Preactivation with IL-12, IL-15, and IL-18 Induces CD25 and a Functional High-Affinity IL-2 Receptor on Human Cytokine-Induced Memory-like Natural Killer.

Slides:

Advertisements

Similar presentations

High-Risk Acute Lymphoblastic Leukemia Cells with bcr-abl and Ink4a/Arf Mutations Retain Susceptibility to Alloreactive T Cells Faith M. Young, Andrew.

Advertisements

Notch signaling induces cytoplasmic CD3ϵ expression in human differentiating NK cells by Magda De Smedt, Tom Taghon, Inge Van de Walle, Greet De Smet,

CD4+CD25+ Immunoregulatory T Cells

The Fifth Epidermal Growth Factor–like Region of Thrombomodulin Alleviates Murine Graft-versus-Host Disease in a G-Protein Coupled Receptor 15 Dependent.

Identification of Stem Cell Transcriptional Programs Normally Expressed in Embryonic and Neural Stem Cells in Alloreactive CD8+ T Cells Mediating Graft-versus-Host.

Juyang Kim, Wongyoung Kim, Hyun J. Kim, Sohye Park, Hyun-A

Krystel Vincent, Marie-Pierre Hardy, Assya Trofimov, Céline M

Volume 136, Issue 4, Pages e3 (April 2009)

Induction of Immunity to Neuroblastoma Early after Syngeneic Hematopoietic Stem Cell Transplantation Using a Novel Mouse Tumor Vaccine Weiqing Jing,

Ping Zhang, Jieying Wu, Divino Deoliveira, Nelson J. Chao, Benny J

Cord Blood–Derived and Peripheral Blood–Derived Cytokine-Induced Killer Cells Are Sensitive to Fas-Mediated Apoptosis Ludovic Durrieu, Mame Massar Dieng,

by Norman Nausch, Ioanna E

Ex Vivo Rapamycin Generates Th1/Tc1 or Th2/Tc2 Effector T Cells With Enhanced In Vivo Function and Differential Sensitivity to Post-transplant Rapamycin.

LBH589 Enhances T Cell Activation In Vivo and Accelerates Graft-versus-Host Disease in Mice Dapeng Wang, Cristina Iclozan, Chen Liu, Changqing Xia, Claudio.

IL-12hi Rapamycin-Conditioned Dendritic Cells Mediate IFN-γ–Dependent Apoptosis of Alloreactive CD4+ T Cells In Vitro and Reduce Lethal Graft-Versus-Host.

Volume 30, Issue 4, Pages (April 2009)

Hans-Peter Raué, Carol Beadling, Jennifer Haun, Mark K. Slifka

Human Progenitor Cells Rapidly Mobilized by AMD3100 Repopulate NOD/SCID Mice with Increased Frequency in Comparison to Cells from the Same Donor Mobilized.

Effects of the NK Cell Recovery on Outcomes of Unmanipulated Haploidentical Blood and Marrow Transplantation for Patients with Hematologic Malignancies

Volume 138, Issue 5, Pages e2 (May 2010)

Mycophenolic Acid Inhibits Natural Killer Cell Proliferation and Cytotoxic Function: A Possible Disadvantage of Including Mycophenolate Mofetil in the.

Human CD4+CD25+ Cells in Combination with CD34+ Cells and Thymoglobulin to Prevent Anti-hematopoietic Stem Cell T Cell Alloreactivity Dolores Mahmud,

Growth and Differentiation Advantages of CD4+OX40+ T Cells from Allogeneic Hematopoietic Stem Cell Transplantation Recipients Takero Shindo, Takayuki.

Low c-Kit Expression Level Induced by Stem Cell Factor Does Not Compromise Transplantation of Hematopoietic Stem Cells Chia-Ling Chen, Katerina Faltusova,

FLT3 ligand administration after hematopoietic cell transplantation increases circulating dendritic cell precursors that can be activated by CpG oligodeoxynucleotides.

CD134-Allodepletion Allows Selective Elimination of Alloreactive Human T Cells without Loss of Virus-Specific and Leukemia-Specific Effectors Xupeng.

IL-21 inhibits T cell IL-2 production and impairs Treg homeostasis

Volume 8, Issue 5, Pages (September 2014)

Virally infected and matured human dendritic cells activate natural killer cells via cooperative activity of plasma membrane-bound TNF and IL-15 by Lazar.

