Advertisement

T cell receptor repertoires within liver allografts are different to those in the peripheral blood

  • Young-Seon Mederacke
    Correspondence
    Corresponding authors. Address: Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Tel.: +49 511 532 6619; Fax: +49 511 532 6998
    Affiliations
    Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Mikalai Nienen
    Affiliations
    Center for Regenerative Therapies, Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin-Brandenburg, Berlin, Germany

    Labor Berlin-Charité Vivantes GmbH, Berlin, Germany
    Search for articles by this author
  • Michael Jarek
    Affiliations
    Department of Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany
    Search for articles by this author
  • Robert Geffers
    Affiliations
    Department of Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany
    Search for articles by this author
  • Katharina Hupa-Breier
    Affiliations
    Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Nina Babel
    Affiliations
    Center for Regenerative Therapies, Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin-Brandenburg, Berlin, Germany

    Medical Department I, University Hospital of the Ruhr-University Bochum, Herne, Germany
    Search for articles by this author
  • Petra Reinke
    Affiliations
    Center for Regenerative Therapies, Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin-Brandenburg, Berlin, Germany

    Center for Advanced Therapies, Charité – Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH) Berlin-Brandenburg, Berlin, Germany
    Search for articles by this author
  • Ingmar Mederacke
    Affiliations
    Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Florian Wolfgang Rudolf Vondran
    Affiliations
    Department of General, Visceral and Transplant Surgery, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Danny Jonigk
    Affiliations
    Institute for Pathology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Heiner Wedemeyer
    Affiliations
    Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
  • Elmar Jaeckel
    Correspondence
    Tel.: +49 511 532 9513; Fax: +49 511 532 6998
    Affiliations
    Department of Gastroenterology, Hepatology & Endocrinology, Hannover Medical School, Hannover, Germany
    Search for articles by this author
Published:December 18, 2020DOI:https://doi.org/10.1016/j.jhep.2020.12.014

      Highlights

      • Specific T cell clones in peripheral blood and in liver allografts were tracked.
      • We observed increased clonality of donor-reactive T cells after liver transplantation.
      • Distinct peripheral and intrahepatic TCR repertoires were identified.
      • Peripheral and intrahepatic TCR repertoires align in acute cellular rejection.

      Summary

      Background & Aims

      T cells are the main mediators of allogeneic immune responses. Specific T cell clones can be tracked by their unique T cell receptor (TCR), but specificity and function remain elusive and have not been investigated in human liver biopsies thus far.

      Methods

      TCR repertoire analysis of CD4+, CD8+, and regulatory T cells of the peripheral blood and liver graft was performed in 7 liver transplant recipients with either stable course (non-rejector, NR), subclinical cellular rejection (SCR), or acute cellular rejection (ACR) during an observation period from pre-transplant to 6 years post-transplant. Furthermore, donor-reactive T cells, identified by their expression of CD154 and glycoprotein A repetitions predominant (GARP) after allogeneic activation, were tracked longitudinally in peripheral blood and within the liver allograft.

      Results

      Although overall clonality of the TCR repertoire did not increase in peripheral blood after liver transplantation, clonality of donor-reactive CD4+ and regulatory T cells increased and these clones accumulated within the liver graft. Surprisingly, the TCR repertoires between the liver graft and the periphery were distinct and showed only limited overlap. Notably, during ACR, TCR repertoires aligned suggesting either graft-specific homing or release of activated T cells from the graft.

      Conclusions

      This is the first study comparing TCR repertoires between liver grafts and blood in patients with NR, SCR, and ACR. Moreover, we attribute specificity and function to a subgroup of intragraft T cell populations. Given the limited overlap between peripheral blood and intragraft repertoires, future studies investigating function and specificities of T cells after liver transplantation should focus on the intragraft immune response.

      Lay summary

      In solid organ transplantation, T cells are key mediators of the recipient’s immune response directed at the transplanted organ. In our study, we characterised the T cell repertoire in a cohort of 7 liver transplant recipients. We demonstrate that donor-specific T cells expand clonally and accumulate in the transplanted liver. Moreover, we show that the composition of T cells in peripheral blood differs from the T cells in the liver allograft, only aligning in the context of acute cellular rejection but not in normal graft or subclinical cellular rejection. This indicates that the intragraft immune response is not mirrored in the peripheral blood. Our findings clarify the importance of protocol liver biopsies in identifying intragraft immune responses for future investigations of allo-directed immune responses.

