Digital image analysis system for the quantification of infiltrates and cell adhesion molecules in inflammatory cardiomyopathy.
BACKGROUND
We attempted to develop a digital image analysis (DIA) system for endomyocardial biopsies (EMBs) to reliably quantify a) biopsy quality, b) immunohistochemically-marked infiltrates, and c) cell adhesion molecules (CAMs) in relation to net heart area (HA) for the semi-automated diagnosis of inflammatory cardiomyopathy (InfCM).


METHODS
140 EMBs from dilated cardiomyopathy (DCM) patients and 14 autopsy heart samples (controls) were immunostained for T-lymphocytes (CD2, CD3, CD4, CD8), beta(2)-integrin+ infiltrates (CD18, LFA-1, Mac-1) and CAMs (immunoglobulin superfamily: ICAM-1, HLA class I, HLA DR, VCAM-1, CD58; selectins: CD62E and CD62P; and the beta(1)-integrin chain CD29). EMB quality was assessed visually on a three-point scale. Infiltrates were quantified visually (per hpf) and by DIA (per mm2 HA). CAM expression was evaluated semiquantitatively and by DIA (area fraction [AF]: stained area relative to HA).


RESULTS
DIA-evaluated HA correlated significantly with the visual assessment of EMB quality. The visual evaluation of both infiltrates and CAMs correlated significantly with the respective DIA-based quantification. DIA-quantified CAM-AF and infiltrates were discriminated by the CAM classification (CAMs+: n=87; 62%) compared to controls. DIA-quantified CAM immunoreactivity correlated significantly with the DIA-quantified counter-receptor+ infiltrates. DIA evaluation of biopsy quality, infiltrates, and CAMs was devoid of inter- and intraobserver variability.


CONCLUSIONS
The DIA system presented here enables standardized and observer-independent assessment of EMB quality and intramyocardial inflammation (density of infiltrates and CAM expression) in DCM biopsies related to HA. Our data confirm that endothelial CAM count and counter-receptor+ immunocompetent infiltration are interdependent pathogenic and diagnostic hallmarks of InfCM.
Noutsias
Michel
M
Department of Cardiology and Pneumonology, University Hospital Benjamin Franklin, Free Univerity of Berlin, Germany. noutsias@zedat.fu-berlin.de
Pauschinger
Matthias
M
Ostermann
Karsten
K
Escher
Felicitas
F
Blohm
Jan-Henrik
JH
Schultheiss
Heinz
H
Kühl
Uwe
U
D000328:Adult; D001706:Biopsy; D017252:CD3 Complex / Q000096:biosynthesis; D009202:Cardiomyopathies / Q000276:immunology* / Q000473:pathology*; D002448:Cell Adhesion; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D007249:Inflammation; D008297:Male; D008875:Middle Aged; D009206:Myocardium / Q000473:pathology*; D010641:Phenotype
D017252:CD3 Complex