In Vivo Radioimmunodetection of Amyloid Deposits in Experimental Amyloidosis
Two rat IgG monoclonal antibodies (MAbs) to mouse AA protein have been used as reagents for an ELISA assay for SAA, for the demonstration of tissue deposits by the immunoperoxidase method, and for the definition of AA polymorphs in solubilized amyloidotic tissue specimens run onto two-dimensional (2D) gels and probed as Western Blots. Both MAbs localize to tissue deposits in vivo when labelled (MAb*) with I125 or I123 and injected into colchicine-pretreated amyloidotic mice, assessed by (a) whole-body autoradiography (WBAR) (b) external photoscanning (c) tissue autoradiography of perfused organs. Serum t 1/2 of MAb* in amyloidotic animals was greatly accelerated compared to controls, and sustained concentration of label was found in spleen, liver, and kidney quantitated by organ counts up to 96 h post injection. MAb* label copurified with amyloid fibrils up to 10-fold over saline-soluble proteins and residue following homogenization in distilled water and acid extraction; purified fibrils contained AA protein and MAb* heavy and light chain visualized on 2D gels and by autoradiography. Animals were also injected with a mixture of the two MAbs, one labeled with Indium111-DTPA (126 μCi), the other with I125 (6 μCi) and serially scanned over a 48 h period. Fractionation of serum taken after injection showed both isotopes to be present as uncomplexed IgG in blood. WBAR was performed at 48 h and at a time at which I125/In111 radioactivity was calculated to be 127/1; autoradiograms were similar and confirmed localization of both isotopes to tissue deposits in amyloidotic animals. These studies provide evidence that MAbs to amyloid subunit proteins may be useful re-agents for the in vivo radioimmunodetection of some of the amyloid diseases.
KeywordsPermeability Formalin Depression Albumin Electrophoresis
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