This study examines the involvement of T cell autocrine hyaluronan (HA) with the immunological synapse (IS) that mediates T cell activation by antigen-presenting cells (APCs). Three-dimensional (3D) confocal images of mouse CD4+ T cells interacting with B cell lymphoma (A20) APCs in vitro showed HA to be primarily on the T cell side of the IS, appearing as a compact mass indenting the T cell nucleus. Similar 3D imaging of CD4+ T cells forming a pseudo-IS on anti-CD3 antibody-coated glass showed HA in the vicinity of the IS in dense masses or complex, arched, columnar structures. Affinity/immunofluorescence labeling studies confirmed the HA masses and columns were cytoplasmic, located beneath the cortical actin layer but outside the nucleus. In T cells forming a pseudo-IS, the HA-binding protein RHAMM was localized to cortical cytoplasm and had limited spatial overlap with cytoplasmic HA. Pre-exposure of T cells to the HA synthesis inhibitor 4-methylumbelliferone (4-MU) or to the HA-binding peptide Pep-1 inhibited IS formation with A20 APCs. Moreover, actin ring development in T cell pseudo-IS was inhibited by pre-exposure to 4-MU, but not by pre-exposure to Pep-1. Collectively, our previous and present studies suggest a complex role for cell surface and cytoplasmic T cell autocrine HA in IS formation and T cell receptor signaling:
DayAJde la MotteCA.Hyaluronan cross-linking: a protective mechanism in inflammation. Trends Immunol. 2005;26(12):637–43.
2.
GarantziotisSSavaniRC.Hyaluronan biology: a complex balancing act of structure, function, location and context. Matrix Biol. 2019;78–79:1–10.
3.
DayAJMilnerCM.TSG-6: a multifunctional protein with anti-inflammatory and tissue-protective properties. Matrix Biol. 2019;78–79:60–83.
4.
TavianatouAGCaonIFranchiMPiperigkouZGalessoDKaramanosNK.Hyaluronan: molecular size-dependent signaling and biological functions in inflammation and cancer. FEBS J. 2019;286(15):2883–908.
5.
JiangDLiangJNoblePW.Hyaluronan as an immune regulator in human diseases. Physiol Rev. 2011;91(1):221–64.
6.
JohnsonPArifAALee-SayerSSMDongY.Hyaluronan and its interactions with immune cells in the healthy and inflamed lung. Front Immunol. 2018;9:Article 2787, 1–8.
7.
NagyNKuipersHFMarshallPLWangEKaberGBollykyPL.Hyaluronan in autoimmunity and immune dysregulation. Matrix Biol. 2019;78–79:292–313.
8.
WinklerCWFosterSCMatsumotoSGPrestonMAXingRBeboBFBanineFBerny-LangMAItakuraAMcCartyOJTShermanLS.Hyaluronan anchored to activated CD44 on central nervous system vascular endothelial cells promotes lymphocyte extravasation in experimental autoimmune encephalomyelitis. J Biol Chem. 2012;287:33237–51.
9.
MutoJMoriokaYYamasakiKKimMGarciaACarlinAFVarkiAGalloRL.Hyaluronan digestion controls DC migration from the skin. J Clin Invest. 2014;124(3):1309–9.
10.
LennonFESingletonPA.Hyaluronan regulation of vascular integrity. Am J Cardiovasc Dis. 2011;1(3):200–13.
11.
HayenWGoebelerMKumarSRiessenRNehlsV.Hyaluronan stimulates tumor cell migration by modulating the fibrin fiber architecture. J Cell Sci. 1999;112(Pt 13):2241–51.
12.
TzircotisGThorneRFIsackeCM.Chemotaxis towards hyaluronan is dependent on CD44 expression and modulated by cell type variation in CD44-hyaluronan binding. J Cell Sci. 2005;118:5119–28.
13.
LongSABucknerJH.CD4+FOXP3+ T regulatory cells in human autoimmunity: more than a numbers game. J Immunol. 2011;187:2061–66.
14.
MummertMEMummertDEdelbaumDHuiFMatsueHTakashimaA.Synthesis and surface expression of hyaluronan by dendritic cells and its potential role in antigen presentation. J Immunol. 2002;169:4322–31.
15.
MahaffeyCLMummertME.Hyaluronan synthesis is required for IL-2-mediated T cell proliferation. J Immunol. 2007;179:8191–99.
16.
GebeJAGoodenMDWorkmanGNagyNBollykyPLWightTNVernonRB.Modulation of hyaluronan synthases and involvement of T cell-derived hyaluronan in autoimmune responses to transplanted islets. Matrix Biol Plus. 2020;9:100052, 1–5.
