Vascular adhesion protein-1, intercellular adhesion molecule-1 and P-Selectin mediate leukocyte binding to ischemic heart in humans
Kimmo Jaakkola, MD*  ,
Sirpa Jalkanen, MD*,
Katja Kaunismäki, MD*,
Esko Vänttinen, MD ,
Pekka Saukko, MD ,
Kalle Alanen, MD||,
Markku Kallajoki, MD||,
Liisa-Maria Voipio-Pulkki, MD and
Marko Salmi, MD*
* National Public Health Institute and MediCity Research Laboratory, University of Turku, Turku, Finland
Department of Medicine, University of Turku, Turku, Finland
Department of Surgery, University of Turku, Turku, Finland
Department of Forensic Medicine, University of Turku, Turku, Finland
|| Department of Pathology, University of Turku, Turku, Finland
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Figure 1 Cardiac expression of endothelial adhesion molecules. a) Blood vessels in normal myocardium are CD31 positive. b) A VCAM-1 positive blood vessel in normal atrium. c) ICAM-2 positive vessels in infarction heart. d) Negative staining of infarcted heart with anti-E-selectin mAb (similar absence of reactivity was also seen with all negative control mAbs, data not shown). In the inset, E-selectin positive vessels in the tonsil are shown. Representative vessels are indicated by arrows. Bar shown in (a) is 100 µm. The bar shown in (b) is 50 µm in all other figures. ICAM = intercellular adhesion molecule; mAb = monoclonal antibody; VCAM = vascular cell adhesion molecule-1.
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Figure 2 Inducibility and scoring of ICAM-1, P-selectin and VAP-1. a) Moderate (score 2) and b) strong (score 3) ICAM-1 positive samples. c) Weak (score 1), (d) moderate (score 2) and (e) strong (score 3) P-selectin positive vessels in heart specimens. f) Moderate (score 2), (g) strong (score 3) and (h) extremely strong (score 4) VAP-1 positive vessels in infarction heart. Representative vessels are indicated by arrows. Bar shown in (a) is 50 µm. ICAM = intercellular adhesion molecule; VAP-1 = vascular adhesion protein-1.
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Figure 3 Intercellular adhesion molecule-1 and P-selectin are induced in ischemic heart, and VAP-1 expression shows ventricular dominance. The expression of (a) ICAM-1, (b) P-selectin and (c) VAP-1 in normal and infarction heart specimens were scored as described in the Methods section. The numbers refer to the numbers of individual acute myocardial infarction and control patients. ICAM-1 = intercellular adhesion molecule-1; VAP-1 = vascular adhesion protein-1.
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Figure 4 Molecular weight alterations of VAP-1 in the heart. After SDS-PAGE and immunoblotting, VAP-1 was immunodetected from untreated and sialidase treated lysates from normal, reperfused and infarcted heart and tonsil as control tissue. To assist size comparisons, all samples were analyzed in parallel lanes of the same gel, and after the scanning two horizontal lines were drawn. In the molecular weight (MW) lane the positions of the markers otherwise not visible in enhanced chemiluminescense are shown. VAP-1 = vascular adhesion protein-1.
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Figure 5 Intercellular adhesion molecule-1, P-selectin and VAP-1 mediate granulocyte adhesion to vessels in infarcted heart. a) In this dark field picture two cardiac vessels (dashed lines) are shown, one with seven bound granulocytes and the other supporting the binding of two granulocytes (arrows). b) Anti-VAP-1, P-selectin and anti-ICAM-antibodies separately or in combination decrease the number of granulocytes adhering to myocardial blood vessels when compared with the specimens treated with nonbinding (set as 100%) control antibodies or without any antibodies (–). ICAM = intercellular adhesion molecule; VAP-1 = vascular adhesion protein-1.
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