Method for the Analysis of the Tumor Microenvironment by Mass Cytometry: Application to Chronic Lymphocytic Leukemia.
Animals
Antigens, CD
/ metabolism
Biomarkers, Tumor
/ antagonists & inhibitors
Flow Cytometry
Immune Checkpoint Inhibitors
/ pharmacology
Immunotherapy
Leukemia, Lymphocytic, Chronic, B-Cell
/ drug therapy
Mice
Programmed Cell Death 1 Receptor
/ metabolism
Tumor Microenvironment
Lymphocyte Activation Gene 3 Protein
chronic lymphocytic leukemia
immunosuppression
lymphoma
mass cytometry
microenvironment
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2020
2020
Historique:
received:
30
06
2020
accepted:
29
09
2020
entrez:
16
11
2020
pubmed:
17
11
2020
medline:
16
6
2021
Statut:
epublish
Résumé
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 analysis. Additionally, we discuss different advantages and pitfalls of this technique.
Identifiants
pubmed: 33193376
doi: 10.3389/fimmu.2020.578176
pmc: PMC7606286
doi:
Substances chimiques
Antigens, CD
0
Biomarkers, Tumor
0
Immune Checkpoint Inhibitors
0
Pdcd1 protein, mouse
0
Programmed Cell Death 1 Receptor
0
Lymphocyte Activation Gene 3 Protein
0
Lag3 protein, mouse
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
578176Informations de copyright
Copyright © 2020 Gonder, Fernandez Botana, Wierz, Pagano, Gargiulo, Cosma, Moussay, Paggetti and Largeot.
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