17.
KakizakiIKojimaKTakagakiKEndoMKannagiRItoMMaruoYSatoHYasudaTMitaSKimataKItanoN.A novel mechanism for the inhibition of hyaluronan biosynthesis by 4-methylumbelliferone. J Biol Chem. 2004;279:33281–89.
18.
NagyNKuipersHFFrymoyerARIshakHDBollykyJBWightTNBollykyPL.4-methylumbelliferone treatment and hyaluronan inhibition as a therapeutic strategy in inflammation, autoimmunity, and cancer. Front Immunol. 2015;6:123.
19.
ValituttiSCoombsDDupréL.The space and time frames of T cell activation at the immunological synapse. FEBS Lett. 2010;584:4851–57.
20.
Ortega-CarrionAVincente-ManzanaresM.Concerning immune synapses: a spatiotemporal timeline. F1000Res. 2016;5:1–11.
21.
BollykyPLEvankoSPWuRPPotter-PerigoSLongSAKinsellaBReijonenHGuebtnerKTengBChanCKBraunKRGebeJANepomGTWightTN.Th1 cytokines promote T-cell binding to antigen-presenting cells via enhanced hyaluronan production and accumulation at the immune synapse. Cell Mol Immunol. 2010;7(3):211–20.
22.
MarshallPLNagyNKaberGBarlowGLRameshAXieBJLindeMHHaddockNLLesterCATranQLde VriesCRHargilAMalkovskiyAVGurevichIMartinezHAKuipersHFYadavaKZhangXEvankoSPGebeJAWangXVernonRBde la MotteCWightTNEnglemanEGKramsSMMeyerEHBollykyPL.Hyaluronan synthesis inhibition impairs antigen presentation and delays transplantation rejection. Matrix Biol. 2021;96:69–86.
VernonRBGoodenMDChanCKWorkmanGObikaMWightTN.Autocrine hyaluronan influences sprouting and lumen formation during HUVEC tubulogenesis in vitro. J Histochem Cytochem. 2021;69(6):415–28.
25.
NaB-RKimH-RPiragyteIOhH-MKwonM-SAkberULeeH-SParkD-SSongWKParkZ-YImS-HRhoM-CHyunY-MKimMJunC-D.TAGLN2 regulates T cell activation by stabilizing the actin cytoskeleton at the immunological synapse. J Cell Biol. 2015;209:143–62.
26.
MummertMEMohamadzadehMMummertDIMizumotoNTakashimaA.Development of peptide inhibitor of hyaluronan-mediated leukocyte trafficking. J Exp Med. 2000;192:769–9.
27.
BárdosTKamathRVMikeczKGlantTT.Anti-inflammatory and chondroprotective effect of TSG-6 (tumor necrosis factor-α-stimulated gene-6) in murine models of experimental arthritis. Am J Pathol. 2001;159:1711–21.
28.
MummertME.Immunologic roles of hyaluronan. Immunol Res. 2005;31:189–205.
29.
WightTNKangIEvankoSPHartenIAChangMYPearceOMTAllenCEFrevertCW.Versican—a critical extracellular matrix regulator of immunity and inflammation. Front Immunol. 2020;11:Article 512, 1–12.
30.
BouchetJdel Río-IñiguezIAlcoverA.Chapter 9: imaging vesicular traffic at the immune synapse. In: BaldariCTDustinML editors. The immune synapse: methods and protocols. Methods in molecular biology, vol 1584. New York: Humana Press; 2017. p. 129–42.
31.
MessamBJTolgCMcCarthyJBNelsonACTurleyEA.RHAMM is a multifunctional protein that regulates cancer progression. Int J Mol Sci. 2021;22:10313.
32.
HardwickCHoareKOwensRHohnHPHookMMooreDCrippsVAustenLNanceDMTurleyEA.Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility. J Cell Biol. 1992;117:1343–50.
33.
SavaniRCWangCYangBZhangSKinsellaMGWightTNSternRNanceDMTurleyEA.Migration of bovine aortic smooth muscle cells after wounding injury: the role of hyaluronan and RHAMM. J Clin Invest. 1995;95(3):1158–68.
34.
EvankoSPParksWTWightTN.Intracellular hyaluronan in arterial smooth muscle cells: association with microtubules, RHAMM, and the mitotic spindle. J Histochem Cytochem. 2004;52(12):1525–35.
35.
MaxwellCAMcCarthyJTurleyE.Cell-surface and mitotic-spindle RHAMM: moonlighting or dual oncogenic functions?J Cell Sci. 2008;121(Pt 7):925–32.
