Treatment of human cerebovascular smooth muscle (HCSM) cells with Aβ1-42 or AβQ22 (a mutant associated with hereditary cerebral hemorrhage with amyloidosis-Dutch type) leads to the formation of amyloid fibrils on the surface of HCSM cells within 24 hours. After three days cell associated APP is increased and cell viability decreased. Using monoclonal antibodies which recognize both FLAPP and APPs or FLAPP alone, Melchor (122.13) was able to demonstrate that most of the increase in cell associated APP was due to APPs. Binding of APPs to fibrillar Aβ was demonstrated by electron microscopy of biotinylated APPs and by a solid phase binding assay akin to the synthaloid assay developed by John Maggio's group.-Dominic Walsh.
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