Regarding heterogeneity, intra-class correlation coefficients (ICC) for CD3+ TILs varied from 0.615 to 0.746 in the central area, and from 0.686 to 0.746 in the invasive area. For both methods the tumor areas of interest (invasive front and central area) were manually delineated by the observer.īased on all sections, the Spearman's correlation coefficients for density estimates varied from 0.9457 to 0.9638 (p < 0.0001), whereas the coefficients for area fraction estimates ranged from 0.9400 to 0.9603 (P < 0.0001). For the image analysis approach an app-based algorithm was developed using Visiopharm Integrator System software. Stereological estimates of the numerical density and area fraction of TILs were obtained using the computer-assisted newCAST stereology system. Serial sections from each of the 129 blocks were immunohistochemically stained for CD3 and CD8, and the slides were scanned.
The purpose of this study was to compare manual stereological estimates of TILs with automatic counts obtained by image analysis, and at the same time investigate the heterogeneity of TILs.įrom 43 patients treated for stage II CC in 2002 three paraffin embedded, tumor containing tissue blocks were selected one of them representing the deepest invasive tumor front. Manual stereological counting is considered the gold standard, but digital pathology with image analysis is preferred due to time efficiency. Investigation of tumor infiltrating lymphocytes (TILs) may be rewarding, but the lack of a standardized analytic technique is a major concern. Precise prognostic and predictive variables allowing improved post-operative treatment stratification are missing in patients treated for stage II colon cancer (CC). 10 Institute of Regional Health Research, University of Southern Denmark, Odense, Denmark.9 Department of Pathology, Vejle Hospital, Beriderbakken 4, 7100, Vejle, Denmark.8 Department of Oncology, Vejle Hospital, Beriderbakken 4, 7100, Vejle, Denmark.7 Danish Colorectal Cancer Center South, Vejle Hospital, Vejle, Denmark.Winsløws Vej 9 A, 1st, 5000, Odense C, Denmark. 6 Research Unit of General Practice, University of Southern Denmark, J.B.5 Visiopharm A/S, Agern Allé 24, 2970, Hoersholm, Denmark.4 Department of Clinical Medicine, Stereological Research Laboratory, Aarhus University, Nørrebrogade 44, 10G, 8000, Aarhus C, Denmark. 3 Department of Pathology, Vejle Hospital, Beriderbakken 4, 7100, Vejle, Denmark. 2 Institute of Regional Health Research, University of Southern Denmark, Odense, Denmark. 1 Danish Colorectal Cancer Center South, Vejle Hospital, Vejle, Denmark.
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