Ruminant B-Lymphocyte Yellow Fluorescent Protein Aggregation Bioassay for Elk Chronic Wasting Disease

The goal of the proposed research is to develop a cell culture model for elk chronic wasting disease (CWD) prion propagation that can be used as a bioassay for detecting CWD. Our goal will be accomplished by bioengineering a bovine B-lymphocyte cell (B-cell) line to surface express elk PrPc fused to yellow fluorescent protein (YFP). Interaction of these B-cells with infectious, mis-folded, protease resistant CWD prion (PrPcwd) will induce conversion of the dispersed YFP-PrPc to aggregated mis-folded YFP-PrPcwd that will be detected by confocal microscopy as aggregated YFP-PrPcwd on the B-cell surface. Once Phase I proof-of-concept has been demonstrated by elk PrPcwd conversion of YFP-PrPc on the B-cell line to YFP-PrPcwd, a stably transfected cell line will be developed and validated as a sensitive, specific, and reproducible bioassay for detecting PrPcwd infected elk and deer tissues and body fluids during Phase II.