Method for the Analysis of the Tumor Microenvironment by Mass Cytometry: Application to Chronic Lymphocytic Leukemia.

Frontiers in Immunology
Susanne GonderAnne Largeot

Abstract

In the past 20 years, the interest for the tumor microenvironment (TME) has exponentially increased. Indeed, it is now commonly admitted that the TME plays a crucial role in cancer development, maintenance, immune escape and resistance to therapy. This stands true for hematological malignancies as well. A considerable amount of newly developed therapies are directed against the cancer-supporting TME instead of targeting tumor cells themselves. However, the TME is often not clearly defined. In addition, the unique phenotype of each tumor and the variability among patients limit the success of such therapies. Recently, our group took advantage of the mass cytometry technology to unveil the specific TME in the context of chronic lymphocytic leukemia (CLL) in mice. We found the enrichment of LAG3 and PD1, two immune checkpoints. We tested an antibody-based immunotherapy, targeting these two molecules. This combination of antibodies was successful in the treatment of murine CLL. In this methods article, we provide a detailed protocol for the staining of CLL TME cells aiming at their characterization using mass cytometry. We include panel design and validation, sample preparation and acquisition, machine set-up, quality control, and ...Continue Reading

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Citations

Sep 5, 2021·Nature Protocols·Marcela Alcántara-Hernández, Juliana Idoyaga
Oct 16, 2021·Frontiers in Cell and Developmental Biology·Carmen-Mariana AaneiLydia Campos Catafal

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Methods Mentioned

BETA
Flow Cytometry
biopsies
the
fluorescence-activated cell sorting
PMA
dissection
flow
FCS
single cell cytometry

Software Mentioned

Fluidigm
Cytosplore
Tableau Prep
TableauPrep
Helios
Bioconductor
Mathlab
cytobank
excel
Graph Pad Prism

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