Ex vivo generation of myeloid-derived suppressor cells that model the tumor immunosuppressive environment in colorectal cancer

TL;DR

This study shows how to generate myeloid-derived suppressor cells in the lab to model immune suppression in colorectal cancer.

Contribution

A novel in vitro method to generate tumor-like myeloid-derived suppressor cells from bone marrow cells using GM-CSF-secreting CT26 conditioned medium.

Findings

Generated granulocytic and monocytic MDSC resembling those in CRC tumors, not spleens of tumor-bearing mice.

MDSC inhibited T-cell responses in vitro, which could be reversed by blocking arginase-1 or iNOS.

In vivo inhibition of arginase-1 or iNOS stimulated T-cell responses and delayed tumor growth.

Abstract

Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of cells that accumulate in tumor-bearing subjects and which strongly inhibit anti-cancer immune responses. To study the biology of MDSC in colorectal cancer (CRC), we cultured bone marrow cells in conditioned medium from CT26 cells, which are genetically modified to secrete high levels of granulocyte-macrophage colony-stimulating factor. This resulted in the generation of high numbers of CD11b+ Ly6G+ granulocytic and CD11b+ Ly6C+ monocytic MDSC, which closely resemble those found within the tumor but not the spleen of CT26 tumor-bearing mice. Such MDSC potently inhibited T-cell responses in vitro, a process that could be reversed upon blocking of arginase-1 or inducible nitric oxide synthase (iNOS). We confirmed that inhibition of arginase-1 or iNOS in vivo resulted in the stimulation of cytotoxic T-cell responses. A delay in tumor growth was observed upon functional repression of both enzymes. These data confirm the role of MDSC as inhibitors of T-cell-mediated immune responses in CRC. Moreover, MDSC differentiated in vitro from bone marrow cells using conditioned medium of GM-CSF-secreting CT26 cells, represent a valuable platform to study/identify drugs that counteract MDSC activities.

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