence: 99%

Paramyosin of canine Onchocerca lupi: usefulness for the diagnosis of a neglected zoonotic disease

et al. 2016

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“…Earlier studies identified the expression of paramyosin in the tegument and on the surface of Taenia solium [24], Schistosoma mansoni , Schistosoma japonicum [25] and Fasciola hepatica [26]. It was observed that the paramyosin of helminth parasites could bind to human collagen [27-29], calgranulin [30], IgG [25,28,29], IgA [31], C1q [32], C8 [33] and C9 [29,33], suggesting that surface-exposed paramyosin may play an important role as a potential modulator of the host immune system.…”

Section: Discussionmentioning

confidence: 99%

Mapping of the complement C9 binding domain on Trichinella spiralis paramyosin

et al. 2014

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BackgroundTrichinellosis is an important foodborne zoonosis that is distributed worldwide. Trichinella spiralis may evade host complement-mediated attack by expressing complement inhibitory proteins, such as paramyosin (Pmy). Previous studies have shown that Trichinella spiralis paramyosin (Ts-Pmy) is able to bind to the human complement component C9 to inhibit the complement activation and protect the parasite from complement-mediated attack. Further determination of the complement-binding domain on Ts-pmy will enable us to better understand the Ts-Pmy’s biofunction in the immune evasion and provide feasible approach to develop epitope-based subunit vaccine against trichinellosis.MethodsThe complement C9 binding region on Ts-Pmy was determined by expression of overlapped fragments of Ts-Pmy and their binding activities to C9. The exact binding site was further narrowed-down to a 14-amino acid peptide at C-terminus using synthesized peptides with different size of amino acid sequence. The C9 complement-binding of the 14-amino acid peptide and its interference in the C9 polymerization and the complement-mediated lysis of rabbit erythrocytes was investigated.ResultsThe protein interaction between human C9 and native Ts-Pmy was further confirmed by immunoprecipitation with T. spiralis lysates. The fragmental expression and C9 binding assays identified that the binding region of Ts-Pmy to C9 is located within 831–885 of Ts-Pmy C-terminus. The exact binding site on Ts-Pmy to C9 was narrowed down to 14 amino acid residues (866Val-879Met) by using different sizes of synthesized peptides. In the presence of the synthesized 14-amino acid peptide, human C9 polymerization and the hemolytic activity of the human complement was inhibited.ConclusionsOur results revealed the precise molecular basis for T. spiralis to produce Ts-Pmy as an immunomodulator to evade the attack of the host complement system as a survival mechanism.

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“…Paramyosin (PMY), commonly known as a muscle protein of invertebrates including D. viviparus , has been considered as a promising vaccine candidate as it is transcribed throughout all developmental stages of bovine lungworms . PMY of D. viviparus displays immunomodulatory functions by binding to collagen and IgG , similar to those described for Taenia crassiceps ‐PMY .…”

Section: Introductionmentioning

confidence: 98%

Vaccination of calves with yeast‐ and bacterial‐expressed paramyosin from the bovine lungworm Dictyocaulus viviparus

Joekel

Hinse

Raulf

et al. 2015

Parasite Immunology

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Summary Previously, vaccination of cattle with Escherichia coli‐expressed bovine lungworm paramyosin (EcPMY) adjuvanted with Quil A resulted in considerable reduction in worm burden and larvae shedding (Strube et al., 2015). To further evaluate the protective potential of PMY, cattle vaccination trials were performed using either E. coli‐ (EcPMY) or Pichia pastoris‐expressed PMY (PpPMY) with different adjuvants (Matrix‐Q™ or Quil A). Combinations EcPMY+Matrix‐Q™ (trial 1), PpPMY+Matrix‐Q™ (trial 2) and PpPMY+Quil A (trial 3) were tested against challenge infections with 2000 Dictyocaulus viviparus larvae. Even though GM worm burden and larvae shedding was lower in almost all vaccinated groups, there were high variations between individuals hampering significant differences. However, in all vaccinated groups, lungworms were significantly shorter compared with those in controls. In vitro stimulation of peripheral blood mononuclear cells (PBMC) with recombinant (r)PMY revealed no significant proliferation following vaccinations or challenge infection. All vaccinated cattle showed a significant rise in specific antibodies, particularly IgG and its subclass IgG1, and detected the native lungworm PMY in immunoblots starting 2 weeks after the first vaccination. The use of a different rPMY‐adjuvant combination or combined vaccination with additional recombinant antigens might be a promising future approach towards a new vaccine against lungworms in cattle.

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Stage-dependent transcriptional changes and characterization of paramyosin of the bovine lungworm Dictyocaulus viviparus

Cited by 10 publications

References 33 publications

Paramyosin of canine Onchocerca lupi: usefulness for the diagnosis of a neglected zoonotic disease

Paramyosin of canine Onchocerca lupi: usefulness for the diagnosis of a neglected zoonotic disease

Mapping of the complement C9 binding domain on Trichinella spiralis paramyosin

Vaccination of calves with yeast‐ and bacterial‐expressed paramyosin from the bovine lungworm Dictyocaulus viviparus

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