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Impact of delayed fixation and decalcification on PD-L1 expression: a comparison of two clones

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Abstract

The bone is a frequent localization for lung non-small cell cancer metastasis; decalcification is required to permit tissue section. Pre-analytical conditions can influence the detection of immunohistochemical markers. The aim of our work is to evaluate PD-L1 expression in samples with delayed fixation and in decalcified tissue with chelating agent or acid at different time. Tumor-expressing PD-L1 and placental tissue were fixed at different times or decalcified with an acid decalcifier or EDTA for different durations. For 22C3 antibody, when tissues were decalcified with DC3, there was a significant decrease in the percentage of tumor cells or placental villi stained which after 4 h (p = 0.035 at 4 h). When EDTA is used for 22C3 antibody, there was a slight decrease in the percentage of stained tumor cells or villi but although there was a trend (p = 0.058 at 20 h), this was never statistically significant. For E1L3N antibody, when tissues were decalcified either with DC3 or EDTA, there was no significant decrease for the proportion of stained tumor cells or placental villi, neither for staining intensity for the first 24 h. The proportion of placental villi and tumor stained or intensity of staining was not significantly lower for any sample after delayed fixation also at 24 h for both PD-L1 clones. Delayed fixation does not affect the proportion of stained cell and intensity with PD-L1 immunohistochemistry. Decalcification also performed with EDTA lower the proportion and intensity of stained cells with PD-L1 immunohistochemistry.

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References

  1. Gandhi L, Rodríguez-Abreu D, Gadgeel S et al (2018) Pembrolizumab plus chemotherapy in metastatic non–small-cell lung cancer. N Engl J Med. https://doi.org/10.1056/NEJMoa1801005

  2. Reck M, Rodríguez-Abreu D, Robinson AG, Hui R, Csőszi T, Fülöp A, Gottfried M, Peled N, Tafreshi A, Cuffe S, O’Brien M, Rao S, Hotta K, Leiby MA, Lubiniecki GM, Shentu Y, Rangwala R, Brahmer JR (2016) Pembrolizumab versus chemotherapy for PD-L1–positive non–small-cell lung cancer. N Engl J Med 375:1823–1833. https://doi.org/10.1056/NEJMoa1606774

    Article  CAS  PubMed  Google Scholar 

  3. Lee AHS, Key HP, Bell JA, Kumah P, Hodi Z, Ellis IO (2014) The effect of delay in fixation on HER2 expression in invasive carcinoma of the breast assessed with immunohistochemistry and in situ hybridisation. J Clin Pathol 67:573–575. https://doi.org/10.1136/jclinpath-2013-201978

    Article  PubMed  Google Scholar 

  4. Qiu J, Kulkarni S, Chandrasekhar R, Rees M, Hyde K, Wilding G, Tan D, Khoury T (2010) Effect of delayed formalin fixation on estrogen and progesterone receptors in breast cancer: a study of three different clones. Am J Clin Pathol 134:813–819. https://doi.org/10.1309/AJCPVCX83JWMSBNO

    Article  CAS  PubMed  Google Scholar 

  5. Schrijver WAME, Van Der Groep P, Hoefnagel LD et al (2016) Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer. Mod Pathol 29:1460–1470. https://doi.org/10.1038/modpathol.2016.116

    Article  CAS  PubMed  Google Scholar 

  6. Confavreux CB, Girard N, Pialat J-B, Bringuier PP, Devouassoux-Shisheboran M, Rousseau JC, Isaac S, Thivolet-Bejui F, Clezardin P, Brevet M (2014) Mutational profiling of bone metastases from lung adenocarcinoma: results of a prospective study (POUMOS-TEC). Bonekey Rep 3(580). https://doi.org/10.1038/bonekey.2014.75

  7. Cree IA, Booton R, Cane P, Gosney J, Ibrahim M, Kerr K, Lal R, Lewanski C, Navani N, Nicholson AG, Nicolson M, Summers Y (2016) PD-L1 testing for lung cancer in the UK: recognizing the challenges for implementation. Histopathology 69:177–186. https://doi.org/10.1111/his.12996