Generation of Highly Cytotoxic Natural Killer Cells for Treatment of Acute Myelogenous Leukemia Using a Feeder-Free, Particle-Based Approach Jeremiah.

Reconstitution of Natural Killer Cells in HLA-Matched HSCT after Reduced-Intensity Conditioning: Impact on Clinical Outcome Caroline Pical-Izard, Roberto.

Combination Therapy Using IL-2 and Anti-CD25 Results in Augmented Natural Killer Cell–Mediated Antitumor Responses William H.D. Hallett, Erik Ames, Maite.

Blocking Activator Protein 1 Activity in Donor Cells Reduces Severity of Acute Graft- Versus-Host Disease through Reciprocal Regulation of IL-17–Producing.

Expansion and Homing of Adoptively Transferred Human Natural Killer Cells in Immunodeficient Mice Varies with Product Preparation and In Vivo Cytokine.

Bone Marrow–Derived Mesenchymal Stromal Cells from Patients with Sickle Cell Disease Display Intact Functionality Elizabeth O. Stenger, Raghavan Chinnadurai,

T Cell–Mediated Rejection of Human CD34+ Cells Is Prevented by Costimulatory Blockade in a Xenograft Model Annie L. Oh, Dolores Mahmud, Benedetta Nicolini,

Quantity and Quality Reconstitution of NKG2A+ Natural Killer Cells Are Associated with Graft-versus-Host Disease after Allogeneic Hematopoietic Cell Transplantation

Clinically Relevant Expansion of Hematopoietic Stem Cells with Conserved Function in a Single-Use, Closed-System Bioprocess Gerard J. Madlambayan, Ian.

Volume 33, Issue 1, Pages (July 2010)

Host Basophils Are Dispensable for Induction of Donor T Helper 2 Cell Differentiation and Severity of Experimental Graft-versus-Host Disease Isao Tawara,

Arming Cytokine-induced Killer Cells With Chimeric Antigen Receptors: CD28 Outperforms Combined CD28–OX40 “Super-stimulation” Andreas A Hombach, Gunter.

Elevation of Intracellular Cyclic AMP in Alloreactive CD4+ T Cells Induces Alloantigen- Specific Tolerance That Can Prevent GVHD Lethality In Vivo Matthew.

Tracking ex vivo-expanded CD4+CD25+ and CD8+CD25+ regulatory T cells after infusion to prevent donor lymphocyte infusion-induced lethal acute graft-versus-host.

Volume 13, Issue 12, Pages (December 2015)

In Situ Activation and Expansion of Host Tregs: A New Approach to Enhance Donor Chimerism and Stable Engraftment in Major Histocompatibility Complex-Matched.

Activated Notch Supports Development of Cytokine Producing NK Cells Which Are Hyporesponsive and Fail to Acquire NK Cell Effector Functions Veronika.

Volume 22, Issue 3, Pages (March 2005)

Volume 24, Issue 9, Pages (September 2016)

The Notch Ligands Jagged2, Delta1, and Delta4 Induce Differentiation and Expansion of Functional Human NK Cells from CD34+ Cord Blood Hematopoietic Progenitor.

CpG-Induced Myeloid CD11b+Gr-1+ Cells Efficiently Suppress T Cell–Mediated Immunoreactivity and Graft-Versus-Host Disease in a Murine Model of Allogeneic.

CD25 expression distinguishes functionally distinct alloreactive CD4+ CD134+ (OX40+) T-cell subsets in acute graft-versus-host disease Philip R Streeter,

Volume 135, Issue 3, Pages (September 2008)

Volume 38, Issue 3, Pages (March 2013)

Volume 17, Issue 2, Pages (February 2009)

Opposing Effects of TGF-β and IL-15 Cytokines Control the Number of Short-Lived Effector CD8+ T Cells Shomyseh Sanjabi, Munir M. Mosaheb, Richard A.

Volume 25, Issue 1, Pages (January 2017)

Volume 29, Issue 4, Pages (October 2008)

Cytokine-Induced Memory-Like Differentiation Enhances Unlicensed Natural Killer Cell Antileukemia and FcγRIIIa-Triggered Responses Julia A. Wagner, Melissa.

Volume 16, Issue 4, Pages (April 2002)

IL-4 and IL-13 Alter Plasmacytoid Dendritic Cell Responsiveness to CpG DNA and Herpes Simplex Virus-1 Jurjen Tel, Ruurd Torensma, Carl G. Figdor, I.

