Programmed death-ligand 1 (PD-L1) expression in cured and not cured testicular and other germ cell tumors (GCT).

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 485-485 ◽  
Author(s):  
Brandon David Bernard ◽  
Laurence Albiges ◽  
Sabina Signoretti ◽  
Jesse Novak ◽  
Michelle S. Hirsch ◽  
...  
Keyword(s):  
Immune Cell ◽  
Therapeutic Option ◽  
Germ Cell Tumors ◽  
Unknown Origin ◽  
Mediastinal Tumors ◽  
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Programmed Death ◽  
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Tumor Types ◽  
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485 Background: PD-L1 is involved in immune regulation and is prognostic in many tumor types. The aim of this study was to assess PD-L1 expression in orchiectomy and metastases (mets) from GCT and to assess for an association with outcome. Methods: Immunohistochemistry (IHC) on whole sections from archival formalin-fixed paraffin-embedded tissue from Dana-Farber Cancer Institute (DFCI) and Gustave Roussy (IGR) was performed using anti-PD-L1 antibody E1L3N. Stained slides were scored semi-quantitatively and assessed independently. At DFCI, PD-L1 was scored as percent of positive tumor cells (TC) and extent of positive immune cell (IC) infiltrate; at IGR, a cut-off of 5% defined positive TC and IC. PD-L1 status was assessed by site, histology and chemotherapy (CT) status. PD-L1 status was correlated with clinical outcome. Most samples at DFCI were pre-CT (74.2%) whereas most at IGR were post-CT (85.4%). Results: IHC from 171 patients with GCT (89 DFCI, 82 IGR) from 1987-2014 was reviewed. The DFCI cohort included 69 orchiectomy, 15 mets and 5 unknown origin with 28 total deaths; the IGR cohort included 26 lung mets, 36 mediastinal or retroperitoneal lymph node mets and 20 primary mediastinal tumors. DFCI had 38 seminoma (S), 50 nonseminoma (NS) and 1 unknown; at IGR, 13 S and 69 NS. In both cohorts, PD-L1 staining in IC was higher in S (DFCI 33/38 moderate (mod)-high (86.8%); IGR 12/13 positive (92.3%)) vs. NS (DFCI 26/50 mod-high (52.0%); IGR 35/68 positive (51.5%)) (DFCI p = 0.003; IGR p = 0.006). At DFCI, 5 samples had PD-L1 positive TC (2 mixed GCT, 2 pure choriocarcinomas (CC) and 1 S); at IGR, 14 had positive TC (all NS, including 11 pure CC). More PD-L1 positive TC were CC than other GCT types (DFCI p = 0.003; IGR p < 0.001). In 36 S orchiectomy samples at DFCI, 0/4 that scored none-mild died and 2/31 (6.5%) that scored mod-high died (p = 1.00); in 30 NS orchiectomy samples, 4/9 (44.4%) that scored none-mild died and 2/16 (12.5%) that scored mod-high died (p = 0.14). Conclusions: Higher PD-L1 status in orchiectomy is not associated with GCT survival. PD-L1 positive TC may serve as a histological marker for CC among NS. Higher PD-L1 expression on IC in S and on TC in CC may point to immunotherapy as a therapeutic option in these tumors.

scholarly journals Deciphering the Immune Microenvironment on A Single Archival Formalin-Fixed Paraffin-Embedded Tissue Section by An Immediately Implementable Multiplex Fluorescence Immunostaining Protocol

Cancers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2449 ◽  
Author(s):  
Adrien Guillot ◽  
Marlene S. Kohlhepp ◽  
Alix Bruneau ◽  
Felix Heymann ◽  
Frank Tacke
Keyword(s):  
Immune Cell ◽  
Simple Procedure ◽  
Parenchymal Cell ◽  
Ffpe Tissue ◽  
Immune Microenvironment ◽  
Tissue Samples ◽  
Tissue Organization ◽  
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Technological breakthroughs have fundamentally changed our understanding on the complexity of the tumor microenvironment at the single-cell level. Characterizing the immune cell composition in relation to spatial distribution and histological changes may provide important diagnostic and therapeutic information. Immunostaining on formalin-fixed paraffin-embedded (FFPE) tissue samples represents a widespread and simple procedure, allowing the visualization of cellular distribution and processes, on preserved tissue structure. Recent advances in microscopy and molecular biology have made multiplexing accessible, yet technically challenging. We herein describe a novel, simple and cost-effective method for a reproducible and highly flexible multiplex immunostaining on archived FFPE tissue samples, which we optimized for solid organs (e.g., liver, intestine, lung, kidney) from mice and humans. Our protocol requires limited specific equipment and reagents, making multiplexing (>12 antibodies) immediately implementable to any histology laboratory routinely performing immunostaining. Using this method on single sections and combining it with automated whole-slide image analysis, we characterize the hepatic immune microenvironment in preclinical mouse models of liver fibrosis, steatohepatitis and hepatocellular carcinoma (HCC) and on human-patient samples with chronic liver diseases. The data provide useful insights into tissue organization and immune–parenchymal cell-to-cell interactions. It also highlights the profound macrophage heterogeneity in liver across premalignant conditions and HCC.

