ABSTRACT
BACKGROUND: There is a demonstrated risk of infection by transmissible spongiform encephalopathies (TSEs) through transfusion from asymptomatic donors. Currently, blood-borne TSE infectivity cannot be detected with a diagnostic test, nor is it likely to be amenable to inactivation; however, its depletion with specific adsorp-tive ligand resins is possible. STUDY DESIGN AND METHODS: Six ligands that bind the prion protein, PrP, were selected by screening large solid-phase combinatorial chemical libraries. The selected resins were placed in columns and challenged with a unit of leukoreduced human red blood cells (RBCs) spiked with hamster brain-derived scrapie infectivity. The performance of each ligand was assessed by comparing the TSE infectivity titer in the RBCs before and after passage through each of five resin columns in series. RESULTS: Four resins were able to reduce infectivity titer by 3 to more than 4 log ID(50) per mL. The reduction was not due to nonspecific matrix interactions since a chemical modification of the most effective ligand completely abolished its ability to bind infectivity (negative control). A small subfraction of the infectivity, 0.01 percent, could not be removed, even upon repeated passage through successive columns. CONCLUSION: If endogenous TSE infectivity in RBCs binds to the ligands in the same proportion as brain-derived infectivity spiked into RBCs, the four most effective ligands would remove 3 to 4 log ID(50) per mL. A follow-up experiment is in progress to test whether endogenous blood-borne infectivity is also reduced.
Subject(s)
Erythrocytes/chemistry , Prion Diseases/prevention & control , Prions/isolation & purification , Animals , Combinatorial Chemistry Techniques , Cricetinae , Erythrocyte Transfusion/methods , Erythrocyte Transfusion/standards , Humans , Ligands , Models, Biological , Prions/blood , Protein Binding , TitrimetryABSTRACT
Public health policy makers need quantitative scientific data to assess the risk to the blood supply posed by transmissible spongiform encephalopathy (TSE) diseases. To this end, our laboratory has developed a model of blood-borne TSE infectivity in hamsters infected with the 263K strain of scrapie, an experimental model of choice for quantitative studies of TSE infectivity. We report here a microsurgical method for cannulation of the carotid artery in the hamster that allows transfusion of a large fraction of the blood volume of the hamster, with virtually no blood loss to the surgical site or exposure to nervous tissue. Animals are minimally affected by the surgery, recover quickly and completely, and survive for their natural lives (as long as 3 years). This procedure has been used to obtain quantitative data on the transmissibility of the TSEs by transfusion, and these findings have informed public health policy on blood donation and blood use.
Subject(s)
Animals, Laboratory/surgery , Blood Transfusion/methods , Carotid Arteries/surgery , Prion Diseases/transmission , Animals , Cricetinae , Prion Diseases/bloodABSTRACT
The infectivity of hamster scrapie strain 263K was measured in platelets isolated from blood pooled from six hamsters with clinical scrapie. The total number of infectious doses present in the blood pool was 220, out of which only 3.5 infectious doses were associated with platelets. A larger proportion of the total infectivity was recovered from the mononuclear leukocyte fraction. This result indicates that platelets are not the source of blood-borne infectivity in transmissible spongiform encephalopathy-infected hamsters.