      Graphical abstract

      Keywords

      To read this article in full you will need to make a payment
      Subscribe to Journal of Hepatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Hara M.
        • Kingsley C.I.
        • Niimi M.
        • Read S.
        • Turvey S.E.
        • Bushell A.R.
        • et al.
        IL-10 is required for regulatory T cells to mediate tolerance to alloantigens in vivo.
        J Immunol. 2001; 166: 3789-3796
      1. Banff schema for grading liver allograft rejection: an international consensus document.
        Hepatology. 1997; 25: 658-663
        • Adams D.H.
        • Sanchez-Fueyo A.
        • Samuel D.
        From immunosuppression to tolerance.
        J Hepatol. 2015; 62: S170-S185
        • Wood K.J.
        • Goto R.
        Mechanisms of rejection: current perspectives.
        Transplantation. 2012; 93: 1-10
        • Todo S.
        • Yamashita K.
        • Goto R.
        • Zaitsu M.
        • Nagatsu A.
        • Oura T.
        • et al.
        A pilot study of operational tolerance with a regulatory T-cell-based cell therapy in living donor liver transplantation.
        Hepatology. 2016; 64: 632-643
        • Mederacke Y.S.
        • Vondran F.W.
        • Kollrich S.
        • Schulde E.
        • Schmitt R.
        • Manns M.P.
        • et al.
        Transient increase of activated regulatory T cells early after kidney transplantation.
        Sci Rep. 2019; 9: 1021
        • Taubert R.
        • Hardtke-Wolenski M.
        • Noyan F.
        • Wilms A.
        • Baumann A.K.
        • Schlue J.
        • et al.
        Intrahepatic regulatory T cells in autoimmune hepatitis are associated with treatment response and depleted with current therapies.
        J Hepatol. 2014; 61: 1106-1114
        • Taubert R.
        • Pischke S.
        • Schlue J.
        • Wedemeyer H.
        • Noyan F.
        • Heim A.
        • et al.
        Enrichment of regulatory T cells in acutely rejected human liver allografts.
        Am J Transpl. 2012; 12: 3425-3436
        • Tran D.
        • Andersson J.
        • Wang R.
        • Ramsey H.
        • Unutmaz D.
        • Shevach E.
        GARP (LRRC32) is essential for the surface expression of latent TGF-beta on platelets and activated FOXP3+ regulatory T cells.
        Proc Natl Acad Sci U S A. 2009; 106: 13445-13450
        • Frentsch M.
        • Arbach O.
        • Kirchhoff D.
        • Moewes B.
        • Worm M.
        • Rothe M.
        • et al.
        Direct access to CD4+ T cells specific for defined antigens according to CD154 expression.
        Nat Med. 2005; 11: 1118-1124
        • Noyan F.
        • Lee Y.
        • Zimmermann K.
        • Hardtke-Wolenski M.
        • Taubert R.
        • Warnecke G.
        • et al.
        Isolation of human antigen-specific regulatory T cells with high suppressive function.
        Eur J Immunol. 2014; 44: 2592-2602
        • van der Burg M.
        • Kreyenberg H.
        • Willasch A.
        • Barendregt B.H.
        • Preuner S.
        • Watzinger F.
        • et al.
        Standardization of DNA isolation from low cell numbers for chimerism analysis by PCR of short tandem repeats.
        Leukemia. 2011; 25: 1467-1470
        • Dziubianau M.
        • Hecht J.
        • Kuchenbecker L.
        • Sattler A.
        • Stervbo U.
        • Rodelsperger C.
        • et al.
        TCR repertoire analysis by next generation sequencing allows complex differential diagnosis of T cell-related pathology.
        Am J Transpl. 2013; 13: 2842-2854
        • Bolotin D.A.
        • Poslavsky S.
        • Mitrophanov I.
        • Shugay M.
        • Mamedov I.Z.
        • Putintseva E.V.
        • et al.
        MiXCR: software for comprehensive adaptive immunity profiling.
        Nat Methods. 2015; 12: 380-381
        • Nazarov V.I.
        • Pogorelyy M.V.
        • Komech E.A.
        • Zvyagin I.V.
        • Bolotin D.A.
        • Shugay M.
        • et al.
        tcR: an R package for T cell receptor repertoire advanced data analysis.
        BMC Bioinformatics. 2015; 16: 175
        • Shugay M.
        • Bagaev D.V.
        • Turchaninova M.A.
        • Bolotin D.A.
        • Britanova O.V.
        • Putintseva E.V.
        • et al.
        VDJtools: unifying post-analysis of T cell receptor repertoires.
        Plos Comput Biol. 2015; 11e1004503
        • Yang G.
        • Ou M.
        • Chen H.
        • Guo C.
        • Chen J.
        • Lin H.
        • et al.
        Characteristic analysis of TCR beta-chain CDR3 repertoire for pre- and post-liver transplantation.
        Oncotarget. 2018; 9: 34506-34519
        • Savage T.M.
        • Shonts B.A.
        • Lau S.
        • Obradovic A.
        • Robins H.
        • Shaked A.
        • et al.
        Deletion of donor-reactive T cell clones after human liver transplant.
        Am J Transpl. 2020; 20: 538-545
        • Han F.F.
        • Fan H.
        • Ren L.L.
        • Wang H.G.
        • Wang C.
        • Ma X.
        • et al.
