Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD‑L1 antibodies

Fujisawa,Takuo; Tsuta,Koji; Yanagimoto,Hiroaki; Yagi,Masao; Suzuki,Kensuke; Nishikawa,Kenji; Takahashi,Masaru; Okada,Hisatake; Nakano,Yasushi; Iwai,Hiroshi

doi:10.3892/mco.2019.1801

Quantitative immunohistochemical assay with novel digital immunostaining for comparisons of PD‑L1 antibodies

Authors:
- Takuo Fujisawa
- Koji Tsuta
- Hiroaki Yanagimoto
- Masao Yagi
- Kensuke Suzuki
- Kenji Nishikawa
- Masaru Takahashi
- Hisatake Okada
- Yasushi Nakano
- Hiroshi Iwai
View Affiliations

Affiliations: Department of Otolaryngology, Head and Neck Surgery, Kansai Medical University, Osaka 573‑1010, Japan, Department of Pathology and Laboratory Medicine, Head and Neck Surgery, Kansai Medical University, Osaka 573‑1010, Japan, Department of Surgery, Head and Neck Surgery, Kansai Medical University, Osaka 573‑1010, Japan, Bio System Development Group, Bio Advanced Technology Division, Corporate R&D Headquarters, Konica‑Minolta, Inc., Tokyo 191‑8511, Japan
Published online on: January 17, 2019 https://doi.org/10.3892/mco.2019.1801
Pages: 391-396
Copyright: © Fujisawa et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

One obstacle in diagnostic pathology is the harmonization of one drug-one diagnostic tests for programmed death ligand‑1 (PD‑L1). There are many challenges in accurate comparisons of diagnostic tests, such as differences in the titer of each antibody, detection system and dynamic range of visualization. Our previously developed digital immunostaining technique is highly sensitive and quantitative with the ability to quantify particles that bind in a one‑to‑one fashion with antibody in each cell. Determining the differences in the titer of each antibody with digital immunostaining may be beneficial for future harmonized analysis. To demonstrate the accuracy of digital immunostaining, the present study compared the number of particles with ELISA and nCounter data from five cell lines. NCI‑H460 exhib‑ited the highest level of PD‑L1 protein, followed by A549, PC‑3, NCI‑H1299, and NCI‑H446 cells. In addition, the PD‑L1 mRNA values determined by nCounter corresponded with the order of the protein levels determined by ELISA. The present study revealed that digital immunostaining for PD‑L1 was highly associated with ELISA and nCounter data. Among the four antibodies tested, the titer of all but SP142 coincided with ELISA and nCounter data. These results indicated that our digital immunostaining technique may be beneficial for future harmonized analysis.

