Restricted accessResearch articleFirst published online 2018-09
Immunosuppression of Human Adipose-Derived Stem Cells on T Cell Subsets via the Reduction of NF-kappaB Activation Mediated by PD-L1/PD-1 and Gal-9/TIM-3 Pathways
Adipose-derived stem cells (ADSCs) are a type of multipotent mesenchymal stem cells with immunosuppressive capacities. However, the underlying mechanisms involved in the inhibitory effects of ADSCs on T cells are not completely elucidated. In this study, human peripheral blood mononuclear cells (PBMCs) stimulated with anti-CD3/CD28 antibody-coated beads were cultured with or without allogeneic ADSCs (ADSC-to-PBMC ratio, 1:5). Surface marker levels, violet-labeled cell proliferation, apoptosis, interferon-gamma (IFN-gamma) production, and nuclear factor-kappaB (NF-kappaB) phosphorylation of CD4+ and CD8+ T cells were detected using flow cytometry. It was observed that ADSCs significantly suppressed the proliferation and IFN-gamma production but enhanced apoptosis of both CD4+ and CD8+ T cells in T cell receptor (TCR)-stimulated PBMCs. The expressions of programmed death-ligand 1 (PD-L1) and galectin 9 (Gal-9) on ADSCs were significantly upregulated and induced during coculture with PBMCs. TCR-stimulated CD4+ and CD8+ T cells cultured with ADSCs had higher expression levels of programmed death-1 (PD-1) and T cell immunoglobulin and mucin-containing protein-3 (TIM-3) than those in cells cultured without ADSCs. Moreover, the suppressive effects of ADSCs on T cells in terms of proliferation and IFN-gamma production were significantly reversed in the presence of anti-PD-L1 and anti-Gal-9 antibodies. Importantly, the phosphorylation of NF-kappaB in CD4+ and CD8+ T cells cocultured with ADSCs was significantly inhibited, and this inhibition was significantly attenuated via the PD-L1 and Gal-9 blockades. In conclusion, human ADSCs perform immunoregulatory functions partially through the inhibition of NF-kappaB activation in T cells via the PD-L1/PD-1 and Gal-9/TIM-3 pathways, which provide new insights into the mechanism of human ADSC-mediated immunomodulation.
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References
1.
CaoF, LiuT, XuY, XuD and FengS. (2015). Culture and properties of adipose-derived mesenchymal stem cells: characteristics in vitro and immunosuppression in vivo. Int J Clin Exp Pathol, 8:7694–7709.
2.
YooKH, JangIK, LeeMW, KimHE, YangMS, EomY, LeeJE, KimYJ, YangSK, et al. (2009). Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol, 259:150–156.
3.
CaruanaG, BertozziN, BoschiE, Pio GriecoM, GrignaffiniE and RaposioE. (2015). Role of adipose-derived stem cells in chronic cutaneous wound healing. Ann Ital Chir, 86:1–4.
4.
Alvaro-GraciaJM, JoverJA, Garcia-VicunaR, CarrenoL, AlonsoA, MarsalS, BlancoF, Martinez-TaboadaVM, TaylorP, et al. (2017). Intravenous administration of expanded allogeneic adipose-derived mesenchymal stem cells in refractory rheumatoid arthritis (Cx611): results of a multicentre, dose escalation, randomised, single-blind, placebo-controlled phase Ib/IIa clinical trial. Ann Rheum Dis, 76:196–202.
5.
PanesJ, Garcia-OlmoD, Van AsscheG, ColombelJF, ReinischW, BaumgartDC, DignassA, NachuryM, FerranteM, et al.; ADMIRE CD Study Group Collaborators (2016). Expanded allogeneic adipose-derived mesenchymal stem cells (Cx601) for complex perianal fistulas in Crohn's disease: a phase 3 randomised, double-blind controlled trial. Lancet, 388:1281–1290.
6.
De FrancescoF, RicciG, D'AndreaF, NicolettiGF and FerraroGA. (2015). Human adipose stem cells: from bench to bedside. Tissue Eng B Rev, 21:572–584.
7.
SchafflerA and BuchlerC. (2007). Concise review: adipose tissue-derived stromal cells—basic and clinical implications for novel cell-based therapies. Stem Cells, 25:818–827.
8.
SpaethEL, KiddS and MariniFC. (2012). Tracking inflammation-induced mobilization of mesenchymal stem cells. Methods Mol Biol, 904:173–190.
9.
EggenhoferE, LukF, DahlkeMH and HoogduijnMJ. (2014). The life and fate of mesenchymal stem cells. Front Immunol, 5:148.
10.
SohniA and VerfaillieCM. (2013). Mesenchymal stem cells migration homing and tracking. Stem Cells Int, 2013:130763.
11.
MeliefSM, ZwagingaJJ, FibbeWE and RoelofsH. (2013). Adipose tissue-derived multipotent stromal cells have a higher immunomodulatory capacity than their bone marrow-derived counterparts. Stem Cells Transl Med, 2:455–463.
12.
NajarM, RaicevicG, Fayyad-KazanH, De BruynC, BronD, ToungouzM and LagneauxL. (2013). Impact of different mesenchymal stromal cell types on T-cell activation, proliferation and migration. Int Immunopharmacol, 15:693–702.
