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. 2015 Jun;185(6):1724–1739. doi: 10.1016/j.ajpath.2015.02.010

Figure 2.

Figure 2

Immunophenotypic traits of human mucin-intrahepatic (IHCCA) and mixed-IHCCA. A: Periodic-acid Schiff (PAS) stain and immunohistochemistry (IHC) for keratin (K)7, EpCAM, LGR5, CD133, CD13, CD90, and α-smooth muscle actin (α-SMA) on human mucin-IHCCAs. Mucin-IHCCAs are diffusely positive for PAS, K7, EpCAM, LGR5, and CD133. By contrast, CD13 is restricted to a small subpopulation of tumor epithelial cells (arrow). CD90 and α-SMA are primarily expressed by tumor stromal cells (arrows). B: PAS stain and IHC for K7, EpCAM, LGR5, CD133, CD13, CD90, and α-SMA on human mixed-IHCCAs. Mixed-IHCCAs are diffusely positive for K7, EpCAM, CD13, and CD133. By contrast, LGR5 is restricted to a small subpopulation of tumor epithelial cells (arrows). CD90 and α-SMA were primarily expressed by tumor stromal cells (arrows). C: Double immunofluorescence (IF) for Desmin (red) and CD133 (green) in mucin-IHCCA; double IF for α-SMA (red) and pan-cytokeratin (Pan-CK; green) in mixed-IHCCAs. Approximately 10% of tumor epithelial cells (CD133+ or Pan-CK+, in green) express mesenchymal markers (desmin or α-SMA, in red) both in mucin- and in mixed-IHCCA. Yellow arrows indicate cells coexpressing epithelial and mesenchymal markers (yellow cells in merged images). Nuclei are shown in blue. Separate channels are shown. Boxed areas are shown magnified in the insets (bottom panels). D: Upper panels: IHC for SNAIL and Twist in human CCA specimens shows the expression of the two antigens within the nuclei of epithelial CCA cells (arrows). Lower panels: Double IF for SNAIL (red) and Pan-CK (green) in human CCA specimens confirms the expression of SNAIL within nuclei of epithelial CCA cells (arrows). Double IF for SNAIL (red) and α-SMA (green) in human CCA specimens shows the presence of CCA cells expressing both α-SMA and SNAIL. Images from mixed-IHCCA samples are displayed. E–J: IHC for CD133, CD13, and CD90 on human nontumor tissue surrounding CCA specimens. CD133 expression (E and F) is restricted to a few cells within Canals of Hering (CoH) and bile ductules (E, arrow) and to peribiliary glands (PBGs) within larger bile ducts (F, arrows). CD90 (G and H) is mostly expressed by stromal cells in the portal spaces (G, arrow) and around PBGs (H, arrows). CD13 (I and J) is diffusely expressed by hepatocytes, cells within CoH/bile ductules (I, yellow arrows), cholangiocytes of interlobular bile ducts (I, red arrow), and PBGs (J, arrow). Original magnification: ×20 (A–C, D, lower panels); ×40 (D, upper panel).