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Open Access Research article

Accumulation profiles of PrPSc in hemal nodes of naturally and experimentally scrapie-infected sheep

Rohana P Dassanayake1*, Thomas C Truscott2, M Özgür Özyiğit3, Dongyue Zhuang2, David A Schneider2 and Katherine I O’Rourke1

Author affiliations

1 Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA, 99164-7040, USA

2 U.S. Department of Agriculture, Animal Disease Research Unit, Agricultural Research Service, Pullman, WA, 99164-6630, USA

3 Department of Pathology, Faculty of Veterinary Medicine, Uludağ University, Görükle, Bursa, 16059, Turkey

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Citation and License

BMC Veterinary Research 2013, 9:82  doi:10.1186/1746-6148-9-82

Published: 19 April 2013

Abstract

Background

In classical scrapie, the disease-associated abnormal isoform (PrPSc) of normal prion protein accumulates principally in the nervous system and lymphoid tissues of small ruminants. Lymph nodes traffic leukocytes via lymphatic and blood vasculatures but hemal nodes lack lymphatic vessels and thus traffic leukocytes only via the blood. Although PrPSc accumulation profiles are well-characterized in ovine lymphoid tissues, there is limited information on such profiles in hemal nodes. Therefore, the objective of this study was to compare the follicular accumulation of PrPSc within hemal nodes and lymph nodes by prion epitope mapping and western blot studies.

Results

Our studies found that PrPSc accumulation in 82% of animals’ abdominal hemal nodes when PrPSc is detected in both mesenteric and retropharyngeal lymph nodes collected from preclinical and clinical, naturally and experimentally (blood transfusion) scrapie-infected sheep representing all three major scrapie-susceptible Prnp genotypes. Abdominal hemal nodes and retropharyngeal lymph nodes were then used to analyze immune cell phenotypes and PrPSc epitope mapping by immunohistochemistry and PrPSc banding patterns by western blot. Similar patterns of PrPSc accumulation were detected within the secondary follicles of hemal nodes and retropharyngeal lymph nodes, where cellular labeling was mostly associated with macrophages and follicular dendritic cells. The pattern of PrPSc accumulation within hemal nodes and retropharyngeal lymph nodes also did not differ with respect to epitope mapping with seven mAbs (N-terminus, n = 4; globular domain, n = 2; C-terminus, n = 1) in all three Prnp genotypes. Western blot analysis of hemal node and retropharyngeal lymph node homogenates revealed identical three banding patterns of proteinase K resistant PrPSc.

Conclusion

Despite the anatomical difference in leukocyte trafficking between lymph nodes and hemal nodes, the follicles of hemal nodes appear to process PrPSc similarly to lymph nodes.

Keywords:
Scrapie; Hemal nodes; Epitope mapping; Sheep; Prions; TSE