13.
YanezR, OviedoA, AldeaM, BuerenJA and LamanaML. (2010). Prostaglandin E2 plays a key role in the immunosuppressive properties of adipose and bone marrow tissue-derived mesenchymal stromal cells. Exp Cell Res, 316:3109–3123.
14.
NajarM, RaicevicG, BoufkerHI, Fayyad KazanH, De BruynC, MeulemanN, BronD, ToungouzM and LagneauxL. (2010). Mesenchymal stromal cells use PGE2 to modulate activation and proliferation of lymphocyte subsets: combined comparison of adipose tissue, Wharton's Jelly and bone marrow sources. Cell Immunol, 264:171–179.
15.
DuffyMM, RitterT, CeredigR and GriffinMD. (2011). Mesenchymal stem cell effects on T-cell effector pathways. Stem Cell Res Ther, 2:34.
16.
SunshineJ and TaubeJM. (2015). PD-1/PD-L1 inhibitors. Curr Opin Pharmacol, 23:32–38.
17.
ZhuC, AndersonAC, Schubart,A, XiongH, ImitolaJ, KhourySJ, ZhengXX, StromTB and KuchrooVK. (2005). The Tim-3 ligand galectin-9 negatively regulates T helper type 1 immunity. Nat Immunol, 6:1245–1252.
18.
GuYZ, XueQ, ChenYJ, YuGH, QingMD, ShenY, WangMY, ShiQ and ZhangXG. (2013). Different roles of PD-L1 and FasL in immunomodulation mediated by human placenta-derived mesenchymal stem cells. Hum Immunol, 74:267–276.
19.
Castro-ManrrezaME, MayaniH, Monroy-GarciaA, Flores-FigueroaE, Chavez-RuedaK, Legorreta-HaquetV, Santiago-OsorioE and MontesinosJJ. (2014). Human mesenchymal stromal cells from adult and neonatal sources: a comparative in vitro analysis of their immunosuppressive properties against T cells. Stem Cells Dev, 23:1217–1232.
20.
TipnisS, ViswanathanC and MajumdarAS. (2010). Immunosuppressive properties of human umbilical cord-derived mesenchymal stem cells: role of B7-H1 and IDO. Immunol Cell Biol, 88:795–806.
21.
YanZ, ZhuansunY, LiuG, ChenR, LiJ and RanP. (2014). Mesenchymal stem cells suppress T cells by inducing apoptosis and through PD-1/B7-H1 interactions. Immunol Lett, 162:248–255.
22.
UngererC, Quade-LyssyP, RadekeHH, HenschlerR, KonigsC, KohlU, SeifriedE and SchuttrumpfJ. (2014). Galectin-9 is a suppressor of T and B cells and predicts the immune modulatory potential of mesenchymal stromal cell preparations. Stem Cells Dev, 23:755–766.
23.
KimSN, LeeHJ, JeonMS, YiT and SongSU. (2015). Galectin-9 is involved in immunosuppression mediated by human bone marrow-derived clonal mesenchymal stem cells. Immune Netw, 15:241–251.
24.
JangIK, YoonHH, YangMS, LeeJE, LeeDH, LeeMW, KimDS and ParkJE. (2014). B7-H1 inhibits T cell proliferation through MHC class II in human mesenchymal stem cells. Transplant Proc, 46:1638–1641.
25.
GerondakisS, FulfordTS, MessinaNL and GrumontRJ. (2014). NF-kappaB control of T cell development. Nat Immunol, 15:15–25.
26.
WeilR and IsraelA. (2004). T-cell-receptor- and B-cell-receptor-mediated activation of NF-kappaB in lymphocytes. Curr Opin Immunol, 16:374–381.
27.
FerrisRL, LuB and KaneLP. (2014). Too much of a good thing? Tim-3 and TCR signaling in T cell exhaustion. J Immunol, 193:1525–1530.
28.
BunnellBA, FlaatM, GagliardiC, PatelB and RipollC. (2008). Adipose-derived stem cells: isolation, expansion and differentiation. Methods, 45:115–120.
29.
DominiciM, Le BlancK, MuellerI, Slaper-CortenbachI, MariniF, KrauseD, DeansR, KeatingA, ProckopD and HorwitzE. (2006). Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy, 8:315–317.
30.
BourinP, BunnellBA, CasteillaL, DominiciM, KatzAJ, MarchKL, RedlH, RubinJP, YoshimuraK and GimbleJM. (2013). Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy, 15:641–648.
31.
PuL, MengM, WuJ, ZhangJ, HouZ, GaoH, XuH, LiuB, TangW, JiangL and LiY. (2017). Compared to the amniotic membrane, Wharton's jelly may be a more suitable source of mesenchymal stem cells for cardiovascular tissue engineering and clinical regeneration. Stem Cell Res Ther, 8:72.
32.
GaoJ, ShiLZ, ZhaoH, ChenJ, XiongL, HeQ, ChenT, RoszikJ, BernatchezC, et al. (2016). Loss of IFN-gamma pathway genes in tumor cells as a mechanism of resistance to anti-CTLA-4 therapy. Cell, 167:397–404.e399.
