Characterization of Vitronectins in Atherosclerotic Lesions

Abstract

Vitronectin is one of the major extracellular matrix proteins that accumulates in atherosclerotic lesions. A monoclonal antibody (EMR1a/212D) specifically stained the extracellular regions in thickened intima which colocalized well with lipid deposition. The antigenic glycoprotein with a molecular weight of 66KDa was revealed to be rabbit vitronectin. When homogenates of WHHL rabbit atheroma were subjected to immunoblot analysis using EMR1a/212D, four molecules with molecular weight 66, 56, 50 and 47KDa were detected.To confirm whether these smaller immunopositive bands were derived from mature vitronectin, another monoclonal antibody (EMR1b/244H) recognizing the polypeptide region of vitronectin was prepared. All four molecules were detected by EMR1b/244H as well as by EMR1a/212D. Two smaller vitronectins (56KDa and 5OKDa) were found in atherosclerotic lesions and increased markedly during the development of atherosclerosis.On the other hand, the vitronectin detected in normal rabbit aorta was mainly of the mature type, while 56KDa and 47KDa forms were not detected. The total amount of the four vitronectins in atherosclerotic lesions was 38.5kigou+5.0 ng/mg wet weight tissue, a value approximately 9.5 fold higher than that found in normal aorta. In conclusion, we found massive accumulation of these vitronectins concomitant with atherosclerotic development in rabbit aorta.

Identification of Human scFvs Targeting Atherosclerotic Lesions

“…A number of reports (25,26) indicate that vitronectin accumulates in atherosclerotic plaques and is strongly expressed after vascular injury. Immunohistological staining of B3.3G on the surface of early lesions and within the plaque, near the necrotic core, where lipids are deposited in more complex plaques, is in good agreement with previously published data (27). Immunoblot experiments have shown that B3.3G detected two bands at ϳ56 and ϳ50 kDa.…”

“…Immunoblot experiments have shown that B3.3G detected two bands at ϳ56 and ϳ50 kDa. Interestingly, Mori et al (27) have previously characterized the 56-and 50-kDa bands as the forms of vitronectin markedly increased in atheromatous aorta in Watanabe heritable hyperlipidemic rabbit. Taken together, these results prove that vitronectin is an attractive target for imaging atherosclerotic lesions.…”

Selective α v β 3 -Receptor Blockade Reduces Macrophage Infiltration and Restenosis After Balloon Angioplasty in the Atherosclerotic Rabbit

“…On frozen sections, the following antibodies were used: LM609, a mouse monoclonal anti-human ␣ v ␤ 3 -integrin, 10 g/mL (Chemicon International Inc) 8 ; mouse anti-human osteopontin, 2.4 g/mL (University of Iowa Hybridoma Bank) 17 ; EMR 1a/212D, a monoclonal anti-rabbit vitronectin (ascites fluid, 1:4000, Dr Tatsuya Takano, Teikyo University, Japan) 18 ; RAM-11, 0.03 g/mL; Rb1/9 and Rb2/3, mouse monoclonal anti-rabbit ICAM-1 (intracellular adhesion molecule-1) and VCAM-1 (vascular cell adhesion molecule-1) antibodies (tissue culture supernatant, 1:10, Dr Myron Cybulsky, University of Toronto) 10 ; and a goat polyclonal anti-rabbit MCP-1 (monocyte chemotactic protein-1) antibody, 5 g/mL (Dr Teizo Yoshimura, National Cancer Institute, Frederick, Md). 19 A Vectastain Elite ABC kit (Vector Labs) was used to detect primary antibodies, and DAB (DAKO) was used for visualization.…”

Arteriosclerosis in the Influx and Intravisceral Arteries of the Liver, Kidney and Lung of WHHL Rabbits.

“…There have been many reports on sclerotic lesions of the coronary artery [7,11,13,16] and aorta [4,5,8,9] until now, but there are no reports on the arterial lesions in the influx and intravisceral arteries of the parenchymatous organs, except for the heart and brain [12], in the WHHL rabbit. In the present study, we investigated histopathologically the arteriosclerotic changes in the influx and intravisceral arteries of the liver, kidney and lung, and clarified the degree and progress level of arteriosclerosis in these parenchymatous organs.…”