title: Effect of nasopharyngeal carcinoma-derived exosomes on human regulatory T cells.
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).
  
  
  CONCLUSIONS
  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.affiliation: 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).
authors.forename: Dhafer
authors.initials: D
authors.lastname: Mrizak
authors.affiliation: 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).
authors.forename: Nathalie
authors.initials: N
authors.lastname: Martin
authors.affiliation: 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).
authors.forename: Clément
authors.initials: C
authors.lastname: Barjon
authors.affiliation: 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).
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Jimenez-Pailhes
authors.affiliation: 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).
authors.forename: Rami
authors.initials: R
authors.lastname: Mustapha
authors.affiliation: 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).
authors.forename: Toshiro
authors.initials: T
authors.lastname: Niki
authors.affiliation: 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).
authors.forename: Joël
authors.initials: J
authors.lastname: Guigay
authors.affiliation: 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).
authors.forename: Véronique
authors.initials: V
authors.lastname: Pancré
authors.affiliation: 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).
authors.forename: Yvan
authors.initials: Y
authors.lastname: de Launoit
authors.affiliation: 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).
authors.forename: Pierre
authors.initials: P
authors.lastname: Busson
authors.affiliation: 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.
authors.forename: Olivier
authors.initials: O
authors.lastname: Moralès
authors.affiliation: 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.
authors.forename: Nadira
authors.initials: N
authors.lastname: Delhem
mesh_terms: D000818:Animals; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002277:Carcinoma / Q000276:immunology / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D054418:Chemokine CCL20 / Q000378:metabolism; D055354:Exosomes / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D064593:Heterografts; D006801:Humans; D016753:Interleukin-10 / Q000276:immunology; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009303:Nasopharyngeal Neoplasms / Q000276:immunology / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000276:immunology
chemical_list: C108355:CCL20 protein, human; C493447:CCL20 protein, mouse; D054418:Chemokine CCL20; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; C508609:IL10 protein, human; C556062:IL10 protein, mouse; C508017:IL2RA protein, human; D053645:Interleukin-2 Receptor alpha Subunit; D053773:Transforming Growth Factor beta1; D016753:Interleukin-10