36.
VeisehMKwonaDHBorowskyADTolgCLeongHSLewiseJDTurleyEABissellMJ.Cellular heterogeneity profiling by hyaluronan probes reveals an invasive but slow-growing breast tumor subset. Proc Natl Acad Sci USA. 2014;111:E1731–39.
37.
ThangavelCBoopathiELiuYHaberAErtelABhardwajAAddyaSWilliamsNCimentSJCotziaPDeanJLSnookAMcNairCPriceMHernandezJRZhaoSGBirbeRMcCarthyJBTurleyEAPientaKJFengFYDickerAPKnudsenKEDenRB.RB loss promotes prostate cancer metastasis. Cancer Res. 2017;77:982–5.
38.
HascallVCMajorsAKDe La MotteCAEvankoSPWangADrazbaJAStrongSAWightTN.Intracellular hyaluronan: a new frontier for inflammation?Biochim Biophys Acta. 2004;1673:3–12.
39.
LeeHPAlisafaeiFAdebawaleKChangJShenoyVBChaudhuriO.The nuclear piston activates mechanosensitive ion channels to generate cell migration paths in confining microenvironments. Sci Adv. 2021;7(2):eabd4058.
40.
KuipersHFRieckMGurevichINagyNButteMJNegrinRSWightTNSteinmanLBollykyPL.Hyaluronan synthesis is necessary for autoreactive T-cell trafficking, activation, and Th1 polarization. Proc Natl Acad Sci USA. 2016;113:339–44.
41.
TrautmannAValituttiS.The diversity of immunological synapses. Curr Opin Immunol. 2003;15(3):249–54.
42.
WeigelPH.Hyaluronan synthase: the mechanism of initiation at the reducing end and a pendulum model for polysaccharide translocation to the cell exterior. Int J Cell Biol. 2015;2015:367579.
43.
HagenfeldDBorkenhagenBSchulzTSchillersHSchumacherUPrehmP.Hyaluronan export through plasma membranes depends on concurrent K+ efflux by K(ir) channels. PLoS ONE. 2012;7(6):e39096.
44.
SkandalisSSKaralisTHeldinP.Intracellular hyaluronan: importance for cellular functions. Semin Cancer Biol. 2020;62:20–30.
45.
TörrönenKNikunenKKärnäRTammiMTammiRRillaK.Tissue distribution and subcellular localization of hyaluronan synthase isoenzymes. Histochem Cell Biol. 2014;141(1):17–31.
46.
SiiskonenHOikariSPasonen-SeppänenSRillaK.Hyaluronan synthase 1: a mysterious enzyme with unexpected functions. Front Immunol. 2015;6:43–11.
47.
EvankoSPWightTN.Intracellular localization of hyaluronan in proliferating cells. J Histochem Cytochem. 1999;47(10):1331–42.
48.
MurphyKTraversPWalportM.Janeway’s immunobiology. 7th ed. New York: Garland Science; Taylor & Frances; 2008.
49.
VarmaRCampiGYokosukaTSaitoTDustinML.T cell receptor-proximal signals are sustained in peripheral microclusters and terminated in the central supramolecular activation cluster. Immunity. 2006;25(1):117–27.
DustinMLGrovesJT.Receptor signaling clusters in the immune synapse. Annu Rev Biophys. 2012;41:543–6.
52.
CheminKBohineustADogniauxSTourretMGuéganSMiroFHivrozC.Cytokine secretion by CD4+ T cells at the immunological synapse requires Cdc42-dependent local actin remodeling but not microtubule organizing center polarity. J Immunol. 2012;189:2159–68.
53.
DinicJRiehlAAdlerJParmrydI.The T cell receptor resides in ordered plasma membrane nanodomains that aggregate upon patching of the receptor. Sci Rep. 2015;5:10082.
54.
NolzJCGomezTSZhuPLiSMedeirosRBShimizuYBurkhardtJKFreedmanBDBilladeauDD.The WAVE2 complex regulates actin cytoskeletal reorganization and CRAC-mediated calcium entry during T cell activation. Curr Biol. 2006;16:24–34.
55.
YiJWuXSCritesTHammerJA3rd. Actin retrograde flow and actomyosin II arc contraction drive receptor cluster dynamics at the immunological synapse in Jurkat T cells. Mol Biol Cell. 2012;23(5):834–52.
56.
Ben-ShmuelAJosephNSabagBBarda-SaadM.Lymphocyte mechanotransduction: the regulatory role of cytoskeletal dynamics in signaling cascades and effector functions. J Leukoc Biol. 2019;105(6):1261–73.
