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Digital image analysis system for the quantification of infiltrates and cell adhesion molecules in inflammatory
Michel Noutsias1, Matthias Pauschinger, Karsten Ostermann
1Department of Cardiology and Pneumonology, University Hospital Benjamin Franklin, Free Univerity of Berlin, Germany. noutsias@zedat.fu-berlin.de
Insights
A new digital image analysis system standardizes endomyocardial biopsy assessment for inflammatory cardiomyopathy. This method reliably quantifies biopsy quality, infiltrates, and cell adhesion molecules, improving diagnostic accuracy.
Area of Science:
- Cardiology
- Digital Pathology
- Immunohistochemistry
Background:
- Developing a digital image analysis (DIA) system for endomyocardial biopsies (EMBs).
- Quantifying biopsy quality, infiltrates, and cell adhesion molecules (CAMs) relative to heart area (HA).
- Aimed at semi-automated diagnosis of inflammatory cardiomyopathy (InfCM).
Purpose of the Study:
- To develop and validate a DIA system for EMB analysis.
- To enable objective quantification of key diagnostic markers for InfCM.
- To reduce observer variability in the assessment of myocardial inflammation.
Main Methods:
- Analyzed 140 EMBs from dilated cardiomyopathy (DCM) patients and 14 controls.
- Used immunohistochemistry for T-lymphocytes, beta(2)-integrin+ infiltrates, and various CAMs.
- Quantified infiltrates and CAM expression using both visual assessment and DIA (per mm2 HA and area fraction).
Main Results:
- DIA assessment of HA correlated with visual biopsy quality.
- Visual and DIA quantifications of infiltrates and CAMs showed significant correlation.
- DIA identified differences in CAMs and infiltrates between InfCM patients and controls.
- DIA evaluation demonstrated no inter- or intraobserver variability.
Conclusions:
- The DIA system provides standardized, observer-independent assessment of EMB quality and intramyocardial inflammation.
- Quantification of infiltrates and CAM expression in DCM biopsies is feasible with DIA.
- Endothelial CAMs and immunocompetent infiltration are interdependent hallmarks of InfCM.
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).
Material/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.
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