scholarly journals Tumor Immune Microenvironment Characterization of Primary Lung Adenocarcinoma and Lymph Node Metastases

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Yuan Zhou ◽  
Xinying Shi ◽  
Huan Chen ◽  
Beibei Mao ◽  
Xue Song ◽  
...  
Keyword(s):  
Immune Cell ◽  
Locally Advanced ◽  
Transcriptome Data ◽  
Immune Microenvironment ◽  
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Primary Lung Adenocarcinoma ◽  
Tumor Immune Microenvironment ◽  
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Ifn Γ ◽  
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Background. The essential roles of the tumor microenvironment (TME) have been recognized during the initiation and progression of primary lung adenocarcinoma (LUAD). The aim of the present study was to delineate the immune landscape in both primary cancer and matched lymph node metastasis from a cohort of locally advanced stage LUAD patients with distinct outcomes. Methods. Formalin-fixed, paraffin-embedded samples were collected from 36 locally advanced LUAD patients. Transcriptome data of the tumor immune microenvironment were resolved using an immune oncology panel RNA sequencing platform. Bioinformatics approaches were used to determine the differentially expressed genes (DEGs), dysregulated pathways, and immune cell fraction between patients with early recurrence (ER) and late recurrence (LR). Results. Here, we showed that in primary cancer tissues, 23 DEGs were obtained between patients with ER and LR. Functional analysis revealed that the LR in LUAD patients may be associated with enriched gene sets belonging to the antigen presentation and MHC protein complex, innate immune response, and IFN-γ signaling pathways. Next, the transcriptome data were adopted to quantify immune cell fractions, indicating that high infiltration of mast cells and neutrophils was correlated with ER. Interestingly, similar findings were observed in metastatic lymph nodes from patients suffering from ER or LR. By analyzing the shared immune features of primary cancers and lymphatic metastases, we unraveled the prognostic value and joint utility of two DEGs, CORO1A and S100A8. Conclusions. In LUAD, the enrichment in antigen presentation, MHC protein complex, and IFN-γ signaling, and low infiltration of neutrophils in primary or metastatic nodules may be indications for a favorable prognosis. Integrated with bioinformatics approaches, transcriptome data of immune-related genes from formalin-fixed, paraffin-embedded (FFPE) samples can effectively profile the landscape of the tumor immune microenvironment and help predict clinical outcomes.

scholarly journals Comprehensive NGS Panel Validation for the Identification of Actionable Alterations in Adult Solid Tumors

2021 ◽  
Vol 11 (5) ◽  
pp. 360
Author(s):  
Paula Martínez-Fernández ◽  
Patricia Pose ◽  
Raquel Dolz-Gaitón ◽  
Arantxa García ◽  
Inmaculada Trigo-Sánchez ◽  
...  
Keyword(s):  
Solid Tumors ◽  
Limit Of Detection ◽  
Copy Number Variants ◽  
Hybridization Capture ◽  
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Ffpe Tumor ◽  
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Tumor Types ◽  
Next Generation Sequencing Ngs ◽  
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The increasing identification of driver oncogenic alterations and progress of targeted therapies addresses the need of comprehensive alternatives to standard molecular methods. The translation into clinical practice of next-generation sequencing (NGS) panels is actually challenged by the compliance of high quality standards for clinical accreditation. Herein, we present the analytical and clinical feasibility study of a hybridization capture-based NGS panel (Action OncoKitDx) for the analysis of somatic mutations, copy number variants (CNVs), fusions, pharmacogenetic SNPs and Microsatellite Instability (MSI) determination in formalin-fixed paraffin-embedded (FFPE) tumor samples. A total of 64 samples were submitted to extensive analytical validation for the identification of previously known variants. An additional set of 166 tumor and patient-matched normal samples were sequenced to assess the clinical utility of the assay across different tumor types. The panel demonstrated good specificity, sensitivity, reproducibility, and repeatability for the identification of all biomarkers analyzed and the 5% limit of detection set was validated. Among the clinical cohorts, the assay revealed pathogenic genomic alterations in 97% of patient cases, and in 82.7%, at least one clinically relevant variant was detected. The validation of accuracy and robustness of this assay supports the Action OncoKitDx’s utility in adult solid tumors.

