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Levels of T-Bet and Eomesodermin Transplantation Is Associated with Reduced Allogeneic Hematopoietic Stem Cell NK Cell Functiona

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Levels of T-Bet and Eomesodermin Transplantation Is Associated with Reduced Allogeneic Hematopoietic Stem Cell NK Cell Functiona NK Cell Functional Impairment after Allogeneic Hematopoietic Stem Cell Transplantation Is Associated with Reduced Levels of T-bet and Eomesodermin This information is current as of September 27, 2021. Federico Simonetta, Amandine Pradier, Carine Bosshard, Stavroula Masouridi-Levrat, Yves Chalandon and Eddy Roosnek J Immunol 2015; 195:4712-4720; Prepublished online 5 October 2015; Downloaded from doi: 10.4049/jimmunol.1501522 http://www.jimmunol.org/content/195/10/4712 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2015/10/03/jimmunol.150152 Material 2.DCSupplemental References This article cites 28 articles, 14 of which you can access for free at: http://www.jimmunol.org/content/195/10/4712.full#ref-list-1 Why The JI? Submit online. by guest on September 27, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2015 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology NK Cell Functional Impairment after Allogeneic Hematopoietic Stem Cell Transplantation Is Associated with Reduced Levels of T-bet and Eomesodermin Federico Simonetta, Amandine Pradier, Carine Bosshard, Stavroula Masouridi-Levrat, Yves Chalandon, and Eddy Roosnek NK cells play a major role in protection against tumor recurrence and infection after allogeneic hematopoietic stem cell trans- plantation (HSCT). It has been shown that NK cell function after HSCT is impaired, but underlying molecular mechanisms are not well-known. In this report we show that the level of T-bet and Eomesodermin (Eomes), two T-box transcription factors regulating lymphocyte effector functions, is strongly reduced in NK cells from HSCT recipients compared with healthy control subjects. Reduction of T-bet and Eomes expression appeared early and persisted for years after HSCT, affecting all peripheral Downloaded from blood NK cells independently of their differentiation status. Reduced T-bet levels in NK cells from allogeneic HSCT recipients significantly correlated with reduced perforin expression. Acute, but not chronic, graft-versus-host disease, as well as CMV reac- tivation, was associated with further downregulation of T-bet expression in NK cells. Lower levels of T-bet expression in NK cells were associated with less favorable outcome after HSCTas a result of increased nonrelapse mortality. Collectively, our results provide a possible molecular explanation for the previously reported functional exhaustion of NK cells after allogeneic HSCT and suggest an impact of the NK transcriptional machinery status on HSCT outcome. The Journal of Immunology, 2015, 195: 4712–4720. http://www.jimmunol.org/ llogeneic hematopoietic stem cell transplantation (HSCT) differentiation (7, 10, 11) and are necessary for maintenance and is a well-established therapeutic modality for a variety differentiation of peripheral NK cells, whereas their deletion in A of hematological malignancies. Unfortunately, delayed mature NK cells results in reversion to an immature phenotype immune reconstitution after HSCT exposes patients to high risk (9). Finally, reduction in T-bet and Eomes occurs during ageing for relapse and infections. NK cells are the first donor-derived (12) or as a consequence of cell exhaustion after lymphopenia lymphocyte subset to recover after HSCT and may therefore play (13). Importantly, low levels of T-bet and Eomes are associated a role in protection against tumors and infections before res- with impairment of NK cell function (13–15) including antitu- toration of T cell immunity (1). However, several reports have moral activity (13, 16). by guest on September 27, 2021 described impaired NK cell function (2, 3) after HSCT and how, In this work, we show that T-bet and Eomes levels are signifi- as a consequence, their capacity to lyse cancer cells is reduced. cantly reduced in NK cells after HSCT and that low T-bet and Currently, underlying molecular mechanisms for this defect are Eomes expression is associated with impaired effector molecules ill-defined. production. Our results identify T-bet and Eomes downregulation T-bet and Eomesodermin (Eomes), two T-box transcription as a previously unraveled molecular defect underlying impaired factors, are master regulators of T cell effector functions including NK function after allogeneic