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Development of an automated PD-L1 immunohistochemistry (IHC) assay for non-small cell lung cancer.

Phillips T, Simmons P, Inzunza HD, Cogswell J, Novotny J, Taylor C, Zhang X - Appl. Immunohistochem. Mol. Morphol. (2015)

Bottom Line: Nivolumab, a fully human IgG4 programmed death 1 (PD-1) immune checkpoint inhibitor antibody, developed by Bristol-Myers Squibb Inc., has activity across non-small cell lung cancer (NSCLC) histologies and is Food and Drug Administration approved for treatment of metastatic squamous NSCLC with progression on or after platinum-based chemotherapy.PD-L1 has been investigated as a potential biomarker to predict clinical response to nivolumab in clinical settings.The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines.

View Article: PubMed Central - PubMed

Affiliation: *Dako North America Inc., Carpinteria ‡Keck School of Medicine of the University of Southern California, Los Angeles, CA †Bristol-Myers Squibb Inc., Princeton, NJ.

ABSTRACT
Nivolumab, a fully human IgG4 programmed death 1 (PD-1) immune checkpoint inhibitor antibody, developed by Bristol-Myers Squibb Inc., has activity across non-small cell lung cancer (NSCLC) histologies and is Food and Drug Administration approved for treatment of metastatic squamous NSCLC with progression on or after platinum-based chemotherapy. PD-L1 has been investigated as a potential biomarker to predict clinical response to nivolumab in clinical settings. We report an automated PD-L1 immunohistochemistry (IHC) assay, which was developed to detect cell surface PD-L1 in formalin-fixed paraffin-embedded human tumor tissue specimens using Dako's Autostainer Link 48. The primary antibody for this assay is a rabbit monoclonal anti-human PD-L1 antibody, clone 28-8. The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines. The specificity of the PD-L1 IHC assay was further evaluated in a collection of 30 normal human tissues. The PD-L1 IHC assay was optimized for high sensitivity and precision in routine application. A pathology scoring and interpretation method specific to nivolumab clinical studies was adopted for the assay. The analytical performance of the assay was validated for application in the determination of PD-L1 status in human NSCLC specimens. The clinical application of the assay and scoring method was further validated in 3 Clinical Laboratory Improvement Amendments certified labs. The assay is currently being investigated in a variety of clinical studies for use as an in vitro diagnostic to select and stratify patients for treatment with the anti-PD-1 therapeutic antibody, nivolumab.

No MeSH data available.


Related in: MedlinePlus

Positive PD-L1 membrane staining in NSCLC tumor tissues illustrating intensity grades (top, ×20) and PD-L1 tumor scores (bottom, ×40). NSCLC indicates non–small cell lung cancer; PD-L1, programmed cell death 1 ligand 1.
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Figure 1: Positive PD-L1 membrane staining in NSCLC tumor tissues illustrating intensity grades (top, ×20) and PD-L1 tumor scores (bottom, ×40). NSCLC indicates non–small cell lung cancer; PD-L1, programmed cell death 1 ligand 1.

Mentions: Results of PD-L1 IHC assay stained slides were interpreted using light microscopy, inspecting the entire section using ×4 objectives and turned to ×10, ×20, and ×40 gradually to examine the PD-L1 staining. Positive PD-L1 staining is defined as complete and/or partial circumferential linear plasma membrane staining at any intensity that can be differentiated from background and diffuse cytoplasmic staining. Granular staining in the cytoplasm was not considered as positive staining although it can be attributed to cytoplasmic PD-L1 protein. Negative staining is defined as no staining of the plasma membrane. Positive PD-L1 was computed for tumor cells in tissues with a minimum of 100 evaluable tumor cells. When applicable, the staining intensity of PD-L1 was graded in a 0 to 3 grading scale; the staining intensity values are 0 (no staining), 1+ (weak), 2+ (moderate), and 3+ (strong) (Fig. 1, top). The percentage of viable tumor cells of the entire specimen that exhibit PD-L1 plasma membrane staining at any intensity was recorded as the PD-L1 tumor score (Fig. 1, bottom). The percentage of positive tumor PD-L1 cells was determined using 1% increment for scores ranging from 0 to 10%. A 10% increment was applied for percentage scores >10% (rounding to the nearest 10%); however, 1% increments was used when applicable. The specimen is considered to be PD-L1 positive if determined to be greater than or equal to the threshold tumor score, or PD-L1 negative if determined to be less than the threshold tumor score. The estimated number of viable tumor cells in the tumor space was used as the denominator for computation of the score.