Immunohistochemistry of CA 125

1998 ◽  
Vol 13 (4) ◽  
pp. 210-215 ◽  
Author(s):  
M. Nap
Keyword(s):  
Unknown Origin ◽  
Daily Practice ◽  
Metastatic Carcinoma ◽  
Ca 125 ◽  
Gynecological Tumors ◽  
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Number Of Publications ◽  
Sensitivity Specificity ◽  
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CA 125 is known as the marker that is most strongly associated with epithelial gynecological tumors. Compared to the number of publications on its use in serum assays, the application in immunohistochemistry is still limited. The availability of many good antibodies that perform well in formalin-fixed paraffin-embedded tissue opens good possibilities for a wider use. Outside the gynecological tract several other structures may react positive for CA 125. Among these are the lung and breast but also the epithelial cells of the conjunctiva and to some extent prostate glandular epithelium. In the fetus reactions can be found in the serosal linings of body cavities but also in the esophagus and skin. In diagnostic pathology CA 125 plays a role in identifying the primary locations of metastatic carcinoma of unknown origin. It is recommended to use CA 125 antibodies not in a solitary setting but in combination with CEA, BRST-2 and Vimentin to discriminate best between the most frequent sites of origin of metastatic carcinoma. Regular analysis of sensitivity/specificity ratios in a balanced population, representing the composition of the patient population seen in daily practice, should be performed to evaluate the position of CA 125 in diagnostic immunohistochemistry.

Programmed death-ligand 1 (PD-L1) expression in small cell carcinoma of bladder.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e16019-e16019
Author(s):  
Angela Sanguino ◽  
Arash Samiei ◽  
Gurleen Pasricha ◽  
Lakshmi Harinath ◽  
Ralph Miller ◽  
...  
Keyword(s):  
Cell Carcinoma ◽  
Tumor Cells ◽  
Small Cell Carcinoma ◽  
Mononuclear Cells ◽  
Checkpoint Inhibitors ◽  
Small Cell ◽  
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Programmed Death ◽  
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e16019 Background: Small cell carcinoma of bladder (SCCB) is a rare but aggressive variant of bladder neoplasm. There is limited insight for risk prognostication and treatment guidance in this entity. Immune checkpoint inhibitors (ICI), anti-PD-1 or anti-PD-L1 antibodies, have been approved for treating urothelial carcinoma, while the evidence of their efficacy in SCCB is lacking. PD-L1 expression in tumor tissue of urothelial cancer has been postulated to correlate with response to ICI but with controversy. We have studied the expression of PD-L1 in SCCB and its association with patient survival. Methods: Nineteen cases of SCCB diagnosed between 2011 and 2017 in a single center were identified. Formalin-fixed paraffin-embedded tumor samples were stained for PD-L1 (Ventana PD-L1 SP142). Cases showing positive stain in 5% or more of tumor cells and tumor stromal mononuclear cells (TSMC) were considered positive. Results: Among 19 cases of SCCB, 4 (21%) stained positive for PD-L1. All 4 cases had strong PD-L1 staining ( > 30%) seen in the TSMC but barely in tumor cells (focal < 5% cells in 2/4 cases). Except for one patient who died from surgery, all remaining 3 patients with positive PD-L1 staining are still alive. Twelve out of 19 SCCB patients developed metastatic disease; 4 of them were treated with ICI. The only responder of the 4 patients had strong PD-L1 expression in TSMC cells. The overall survival for patients with positive PD-L1 staining was 41 months versus 14 months for those with negative staining (p = 0.09). Age, pathologic stage and treatments were similar between the two groups. Conclusions: In our study, PD-L1 expression was seen in 21% of tumor samples from patients with SCCB, mostly in TSMC, but minimal in tumor cells. The strong expression of PD-L1 in TSMC correlates with a trend of improved survival and potential response to ICI in SCCB. PD-L1 expression in TSMC, rather than tumor cells, could be used as a marker for prognosis in SCCB.