33.
RubtsovY, GoryunovK, RomanovA, SuzdaltsevaY, SharonovG and TkachukV. (2017). Molecular mechanisms of immunomodulation properties of mesenchymal stromal cells: a new insight into the role of ICAM-1. Stem Cells Int, 2017:6516854.
34.
BlazquezR, Sanchez-MargalloFM, de la RosaO, DalemansW, AlvarezV, TarazonaR and CasadoJG. (2014). Immunomodulatory potential of human adipose mesenchymal stem cells derived exosomes on in vitro stimulated T cells. Front Immunol, 5:556.
35.
MatulaZ, NemethA, LorinczP, SzepesiA, BrozikA, BuzasEI, LowP, NemetK, UherF and UrbanVS. (2016). The role of extracellular vesicle and tunneling nanotube-mediated intercellular cross-talk between mesenchymal stem cells and human peripheral T cells. Stem Cells Dev, 25:1818–1832.
36.
ChinnaduraiR, CoplandIB, PatelSR and GalipeauJ. (2014). IDO-independent suppression of T cell effector function by IFN-gamma-licensed human mesenchymal stromal cells. J Immunol, 192:1491–1501.
37.
KronsteinerB, WolbankS, PeterbauerA, HacklC, RedlH, van GriensvenM and GabrielC. (2011). Human mesenchymal stem cells from adipose tissue and amnion influence T-cells depending on stimulation method and presence of other immune cells. Stem Cells Dev, 20:2115–2126.
38.
GormanJV and ColganJD. (2014). Regulation of T cell responses by the receptor molecule Tim-3. Immunol Res, 59:56–65.
39.
KiesslingMK, LinkeB, BrechmannM, SussD, KrammerPH and GulowK. (2010). Inhibition of NF-kappaB induces a switch from CD95L-dependent to CD95L-independent and JNK-mediated apoptosis in T cells. FEBS Lett, 584:4679–4688.
40.
ChengJ, PhongB, WilsonDC, HirschR and KaneLP. (2011). Akt fine-tunes NF-kappaB-dependent gene expression during T cell activation. J Biol Chem, 286:36076–36085.
41.
ParryRV, ChemnitzJM, FrauwirthKA, LanfrancoAR, BraunsteinI, KobayashiSV, LinsleyPS, ThompsonCB and RileyJL. (2005). CTLA-4 and PD-1 receptors inhibit T-cell activation by distinct mechanisms. Mol Cell Biol, 25:9543–9553.
42.
Di NicolaM, Carlo-StellaC, MagniM, MilanesiM, LongoniPD, MatteucciP, GrisantiS and GianniAM. (2002). Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood, 99:3838–3843.
43.
DaviesLC, HeldringN, KadriN and Le BlancK. (2017). Mesenchymal Stromal cell secretion of programmed death-1 ligands regulates T cell mediated immunosuppression. Stem Cells, 35:766–776.
44.
BenvenutoF, FerrariS, GerdoniE, GualandiF, FrassoniF, PistoiaV, MancardiG and UccelliA. (2007). Human mesenchymal stem cells promote survival of T cells in a quiescent state. Stem Cells, 25:1753–1760.
45.
CuerquisJ, Romieu-MourezR, FrancoisM, RoutyJP, YoungYK, ZhaoJ and EliopoulosN. (2014). Human mesenchymal stromal cells transiently increase cytokine production by activated T cells before suppressing T-cell proliferation: effect of interferon-gamma and tumor necrosis factor-alpha stimulation. Cytotherapy, 16:191–202.
46.
AggarwalS and PittengerMF. (2005). Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood, 105:1815–1822.
47.
TseWT, PendletonJD, BeyerWM, EgalkaMC and GuinanEC. (2003). Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation, 75:389–397.
48.
ChangCJ, YenML, ChenYC, ChienCC, HuangHI, BaiCH and YenBL. (2006). Placenta-derived multipotent cells exhibit immunosuppressive properties that are enhanced in the presence of interferon-gamma. Stem Cells, 24:2466–2477.
49.
ShiD, LiaoL, ZhangB, LiuR, DouX, LiJ, ZhuX, YuL, ChenD and ZhaoRC. (2011). Human adipose tissue-derived mesenchymal stem cells facilitate the immunosuppressive effect of cyclosporin A on T lymphocytes through Jagged-1-mediated inhibition of NF-kappaB signaling. Exp Hematol, 39:214–224.e211.
50.
BragdonB, BurnsR, BakerAH, BelkinaAC, DenisEF Morgan, GV, GerstenfeldLC, SchlezingerJJ and SchlezingerJJ. (2015). Intrinsic sex-linked variations in osteogenic and adipogenic differentiation potential of bone marrow multipotent stromal cells. J Cell Physiol, 230:296–307.
51.
YaoW, LayYE, KotA, LiuR, ZhangH, ChenH, LamK and LaneNE. (2016). Improved mobilization of exogenous mesenchymal stem cells to bone for fracture healing and sex difference. Stem Cells, 34:2587–2600.
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