The role of a putative microfibrillar protein (80 kDa) in abdominal aortic aneurysm disease.
BACKGROUND
We previously have reported the partial amino acid sequence of a putative aortic autoantigen in patients with abdominal aortic aneurysm (AAA) disease that has homologies with an elastin-associated microfibrillar protein found in aorta of pigs. This study was conducted to further define the role that microfibrillar proteins may play as autoantigens in AAA disease.


METHODS
An extraction procedure was performed on AAA tissue using high concentrations of guanidinium hydrochloride (GuHCl) under reducing conditions. The microfibrillar extract was then probed with immunoglobulin (Ig) G isolated with Protein A from phosphate-buffered saline (PBS) extracts of 10 AAA specimens and 6 atherosclerotic, nonaneurysmal aortas. Immunoblotting was also performed with serum IgG from 9 AAA patients and 9 normal control patients. Immunohistochemistry using goat anti-human IgG (Fc-specific) on AAA tissue and AAA wall IgG on normal aorta were also performed.


RESULTS
Eight of 10 AAA wall IgG reacted with an 80-kDa protein from the aortic microfibrillar extract, compared to 0 out of 6 atherosclerotic wall IgG (P = 0.0035, Fischer's Exact Test). Staining of the 80-kDa band appeared to increase with progressive additions of GuHCl, up to extract SKGCGC. Immunoblotting using serum IgG from 9 AAA patients and 9 normal control patients on the aneurysm microfibrillar extracts revealed no reactive bands. Immunohistochemistry using IgG from AAA wall showed the localization of the antibodies to the adventitial connective tissue matrix, mainly collagen fibers.


CONCLUSIONS
These observations suggest that a collagen-associated protein, extractable by a microfibrillar extraction procedure from aortic aneurysm tissue, may be among the targets of an autoimmune response in AAA disease.
D001012:Aorta, Abdominal / Q000737:chemistry*; D017544:Aortic Aneurysm, Abdominal / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology; D003285:Contractile Proteins / Q000032:analysis*; D016326:Extracellular Matrix Proteins; D005656:Fungal Proteins; D006360:Heat-Shock Proteins / Q000032:analysis*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D000072260:RNA Splicing Factors
D001324:Autoantigens; D003285:Contractile Proteins; D016326:Extracellular Matrix Proteins; D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D007074:Immunoglobulin G; D000072260:RNA Splicing Factors; C016598:microfibrillar protein