    Article  PubMed  Google Scholar 

  8. Miettinen M (2012) A simple method for generating multitissue blocks without special equipment. Appl Immunohistochem Mol Morphol 20:410–412. https://doi.org/10.1097/PAI.0b013e318245c82f

    Article  PubMed  Google Scholar 

  9. R Core Team (2018) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org 

  10. Confavreux CB, Pialat J-B, Bellière A, Brevet M, Decroisette C, Tescaru A, Wegrzyn J, Barrey C, Mornex F, Souquet PJ, Girard N (2018) Bone metastases from lung cancer: a paradigm for multidisciplinary onco-rheumatology management. Joint Bone Spine 86:185–194. https://doi.org/10.1016/j.jbspin.2018.03.005

    Article  CAS  PubMed  Google Scholar 

  11. Parra ER, Villalobos P, Mino B, Rodriguez-Canales J (2018) Comparison of different antibody clones for immunohistochemistry detection of programmed cell death ligand 1 (PD-L1) on non-small cell lung carcinoma. Appl Immunohistochem Mol Morphol 26:83–93. https://doi.org/10.1097/PAI.0000000000000531

    Article  CAS  PubMed  Google Scholar 

  12. Adam J, Le Stang N, Rouquette I et al (2018) Multicenter harmonization study for PD-L1 IHC testing in non-small-cell lung cancer. Ann Oncol 29:953–958. https://doi.org/10.1093/annonc/mdy014

    Article  CAS  PubMed  Google Scholar 

  13. Cogswell J, Inzunza HD, Wu Q, Feder JN, Mintier G, Novotny J, Cardona DM (2017) An analytical comparison of Dako 28-8 PharmDx assay and an E1L3N laboratory-developed test in the immunohistochemical detection of programmed death-ligand 1. Mol Diagn Ther 21:85–93. https://doi.org/10.1007/s40291-016-0237-9

    Article  CAS  PubMed  Google Scholar 

  14. Sholl LM, Aisner DL, Allen TC, Beasley MB, Borczuk AC, Cagle PT, Capelozzi V, Dacic S, Hariri L, Kerr KM, Lantuejoul S, Mino-Kenudson M, Raparia K, Rekhtman N, Roy-Chowdhuri S, Thunnissen E, Tsao MS, Yatabe Y, for the members of the Pulmonary Pathology Society (2016) Programmed death ligand-1 immunohistochemistry— a new challenge for pathologists: a perspective from members of the pulmonary pathology society. Arch Pathol Lab Med 140:341–344. https://doi.org/10.5858/arpa.2015-0506-SA

    Article  CAS  PubMed  Google Scholar 

  15. Forest F, Stachowicz M-L, Casteillo F, Karpathiou G, Gouzy-Grosjean F, Guilaubey C, Cottier M, Beal J, Clemenson A, Péoc’h M (2017) EGFR , KRAS , BRAF and HER2 testing in metastatic lung adenocarcinoma: value of testing on samples with poor specimen adequacy and analysis of discrepancies. Exp Mol Pathol 103:306–310. https://doi.org/10.1016/j.yexmp.2017.11.013

    Article  CAS  PubMed  Google Scholar 

  16. Khoury T, Sait S, Hwang H, Chandrasekhar R, Wilding G, Tan D, Kulkarni S (2009) Delay to formalin fixation effect on breast biomarkers. Mod Pathol 22:1457–1467. https://doi.org/10.1038/modpathol.2009.117

    Article  CAS  PubMed  Google Scholar 

  17. Khoury T (2018) Delay to formalin fixation (cold ischemia time) effect on breast cancer molecules. Am J Clin Pathol 149:275–292. https://doi.org/10.1093/ajcp/aqx164

