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

Published online by Cambridge University Press:  04 July 2006

A. J. NISBET ,
A. MACKELLAR ,
H. W. WRIGHT ,
G. P. BRENNAN ,
K. Y. CHUA ,
N. CHEONG ,
J. E. THOMAS  and
J. F. HUNTLEY
A. J. NISBET
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland
A. MACKELLAR
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland
H. W. WRIGHT
Affiliation:
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland
G. P. BRENNAN
Affiliation:
School of Biology and Biochemistry, Queen's University Belfast, Lisburn Road Belfast BT9 7BL, Northern Ireland
K. Y. CHUA
Affiliation:
National University of Singapore, Yong Loo Lin School of Medicine, Department of Paediatrics, Lower Kent Ridge, Singapore 119074
N. CHEONG
Affiliation:
National University of Singapore, Yong Loo Lin School of Medicine, Department of Paediatrics, Lower Kent Ridge, Singapore 119074
J. E. THOMAS
Affiliation:
Institute of Molecular and Cellular Biology, Miall Building, University of Leeds, Leeds LS2 9JT, UK
J. F. HUNTLEY
Affiliation:
Institute of Molecular and Cellular Biology, Miall Building, University of Leeds, Leeds LS2 9JT, UK
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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.

Keywords

PsoroptesallergenDer p 10Der p 11tropomyosinparamyosin
Type
Research Article
Information
Parasitology , Volume 133 , Issue 4 , October 2006 , pp. 515 - 523
Copyright
© 2006 Cambridge University Press

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