Gregori, L. et al. Reduction in infectivity of endogenous transmissible spongiform encephalopathies present in blood by adsorption to selective affinity resins. Lancet 368, 2226-2230

Veterans Affairs Maryland Health Care System, VA Medical Center, University of Maryland at Baltimore, MD 21201, USA.
The Lancet (Impact Factor: 45.22). 01/2007; 368(9554):2226-30. DOI: 10.1016/S0140-6736(06)69897-8
Source: PubMed


Transmissible spongiform encephalopathies (TSE) can be contracted through blood transfusion. Selective adsorption of the causative agent from donated blood might be one of the best ways of managing this risk. In our study, affinity resin L13, which reduces brain-derived infectivity spiked into human red blood cell concentrate by around 4 log(10)ID(50), and its equivalent, L13A, produced on a manufacturing scale, were assessed for their ability to remove TSE infectivity endogenously present in blood.
500 mL of scrapie-infected hamster whole blood was leucoreduced at full scale before passage through the affinity resins. Infectivity of whole blood, leucoreduced whole blood (challenge), and the recovered blood from each flow-through was measured by limiting dilution titration.
Leucoreduction removed 72% of input infectivity. 15 of 99 animals were infected by the challenge, whereas none of the 96 or 100 animals inoculated with the final flow-throughs from either resin developed the disease after 540 days. The limit of detection of the bioassay was 0.2 infectious doses per mL. The overall reduction of the challenge infectivity was more than 1.22 log10ID. The results showed removal of endogenous TSE infectivity from leucoreduced whole blood by affinity ligands. The same resins adsorb normal and abnormal prion protein from human infections with variant, sporadic, and familial Creutzfeldt-Jakob disease, in the presence of blood components.
TSE affinity ligands, when incorporated into appropriate devices, can be used to mitigate the risks from TSE-infected blood, blood products, and other materials exposed to TSE infectivity.

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    • "On the basis of the relative infectious titre in plasma, as assessed in rodent models, LD has been considered as a potentially beneficial at reducing the risk of v-CJD transmission associated with RBCs transfusion but this risk redution was insufficient measure [15]. "
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    ABSTRACT: The identification in the UK of 4 v-CJD infected patients thought to be due to the use of transfused Red Blood Cell units prepared from blood of donors incubating v-CJD raised major concerns in transfusion medicine. The demonstration of leucocyte associated infectivity using various animal models of TSE infection led to the implementation of systematic leuco-depletion (LD) of Red Blood cells concentrates (RBCs) in a number of countries. In the same models, plasma also demonstrated a significant level of infectivity which raised questions on the impact of LD on the v-CJD transmission risk. The recent development of filters combining LD and the capture of non-leucocyte associated prion infectivity meant a comparison of the benefits of LD alone versus LD/prion-reduction filters (LD/PR) on blood-borne TSE transmission could be made. Due to the similarity of blood/plasma volumes to human transfusion medicine an experimental TSE sheep model was used to characterize the abilities of whole blood, RBCs, plasma and buffy-coat to transmit the disease through the transfusion route. The impact of a standard RBCs LD filter and of two different RBCs LD/PR prototype filters on the disease transmission was then measured. Homologous recipients transfused with whole-blood, buffy-coat and RBCs developed the disease with 100% efficiency. Conversely, plasma, when intravenously administered resulted in an inconstant infection of the recipients and no disease transmission was observed in sheep that received cryo-precipitated fraction or supernatant obtained from infectious plasma. Despite their high efficacy, LD and LD/PR filtration of the Red Blood Cells concentrate did not provide absolute protection from infection. These results support the view that leuco-depletion strongly mitigates the v-CJD blood borne transmission risk and provide information about the relative benefits of prion reduction filters.
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    • "In an earlier study, we described the detection of PrP aggregates with high sensitivity in brain homogenate of BSE cattle, and in a small number of cerebrospinal fluid samples from BSE cattle [26]. According to the literature, infectivity in blood - even in symptomatic experimental hamsters - is as low as 10 infectious units per ml [33], [34]. In BSE-afflicted cattle infectivity is absent from the lymphatic system and has never been reported in blood [35], [36]. "
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    ABSTRACT: Prion diseases are transmissible neurodegenerative diseases affecting humans and animals. The agent of the disease is the prion consisting mainly, if not solely, of a misfolded and aggregated isoform of the host-encoded prion protein (PrP). Transmission of prions can occur naturally but also accidentally, e.g. by blood transfusion, which has raised serious concerns about blood product safety and emphasized the need for a reliable diagnostic test. In this report we present a method based on surface-FIDA (fluorescence intensity distribution analysis), that exploits the high state of molecular aggregation of PrP as an unequivocal diagnostic marker of the disease, and show that it can detect infection in blood. To prepare PrP aggregates from blood plasma we introduced a detergent and lipase treatment to separate PrP from blood lipophilic components. Prion protein aggregates were subsequently precipitated by phosphotungstic acid, immobilized on a glass surface by covalently bound capture antibodies, and finally labeled with fluorescent antibody probes. Individual PrP aggregates were visualized by laser scanning microscopy where signal intensity was proportional to aggregate size. After signal processing to remove the background from low fluorescence particles, fluorescence intensities of all remaining PrP particles were summed. We detected PrP aggregates in plasma samples from six out of ten scrapie-positive sheep with no false positives from uninfected sheep. Applying simultaneous intensity and size discrimination, ten out of ten samples from scrapie sheep could be differentiated from uninfected sheep. The implications for ante mortem diagnosis of prion diseases are discussed.
    Full-text · Article · May 2012 · PLoS ONE
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    • "While infection of the lymphoid tissue has no apparent deleterious effects on the health of the affected individual, it is likely to be a significant reservoir of infectivity, which may be spread through blood and blood products. Indeed, infectivity in blood was recently demonstrated in both scrapie-and BSE-infected sheep (Houston et al., 2000, 2008; Hunter et al., 2002) , in deer with CWD (Mathiason et al., 2009), and in rodent models of prion disease (Diringer, 1984; Gregori et al., 2006a,b; Gregori and Rohwer, 2007; Holada et al., 2002; Manuelidis et al., 1978). In addition, blood transfusion or blood products have been implicated in the transmission of vCJD to four human recipients (Hewitt, 2006; HPA, 2007; Llewelyn et al., 2004; Peden et al., 2004; Wroe et al., 2006). "
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    ABSTRACT: Recently, we reported that PrP(Sc), a surrogate marker for prion disease, is associated with the cellular fraction of blood from scrapie-infected sheep using a ligand-based immunoassay. In the study reported here, we found that a subset of peripheral blood mononuclear cells is most likely to sequester PrP(Sc) during both the preclinical phase of disease and at clinical end point. These cells had a cell surface phenotype of MHC class II DQ(+), surface immunoglobulin(+), CD11b(+), CD11c(+), CD21(+/)(-), which is consistent with a subpopulation of B cells. What role these cells play in the pathogenesis of scrapie is unclear, but they may contribute to the trafficking of prions to the spleen during early pathogenesis of the disease. Furthermore, tests for preclinical diagnostics could be further improved by targeting these cells.
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