    Article  CAS  PubMed  Google Scholar 

  18. Nakamura Y, Kobayashi T, Nishii Y, Suzuki Y, Saiki H, Ito K, Watanabe F, Nishihama K, Yasuma T, D’Alessandro-Gabazza CN, Katsuta K, Fujimoto H, Gabazza EC, Taguchi O, Hataji O (2018) Comparable immunoreactivity rates of PD-L1 in archival and recent specimens from non-small cell lung cancer. Thorac Cancer 9:1476–1482. https://doi.org/10.1111/1759-7714.12861

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Herbst RS, Baas P, Perez-Gracia JL, Felip E, Kim DW, Han JY, Molina JR, Kim JH, Dubos Arvis C, Ahn MJ, Majem M, Fidler MJ, Surmont V, de Castro G Jr, Garrido M, Shentu Y, Emancipator K, Samkari A, Jensen EH, Lubiniecki GM, Garon EB (2019) Use of archival versus newly collected tumor samples for assessing PD-L1 expression and overall survival: an updated analysis of KEYNOTE-010 trial. Ann Oncol 30:281–289. https://doi.org/10.1093/annonc/mdy545

    Article  CAS  PubMed  Google Scholar 

  20. Hirsch FR, McElhinny A, Stanforth D, Ranger-Moore J, Jansson M, Kulangara K, Richardson W, Towne P, Hanks D, Vennapusa B, Mistry A, Kalamegham R, Averbuch S, Novotny J, Rubin E, Emancipator K, McCaffery I, Williams JA, Walker J, Longshore J, Tsao MS, Kerr KM (2017) PD-L1 immunohistochemistry assays for lung cancer: results from phase 1 of the blueprint PD-L1 IHC assay comparison project. J Thorac Oncol 12:208–222. https://doi.org/10.1016/j.jtho.2016.11.2228

    Article  PubMed  Google Scholar 

  21. t Hart NA, van der Starre J, Vonk JM, Timens W (2018) Essential preanalytics in PD-L1 immunocytochemistry. Histopathology 74:0–3. https://doi.org/10.1111/his.13717

    Article  Google Scholar 

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Acknowledgments

The authors thank Philippe Cosmo from Centre de Ressources Biologiques de CHU Saint-Etienne (BRIF no. BB-0033–00041) for his assistance in data retrieval. We thank Olivier Tiffet, MD, PhD; Arnaud Patoir, MD; Abdulrazzaq Sulaiman, MD, PhD; and Eric Parietti, MD, for providing surgical samples.

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Authors and Affiliations

  1. Department of Pathology, University Hospital of Saint Etienne, North Hospital, Avenue Albert Raimond, 42055, Saint Etienne CEDEX 2, France

    Fabien Forest, Gaelle Cote, David Laville, Vanessa Da Cruz, Pierre Dal Col, Florian Camy, Mousa Mobarki, Alix Clemenson, Violaine Yvorel & Michel Péoc’h

  2. Corneal Graft Biology, Engineering and Imaging Laboratory, BiiGC, EA2521, Federative Institute of Research in Sciences and Health Engineering, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France

    Fabien Forest & Michel Péoc’h

  3. Department of Pathology, Faculty of Medicine, Jazan University, Jizan, Saudi Arabia

    Mousa Mobarki

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  1. Fabien Forest
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  2. Gaelle Cote
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Contributions

FF and GC wrote the manuscript. FF and GC designed the study. FF, GC, DL, VDC, PDC, MM, AC, VY, and MP edited and reviewed the manuscript. FF and GC analyzed the data. FF, GC, DL, VDC, PDC, MM, AC, VY, and MP collected the data. All authors gave final approval for publication. FF takes full responsibility for the work as a whole, including the study design, access to data, and the decision to submit and publish the manuscript.

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Correspondence to Fabien Forest.

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Local ethics committee approved the study (IRBN172018/CHUSTE). This study was performed according to the standards of French law.

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The authors declare that they have no conflict of interest.

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Forest, F., Cote, G., Laville, D. et al. Impact of delayed fixation and decalcification on PD-L1 expression: a comparison of two clones. Virchows Arch 475, 693–699 (2019). https://doi.org/10.1007/s00428-019-02613-w

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  • DOI: https://doi.org/10.1007/s00428-019-02613-w

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