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Neoangiogenesis, T-lymphocyte infiltration, and heat shock protein-60 are biological hallmarks of an immunomediated inflammatory process in end-stage calcified aortic valve stenosis

Annamaria Mazzone, MD^*,*, Maria Carmela Epistolato, BSc, Raffaele De Caterina, MD, PhD, Simona Storti, BSc, Simona Vittorini, PhD^*, Silverio Sbrana, MD, PhD^*, Jacopo Gianetti, MD, PhD^*, Stefano Bevilacqua, MD^*, Mattia Glauber, MD^*, Andrea Biagini, MD^* and Piero Tanganelli, MD

^* CNR Institute of Clinical Physiology, Ospedale Pasquinucci, Massa, Italy
Department of Pathology, University of Siena, Siena, Italy
Chair of Cardiology, "G. d'Annunzio" University, Chieti, Italy

View larger version (97K): [in a new window]   Figure 1 (a) The majority of calcified stenotic valves studied, as in this case, are occupied by calcium deposits (violet area, hematoxylin-eosin [HE]). (b) Case 12: areas with ossification and bone marrow formation were observed in a calcified aortic valve leaflet (HE). (c) Sample of normal aortic valve from control patient showing slight thickening and fibrosis of the valve leaflet (HE).

View larger version (106K): [in a new window]   Figure 2 (a) In a calcified aortic valve, abundant inflammatory infiltrates are observed with small thin-walled neovessels, prevalently composed of T-lymphocytes, plasma cells, and macrophages (hematoxylin-eosin [HE]). (b) In a calcified aortic valve, thick-walled neovessels are observed in a fibrotic area devoid of inflammatory infiltrates (HE). (c, d) Immunohistochemical positivity for intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) (stained dark brown) is observed in the endothelial cells of thin-walled neovessels 3,3'-diaminobenzidine.

View larger version (111K): [in a new window]   Figure 3 Gel electrophoresis of two samples from patients with calcified aortic valve disease (Cases 13 and 19) after reverse transcription (RT)-polymerase chain reaction. Both glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and heat shock protein-60 (hsp60) indicate their respective amplification products. In the "No RT" lane, retrotranscription after the DNase treatment of the samples was omitted.