Browsing by Subject "Amyloid"

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    Anti-l-light chain-peptide antibodies are suitable for the immunohistochemical classification of AL amyloid
    (Murcia : F. Hernández, 2007) Kuçi, H.; Ebert, M.; Röcken, C.
    We aimed to test whether antibodies raised against recombinant peptides corresponding to the variable region of immunoglobulin light chains are suitable for the immunohistochemical classification of amyloid. The Entrez database of the National Center for Biotechnology Information (NCBI) was searched for all protein sequence entries which met the search criteria “amyloid” and “lambda light chain”. Sixty-four different l-light chain-derived amyloid protein sequences were retrieved, aligned and categorized into the V region subgroups of l-light chain detailed by the NCBI, i.e. subgroup I (21 protein sequences), II (14), III (6), IV (1), V (1) and VI (21). V region subgroup I was chosen for epitope sequence selection and two rabbits were immunized with the following peptides: NH2- ISCSGSSSNIGSNTV-CONH2 and NH2-QRPSG VPDRFSGSKSGTS-CONH2. Sensitivity and specificity of the IgG-purified antibodies was tested by Western blotting using amyloid A- (AA), ALl- and ALk-amyloid proteins, and by immunohistochemistry on tissue microarrays with 110 different amyloid containing tissue samples obtained at autopsy from 22 patients, and on 27 biopsy specimens from a series of 24 patients. Our peptide antibodies specifically stained AL amyloid l- light chain-origin, in both Western blots and formalinthat peptide-antibodies directed against immunoglobulin- derived l-light chain proteins can be applied for the immunohistochemical classification of amyloid. This offers the opportunity to generate a large set of anti- l-light chain protein-antibodies for the immunohistochemical classification of amyloid independently from native human tissue sources.
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    Transgenic mice overexpressing both amyloid ß-protein and perlecan in pancreatic acinar cells
    (Murcia : F. Hernández, 2004) Fukuchi, K.; Hart, M.; Yan, Z.; Hassell, J.R.; Li, L.
    Heparan sulfate proteoglycans such as perlecan are thought to facilitate amyloid fibril formation. Tg3695 mice overexpress perlecan core protein in many tissues including the brain and pancreas. Tg13592 mice overexpress the signal plus 99-amino acid carboxyl terminal sequences (C99) of amyloid ß-protein precursor in multiple tissues and develop amyloid deposits in the pancreas. To investigate a role of perlecan in ß-amyloidosis, we established doubly transgenic mice by crossing the two lines of transgenic mice. The expression levels of the two transgenes remained unchanged in the brain and pancreas and the doubly transgenic mice did not develop amyloid deposits in the brain up to 19-months of age. Amyloid load detected by thioflavine S in the pancreas of the doubly transgenic mice was not significantly different from that in the transgenic littermates expressing only C99. Amyloid load in the pancreas increased during aging. We found a positive correlation between the Aß-immunoreactive (non-fibrillar and fibrillar) and thioflavine S-positive (fibrillar) Aß deposits in the single (C99) but not doubly transgenic mice. Our results suggest that perlecan does not independently influence amyloid formation in the pancreas of the transgenic mice and that there may be other factors that may modulate amyloid formation together with perlecan.