1. A prion disease initiation model based on a credible candidate for protein X enables detection of human exposure to BSE, in serum. Bergmann J, Bergmann R, Janetzky B, Preddie E Altegen, Inc.

2. PrP gene induced expression constitutive expression PrP C prionin multi- factorial events ( unknown ) prin gene disease initiation Altegen, Inc.

3. Prionin genes are present in all mammals investigated so far Prionins have unique species-specific antigenic epitopes Pure, synthetic bovine prionin converts human native PrP in a cross-species manner, and recombinant bovine PrP, to conformers with a 27-29 kDa PK- resistant core under physiological conditions within minutes of contact, in a test tube We suggest that prionins are protein-X, the illusive converting factor in TSE, and add that prionins play a role in TSE initiation The model which follows shows how prionins provide means for the detection of human exposure to TSE humans deerrabbit cattle moosehamster sheep elkhorse mouse (atypical) human PK-resistant PrP PrP 5 15 25 min human PrP rec. PrP 30‘ 90‘ 30‘ 29 k - Altegen, Inc.

4. PrP gene induced expression constitutive expression PrP C prionin prin gene disease initiation immune modulation Altegen, Inc. Detection by a specific antibody-trap ELISA

5. 1 prionin gene is induced 2 the expressed prionin elicits an immune response, inhibiting disease symptoms 3 the immune response declines with time in some cases 4 released from immune control, prionins interact with PrP and convert it to PrP Sc 5 prionins are chaperoned to the brain complexed to PrP Sc ; in the brain, the complex dissociates at the neuronal membrane 6 converted PrP (PrP Sc ) is deposited extra-neuronally 7 prionins enter the neuronal membranes and initiate neurodegeneration Altegen, Inc. immune modulation human prionin (prinH) constitutive expression escape PrP Sc depos. TSE initiation model for vCJD PrP C induced expression con- version PrP geneprin gene cellular membranes neuro- degener. 4 6 7 1 5 multi- factorial events 2 toxic inter- mediate 3 disease initiation

6. Altegen, Inc. bovine prionin (prinB) external source immune modulation human prionin (prinH) constitutive expression escape PrP Sc depos. toxic inter- mediates TSE initiation model for vCJD PrP C induced expression con- version PrP geneprin gene cellular membranes neuro- degener. 3 4 6 7 1 5 multi- factorial events 2 immune modulation escape 2a 3 1a external source disease initiation 1 prionin gene is induced 2 the expressed prionin elicits an immune response, inhibiting disease symptoms 3 the immune response declines with time in some cases 4 released from immune control, prionins interact with PrP and convert it to PrP Sc 5 prionins are chaperoned to the brain complexed to PrP Sc ; in the brain, the complex dissociates at the neuronal membrane 6 converted PrP (PrP Sc ) is deposited extra-neuronally 7 prionins enter the neuronal membranes and initiate neurodegeneration

7. OD 495nm 0.5 – - 0.4 – - 0.3 – - 0.2 – - 0.1 – - 0.0 – anti-prinB anti-prinH 5 months 7 months 20 months anti-prionin antibodies in a suspected vCJD patient Altegen, Inc. 5 months 7 months 20 months

8. Anti-bovine prionin antibody in blood bank samples number of samples number of positives 3000 - 2000 - 1000 - 0 - - 20 - 10 - 0 2883 - 571 - - 0 - 4 2003 country 1 1996-8 country 2 Altegen, Inc.

9. Conclusion Prionins are TSE-related proteins Transmitted (contaminating) prionins elicit an immune response Anti-prionin ELISAs can detect this immune response in blood Endogenous prionins are related to TSE-initiation They elicit an auto-immune response Anti-prionin ELISAs can detect the auto-immune response in blood We suggest that the anti-prionin ELISA should be used routinely to test blood donations for exposure to TSE

10. Thank you for your attention Altegen, Inc.