Accelerated, spleen-based titration of variant Creutzfeldt-Jakob disease infectivity in transgenic mice expressing human prion protein with sensitivity comparable to that of survival time bioassay.
The dietary exposure of the human population to the prions responsible for the bovine spongiform encephalopathy (BSE) epizooty has led to the emergence of variant Creutzfeldt-Jakob disease (vCJD). This fatal, untreatable neurodegenerative disorder is a growing public health concern because the prevalence of the infection seems much greater than the disease incidence and because secondary transmission of vCJD by blood transfusion or use of blood products has occurred. A current limitation in variant CJD risk assessment is the lack of quantitative information on the infectivity of contaminated tissues. To address this limitation, we tested the potential of a transgenic mouse line overexpressing human prion protein (PrP), which was previously reported to propagate vCJD prions. Endpoint titration of vCJD infectivity in different tissues was evaluated by two different methods: (i) the "classical" bioassay, based on the appearance of clinical symptoms and the detection of pathological prion protein in tissues of the inoculated mouse, and (ii) a shortened bioassay based on the detection of the protein in the mouse spleen at defined time points. The two methods proved equally sensitive in quantifying infectivity, even after very-low-dose inoculation of infected material, but the time schedule was shortened from ~2.5 years to ~1 year with the spleen bioassay. Compared to the "gold-standard" RIII model routinely used for endpoint titration of vCJD/BSE prions, either method improved the sensitivity by >2 orders of magnitude and allowed reevaluating the infectious titer of spleen from a vCJD individual at disease end stage to >1,000-fold-higher values.


OBJECTIVE
Here, we provide key reevaluation of the infectious titer of variant CJD brain and spleen tissues. The highly sensitive, accelerated spleen-based assay should thus constitute a key advance for variant CJD epidemiological and risk assessment purposes and should greatly facilitate future titration studies, including, for example, those aimed at validating decontamination procedures. The overlooked notion that the lymphoid tissue exhibits a higher capacity than the brain to replicate prions even after low-dose infection raises new questions about the molecular and/or cellular determinant(s) involved, a key issue regarding potent silent carriers of variant CJD in the lymphoid tissue.
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Sophie
S
Halliez
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Fabienne
F
Reine
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Laetitia
L
Herzog
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Emilie
E
Jaumain
INSERM, Equipe maladie d'Alzheimer et maladies à Prions, CRicm, UMRS 975, CNRS UMR7225, UPMC. R., Hôpital de la Salpêtrière, Paris, France InVS, Centre national de référence des Agents Transmissibles Non Conventionnels, Hôpital de la Salpêtrière, Paris, France AP-HP, Laboratoire de Neuropathologie, Hôpital de la Salpêtrière, Paris, France.
Stéphane
S
Haïk
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Human
H
Rezaei
INRA, UMR1313, Génétique Animale et Biologie Intégrative, Jouy-en-Josas, France.
Jean-Luc
JL
Vilotte
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France.
Hubert
H
Laude
INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moléculaires, Jouy-en-Josas, France vincent.beringue@jouy.inra.fr.
Vincent
V
Béringue
D000818:Animals; D001681:Biological Assay; D019411:Clinical Laboratory Techniques / Q000379:methods*; D007562:Creutzfeldt-Jakob Syndrome / Q000175:diagnosis*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D011328:Prions / Q000032:analysis*; D012680:Sensitivity and Specificity; D013154:Spleen / Q000737:chemistry*; D013997:Time Factors
D011328:Prions