Role of lymph-borne cells in the early stages of scrapie agent dissemination from the skin.

Abstract

Scrapie is a natural transmissible spongiform encephalopathy (TSE) of sheep, infecting the animal via the gastrointestinal tract or the skin. This project tested the hypotheses that lymph-borne cells (especially dendritic cells) are crucial for the systemic dissemination of the infectious agent from the site of infection in the skin, that PrP genotype affects PrPSC association with dendritic cells and that PrPSC carriage by cells affects their expression of cytokines. Skin, of scrapie-susceptible VRQ/ARR and scrapie-resistant ARR/ARR PrP genotypes, was scarified with FITC-labelled PrPSC. Pseudoafferent lymphatic cannulation was then used to monitor the presence of FITC-PrPSC over time in different lymph cell populations and plasma in the draining afferent lymphatics. The major observation was that PrPSC did not associate significantly with any lymphocyte or dendritic cell population in the 5 days following PrPSC scarification. The only cells seen to associate with PrPSC were neutrophils. Furthermore, despite the quantity of PrPSC used for scarification being equivalent to a standard infectious dose (the VRQ/ARR sheep dying at approximately 260 days) the only PrP found in afferent lymph during the 0-5-day period was proteinase K sensitive (i.e. soluble PrPC). No differences were observed between the PrP genotypes. Analysis of the effects of PrPSC scarification of cellular cytokine mRNA expression (by a nuclease protection assay) showed raised levels of IL-1beta and IL-8 in the susceptible VRQ/ARR group and raised levels of IFNgamma in the resistant ARR/ARR animals.