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Tumor Phenotyping and Immune Cell Regulation using Multiplex IHC

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Elena Baranova1, Naomi Defort1, Domenico Lazzaro1, Amanda Finan1, Morgan Mathieu2, Renaud Burrer1
1 Histalim, Montpellier, France
2 Enzo Life Sciences, Lausen, Switzerland

Featured Product: MULTIVIEW® PLUS (MOUSE-HRP/RABBIT-AP) IHC Kit, POLYVIEW® PLUS (RABBIT-HRP) Reagent, HIGHDEF® Black IHC Chromogen (HRP), HIGHDEF® Blue IHC Chromogen (HRP), HIGHDEF® Red IHC Chromogen (HRP), HIGHDEF® Yellow IHC Chromogen (HRP), HIGHDEF® IHC Mount


INTRODUCTION

For over a decade, Histalim has provided services in histology, cytology, and molecular biology with extensive experience in immuno-oncology and immune therapies. With broad experience in neurosciences, toxicity assessment, therapeutic antibodies characterization, evaluation of implantable medical devices, and more. Histalim focuses on providing each client with the highest quality service. Histalim’s scientific experts and European network of histopathology experts along with top of the line histological and image processing techniques and analysis capabilities enable Histalim to efficiently meet each client’s specific need.

Immunohistochemistry (IHC) is important for pathologists and researchers in the diagnosis of tumors and analysis of the tissue environment. A technique which allows for maximal data per tissue section is very appealing when patient tissue is limited. Multiplex IHC allows for the evaluation of co-expression and spatial organization of multiple targets within preserved tissue architecture. This is particularly important in tumor immunology where it is critical to phenotype the immune and cancer cells in the context of the tumor microenvironment.

The MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC kit from Enzo Life Sciences is a non-biotin, one-step detection system suitable for demonstrating the expression of multiple antigens on tissue sections. It enables faster and more consistent staining procedures than traditional two-step methods using biotin and avidin/streptavidin conjugates. Indeed, mouse antibodies are directly tagged with HRP and rabbit antibodies with AP at the same time, all with significantly lower background staining.

The main objective of this study was to validate the use of the MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC kit and the HIGHDEF® IHC chromogens to detect multiple markers on human tissue sections. To that effect, two panels composed of four markers each were chosen. The first panel includes phenotypic markers commonly used in the diagnosis of lung cancer (CK5/6, Napsin A, p63, and TTF-1) while the second panel focusses on the characterization of the immune cell response and its regulation (CD3, CD8, PD-1, and PD-L1). The intensity, sensitivity, and specificity obtained with both panels demonstrated the capacity of Enzo’s detection reagents and chromogens in multiplex protocols.

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