View Figures

View References

1	Gonda K, Miyashita M, Watanabe M, Takahashi Y, Goda H, Okada H, Nakano Y, Tada H, Amari M and Ohuchi N: Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles. Biochem Biophys Res Commun. 426:409–414. 2012. View Article : Google Scholar : PubMed/NCBI
2	Gonda K, Watanabe M, Tada H, Miyashita M, Takahashi-Aoyama Y, Kamei T, Ishida T, Usami S, Hirakawa H, Kakugawa Y, et al: Quantitative diagnostic imaging of cancer tissues by using phosphor-integrated dots with ultra-high brightness. Sci Rep. 7:75092017. View Article : Google Scholar : PubMed/NCBI
3	Yamaki S, Yanagimoto H, Tsuta K, Ryota H and Kon M: PD-L1 expression in pancreatic ductal adenocarcinoma is a poor prognostic factor in patients with high CD8+ tumor-infiltrating lymphocytes: Highly sensitive detection using phosphor-integrated dot staining. Int J Clin Oncol. 22:726–733. 2017. View Article : Google Scholar : PubMed/NCBI
4	Topalian SL, Taube JM, Anders RA and Pardoll DM: Mechanism-driven biomarkers to guide immune checkpoint blockade in cancer therapy. Nat Rev Cancer. 16:275–287. 2016. View Article : Google Scholar : PubMed/NCBI
5	Topalian SL, Hodi FS, Brahmer JR, Gettinger SN, Smith DC, McDermott DF, Powderly JD, Carvajal RD, Sosman JA, Atkins MB, et al: Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Engl J Med. 366:2443–2454. 2012. View Article : Google Scholar : PubMed/NCBI
6	Reck M, Rodríguez-Abreu D, Robinson AG, Hui R, Csőszi T, Fülöp A, Gottfried M, Peled N, Tafreshi A, Cuffe S, et al: KEYNOTE-024 Investigators: Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer. N Engl J Med. 375:1823–1833. 2016. View Article : Google Scholar : PubMed/NCBI
7	Hirsch FR, McElhinny A, Stanforth D, Ranger-Moore J, Jansson M, Kulangara K, Richardson W, Towne P, Hanks D, Vennapusa B, et al: 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. 2017. View Article : Google Scholar : PubMed/NCBI
8	Chen G, Gharib TG, Huang CC, Taylor JM, Misek DE, Kardia SL, Giordano TJ, Iannettoni MD, Orringer MB, Hanash SM, et al: Discordant protein and mRNA expression in lung adenocarcinomas. Mol Cell Proteomics. 1:304–313. 2002. View Article : Google Scholar : PubMed/NCBI
9	Skaland I, Nordhus M, Gudlaugsson E, Klos J, Kjellevold KH, Janssen EA and Baak JP: Evaluation of 5 different labeled polymer immunohistochemical detection systems. Appl Immunohistochem Mol Morphol. 18:90–96. 2010. View Article : Google Scholar : PubMed/NCBI
10	Kataoka K, Shiraishi Y, Takeda Y, Sakata S, Matsumoto M, Nagano S, Maeda T, Nagata Y, Kitanaka A, Mizuno S, et al: Aberrant PD-L1 expression through 3′-UTR disruption in multiple cancers. Nature. 534:402–406. 2016. View Article : Google Scholar : PubMed/NCBI
11	Garon EB, Rizvi NA, Hui R, Leighl N, Balmanoukian AS, Eder JP, Patnaik A, Aggarwal C, Gubens M, Horn L, et al KEYNOTE-001 Investigators, : Pembrolizumab for the treatment of non-small-cell lung cancer. N Engl J Med. 372:2018–2028. 2015. View Article : Google Scholar : PubMed/NCBI
12	Mori H, Kubo M, Yamaguchi R, Nishimura R, Osako T, Arima N, Okumura Y, Okido M, Yamada M, Kai M, et al: The combination of PD-L1 expression and decreased tumor-infiltrating lymphocytes is associated with a poor prognosis in triple-negative breast cancer. Oncotarget. 8:15584–15592. 2017. View Article : Google Scholar : PubMed/NCBI
13	Ameratunga M, Asadi K, Lin X, Walkiewicz M, Murone C, Knight S, Mitchell P, Boutros P and John T: PD-L1 and tumor infiltrating lymphocytes as prognostic markers in resected NSCLC. PLoS One. 11:e01539542016. View Article : Google Scholar : PubMed/NCBI
14	Twyman-Saint Victor C, Rech AJ, Maity A, Rengan R, Pauken KE, Stelekati E, Benci JL, Xu B, Dada H, Odorizzi PM, et al: Radiation and dual checkpoint blockade activate non-redundant immune mechanisms in cancer. Nature. 520:373–377. 2015. View Article : Google Scholar : PubMed/NCBI
15	McLaughlin J, Han G, Schalper KA, Carvajal-Hausdorf D, Pelekanou V, Rehman J, Velcheti V, Herbst R, LoRusso P and Rimm DL: Quantitative assessment of the heterogeneity of PD-L1 expression in non-small-cell lung cancer. JAMA Oncol. 2:46–54. 2016. View Article : Google Scholar : PubMed/NCBI