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Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N

BACKGROUND: Programmed Death Ligand 1 (PD-L1) is an immune modulating protein expressed on the surface of various inflammatory cells, including T Cells, B Cells, dendritic cells, and macrophages. PD-L1 represents an important diagnostic target; expression of PD-L1 on the surface of tumor cells, or w...

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Autores principales: Smith, Jacquelyn, Robida, Mark D., Acosta, Krista, Vennapusa, Bharathi, Mistry, Amita, Martin, Greg, Yates, Alton, Hnatyszyn, H. James
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4870735/
https://www.ncbi.nlm.nih.gov/pubmed/27189072
http://dx.doi.org/10.1186/s13000-016-0494-2
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author Smith, Jacquelyn
Robida, Mark D.
Acosta, Krista
Vennapusa, Bharathi
Mistry, Amita
Martin, Greg
Yates, Alton
Hnatyszyn, H. James
author_facet Smith, Jacquelyn
Robida, Mark D.
Acosta, Krista
Vennapusa, Bharathi
Mistry, Amita
Martin, Greg
Yates, Alton
Hnatyszyn, H. James
author_sort Smith, Jacquelyn
collection PubMed
description BACKGROUND: Programmed Death Ligand 1 (PD-L1) is an immune modulating protein expressed on the surface of various inflammatory cells, including T Cells, B Cells, dendritic cells, and macrophages. PD-L1 represents an important diagnostic target; expression of PD-L1 on the surface of tumor cells, or within tumor-associated immune cells, is an important predictor of likely response to targeted therapies. In this study, we describe the optimization of immunohistochemistry (IHC) assays using two PD-L1 antibodies (SP263 and E1L3N) and compare the performance of the optimized assays. METHODS: Fully automated immunohistochemical assays were optimized for the VENTANA PD-L1 (SP263) Rabbit Monoclonal Antibody and the PD-L1 (E1L3N®) XP® Rabbit mAb using instruments and detection chemistries from Ventana Medical Systems, Inc. (“SP263 assay” and “E1L3N assay,” respectively). Tissue microarrays (TMAs) containing formalin fixed paraffin embedded (FFPE) non-small cell lung cancer (NSCLC) cases were used for the optimization and comparison staining. H scores were used for membrane scoring whereas percent positivity was used for tumor-associated immune cell scoring. RESULTS: One-hundred NSCLC TMA case cores each stained with the SP263 and E1L3N assays were evaluated by two pathologists in a blinded study. Analysis of these specimens showed that the SP263 assay was more sensitive and had a wider dynamic range than the E1L3N assay. For sensitivity, many cases were found to be negative for membrane staining with the E1L3N assay, but had measurable staining with the SP263 assay. Dynamic range was demonstrated by the SP263 assay having well-distributed H scores while the E1L3N assay had a significantly higher proportion of cases clustered in the lowest H score bins. For tumor-associated immune cell staining, the two assays identified similar amounts of cells staining in each case, but the SP263 assay gave overall darker staining. CONCLUSION: Since PD-L1 status is important for targeted therapies, having a specific and accurate diagnostic test is crucial for identifying patients who could benefit from these treatments. Due to its staining intensity, scoring range, and pathologist preference, the SP263 IHC assay has been deemed superior to the E1L3N IHC assay. Future clinical utility remains to be determined. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13000-016-0494-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-48707352016-05-19 Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N Smith, Jacquelyn Robida, Mark D. Acosta, Krista Vennapusa, Bharathi Mistry, Amita Martin, Greg Yates, Alton Hnatyszyn, H. James Diagn Pathol Research BACKGROUND: Programmed Death Ligand 1 (PD-L1) is an immune modulating protein expressed on the surface of various inflammatory cells, including T Cells, B Cells, dendritic cells, and macrophages. PD-L1 represents an important diagnostic target; expression of PD-L1 on the surface of tumor cells, or within tumor-associated immune cells, is an important predictor of likely response to targeted therapies. In this study, we describe the optimization of immunohistochemistry (IHC) assays using two PD-L1 antibodies (SP263 and E1L3N) and compare the performance of the optimized assays. METHODS: Fully automated immunohistochemical assays were optimized for the VENTANA PD-L1 (SP263) Rabbit Monoclonal Antibody and the PD-L1 (E1L3N®) XP® Rabbit mAb using instruments and detection chemistries from Ventana Medical Systems, Inc. (“SP263 assay” and “E1L3N assay,” respectively). Tissue microarrays (TMAs) containing formalin fixed paraffin embedded (FFPE) non-small cell lung cancer (NSCLC) cases were used for the optimization and comparison staining. H scores were used for membrane scoring whereas percent positivity was used for tumor-associated immune cell scoring. RESULTS: One-hundred NSCLC TMA case cores each stained with the SP263 and E1L3N assays were evaluated by two pathologists in a blinded study. Analysis of these specimens showed that the SP263 assay was more sensitive and had a wider dynamic range than the E1L3N assay. For sensitivity, many cases were found to be negative for membrane staining with the E1L3N assay, but had measurable staining with the SP263 assay. Dynamic range was demonstrated by the SP263 assay having well-distributed H scores while the E1L3N assay had a significantly higher proportion of cases clustered in the lowest H score bins. For tumor-associated immune cell staining, the two assays identified similar amounts of cells staining in each case, but the SP263 assay gave overall darker staining. CONCLUSION: Since PD-L1 status is important for targeted therapies, having a specific and accurate diagnostic test is crucial for identifying patients who could benefit from these treatments. Due to its staining intensity, scoring range, and pathologist preference, the SP263 IHC assay has been deemed superior to the E1L3N IHC assay. Future clinical utility remains to be determined. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13000-016-0494-2) contains supplementary material, which is available to authorized users. BioMed Central 2016-05-18 /pmc/articles/PMC4870735/ /pubmed/27189072 http://dx.doi.org/10.1186/s13000-016-0494-2 Text en © Smith et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Smith, Jacquelyn
Robida, Mark D.
Acosta, Krista
Vennapusa, Bharathi
Mistry, Amita
Martin, Greg
Yates, Alton
Hnatyszyn, H. James
Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title_full Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title_fullStr Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title_full_unstemmed Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title_short Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N
title_sort quantitative and qualitative characterization of two pd-l1 clones: sp263 and e1l3n
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4870735/
https://www.ncbi.nlm.nih.gov/pubmed/27189072
http://dx.doi.org/10.1186/s13000-016-0494-2
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