Immunohistochemistry is a Reliable Screening Tool for Identification of ALK Rearrangement in Non–Small-Cell Lung Carcinoma and is Antibody Dependent

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Immunohistochemistry is a Reliable Screening Tool for Identification of ALK Rearrangement in Non–Small-Cell Lung Carcinoma and is Antibody Dependent Chris M.J. Conklin, MD, Kenneth J. Craddock, MD, Cherry Have, BSc, MLT, Janessa Laskin, MD, Christian Couture, MD, Diana N. Ionescu, MD Journal of Thoracic Oncology Volume 8, Issue 1, Pages 45-51 (January 2013) DOI: 10.1097/JTO.0b013e318274a83e Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Semiquantitative immunohistochemical staining using an antibody to ALK (D5F3 by Cell Signaling [Billerica, MA] with ADVANCE detection), 400x original magnification. A, IHC score 3+ showing intense granular cytoplasmic staining. B, IHC score 2+ showing moderate smooth cytoplasmic staining (without granularity seen in score 3+). C, IHC score 1+ showing faint, barely discernible cytoplasmic staining in any tumor cells. D, IHC score 0/negative showing no staining. ALK, anaplastic lymphoma receptor tyrosine kinase; IHC, immunohistochemistry Journal of Thoracic Oncology 2013 8, 45-51DOI: (10.1097/JTO.0b013e318274a83e) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Dual-color break-apart FISH performed using the commercially available ALK probe (Vysis LSI ALK Dual Color, Break Apart Rearrangement Probe, Abbott Molecular, Abbott, IL) on case 11. ALK break-apart FISH showed a straightforward break-apart signal configuration (1 red, 1 green, 1 fusion [1R1G1F]) in 51% of the nuclei, indicating ALK rearrangement. Splitting of the red and green signals (shown with arrows) indicates ALK rearrangement. ALK, anaplastic lymphoma receptor tyrosine kinase; NSCLC, non–small-cell lung carcinoma; FISH, fluorescence in situ hybridization. Journal of Thoracic Oncology 2013 8, 45-51DOI: (10.1097/JTO.0b013e318274a83e) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Diagnostic screening algorithm using ALK-immunohistochemistry (5A4 by Novocastra or D5F3 by Cell Signaling with ADVANCE detection), and FISH for confirmation in NSCLC. Only cases showing any degree of ALK-expression by IHC are tested by FISH. ALK, anaplastic lymphoma receptor tyrosine kinase; FISH, fluorescence in situ hybridization; NSCLC, non–small-cell lung cancer; IHC, immunohistochemistry. Journal of Thoracic Oncology 2013 8, 45-51DOI: (10.1097/JTO.0b013e318274a83e) Copyright © 2012 International Association for the Study of Lung Cancer Terms and Conditions