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Molecular characterization, expression and localization of tropomyosin and paramyosin immunodominant allergens from sheep scab mites (Psoroptes ovis)

  • A. J. NISBET (a1), A. MACKELLAR (a1), H. W. WRIGHT (a1), G. P. BRENNAN (a2), K. Y. CHUA (a3), N. CHEONG (a3), J. E. THOMAS (a4) and J. F. HUNTLEY (a4)...

Abstract

cDNAs encoding the immunodominant allergens tropomyosin and paramyosin were amplified from RNA extracted from the sheep scab mite Psoroptes ovis. The tropomyosin cDNA contained an open reading frame (ORF) of 852 bp which encoded a predicted protein with 98% and 97% identity to the house dust mite allergens Der f 10 and Der p 10 respectively. The complete paramyosin ORF generated by RT-PCR was 2625 bp in length and encoded an 875aa predicted protein of 102·6 kDa with 97%, 95% and 89% identity to the paramyosins of Dermatophagoides pteronyssinus (Der p 11), Sarcoptes scabiei and Blomia tropicalis (Blo t 11) respectively. Full length tropomyosin and truncated and full-length paramyosin were expressed as recombinant proteins. IgG and IgE in sera from sheep with a 6-week duration primary infestation of P. ovis did not detect either full-length or truncated recombinant paramyosin. IgG in both infested and naïve sheep sera detected recombinant tropomyosin, suggesting cross-reactivity to tropomyosin and to other invertebrate species to which the sheep may have been exposed. Staining with antibodies directed against tropomyosin and paramyosin was observed throughout sections of P. ovis. Staining was especially prevalent in the anterior sections of the mites, possibly associated with locomotory muscles in this region.

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Corresponding author

Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland. Tel: +44 (0)131 445 5111. Fax: +44 (0)131 445 6111. E-mail: Alasdair.Nisbet@moredun.ac.uk

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REFERENCES

Brown, J. H., Kim, K. H., Jun, G., Greenfield, N. J., Dominguez, R., Volkmann, N., Hitchcock-DeGregori, S. E. and Cohen, C. ( 2001). Deciphering the design of the tropomyosin molecule. Proceedings of the National Academy of Sciences, USA 98, 84968501.
Cohen, C. and Parry, D. A. ( 1990). Alpha-helical coiled coils and bundles: how to design an alpha-helical protein. Proteins 7, 115.
Ferreira, C. A., Barbosa, M. C., Silveira, T. C., Valenzuela, J. G., Da Silva Vaz Jr., I. and Masuda, A. ( 2002). cDNA cloning, expression and characterization of a Boophilus microplus paramyosin. Parasitology 125, 265274.
Gobert, G. N. and McManus, D. P. ( 2005). Update on paramyosin in parasitic worms. Parasitology International 54, 101107.
Harrison, R. A. and Bianco, A. E. ( 2000). DNA immunization with Onchocerca volvulus genes, Ov-tmy-1 and OvB20: serological and parasitological outcomes following intramuscular or GeneGun delivery in a mouse model of onchocerciasis. Parasite Immunology 22, 249257.
Hartmann, S., Adam, R., Marti, T., Kirsten, C., Seidinger, S. and Lucius, R. ( 1997). A 41-kDa antigen of the rodent filaria Acanthocheilonema viteae with homologies to tropomyosin induces host-protective immune responses. Parasitology Research 83, 390393.
Hitchcock-DeGregori, S. E., Song, Y. and Greenfield, N. J. ( 2002). Functions of tropomyosin's periodic repeats. Biochemistry 41, 1503615044.
Huntley, J. F., Machell, J., Nisbet, A. J., Van den Broek, A., Chua, K. Y., Cheong, N., Hales, B. and Thomas, W. R. ( 2004). Identification of tropomyosin, paramyosin and apolipophorin/vitellogenin as three major allergens of the sheep scab mite, Psoroptes ovis. Parasite Immunology 26, 335342.
Jacober, P. H., Ochs, H., Torgerson, P. R., Schnyder, M. and Deplazes, P. ( 2006). A method for sheep scab control by applying selective treatment based on flock serology. Veterinary Parasitology 136, 373378.
Jeanmougin, F., Thompson, J. D., Gouy, M., Higgins, D. G. and Gibson, T. J. ( 1998). Multiple sequence alignment with Clustal X. Trends in Biochemical Sciences 10, 403405.
Kenyon, F., Welsh, M., Parkinson, J., Whitton, C., Blaxter, M. L. and Knox, D. P. ( 2003). Expressed sequence tag survey of gene expression in the scab mite Psoroptes ovis – allergens, proteases and free-radical scavengers. Parasitology 126, 451460.
Lonneux, J.-F., Nguyen, T. Q., Hollanders, W. and Losson, B. J. ( 1996). A sandwich enzyme-linked immunosorbent assay to study the antibody response against Psoroptes ovis in cattle. Veterinary Parasitology 64, 299314.
Mattsson, J. G., Ljunggren, E. L. and Bergstrom, K. ( 2001). Paramyosin from the parasitic mite Sarcoptes scabiei: cDNA cloning and heterologous expression. Parasitology 122, 555562.
Nisbet, A. J. and Huntley, J. F. ( 2006). Progress and opportunities in the development of vaccines against mites, fleas and myiasis-causing flies of veterinary importance. Parasite Immunology 28, 165172.
Page, R. D. M. ( 1996). TreeView: an application to display phylogenetic trees on personal computers. Computer Applications in the Biological Sciences 12, 357358.
Pearce, E. J., James, S. L., Hieny, S., Lanar, D. E. and Sher, A. ( 1988). Induction of protective immunity against Schistosoma mansoni by vaccination with schistosome paramyosin (Sm97), a nonsurface parasite antigen. Proceedings of the National Academy of Sciences, USA 85, 56785682.
Pettit, D., Smith, W. D., Richardson, J. and Munn, E. A. ( 2000). Localisation and characterisation of ovine immunoglobulin within the sheep scab mite, Psoroptes ovis. Veterinary Parasitology 89, 231239.
Ramos, J. D. A., Teo, A. S. M., Lee, B. W., Cheong, N. and Chua, K. Y. ( 2004). DNA immunization for the production of monoclonal antibodies to Blo t 11, a paramyosin homolog from Blomia tropicalis. Allergy 59, 539547.
Ramirez, B. L., Kurtis, J. D., Wiest, P. M., Arias, P., Aligui, F., Acosta, L., Peters, P. and Olds, G. R. ( 1996). Paramyosin: a candidate vaccine antigen against Schistosoma japonicum. Parasite Immunology 18, 4952.
Reese, G., Jeoung, B. J., Daul, C. B. and Lehrer, S. B. ( 1997). Characterisation of recombinant shrimp allergen Pen a 1 (tropomyosin). International Archives of Allergy and Immunology 113, 240242.
Thomas, W. R. and Smith, W. ( 1998). An update on allergens. House-dust-mite allergens. Allergy 53, 821832.
Van den Broek, A. H., Huntley, J. F., Machell, J., Taylor, M., Bates, P., Groves, B. and Miller, H. R. P. ( 2000). Cutaneous and systemic responses during primary and challenge infestations of sheep with the sheep scab mite, Psoroptes ovis. Parasite Immunology 22, 407414.
Van den Broek, A. H., Huntley, J. F., Machell, J., Taylor, M. and Miller, H. R. P. ( 2003). Temporal pattern of isotype-specific antibody responses in primary and challenge infestations of sheep with Psoroptes ovis – the sheep scab mite. Veterinary Parasitology 111, 217230.

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