Volume 26, Issue 4, Pages (April 2007)

Volume 32, Issue 6, Pages (June 2010)

Volume 32, Issue 1, Pages (January 2010)

Volume 131, Issue 6, Pages (December 2006)

Volume 22, Issue 8, Pages (February 2018)

IL-33, IL-25, and TSLP induce a distinct phenotypic and activation profile in human type 2 innate lymphoid cells by Ana Camelo, Guglielmo Rosignoli, Yoichiro.

Notch 1 Signaling Regulates Peripheral T Cell Activation

by Martin Felices, Behiye Kodal, Peter Hinderlie, Michael F

ALT-803 stimulates proliferation and activation of human NK cells and T cells in vitro. ALT-803 stimulates proliferation and activation of human NK cells.

Volume 13, Issue 11, Pages (December 2015)

Presentation transcript:

Preactivation with IL-12, IL-15, and IL-18 Induces CD25 and a Functional High-Affinity IL-2 Receptor on Human Cytokine-Induced Memory-like Natural Killer Cells Jeffrey W. Leong, Julie M. Chase, Rizwan Romee, Stephanie E. Schneider, Ryan P. Sullivan, Megan A. Cooper, Todd A. Fehniger Biology of Blood and Marrow Transplantation Volume 20, Issue 4, Pages 463-473 (April 2014) DOI: 10.1016/j.bbmt.2014.01.006 Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 CD25 expression is induced on human NK cells by combined cytokine activation. Freshly purified human NK cells were stained for baseline CD25 expression and activated overnight with the following cytokines (1 ng/mL IL-15, LD15; 1 nM IL-2; 100 ng/mL IL-15; 10 ng/mL IL-12; 50 ng/mL IL-18) or combinations of the cytokines as indicated. After culture for 16 hours, cells were washed extensively and cultured for an additional 3 or 7 days in LD15 for all later time points. (A) Cell surface CD25 is induced after activation with IL-12, IL-15, and IL-18, compared with LD15, on NK cells. Representative flow histograms at baseline, 16 hours, 3 days, or 7 days after the initiation of IL-12, IL-15, and IL-18 preactivation or culture in LD15, demonstrating the marked increase and prolonged expression of CD25. (B and C) CD25 percentage (B) and mean fluorescence intensity (MFI) (C) of NK cells at various time points after cytokine activation. Summary data represent n = 6 donors (3 independent experiments). Data shown are mean ± SEM of percentage or the MFI of CD25+ NK cells. Significance is shown comparing each time point versus fresh (baseline) NK cells, and was calculated by 1-way ANOVA. These data show that although individually IL-2, IL-12, IL-15, and IL-18 induce CD25+ CD56dim NK cells, the combinations of IL-15+IL-18 and IL-12+IL-18 result in higher per cell CD25 expression. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Picomolar concentrations of IL-2 induce phospho-STAT5 expression in preactivated NK cells. Purified NK cells were preactivated with LD15 (control), IL-12+IL-18, or IL-15+IL-18, washed extensively, and replated in cytokine-free media for 2 days to allow recovery to a resting state. IL-2 was added at the indicated concentrations for 15 minutes and levels of phospho-STAT5 were analyzed by intracellular flow cytometry. Shown is 1 representative donor (A), or summary data of the mean ± SEM percentage pSTAT5 positive CD56dim NK cells from n = 3 donors and experiments (B). Preactivated NK cells respond to 10 and 100 pM IL-2 with enhanced STAT5 phosphorylation over control. Significance was calculated by ANOVA. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Induced CD25 on preactivated NK cells results in a functional high-affinity IL-2Rαβγ that signals for enhanced IFN-γ production. Purified NK cells were activated for 16 hours with IL-12+IL-18 or IL-15+IL-18, washed extensively, and replated in cytokine-free media for 2 days to allow recovery to a resting state. IL-2 was then added at the indicated doses in combination with IL-12 (1.25 ng/mL). After 6 hours, IFN-γ was measured by intracellular flow cytometry. (A) Representative flow cytometric plots from 1 preactivated donor depicting dose-dependent IFN-γ production costimulated by picomolar IL-2. (B) Representative flow plots from IL-15+IL-18 preactivated NK cells that were subgated based on CD25 expression and IFN-γ expression with stimulation by IL-12 + 1 or 100 pM IL-2. (C) Summary data of preactivated CD56dim NK cell IFN-γ production