Immunolocalization of proteoglycan types in aortas of pigeons with spontaneous or diet-induced atherosclerosis.

The location of different proteoglycan (PG) types in the developing atherosclerotic lesion was examined by the use of monoclonal antibodies directed toward specific epitopes on distinct PG types. Sections of aorta were prepared from young White Carneau pigeons fed an atherogenic diet to induce lesions rich in lipid-laden macrophages and from older pigeons that had naturally-occurring atherosclerotic lesions. Monoclonal antibodies (MAb) 3-B-3, 9-A-2, or 2-B-6 and 5-D-4, recognizing delta Di-6S generated from chondroitin 6-sulfate (C6S) PG; delta Di-4S generated from dermatan sulfate (DS) PG and from chondroitin 4-sulfate (C4S); and sulfated poly N-acetyllactosamine sequences common to keratan sulfate (KS), respectively, were used to localize PG types by indirect immunofluorescence. In normal aorta, C6S PG was localized primarily in the media and showed a fluorescent gradient (inner media greater than outer media greater than intima greater than adventitia). In the atherosclerotic plaque, major immunoreactivity was observed using MAb 9-A-2 or 2-B-6, whereas lesser amounts were observed with 3-B-3. Patterns of immunoreactivity differed; 9-A-2 or 2-B-6 appeared to be associated with cells whereas 3-B-3 appeared to be intercellular. Although normal aorta was negative for antibody 5-D-4, recognizing KS, atherosclerotic plaques were consistently positive for this antibody. The pattern of 5-D-4 reactivity appeared to be intercellular. Except for immediately below the lesion, no reactive product using 5-D-4 was observed in the media. No major differences in distribution of PG were observed between naturally-occurring or cholesterol-induced fibrous plaques. These results indicate that both 4-sulfated PG and a previously undescribed KS glycoconjugate are major components of the atherosclerotic lesion.