1179. Deep spatial immuno-profiling through high biomarker colocalization in FFPE tumor tissue samplesAbdul Mohammed, Chineme Onwubueke, and Maël ManesseUltivue, Inc. 763D Concord Avenue, Cambridge, MA 02138 • +1-617-945-2662 • www.ultivue.com • contact@ultivue.com • Twitter: @Ultivue

The recent emergence of multiplexed immunohistochemistry(IHC) has the potential to revolutionize immuno-oncology andpathology research as it enables the identification of complex cellsubtypes and their interactions in the tumor environment.Comprehensive classification of the different cell types inimmuno-oncology presents a unique challenge as many of therelevant biomarkers are common to large subsets of cell typeswithin a particular cell class. Therefore, a key feature for accuratephenotypic classification is the ability to detect a high number ofcolocalized biomarkers. In this study, we demonstrate how theInSituPlex® technology enables researchers to reliably label anddetect phenotypes with markers expressed in the samecompartment (membrane, cytoplasmic, or nuclear) on single cellsof a NSCLC tissue section stained with 16 different markers. Formore information, refer to poster #1183.

Conclusions

Introduction

PD-L1 Panel APC Panel

High co-expression and deep phenotyping in a single NSCLC tissue section

For Research Use Only. Not for use in diagnostic procedures.Tissue samples were procured from commercial vendors for this research study.Ultivue®, InSituPlex®, UltiMapper™ are either registered trademarks or trademarks of Ultivue in the United States and/or other countries. All other trademarks are property of their respective owners.

In this poster, we demonstrate research of the identification ofphenotypes based on the co-expression and colocalization ofbiomarkers on the same cells in a lung tumor section. Analysis ofthe high dimensional, spatially resolved data obtained from a 16-plex assay provided phenotypic information of differentlymphocytes, macrophages, dendritic cells, antigen-presentingcells, and tumor cell populations with multiple colocalizedbiomarkers.

Overlay CD3 CD8

Cytotoxic T cells

Overlay CD3 CD4

Helper T cellsOverlay CD3 CD4 FOXP3

Naïve regulatory T cells

Overlay CD3 CD8 GrB

Cytotoxic T cells expressing Granzyme B

Overlay CD3 CD4 CD45RO

Memory helper T cells

Overlay CD11c

Dendritic cells B cellsOverlay CD20

Overlay CD3 CD4 CD45RO FOXP3

Memory regulatory T cells

Overlay CD3 CD8 CD45RO LAG3 PD-1

Exhausted memory cytotoxic T cells

Overlay CD3 CD8 CD45RO

Memory cytotoxic T cells

Overlay CD68 CD163

M2 macrophagesCD8 regulatory T cells

Overlay CD3 CD8 FOXP3

Overlay CD68 CD163 PD-L1

Immunosuppressive M2 macrophages

Overlay CD68 MHCII

Antigen-presenting M1 macrophages

APC Panel (ROI)

CK Ki67Overlay

Proliferating tumor cells

Proliferating immune evading tumor cells

CK PD-L1 Ki67Overlay

DAPICD3CD4CD8CD11cCD20CD45ROCD68CD163FoxP3GranzymeBKi67LAG3MHCIIPD-1PD-L1Cytokeratin

Proliferating exhausted memory T cellsCD3 CD45RO PD-1 Ki67Overlay

Overlay CD3 CD8 GrB

NK cells expressing Granzyme B

TumorCells

ImmuneCells