HSCT. cytotoxicity and IFN-g production (4, 5). Moreover, recent reports have shown that these two transcription factors also regulate Materials and Methods maturation and function of NK cells. Murine (6) and human (7, 8) Study design, patients, and control subjects NK cells express T-bet and Eomes constitutively, and mice lacking both T-bet and Eomes are deprived of NK cells (9). Moreover, the Peripheral blood samples were obtained from 72 patients at different time two T-box transcription factors are modulated during NK cell points after allogeneic HSCT. Blood samples from 44 healthy blood donors from the Geneva University Hospitals blood transfusion center served as control. Written, informed consent was provided by all healthy donors and HSCT recipients enrolled, and the study was approved by the ethics Division of Hematology, Department of Medical Specialties, Geneva University committee of the Geneva University Hospitals. Hospitals, University of Geneva, CH-1211 Geneva 14, Switzerland Clinical characteristics of patients are summarized in Table I. Median age at HSCT was 50 y (range 18–70 y). The analysis was performed at Received for publication July 7, 2015. Accepted for publication September 11, 2015. a median of 8 mo after transplantation (range 1–143 mo). One third of This work was supported by grants from the Swiss National Science Foundation and patients was female (24 patients, 33%) and two thirds were male (48 Swiss Cancer Research (E.R.). patients, 67%). Indications for allogeneic HSCT included acute myeloid Address correspondence and reprint requests to Dr. Federico Simonetta, Division of leukemia (33 patients, 46%), acute lymphocytic leukemia (11 patients, Hematology, Department of Medical Specialties, Geneva University Hospitals, Rue 15%), lymphoma (8 patients, 11%), myelodysplastic syndrome (8 patients, Gabrielle-Perret-Gentil 4, CH-1211 Geneva 14, Switzerland. E-mail address: 11%), myeloproliferative syndrome (6 patients, 8%), multiple myeloma (3 [email protected] patients, 4%), chronic myeloid leukemia (2 patients, 3%), or aplastic The online version of this article contains supplemental material. anemia (1 patient). Most patients received peripheral blood stem cell (PBSC) grafts (64 Abbreviations used in this article: GvHD, graft-versus-host disease; HC, healthy control subject; HSCT, hematopoietic stem cell transplantation; MFIR, median fluo- patients, 91%), whereas 6 patients (8%) received bone marrow and 1 patient rescence intensity ratio; NRM, nonrelapse mortality; PBSC, peripheral blood stem received cord blood. Thirty-five patients (49%) received grafts from cell. an HLA-identical sibling and 25 patients (34%) from an HLA-matched unrelated donor, whereas 12 (17%) patients received grafts from an Copyright Ó 2015 by The American Association of Immunologists, Inc. 0022-1767/15/$25.00 HLA-mismatched relative or unrelated donor. Myeloablative conditioning www.jimmunol.org/cgi/doi/10.4049/jimmunol.1501522 The Journal of Immunology 4713 regimen was used in 34 (47%) patients, whereas 38 (53%) patients received Table I. Clinical characteristics of HSCT recipients reduced intensity conditioning. Almost half of the patients (35 patients, 49%) received partially T cell–depleted grafts consisting of T cell depletion Clinical Characteristics Patients (n =72) by alemtuzumab in the bag with T cell add-back at day 1 (17). Graft-versus-host disease (GvHD) prophylaxis consisted of cyclosporine Age (range), y (for 3 mo duration in the absence of GvHD in the case of partial T cell Age at transplant 50 (18–70) depletion and for 6 mo for T cell–repleted graft recipients) in combina- Age at analysis 51 (20–72) tion with either methotrexate, in case of myeloablative conditioning reg- Time from transplant (range), mo 8 (1–143) imen, or mycophenolate mofetil for patients transplanted after reduced Sex, n (%) intensity conditioning. T cell–depleted graft recipients also received Male 48 (67) methylprednisolone on days 22 and 21. Acute or chronic GvHD was Female 24 (33) treated with corticosteroids alone or in combination with mycophenolate Primary disease, n (%) mofetil and/or cyclosporine. Patients received antiviral prophylaxis with AML 33 (46) valacyclovir for at least 2 y after transplantation, which was discontinued ALL 11 (15) in the absence of active viral replication or GvHD. Lymphoma 8 (11) MDS 8 (11) FACS analysis MPS 6 (8) Multiple myeloma 3 (4) PBMCs were isolated from anticoagulated blood by Ficoll density gradient CML 2 (3) centrifugation. PBMCs were analyzed by flow cytometry.
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