Faculty Bibliography

Several mutations within the Abeta sequence of the APP gene are associated with autosomal dominant cerebral amyloid angiopathy. An Asp to Asn mutation at position 23 of Abeta due to a single nucleotide change at codon 694 of the APP gene causes early onset dementia with leukoencephalopathy and cortical calcification in the Iowa kindred. Severe cerebral amyloid angiopathy and cortical pre-amyloid deposits as well as widespread neurofibrillary tangles are the main neuropathological features of the disease. We have extracted the deposited Abeta species from affected brain areas and biochemically analyzed them using a combination of immunoprecipitation, mass spectrometry, amino acid sequence and western blot analysis. Corroborating previous histological data, western blots with C-terminal specific antibodies revealed an extensive accumulation of Abetax-40 in cortical lesions whereas Abetax-42 was only a minor component. Amino acid sequence of the formic-acid soluble extracts showed high degree of N-terminal heterogeneity, being Abeta starting at position 2 (Ala) the predominant species followed by Abeta4 (Phe) and Abeta1 (Asp) in a 4:2:1 ratio, respectively. IP/mass spect confirmed the presence of Abeta1-40, Abeta2-40 and Abeta4-40 as well as minor components starting at Asp1 and Ala2 but ending at Gly38. Interestingly, amino acid sequence analysis demonstrated the presence of both Asp and Asn at position 23 of the Abeta sequence at a 1:4 ratio, respectively, indicating that the deposited amyloid is a mixture of mutant and wild-type Abeta. Whether one of them is an innocent bystander being recruited by the other (conformational mimicry) or both mutated and non-mutated Abeta peptides are important for the amyloidogenesis process in the Iowa family is being currently investigated

Chromosome 13 dementias, familial British dementia (FBD) and familial Danish dementia (FDD), are associated with neurodegeneration and cerebrovascular amyloidosis, with striking neuropathological similarities to Alzheimer's disease (AD). Despite the structural differences among the amyloid subunits (ABri in FBD, ADan in FDD, and Abeta in AD), these disorders are all characterized by the presence of neurofibrillary tangles and parenchymal and vascular amyloid deposits co-localizing with markers of glial activation, suggestive of local inflammation. Proteins of the complement system and their pro-inflammatory activation products are among the inflammation markers associated with AD lesions. Immunohistochemistry of FBD and FDD brain sections demonstrated the presence of complement activation components of the classical and alternative pathways as well as the neo-epitope of the membrane attack complex. Hemolytic experiments and enzyme-linked immunosorbent assays specific for the activation products iC3b, C4d, Bb, and C5b-9 indicated that ABri and ADan are able to fully activate the complement cascade at levels comparable to those generated by Abeta1-42. ABri and ADan specifically bound C1q with high affinity and formed stable complexes in physiological conditions. Activation proceeds approximately 70-75% through the classical pathway while only approximately 25-30% seems to occur through the alternative pathway. The data suggest that the chronic inflammatory response generated by the amyloid peptides in vivo might be a contributing factor for the pathogenesis of FBD and FDD and, in more general terms, to other neurodegenerative conditions

Tumoral monoclonal immunoglobulin (Ig) light chain non-fibrillar deposits ('aggregomas'), which can be considered analogous to solitary light chain amyloidomas, are a rare presenting feature of B-cell dyscrasias. It is not certain if they are truly localized or if in reality they represent an initial expression of a silent systemic non-amyloid light chain deposition disease (LCDD). This report describes three patients, two of whom presented with cervical masses and the third with a solitary lung nodule, each comprising granular aggregates of monoclonal kappa light chain. Extracted deposits from the lymph node of one patient were shown by N-terminal amino acid sequence analysis to belong to the variable-region kappa I (Vkappa I) light chain subgroup, the first reported kappa-LCDD protein encoded by the L9 gene and the first report of an expressed protein related to this gene. Extracted deposits from the lung nodule of the second patient belonged to the Vkappa IV light chain subgroup encoded by the B3 germ line gene. The N-terminal amino acid sequences of the light chains from the aggregomas were compared with the related germ line sequences and to the N-terminal amino acid sequences of the nine other known kappa-LCDD light chains reported thus far from patients with systemic LCDD