Abstract
The integrative analysis of tumor immune microenvironment (TiME) components, their interactions and their microanatomical distribution is mandatory to better understand tumor progression. Imaging Mass Cytometry (IMC) is a high dimensional tissue imaging system which allows the comprehensive and multiparametric in situ exploration of tumor microenvironments at a single cell level. We describe here the design of a 39-antibody IMC panel for the staining of formalin-fixed paraffin-embedded human tumor sections. We also provide an optimized staining procedure and details of the experimental workflow. This panel deciphers the nature of immune cells, their functions and their interactions with tumor cells and cancer-associated fibroblasts as well as with other TiME structural components known to be associated with tumor progression like nerve fibers and tumor extracellular matrix proteins. This panel represents a valuable innovative and powerful tool for fundamental and clinical studies that could be used for the identification of prognostic biomarkers and mechanisms of resistance to current immunotherapies.
Highlights
The tumor immune microenvironment (TiME) is characterized by complex interactions of immune cells with other heterogeneous cellular and acellular components of this ecosystem
We describe here the development of a 39-antibody panel that can be used in Imaging Mass Cytometry (IMC) to stain a single formalin-fixed paraffinembedded (FFPE) human tissue section
To design the IMC panel, adapted to the characterization of human TiME, we first selected the cellular and acellular components of this environment that are known to be associated with tumor progression and described to modulate the immune response
Summary
The tumor immune microenvironment (TiME) is characterized by complex interactions of immune cells with other heterogeneous cellular and acellular components of this ecosystem Their crosstalk directly and indirectly contributes to tumor progression and immune surveillance evasion [1,2,3,4]. Both innate and adaptive immune cells participate in tumor development via active antitumoral and immunosuppressive protumor functions. These anti- and protumor immune responses are modulated by contextual signals from other TiME actors [5]. Active cancer-associated fibroblasts (CAFs) exhibit increased expression of
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