Optimized immunohistochemistry using the D5F3 antibody provides a reliable test for identification of ALK-positive lung adenocarcinomas
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We used optimized immunohistochemistry (IHC) with the D5F3 antibody for detection of tumours in a prospective study of 307 pulmonary adenocarcinomas. Cases positive by IHC (1+, 2+, 3+) were further investigated by fluorescent in situ hybridization (FISH). Of 307 cases, 22 (7.2%) were moderately intensely positive (2+/3+); 18 of these (82%) were also positive by FISH. Of the four IHC-positive/FISH-negative cases, one was unsuitable for FISH and three had abnormalities of the ALK gene. All cases with weak reactivity with D5F3 (1+) were FISH-negative. The FISH positive/IHC-positive cases with moderately intense reactivity had the typical clinicopathologic features of ALK-positive patients (younger age, p < 0.01; higher frequency in metastatic sites, p < 0.01; cribriform/mucinous/signet histology, p < 0.01; stage IV disease, p < 0.01). In conclusion, our findings indicate that optimized IHC using the D5F3 antibody provides a reliable and inexpensive test for identification of ALK-positive adenocarcinomas. Inclusion of this information in the pathology report at the time of the histological diagnosis might significantly shorten time to treatment.
KeywordsLung Pulmonary Adenocarcinoma ALK Immunohistochemistry D5F3
This work has been supported by a grant from “Sapienza” University, Rome, Italy.
Compliance with ethical standards
The study was conducted in accordance with the informed consent law of Italy.
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Soda M, Choi YL, Enomoto M, Takada S, Yamashita Y, Ishikawa S, Fujiwara S, Watanabe H, Kurashina K, Hatanaka H, Bando M, Ohno S, Ishikawa Y, Aburatani H, Niki T, Sohara Y, Sugiyama Y, Mano H (2007) Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer. Nature 448:561–566CrossRefPubMedGoogle Scholar
- 3.Rodig SJ, Mino-Kenudson M, Dacic S, Yeap BY, Shaw A, Barletta JA, Stubbs H, Law K, Lindeman N, Mark E, Janne PA, Lynch T, Johnson BE, Iafrate AJ, Chirieac LR (2009) Unique clinicopathologic features characterize ALK-rearranged lung adenocarcinoma in the western population. Clin Cancer Res 15:5216–5223CrossRefPubMedPubMedCentralGoogle Scholar
- 4.Kwak EL, Bang YJ, Camidge DR, Shaw AT, Solomon B, Maki RG, Ou SH, Dezube BJ, Jänne PA, Costa DB, Varella-Garcia M, Kim WH, Lynch TJ, Fidias P, Stubbs H, Engelman JA, Sequist LV, Tan W, Gandhi L, Mino-Kenudson M, Wei GC, Shreeve SM, Ratain MJ, Settleman J, Christensen JG, Haber DA, Wilner K, Salgia R, Shapiro GI, Clark JW, Iafrate AJ (2010) Anaplastic lymphoma kinase inhibition in non-small-cell lung cancer. N Engl J Med 363:1693–1703CrossRefPubMedPubMedCentralGoogle Scholar
- 7.Selinger CI, Rogers TM, Russell PA, O'Toole S, Yip P, Wright GM, Wainer Z, Horvath LG, Boyer M, McCaughan B, Kohonen-Corish MR, Fox S, Cooper WA, Solomon B (2013) Testing for ALK rearrangement in lung adenocarcinoma: a multicenter comparison of immunohistochemistry and fluorescent in situ hybridization. Mod Pathol 26:1545–1553CrossRefPubMedGoogle Scholar
- 8.Le Quesne J, Maurya M, Yancheva SG, O'Brien M, Popat S, Wotherspoon AC, de Castro DG, Nicholson AG (2014) A comparison of immunohistochemical assays and FISH in detecting the ALK translocation in diagnostic histological and cytological lung tumor material. J Thorac Oncol 9:769–774CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Alì G, Proietti A, Pelliccioni S, Niccoli C, Lupi C, Sensi E, Giannini R, Borrelli N, Menghi M, Chella A, Ribechini A, Cappuzzo F, Melfi F, Lucchi M, Mussi A, Fontanini G (2014) ALK rearrangement in a large series of consecutive non-small cell lung cancers: comparison between a new immunohistochemical approach and fluorescence in situ hybridization for the screening of patients eligible for crizotinib treatment. Arch Pathol Lab Med 138:1449–1458CrossRefPubMedGoogle Scholar
- 10.Wynes MW, Sholl LM, Dietel M, Schuuring E, Tsao MS, Yatabe Y, Tubbs RR, Hirsch FR (2014) An international interpretation study using the ALK IHC antibody D5F3 and a sensitive detection kit demonstrates high concordance between ALK IHC and ALK FISH and between evaluators. J Thorac Oncol 9:631–638CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Marchetti A, Di Lorito A, Pace MV, Iezzi M, Felicioni L, D'Antuono T, Filice G, Guetti L, Mucilli F, Buttitta F (2016) ALK protein analysis by IHC staining after recent regulatory changes: a comparison of two widely used approaches, revision of the literature, and a new testing algorithm. J Thorac Oncol 11:487–495CrossRefPubMedGoogle Scholar
- 12.Department of Health and Human Services. Letter to Ventana Medical Systems on approval of premarket authorization application for the VENTANA ALK (D5F3) CDx Assay. http://www.accessdata.fda.gov/cdrh_docs/pdf14/P140025a.pdf. Accessed June 1, 2015.
- 13.Italian Medicines Agency. [Eligibility data sheet for crizotinib treatment.] http://www.agenziafarmaco.gov.it/sites/default/files/Xalkori_NSCLC_20150219.zip [in Italian]. Accessed February 1, 2015.