Published OnlineFirst October 25, 2017; DOI: 10.1158/0008-5472.CAN-17-1416 Cancer Tumor Biology and Immunology Research Type I IFN Receptor Signaling Controls IL7-Dependent Accumulation and Activity of Protumoral IL17A-Producing gdT Cells in Breast Cancer Emmanuel C. Patin1, Daphnee Soulard1,Sebastien Fleury2,3, Maya Hassane1,4, David Dombrowicz2,3, Christelle Faveeuw1,Francois¸ Trottein1, and Christophe Paget1,5 Abstract The protumoral activity of gdT17 cells has recently emerged tumorbedofapeculiarsubsetofgdT17 cells displaying a À in a wide variety of solid malignancies, including breast cancer. CD27 CD3bright phenotype (previously associated with the þ These cells exert their detrimental functions by promoting invariant Vg6Vd1 TCR). Interestingly, this effect was indirect tumor growth, angiogenesis, and subsequent metastasis devel- and partially relied on the IFNAR1-dependent control of IL7 opment. However, the intratumoral factors that regulate the secretion, a factor that triggers proliferation and activating biology of gdT17cells within the tumor microenvironment are functions of deleterious gdT17 cells. Our work therefore iden- less well understood. Here, using two experimental models of tifies a key role of the type I IFN/IL7 axis in the regulation of breast cancer, we reinforced the concept that tumor-infiltrating intratumoral gdT17-cell functions and in the development of gdT17 cells are endowed with protumoral functions, which primary breast tumor growth and metastasis. promote tumor progression and metastasis development. More Significance: Tumor-derived IL7 can represent a therapeutic importantly, we demonstrated a critical role for type I IFN target to prevent accumulation of immune cells endowed with signaling in controlling the preferential accumulation in the potent protumoral activities. Cancer Res; 78(1); 195–204. Ó2017 AACR. Introduction Among these cell subsets, tumor-infiltrating IL17A-producing gdT(gdT17) cells represent interesting candidates (3). Mouse Despite significant advances in treatment and earlier detection, gdT17 cells are characterized by the constitutive expression of cancer remains one of the most devastating diseases in industri- the orphan nuclear receptor RORgt (4) and the absence of the alized countries. Over the last decades, immunology has gained CD27 marker, a thymic regulator that conditions the cytokine bias considerable attention in the complex discipline of oncology. (IFNg vs. IL17A production) of gdT cells in the periphery (5). Indeed, it is now well established that leukocyte composition Moreover, many reports indicate that TCR chain usage is linked within tumors is considered significant for patient prognosis in a þ þ with specialized functions (6). In this context, Vg4 and Vg6 T wide variety of cancers, and therefore some immune populations cells have been described to preferentially produce IL17 (7); a are highly regarded by clinicians as new biomarkers for prognosis phenomenon that primarily relies on the acquisition of a specific and therapeutic success (1, 2). functional programming during their development in fetal thy- mus (8). Immune system evolves to avoid functional redundan- cies, and therefore it is likely that gdT17 cells bearing either Vg4or Vg6 TCR have unique properties. We recently reported that bright 1Universite de Lille, CNRS, INSERM, CHU Lille, Institut Pasteur de Lille, U1019-UMR levels of CD3 signals on gdT cells can be used as a surrogate marker 8204-CIIL-Center for Infection and Immunity of Lille, France. 2Universite de Lille, to specifically analyze the prototypic IL17 producer subset þ INSERM, Institut Pasteur de Lille, CHU Lille, U1011, EGID, Lille, France. 3European Vg6Vd1 T cells (9, 10). Genomic Institute of Diabetes, Lille, France. 4Laboratoire Microbiologie Santeet Over the past decade, numerous reports have consensually Environnement, Ecole doctorale en Sciences et Technologies/Faculte de Sante attributed protumoral functions to gdT17 cells in various exper- 5 Publique, Universite Libanaise, Tripoli, Liban. Universite de Tours, INSERM, imental transplantable or inducible models of cancer including Centre d'Etude des Pathologies Respiratoires (CEPR), UMR 1100, Tours, France. liver and lung carcinomas (11, 12), melanoma (13), breast cancer Note: Supplementary data for this article are available at Cancer Research (14), fibrosarcoma (15), and peritoneal/ovarian cancer (16). Online (http://cancerres.aacrjournals.org/). Similar observations have also been reported in tumor-bearing Current address for E.C. Patin: Retroviral Immunology, The Francis Crick patients such as colorectal cancer (17). Moreover, the frequency of Institute, London, United Kingdom. infiltrating gdT cells in breast cancer patients is a relevant