NR AGUI
AU Krasemann,S.; Groschup,M.H.; Hunsmann,G.; Bodemer,W.
TI Induction of antibodies against human prion proteins (PrP) by DNA-mediated immunization of PrP0/0 mice
QU Journal of Immunological Methods 1996 Dec 15; 199(2): 109-18
PT journal article
AB Prion diseases are neurodegenerative disorders, affecting humans and animals. The human diseases include kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). To generate monospecific antisera against human prion proteins we have immunized mice with DNA coding for different human prion proteins. We constructed immunization vectors expressing individual genotypes of either the cellular prion gene (PRNP) or mutant forms under appropriate promoters. This approach avoids the preparation of infectious material for immunization. To circumvent immunological tolerance prion protein-deficient PrP0/0 mice were used for the DNA-mediated immunization. Thereby monospecific sera were raised capable of specifically precipitating in vitro synthesized human prion proteins. With prion protein-specific peptide ELISAs, we found that antibodies are predominantly directed against the octapeptide repeat region and to a lesser extent to regions comprising the signal peptide, the neurotoxic domain or the GPI anchor. In contrast, prion gene-positive (PrP+/+) BALB/c mice immunized under the same experimental conditions as the PrP0/0 mice did not respond with antibody formation against the human prion protein. This is the first report clearly showing that immune competent prion protein-deficient mice react with a vigorous polyclonal immune response after DNA-mediated immunization with human prion gene sequences.
IN Die Autoren setzten die kodierenden Regionen verschiedener Varianten des menschlichen Prionproteingens in Expressionsvektoren ein und sorgten für eine Expression in Mäusen, die teilweise keine eigenen Prionproteine besaßen. Daraufhin produzierten die Mäuse ohne eigene Prionproteine monospezifische Antisera, welche spezifisch in vitro synthetisierte menschliche Prionproteine präzipitieren. Mit Peptiden stellten die Autoren fest, dass die Antikörper hauptsächlich die Oktapeptidregion, zum Teil auch das Signalpeptid, die neurotoxische Domaine oder den GPI-Anker erkannten. Die Mäuse mit eigenen Prionproteinen produzierten hingegen keine Antikörper gegen menschliche Prionproteine.
ZR 22
MH Amino Acid Sequence; Animal; Antibody Formation; DNA/*immunology; Genes, Structural; Human; Mice; Mice, Mutant Strains; Molecular Sequence Data; Peptides/immunology; Prions/genetics/*immunology; Sequence Alignment; Support, Non-U.S. Gov't; Vaccines, Synthetic/immunology
AD German Primate Centre, Department of Virology and Immunology, Göttingen, Germany.
SP englisch
PO Niederlande