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

Rohana P Dassanayake, Thomas C Truscott, M Özgür Özyiğit, Dongyue Zhuang, David A Schneider, Katherine I O’Rourke
BMC veterinary research 2013 v.9 no.1 pp. 82
PrPSc proteins, Western blotting, abdominal cavity, bioaccumulation, blood transfusion, dendritic cells, genotype, immunohistochemistry, leukocytes, lymph nodes, nervous system, phenotype, scrapie, sheep
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.