Immunohistochemistry for the Prion Protein: Comparison of Different Monoclonal Antibodies in Human Prion Disease Subtypes

Abstract

Demonstration of the abnormal form of the prion protein (PrP) in the brain confirms the diagnosis of human prion disease (PrD). Using immunohistochemistry, we have compared ten monoclonal antibodies in PrD subtypes including sporadic and variant Creutzfeldt-Jakob disease (CJD), fatal familial insomnia, Alzheimer's disease (AD), and control brains. CJD subgroups were determined using Western blot analysis for the protease-resistant PrP type in combination with sequencing to determine the genotype at the methionine/valine polymorphism at codon 129 of the prion protein gene. None of the antibodies labeled given subgroups exclusively, but the intensity of immunoreactivity varied among morphologically distinct types of deposit. Fine granular or synaptic PrP deposits stained weakly or not at all with antibodies against the N-terminus of PrP, and were visible in one case only with 12F10 and SAF54. Coarser and plaque type deposits were immunolabeled with all antibodies. The immunostaining patterns appear characteristic for the disease subgroups. Labeling of certain neurons in all cases irrespective of disease, and staining at the periphery and/or throughout the senile plaques of AD patients were also noted. Antibodies such as 6H4 and 12F10 failed to give this type of labeling and are therefore less likely to recognise non-pathological PrP material in immunohistochemistry. Brain Pathol 2002;12:1-11 Recently a molecular classification of sporadic Creutzfeldt-Jakob disease (sCJD) suggested two major types of protease-resistant PrP (PrP Res ), type-1 and type-

Human prion diseases: from antibody screening to a standardized fast immunodiagnosis using automation

“…By comparison, the 12F10 automated method was, in our hands, more sensitive than the other procedures using the reference 3F4 monoclonal antibody, which is not suitable for this automated system. Our results confirm and extend the results of Kovacs et al, 25 who identified 12F10 as a potentially useful antibody for immunodiagnosis, owing to a reduction in the number of pretreatments and a speeding up of the procedure using a fast immunodiagnosis automate. In addition, we demonstrated the usefulness of this procedure in a very large panel of patients affected by a high diversity of human prion disorders.…”

Subcellular Localization of Disease-Associated Prion Protein in the Human Brain

“…2 Due to the lack of unequivocally conformation-specific antibodies, immunohistochemical differentiation of PrP C from PrP Sc has remained a matter of debate and creates a barrier for the evaluation of disease process. However, detailed immunohistochemical descriptions define patterns which are specific for disease, 3 thus the term disease-associated PrP (labeled PrP TSE in the present study) is suitable to label conformationally uncharacterized deposits of PrP observed only in priondiseased brains by immunohistochemistry.…”

“…3 The first is characterized by a diffuse fine-granular distribution of anti-PrP TSE immunoreactivity resembling the distribution of the presynaptic protein synaptophysin (hence the term "synaptic pattern"). Presynaptic domains appear to be the privileged site of PrP C , as has been confirmed by double labeling with synaptophysin.…”

“…We used the monoclonal anti-PrP antibody 12F10 that was previously shown to be the most reliable in detecting disease-associated PrP. 3 Sections were incubated overnight at room temperature with each antibody. The Netherlands) for other antibodies.…”

No association of prion protein gene polymorphisms with Alzheimer's disease in Korean population

“…A in AD and PrP Sc in CJD can coexist. Prion protein was localized to senile plaques in the aged and AD brain and promoted plaque formation in 3 month-old bigenic mice carrying mutant human APP and prion protein genes (Kovacs et al, 2002;Schwarze-Eicker et al, 2005). Also, both diseases show a consistent pattern of cortical degeneration (reviewed in Forman et al, 2004;Armstrong et al, 2005).…”