Isolation, characterization and immunolocalization of a globin-like antigen from <i>Ostertagia ostertagi</i> adults

Graaf, Dirk C. de; Berghen, P.; Moëns, Luc; Marez, Tine De; Raes, Stefaan; Blaxter, Mark; Vercruysse, J.

doi:10.1017/s0031182000066282

Cited by 9 publications

(4 citation statements)

References 25 publications

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“…It belongs to the transthyretin-like nematode-specific protein family (52 members in C. elegans) with weak similarity to transthyretin, a thyroid hormone transport protein in vertebrates. Interestingly, two other spots (179 and (33) and Trichostrongylus colubriformis (34) and have been detected in H. contortus (35) and Ostertagia ostertagia (36). The secreted form is present in the cuticle of the parasite and might be involved in oxygen transport for muscular activity of the worm, because nematode globins have higher affinity for oxygen than the host globins (100 and 1000 times higher in N. brasiliensis and Ascaris suum, respectively) (37,38).…”

Section: Variation In and Validation Of Vaccine Candidates Hc15 Hc24mentioning

confidence: 99%

Comprehensive Analysis of the Secreted Proteins of the Parasite Haemonchus contortus Reveals Extensive Sequence Variation and Differential Immune Recognition

Yatsuda

Krijgsveld

Cornelissen

et al. 2003

Journal of Biological Chemistry

194

164

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Haemonchus contortus is a nematode that infects small ruminants. It releases a variety of molecules, designated excretory/secretory products (ESP), into the host. Although the composition of ESP is largely unknown, it is a source of potential vaccine components because ESP are able to induce up to 90% protection in sheep. We used proteomic tools to analyze ESP proteins and determined the recognition of these individual proteins by hyperimmune sera. Following two-dimensional electrophoresis of ESP, matrix-assisted laser desorption ionization time-of-flight and liquid chromatographytandem mass spectrometry were used for protein identification. Few sequences of H. contortus have been determined. Therefore, the data base of expressed sequence tags (dbEST) and a data base consisting of contigs from Haemonchus ESTs were also consulted for identification. Approximately 200 individual spots were observed in the two-dimensional gel. Comprehensive proteomics analysis, combined with bioinformatic search tools, identified 107 proteins in 102 spots. The data include known as well as novel proteins such as serine, metallo-and aspartyl proteases, in addition to H. contortus ESP components like Hc24, Hc40, Hc15, and apical gut GA1 proteins. Novel proteins were identified from matches with H. contortus ESTs displaying high similarity with proteins like cyclophilins, nucleoside diphosphate kinase, OV39 antigen, and undescribed homologues of Caenorhabditis elegans. Of special note is the finding of microsomal peptidase H11, a vaccine candidate previously regarded as a "hidden antigen" because it was not found in ESP. Extensive sequence variation is present in the abundant Hc15 proteins. The Hc15 isoforms are differentially recognized by hyperimmune sera, pointing to a possible specific role of Hc15 in the infectious process and/or in immune evasion. This concept and the identification of multiple novel immune-recognized components in ESP should assist future vaccine development strategies.

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Section: Variation In and Validation Of Vaccine Candidates Hc15 Hc24mentioning

confidence: 99%

Comprehensive Analysis of the Secreted Proteins of the Parasite Haemonchus contortus Reveals Extensive Sequence Variation and Differential Immune Recognition

Yatsuda

Krijgsveld

Cornelissen

et al. 2003

Journal of Biological Chemistry

194

164

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“…Two antigens were recently cloned. This, however, did not result in a reliable ELISA based on a recombinant product (7).…”

mentioning

confidence: 93%

Use of cloned excretory/secretory low-molecular-weight proteins of Cooperia oncophora in a serological assay

et al. 1997

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The potential of Cooperia oncophora excretory/secretory (ES) proteins as antigens in a serological assay which aims to establish exposure levels in cattle was assessed. ES proteins were analyzed by one-and two-dimensional polyacrylamide gel electrophoresis and immunoblotting. The N-terminal domains of two ES proteins were sequenced, and the corresponding cDNAs were cloned. Two cDNAs, designated CoES14.0 and CoES14.2, were expressed in Escherichia coli. The recombinant proteins were tested in an indirect enzymelinked immunosorbent assay (ELISA) in which crude worm antigen (CWA) was used as a reference standard. In total, 67 reference serum samples were used: 27 negative serum samples, 29 C. oncophora-specific serum samples, 7 Dictyocaulus viviparus-specific serum samples, and 4 Ostertagia ostertagi-specific serum samples. This showed respective sensitivities and specificities of 17 and 84%, 0 and 100%, and 100 and 100% by the ELISAs with the three different types of proteins (CWA, CoES14.0, and CoES14.2, respectively). Since the CoES14.2 ELISA had the best sensitivity and specificity with reference sera, its specificity was further validated in an antigen inhibition ELISA. In this assay CoES14.2 and CWA preparations of C. oncophora, Cooperia curticei, O. ostertagi, Nematodirus helvetianus, Fasciola hepatica, D. viviparus, Haemonchus placei, and Trichostrongylus colubriformus were used as competitor antigens. This experiment showed that only the homologous antigens C. oncophora CWA and CoEs14.2 resulted in 100% inhibition. The CWA preparations of all other nematodes did not affect the ELISA, even if concentrations of 250 times the 50% inhibitory concentration of C. oncophora CWA were used. These results indicate that CoES14.2 does not share cross-reactive epitopes with heterologous CWAs. Finally, we tested the CoES14.2 ELISA with sequential serum samples from naturally infected groups of animals. The optical density values that were obtained correlated well with exposure levels based on cumulative egg excretion. Thus, the CoES14.2 ELISA seems to be a very sensitive tool for estimating exposure levels in cattle.

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“…Insoluble fragments were removed by centrifugation (25 000 g, 4°C) and the clear supernatant fraction was frozen at -20°C (De Graaf et al, 1996).…”

Section: Isolation Of Somatic Proteins Of S Obvelatamentioning

confidence: 99%

The presence of Syphacia obvelata in laboratory mice (BALB/c) — parasite antigens in immune response

Perec

Okulewicz

2006

Helminthologia

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203 SummaryIn conventional mice colonies, mouse pinworm, Syphacia obvelata is found very often. Several studies indicate that infection with this parasite can modulate the immune system of the host and can affect experimental final results. The aim of our study was to investigate the most immunogenic proteins of S. obvelata inducing both local and systemic immune response in naturally infected laboratory mice. Protein extracts of S. obvelata were analysed by Western blotting to examine their antigenic character. The antigens were probed with serum and mucosa of S. obvelata naturally infected mice. Surface and somatic antigens were recognized by serum and mucosal IgG, IgA and IgM antibodies. The most immunogenic and dominant proteins were observed. Proteins of Mw ~ 70, 65 and 48 kDa showed the most evident reaction with serum and mucosa antibodies of infected animals. Surface and somatic antigens of nematode S. obvelata eliciting immune response in laboratory mice may be useful in development of a diagnostic test which could be applied for the infection control prior the experiments.

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Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults

Cited by 9 publications

References 25 publications

Comprehensive Analysis of the Secreted Proteins of the Parasite Haemonchus contortus Reveals Extensive Sequence Variation and Differential Immune Recognition

Comprehensive Analysis of the Secreted Proteins of the Parasite Haemonchus contortus Reveals Extensive Sequence Variation and Differential Immune Recognition

Use of cloned excretory/secretory low-molecular-weight proteins of Cooperia oncophora in a serological assay

The presence of Syphacia obvelata in laboratory mice (BALB/c) — parasite antigens in immune response

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