scholarly journals Gene expression analysis in formalin fixed paraffin embedded melanomas is associated with density of corresponding immune cells in those tissues

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Minyoung Kwak ◽  
Gulsun Erdag ◽  
Craig L. Slingluff
Keyword(s):  
Gene Expression ◽  
Immune Cells ◽  
Expression Profiling ◽  
Gene Expression Analysis ◽  
Immune Cell ◽  
Single Gene ◽  
Cell Marker ◽  
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Abstract Immune cell infiltrates in melanoma have important prognostic value. Gene expression analysis may simultaneously quantify numbers and function of multiple immune cell subtypes in formalin-fixed paraffin-embedded (FFPE) tissues. Prior studies report single gene expression can represent individual immune cell subtypes, but this has not been shown in FFPE melanomas. We hypothesized that gene expression profiling of human melanomas using a new RNA expression technology in FFPE tissue would correlate with the same immune cells identified by immunohistochemistry (IHC). This retrospective study included melanoma specimens analyzed by IHC on tumor tissue microarray (TMA) cores and by gene expression profiling with EdgeSeq Immuno-Oncology Assay using qNPA technology on the corresponding tumors. Standardized gene expression levels were analyzed relative to enumerated cells by IHC using Spearman rank test to calculate r-values. Multivariate analysis was performed by Kruskal–Wallis test. 119 melanoma specimens had both IHC and gene expression information available. There were significant associations between the level of gene expression and its quantified IHC cell marker for CD45+, CD3+, CD8+, CD4+, and CD20+ cells (all p < 0.001). There were also significant associations with exhaustion markers FoxP3+, PD-1+, and PD-L1+ (all p ≤ 0.0001). This new qNPA technology is useful to quantify intratumoral immune cells on FFPE specimens through RNA gene expression in metastatic melanoma. As previous studies have shown on other solid human tumors, we also confirm that the expression level of a single gene may be used to represent a single IHC immune cell marker in melanoma.

Impact of Preanalytical Factors on the Measurement of Tumor Tissue Biomarkers Using Immunohistochemistry

2021 ◽  
pp. 002215542199560
Author(s):  
Aditi Bagchi ◽  
Zachary Madaj ◽  
Kelly B. Engel ◽  
Ping Guan ◽  
Daniel C. Rohrer ◽  
...  
Keyword(s):  
Tumor Tissue ◽  
Cancer Biomarkers ◽  
Staining Intensity ◽  
Ffpe Tissue ◽  
Kidney Tumor ◽  
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Formalin Fixed Paraffin Embedded ◽  
Tumor Types ◽  
Formalin Fixed ◽  
Tissue Biomarkers

Analysis of formalin-fixed paraffin-embedded (FFPE) tissue by immunohistochemistry (IHC) is commonplace in clinical and research laboratories. However, reports suggest that IHC results can be compromised by biospecimen preanalytical factors. The National Cancer Institute’s Biospecimen Preanalytical Variables Program conducted a systematic study to examine the potential effects of delay to fixation (DTF) and time in fixative (TIF) on IHC using 24 cancer biomarkers. Differences in IHC staining, relative to controls with a DTF of 1 hr, were observed in FFPE kidney tumor specimens after a DTF of ≥2 hr. Reductions in H-score and/or staining intensity were observed for c-MET, p53, PAX2, PAX8, pAKT, and survivin, whereas increases were observed for RCC1, EGFR, and CD10. Prolonged TIF of 72 hr resulted in significantly reduced H-scores of CD44 and c-Met in kidney tumor specimens, compared with controls with 12-hr TIF. An elevated probability of altered staining intensity due to DTF was observed for nine antigens, whereas for prolonged TIF an elevated probability was observed for one antigen. Results reported here and elsewhere across tumor types and antigens support limiting DTF to ≤1 hr when possible and fixing tissues in formalin for 12–24 hr to avoid confounding effects of these preanalytical factors on IHC.

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