Analysis by one-dimensional (1-D) SDS-PAGE/Western blotting of whole mite extract (larval and adult stages) with sheep sera taken 0-84 days after infection with the sheep scab mite, Psoroptes ovis revealed a progressive IgE antibody response, with a dominant high molecular weight allergen (MW 180 kDa) detected early during infection. Further analysis by two-dimensional (2-D) SDS-PAGE/Western blotting with post-infection sera, revealed a more complex picture with numerous (> 20) IgE reactive spots. Trypsin digest and Maldi-ToF analyses of these spots identified two house dust mite allergen homologues, namely tropomyosin (Der p 10) and paramyosin (Der p 11), and analysis of a third spot indicated an apolipoprotein-like IgE reactive protein (Der p 14). Further 1-D and 2-D SDS/Western blotting, with specific antibodies to the house dust mite allergens Der p 10, 11, and to the IgE reactive peptide of the high molecular weight house dust mite allergen, Der p 14 (vitellogenin/apolipophorin), provided firm evidence for the presence of these three allergens in extracts of the Psoroptes mite. These studies show for the first time that homologues of the house dust mite 10, 11 and 14 group allergens are represented as immunodominant allergens of the sheep scab mite, and may represent important vaccine candidates.
Adult Psoroptes ovis were successively extracted in saline, 1% Tween, 1% Triton and 8 m urea +0.1% CHAPS. The Triton extract was separated into fractions which did or did not bind to ConA lectin. Using QuilA as adjuvant, both Triton fractions and the saline, Tween and urea extracts were tested separately as candidate protective antigens against a P. ovis challenge infestation in sheep. All induced circulating antibody responses, but the saline and Tween extracts also stimulated significant protective effects in those sheep which developed active disease. Compared to control sheep injected with adjuvant only, these immunized animals had more than two and seven-fold reductions in mean lesion areas and mite numbers, respectively.
Regulatory T cells (Tregs) play a central role in maintenance of immune homeostasis by controlling harmful immune responses to inappropriate antigens and are thought to play a key role in modulating hypersensitivity reactions. Infestation of sheep with Psoroptes ovis results in a pronounced cutaneous hypersensitivity-type response, which appears to be crucial for mite survival. We hypothesize that (i) Tregs are involved in sheep scab lesions and (ii) Treg responses may crucially affect lesion development and subsequent mite survival. Foxp3 is a key transcription factor required for generation and maintenance of Tregs in rodents and humans, and is the most widely used marker for Tregs in these species. In this study, we sequence ovine foxp3 and show that it exhibits a high degree of homology with foxp3 from other species. Using a validated immunohistochemical staining technique, we demonstrate that infestation of sheep with P. ovis results in an influx of Foxp3(+) T cells into the skin. Future work will investigate the regulatory function of ovine Foxp3(+) T cells and determine whether the quality of the Treg response to P. ovis plays a role in individual susceptibility to the mite.
Pododermatitis is common in dogs and has numerous and diverse causes. The clinical presentation, approach to diagnosis and management of the more common and important causes of canine pododermatitis are considered in this article. The more common causes dealt with are atopic dermatitis, cutaneous adverse food reactions and trombiculidiasis. Although occurring less frequently, important conditions such as demodicosis, deep interdigital pyoderma, pemphigus foliaceus, symmetrical (lupoid) onychodystrophy and superficial necrolytic dermatitis are also covered.
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