Immunohistochemical localization of heat shock protein-70 in normal-appearing and atherosclerotic specimens of human arteries.

Heat shock proteins (HSPs) are synthesized by cells under metabolic stress and are known to enhance a cell's ability to survive life-threatening stress. The authors have begun to examine HSPs in the context of human atherosclerosis. This study demonstrated immunohistochemically the presence of HSP-70 in human and rabbit arteries, and its distribution in relation to necrosis and lipid accumulation, as well as vascular smooth muscle cells and macrophages, in human atherosclerotic plaques. Advanced lesions from 10 human carotid endarterectomy specimens were compared with 11 human aortic specimens from autopsy and 8 rabbit aortas. The immunostaining procedure used a mouse monoclonal antibody specific for the inducible form of HSP-70. Normal rabbit aortas were tested for changes in HSP-70 up to 24 hours after removal, and were used as controls for the human aortas. Representative plaques were examined for lipid content by osmium staining, and for smooth muscle cell and macrophage components using cell-specific monoclonal antibodies followed by immunostaining. The results indicated that HSP-70 was present in human and rabbit arteries and remained unchanged in distribution or concentration up to 15 hours after death. HSP-70 was present weakly throughout the media of normal-appearing arterial specimens. In contrast, HSP-70 was concentrated in the central portions of more thickened atheromas around sites of necrosis and lipid accumulation. Macrophages were coincident with these areas and were observed to be lipid-loaded. In contrast, patches of smooth muscle cells were observed in very complicated plaques, but without consistent association with necrosis or increased HSP-70; plaque smooth muscle cells also were observed to contain lipid. Large, relatively avascular and collagenous areas of plaque also were occasionally positive for HSP-70 staining. The results support the hypothesis that elevated HSPs indicate which plaque cells, particularly macrophages, are more stressed in the depth of atheroma, especially in association with necrosis, and should prompt further investigation of the significance of HSP accumulation to the evolution of atherosclerotic plaques.