predic- Corresponding Author: Christophe Paget, Institut Pasteur de Lille, 1 rue du Pr tive marker of clinical outcome; associated with poor prognosis Calmette, Lille 59000, France. Phone: 333-2087-7339; E-mail: and relapses (18). gdT17 cells exert their detrimental functions [email protected] through interactions with neighboring cells from the tumor doi: 10.1158/0008-5472.CAN-17-1416 environment, including suppressive immune cells (e.g., mye- Ó2017 American Association for Cancer Research. loid-derived suppressor cells, regulatory T cells, M2 macrophages) www.aacrjournals.org 195 Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 2018 American Association for Cancer Research. Published OnlineFirst October 25, 2017; DOI: 10.1158/0008-5472.CAN-17-1416 Patin et al. and endothelial cells. This ultimately leads (i) to the inhibition of conditions. Animal studies have been conducted in accordance þ the protective CD8 T-cell response and (ii) to the initiation with an Institutional Animal Care and Use Committee. All animal of angiogenesis, subsequent increase of tumor growth, and metas- work conformed to the French "Comite d'Ethique en Experimen- tasis development. tation Animale" (C.E.E.A. 75; #00357.01) and committee The factors that regulate gdT17 cell accumulation and activity guidelines. within the tumor have been mainly attributed to Th17-driving cytokines such as IL1b, IL6, IL23, and TGFb (11, 15). IL7 has been Reagents recently involved in selective expansion and functions of IL17A- Monoclonal antibodies against mouse CD45 (APCCy7- or producing innate-like T cells, including type I natural killer T AF700-conjugated), CD3 (Pacific Blue-conjugated), TCRd (NKT)17 and gdT17 cells (19, 20). Interestingly, the level of IL7 (PerCpCy5.5-conjugated), TCRb (FITC-conjugated), Vg1 (APC- increases with tumor progression in peritoneal exudates from conjugated), Vg4 (PECy7-conjugated), CD27 (PECy7-conjugat- ovarian cancer-bearing mice (16). Moreover, levels of IL7 have ed), CD127 (FITC- or PECy7-conjugated), IL17A (PE-conjugat- been associated with poor prognosis in patients with prostate ed), CD69 (AF700-conjugated), CD44 (PE-conjugated), and cancer (21, 22) and positively correlated with tumor aggres- appropriated isotype controls were purchased from BioLegend, siveness in patients with breast cancers (23). In view of this, it BD Pharmingen, and eBioscience. PBS-57 glycolipid-loaded and is tempting to speculate that the putative regulatory functions of -unloaded control CD1d tetramers (APC-conjugated) were from tumor-derived IL7 on gdT17 cell biology can somehow explain the National Institute of Allergy and Infectious Diseases Tetramer the phenotypes stated above. Facility (Emory University, Atlanta, GA). Propidium iodide was Type I IFNs are composed of several proteins (IFNa, IFNb, purchased from BD Pharmingen. Mouse monoclonal anti-IL17A and atypical IFNs) that exert major antimicrobial functions (24). (clone 17F3) and anti-IL7 (clone M25) antibodies and their There is also accumulating evidence, indicating that type I IFNs are respective isotype controls [MOPC-21 (mouse IgG1) for anti- pivotal cytokines in cancer immunosurveillance and response to IL17A and MCP-11 (mouse IgG2b) for anti-IL7] were purchased anticancer therapies (25). Type I IFNs have been shown to regulate from Bio X Cell. Recombinant mouse IFNb and IL7 were pur- gdT-cell response. Although type I IFNs increase activity of cyto- chased from R&D Systems. Recombinant mouse IL1b and IL23 toxic/IFNg–producing gdT cells (26, 27), they constrain IL17 were purchased from Peprotech. Phorbol 12-myristate 13-acetate production by gdT17 cell subsets in various models of infection (PMA) and ionomycin were from Sigma-Aldrich. (28–30). For instance, this effect is associated with the suppressive activity of type I IFN-induced IL27 (31–36). However, the regu- Mouse tumor models latory role of type I IFNs on gdT17-cell activity in cancer is The C57BL/6 mouse breast carcinoma cell line AT-3 was currently unknown. obtained from Trina Stewart (Griffith University, Brisbane, Aus- We have here analyzed the role and regulatory mechanisms of tralia; ref. 38), 4T1.2 tumor cells were kindly provided by Robin gdT17 cells in cancer immunosurveillance using two syngeneic Anderson (Peter MacCallum Cancer Centre, Melbourne, Austra- mouse models of breast cancer. Similar to the human disease, lia). Cells were cultured and maintained in DMEM supplemented both cell lines enable growth of highly metastatic primary tumors with 10% FCS, 2mmol/L L-glutamine, penicillin/streptomycin (37). We demonstrate that gdT cells are a prime source of IL17A and sodium pyruvate and 55nmol/L 2-mercaptoethanol. For within the tumor and that T-cell receptor (TCR) d-deficient mice as mouse cancer models, mice