Dhafer Mrizak1, Nathalie Martin1, Clément Barjon1, Anne-Sophie Jimenez-Pailhes1, Rami Mustapha1, Toshiro Niki1, Joël Guigay1, Véronique Pancré1, Yvan de Launoit1, Pierre Busson1, Olivier Moralès2, Nadira Delhem2. 1. CNRS UMR 8161, Institut de Biologie de Lille, Université de Lille, Institut Pasteur de Lille, IFR142, Lille, France (DM, NM, RM, VP, YdL, OM, ND); Université Paris-sud, CNRS UMR 8126 and Institut Gustave Roussy, Villejuif, France (CB, ASJP, PB); GalPharma Co., Ltd. 884-3-302, Fuseishi-Cho, Takamatsu-shi, Kagawa 761-8071 Japan (TN); Department of Immunology, Kagawa University. 1750-1 Ikenobe, Miki-Cho, Kagawa 761-0793 Japan (TN); Department of head and Neck Oncology, Institut Gustave Roussy, Villejuif, France (JG). 2. CNRS UMR 8161, Institut de Biologie de Lille, Université de Lille, Institut Pasteur de Lille, IFR142, Lille, France (DM, NM, RM, VP, YdL, OM, ND); Université Paris-sud, CNRS UMR 8126 and Institut Gustave Roussy, Villejuif, France (CB, ASJP, PB); GalPharma Co., Ltd. 884-3-302, Fuseishi-Cho, Takamatsu-shi, Kagawa 761-8071 Japan (TN); Department of Immunology, Kagawa University. 1750-1 Ikenobe, Miki-Cho, Kagawa 761-0793 Japan (TN); Department of head and Neck Oncology, Institut Gustave Roussy, Villejuif, France (JG). nadira.delhem@ibl.cnrs.fr olivier.morales@ibl.cnrs.fr.
Abstract
BACKGROUND: Regulatory T cells (Treg) and tumor-exosomes are thought to play a role in preventing the rejection of malignant cells in patients bearing nasopharyngeal carcinoma (NPC). METHODS: Treg recruitment by exosomes derived from NPC cell lines (C15/C17-Exo), exosomes isolated from NPC patients' plasma (Patient-Exo), and CCL20 were tested in vitro using Boyden chamber assays and in vivo using a xenograft SCID mouse model (n = 5), both in the presence and absence of anti-CCL20 monoclonal antibodies (mAb). Impact of these NPC exosomes (NPC-Exo) on Treg phenotype and function was determined using adapted assays (FACS, Q-PCR, ELISA, and MLR). Experiments were performed in comparison with exosomes derived from plasma of healthy donors (HD-Exo). The Student's t test was used for group comparisons. All statistical tests were two-sided. RESULTS: CCL20 allowed the intratumoral recruitment of human Treg. NPC-Exo also facilitated Treg recruitment (3.30 ± 0.34 fold increase, P < .001), which was statistically significantly inhibited (P < .001) by an anti-CCL20 blocking mAb. NPC-Exo also recruited conventional CD4(+)CD25(-) T cells and mediated their conversion into inhibitory CD4(+)CD25(high) cells. Moreover, NPC-Exo enhanced (P = .0048) the expansion of human Treg, inducing the generation of Tim3(Low) Treg with increased expression of CD25 and FOXP3. Finally, NPC-Exo induced an overexpression of cell markers associated with Treg phenotype, properties and recruitment capacity. For example, GZMB mean fold change was 21.45 ± 1.75 (P < .001). These results were consistent with a stronger suppression of responder cells' proliferation and the secretion of immunosuppressive cytokines (IL10, TGFB1). CONCLUSION: Interactions between NPC-Exo and Treg represent a newly defined mechanism that may be involved in regulating peripheral tolerance by tumors and in supporting immune evasion in human NPC.
BACKGROUND: Regulatory T cells (Treg) and tumor-exosomes are thought to play a role in preventing the rejection of malignant cells in patients bearing nasopharyngeal carcinoma (NPC). METHODS: Treg recruitment by exosomes derived from NPC cell lines (C15/C17-Exo), exosomes isolated from NPC patients' plasma (Patient-Exo), and CCL20 were tested in vitro using Boyden chamber assays and in vivo using a xenograft SCIDmouse model (n = 5), both in the presence and absence of anti-CCL20 monoclonal antibodies (mAb). Impact of these NPC exosomes (NPC-Exo) on Treg phenotype and function was determined using adapted assays (FACS, Q-PCR, ELISA, and MLR). Experiments were performed in comparison with exosomes derived from plasma of healthy donors (HD-Exo). The Student's t test was used for group comparisons. All statistical tests were two-sided. RESULTS:CCL20 allowed the intratumoral recruitment of human Treg. NPC-Exo also facilitated Treg recruitment (3.30 ± 0.34 fold increase, P < .001), which was statistically significantly inhibited (P < .001) by an anti-CCL20 blocking mAb. NPC-Exo also recruited conventional CD4(+)CD25(-) T cells and mediated their conversion into inhibitory CD4(+)CD25(high) cells. Moreover, NPC-Exo enhanced (P = .0048) the expansion of human Treg, inducing the generation of Tim3(Low) Treg with increased expression of CD25 and FOXP3. Finally, NPC-Exo induced an overexpression of cell markers associated with Treg phenotype, properties and recruitment capacity. For example, GZMB mean fold change was 21.45 ± 1.75 (P < .001). These results were consistent with a stronger suppression of responder cells' proliferation and the secretion of immunosuppressive cytokines (IL10, TGFB1). CONCLUSION: Interactions between NPC-Exo and Treg represent a newly defined mechanism that may be involved in regulating peripheral tolerance by tumors and in supporting immune evasion in human NPC.
Authors: Shannon M Clayton; Joehleen A Archard; Joseph Wagner; D Gregory Farwell; Arnaud F Bewley; Angela Beliveau; Andrew Birkeland; Shyam Rao; Marianne Abouyared; Peter C Belafsky; Johnathon D Anderson Journal: Stem Cells Dev Date: 2020-01-30 Impact factor: 3.272