with IL-12 combined with various doses of IL-2, shown as mean ± SEM normalized IFN-γ percentage (n = 4 donors). (D) In a separate assay, cells were pretreated for 1 hour with either an anti-CD25–blocking antibody or IgG1 isotype control, followed by stimulation with 1.25 ng/mL IL-12 and 10-fold dilutions of IL-2. In (B,C), data are shown with the maximum IFN-γ positive percentage in each donor (n = 4) normalized to 100%, as absolute IFN-γ+ percentages between donors are variable (range of maximum %IFN-γ+:14% to 66% for IL-12+IL-18 preactivated cells, 28% to 86% for IL-15+IL-18 preactivated cells). Significance was calculated by ANOVA. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Low concentrations of IL-2 enhance preactivated CD25+ CD56dim NK cell killing of K562 leukemia target cells. Flow-sorted CD56dim NK cells were preactivated with LD15, IL-12+IL-18, or IL-15+IL-18, washed, and replated in cytokine-free media for 2 days. Cells were then incubated for 1 hour with an isotype or anti-CD25–blocking antibody before activation with dilutions of IL-2. After 24 hours, cells were harvested and plated in a 4-hour in vitro flow-based killing assay with CFSE-labeled K562 cells. Shown is mean ± SEM percent specific killing at a 4:1 effector to target ratio preincubated with (A) an isotype control or (B,C) an anti-CD25– blocking antibody. (D,E) Killing by preactivated CD56dim NK cells is NK cell dose- (effector to target ratio) dependent, shown at 10 pM IL-2, with anti-CD25 or isotype control preincubation. Data are summarized from 3 to 5 donors in 2 independent experiments. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 Low concentrations of IL-2 stimulate the proliferation of preactivated CD56dim NK cells. CFSE-labeled, flow-sorted CD56dim NK cells were preactivated with low-dose IL-15, IL-12+IL-18, or IL-15+IL-18 for 16 hours, washed extensively, and blocked with an isotype or anti-CD25–blocking antibody. Dilutions of IL-2 were then added, with subsequent media, cytokine, and blocking antibody additions occurring every other day. After 7 days, cells were harvested and analyzed for CFSE dilution by flow cytometry. Analysis was restricted to live cells by 7-AAD exclusion. (A) Data shown represent histograms from 1 donor. Both isotype-blocked (open histograms) and anti-CD25–blocked (shaded histograms) NK cell proliferation are shown. (B and C) Summary data depicting the variability of proliferation between IL-12+IL-18 or IL-15+IL-18 activated CD56dim NK cells from different donors and the dependence of picomolar IL-2 mediated proliferation on CD25/IL-2Rαβγ. Significance was assessed by 2-way ANOVA. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 6 Exogenous IL-2 supports CIML NK cells in a NSG xenograft model. (A) Purified human NK cells were preactivated with IL-12, IL-15, and IL-18 (CIML) or low-dose IL-15 only (control) for 16 hours, washed, and identical numbers of NK cells were transferred into NSG mice, followed by injections of 75,000 IU i.p. of rhIL-2 every other day. After 7 days, mice were assessed for human NK cell content and CIML NK cell recall responses. (B) Representative flow plots from the peripheral blood of mice injected with the same number of CIML or control NK cells from the same donor 7 days earlier. Human CD45+ cells are CD56+CD3- NK cells with preserved CD56bright and CD56dim NK cells subsets. (C) Summary data from 3 different NK cell donors transferred into a total of 12 NSG mice demonstrating superior engraftment of CIML, compared with control NK cells. The ratio of human to mouse CD45+ cells, representing the relative abundance of human NK cells, is used to control for differences in blood volume obtained. (D) Mouse splenocytes containing adoptively transferred human NK cells were evaluated for a recall IFN-γ response after IL-12+IL-15 restimulation, demonstrating that CIML NK cells exhibit preserved, enhanced functionality in this xenograft model. Significance was assessed by t-test, with *P < .05, **P < .01, and ***P < .001. Biology of Blood and Marrow Transplantation 2014 20, 463-473DOI: (10.1016/j.bbmt.2014.01.006) Copyright © 2014 American Society for Blood and Marrow Transplantation Terms and Conditions