Development of an automated PD-L1 immunohistochemistry (IHC) assay for non-small cell lung cancer.

Phillips T, Simmons P, Inzunza HD, Cogswell J, Novotny J, Taylor C, Zhang X - Appl. Immunohistochem. Mol. Morphol. (2015)

Positive PD-L1 membrane staining in NSCLC tumor tissues illustrating intensity grades (top, ×20) and PD-L1 tumor scores (bottom, ×40). NSCLC indicates non–small cell lung cancer; PD-L1, programmed cell death 1 ligand 1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561627&req=5

Figure 1: Positive PD-L1 membrane staining in NSCLC tumor tissues illustrating intensity grades (top, ×20) and PD-L1 tumor scores (bottom, ×40). NSCLC indicates non–small cell lung cancer; PD-L1, programmed cell death 1 ligand 1.
Mentions: Results of PD-L1 IHC assay stained slides were interpreted using light microscopy, inspecting the entire section using ×4 objectives and turned to ×10, ×20, and ×40 gradually to examine the PD-L1 staining. Positive PD-L1 staining is defined as complete and/or partial circumferential linear plasma membrane staining at any intensity that can be differentiated from background and diffuse cytoplasmic staining. Granular staining in the cytoplasm was not considered as positive staining although it can be attributed to cytoplasmic PD-L1 protein. Negative staining is defined as no staining of the plasma membrane. Positive PD-L1 was computed for tumor cells in tissues with a minimum of 100 evaluable tumor cells. When applicable, the staining intensity of PD-L1 was graded in a 0 to 3 grading scale; the staining intensity values are 0 (no staining), 1+ (weak), 2+ (moderate), and 3+ (strong) (Fig. 1, top). The percentage of viable tumor cells of the entire specimen that exhibit PD-L1 plasma membrane staining at any intensity was recorded as the PD-L1 tumor score (Fig. 1, bottom). The percentage of positive tumor PD-L1 cells was determined using 1% increment for scores ranging from 0 to 10%. A 10% increment was applied for percentage scores >10% (rounding to the nearest 10%); however, 1% increments was used when applicable. The specimen is considered to be PD-L1 positive if determined to be greater than or equal to the threshold tumor score, or PD-L1 negative if determined to be less than the threshold tumor score. The estimated number of viable tumor cells in the tumor space was used as the denominator for computation of the score.

Bottom Line: Nivolumab, a fully human IgG4 programmed death 1 (PD-1) immune checkpoint inhibitor antibody, developed by Bristol-Myers Squibb Inc., has activity across non-small cell lung cancer (NSCLC) histologies and is Food and Drug Administration approved for treatment of metastatic squamous NSCLC with progression on or after platinum-based chemotherapy.PD-L1 has been investigated as a potential biomarker to predict clinical response to nivolumab in clinical settings.The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines.

View Article: PubMed Central - PubMed

Affiliation: *Dako North America Inc., Carpinteria ‡Keck School of Medicine of the University of Southern California, Los Angeles, CA †Bristol-Myers Squibb Inc., Princeton, NJ.

ABSTRACT
Nivolumab, a fully human IgG4 programmed death 1 (PD-1) immune checkpoint inhibitor antibody, developed by Bristol-Myers Squibb Inc., has activity across non-small cell lung cancer (NSCLC) histologies and is Food and Drug Administration approved for treatment of metastatic squamous NSCLC with progression on or after platinum-based chemotherapy. PD-L1 has been investigated as a potential biomarker to predict clinical response to nivolumab in clinical settings. We report an automated PD-L1 immunohistochemistry (IHC) assay, which was developed to detect cell surface PD-L1 in formalin-fixed paraffin-embedded human tumor tissue specimens using Dako's Autostainer Link 48. The primary antibody for this assay is a rabbit monoclonal anti-human PD-L1 antibody, clone 28-8. The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines. The specificity of the PD-L1 IHC assay was further evaluated in a collection of 30 normal human tissues. The PD-L1 IHC assay was optimized for high sensitivity and precision in routine application. A pathology scoring and interpretation method specific to nivolumab clinical studies was adopted for the assay. The analytical performance of the assay was validated for application in the determination of PD-L1 status in human NSCLC specimens. The clinical application of the assay and scoring method was further validated in 3 Clinical Laboratory Improvement Amendments certified labs. The assay is currently being investigated in a variety of clinical studies for use as an in vitro diagnostic to select and stratify patients for treatment with the anti-PD-1 therapeutic antibody, nivolumab.

No MeSH data available.


Related in: MedlinePlus