In-Depth Myeloid Cell Characterization in the Murine Syngeneic CT.26 Colon Carcinoma Model by 10-Color Flow Cytometry

The efficacy of immune-modulating anti-cancer therapeutic antibodies that have been FDA-approved in recent years, such as anti-CTLA-4 and anti-PD-1, has driven growing interest in methods that provide a mechanistic understanding of drug function. Development of new mono- and combination therapies with immune-modulatory effects requires more powerful immunophenotyping techniques capable of in depth cell characterization.

During AACR’s Annual Meeting 2016, Matt Thayer, Sr. Assistant Scientist, In Vitro Operations, presented his poster session and spoke about his work with using the CT.26 murine syngeneic colorectal cancer model to develop a 10-color flow cytometry antibody panel. This panel focuses on the identification of tumor-infiltrating immune cell subsets derived from myeloid lineage precursors utilizing the high-throughput-capable 4-laser, 14-color Attune™ NxT Flow Cytometer.

The panel includes a combination of antibodies against CD45, CD3, CD19, CD49b, CD335, CD11b, CD11c, Ly-6G, Ly-6C, F4/80, and CD115. By excluding cells of lymphoid lineage, we can show that this panel facilitates analysis of natural killer (NK) cells and myeloid derived cells including macrophages, neutrophils, dendritic cells (DCs), and monocytic or granulocytic myeloid-derived suppressor cell (mMDSC and gMDSC) subsets in tumor and peripheral blood.

In addition, this antibody combination allows for a more complete analysis of MDSC cells which can differentially express several disease-relevant myeloid specific markers including Ly-6G, Ly-6C, F4/80, CD11c, and CD115. This panel was also utilized to characterize changes in the myeloid subset between control and anti-PD-L1 treated mice.

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