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

// Ines Dufait 1,2 , Julia Katharina Schwarze 1 , Therese Liechtenstein 3,4 , Wim Leonard 1 , Heng Jiang 1 , David Escors 3,4 , Mark De Ridder 1,* and Karine Breckpot 2,* 1 UZ Brussel, Department of Radiotherapy, Vrije Universiteit Brussel, Brussels, Belgium 2 Laboratory of Molecular and Cellular Therapy, Vrije Universiteit Brussel, Brussels, Belgium 3 Navarrabiomed-Fundacion Miguel Servet, Immunomodulation group, Pamplona, Spain 4 Division of Infection and Immunity, University College London, London, UK * These senior authors contributed equally to this work Correspondence to: Karine Breckpot, email: // Keywords : MDSC, CRC, arginase-1, inducible nitric oxide synthase, GM-CSF Received : February 18, 2015 Accepted : March 11, 2015 Published : March 29, 2015 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.