        Profiling the pattern of human TRB/IGH-CDR3 repertoire in liver transplantation patients via high-throughput sequencing analysis.
        Scand J Immunol. 2020; e12912
        • San Segundo D.
        • Ruiz J.C.
        • Fernandez-Fresnedo G.
        • Izquierdo M.
        • Gomez-Alamillo C.
        • Cacho E.
        • et al.
        Calcineurin inhibitors affect circulating regulatory T cells in stable renal transplant recipients.
        Transpl Proc. 2006; 38: 2391-2393
        • Presser D.
        • Sester U.
        • Mohrbach J.
        • Janssen M.
        • Kohler H.
        • Sester M.
        Differential kinetics of effector and regulatory T cells in patients on calcineurin inhibitor-based drug regimens.
        Kidney Int. 2009; 76: 557-566
        • Demirkiran A.
        • Kok A.
        • Kwekkeboom J.
        • Kusters J.G.
        • Metselaar H.J.
        • Tilanus H.W.
        • et al.
        Low circulating regulatory T-cell levels after acute rejection in liver transplantation.
        Liver Transpl. 2006; 12: 277-284
        • Stenard F.
        • Nguyen C.
        • Cox K.
        • Kambham N.
        • Umetsu D.T.
        • Krams S.M.
        • et al.
        Decreases in circulating CD4+CD25hiFOXP3+ cells and increases in intragraft FOXP3+ cells accompany allograft rejection in pediatric liver allograft recipients.
        Pediatr Transpl. 2009; 13: 70-80
        • He Q.
        • Fan H.
        • Li J.Q.
        • Qi H.Z.
        Decreased circulating CD4+CD25highFoxp3+ T cells during acute rejection in liver transplant patients.
        Transpl Proc. 2011; 43: 1696-1700
        • Furukawa A.
        • Wisel S.A.
        • Tang Q.
        Impact of immune-modulatory drugs on regulatory T cell.
        Transplantation. 2016; 100: 2288-2300
        • Kubota N.
        • Sugitani M.
        • Takano S.
        • Sheikh A.
        • Takayama T.
        • Haga H.
        • et al.
        Correlation between acute rejection severity and CD8-positive T cells in living related liver transplantation.
        Transpl Immunol. 2006; 16: 60-64
        • Taner T.
        • Gustafson M.P.
        • Hansen M.J.
        • Park W.D.
        • Bornschlegl S.
        • Dietz A.B.
        • et al.
        Donor-specific hypo-responsiveness occurs in simultaneous liver-kidney transplant recipients after the first year.
        Kidney Int. 2018; 93: 1465-1474
        • Kamada N.
        • Wight D.G.
        Antigen-specific immunosuppression induced by liver transplantation in the rat.
        Transplantation. 1984; 38: 217-221
        • Praseedom R.K.
        • McNeil K.D.
        • Watson C.J.
        • Alexander G.J.
        • Calne R.Y.
        • Wallwork J.
        • et al.
        Combined transplantation of the heart, lung, and liver.
        Lancet. 2001; 358: 812-813
        • Calne R.Y.
        • White H.J.
        • Binns R.M.
        • Herbertson B.M.
        • Millard P.R.
        • Pena J.
        • et al.
        Immunosuppressive effects of the orthotopically transplanted porcine liver.
        Transpl Proc. 1969; 1: 321-324
        • Crispe I.N.
        • Giannandrea M.
        • Klein I.
        • John B.
        • Sampson B.
        • Wuensch S.
        Cellular and molecular mechanisms of liver tolerance.
        Immunol Rev. 2006; 213: 101-118
        • Crispe I.N.
        • Mehal W.Z.
        Strange brew: T cells in the liver.
        Immunol Today. 1996; 17: 522-525
        • Norris S.
        • Collins C.
        • Doherty D.G.
        • Smith F.
        • McEntee G.
        • Traynor O.
        • et al.
        Resident human hepatic lymphocytes are phenotypically different from circulating lymphocytes.
        J Hepatol. 1998; 28: 84-90
        • Lim C.J.
        • Lee Y.H.
        • Pan L.
        • Lai L.
        • Chua C.
        • Wasser M.
        • et al.
        Multidimensional analyses reveal distinct immune microenvironment in hepatitis B virus-related hepatocellular carcinoma.
        Gut. 2019; 68: 916-927
        • Shoukry N.H.
        • Grakoui A.
        • Houghton M.
        • Chien D.Y.
        • Ghrayeb J.
        • Reimann K.A.
        • et al.
        Memory CD8+ T cells are required for protection from persistent hepatitis C virus infection.
        J Exp Med. 2003; 197: 1645-1655
        • Pallett L.J.
        • Davies J.
        • Colbeck E.J.
        • Robertson F.
        • Hansi N.
        • Easom N.J.W.
        • et al.
        IL-2(high) tissue-resident T cells in the human liver: sentinels for hepatotropic infection.
        J Exp Med. 2017; 214: 1567-1580
        • Fernandez-Ruiz D.
        • Ng W.Y.
        • Holz L.E.
        • Ma J.Z.
        • Zaid A.
        • Wong Y.C.
        • et al.
        Liver-resident memory CD8(+) T cells form a front-line defense against malaria liver-stage infection.
        Immunity. 2016; 45: 889-902
        • Tse S.W.
        • Radtke A.J.
        • Espinosa D.A.
        • Cockburn I.A.
        • Zavala F.
        The chemokine receptor CXCR6 is required for the maintenance of liver memory CD8(+) T cells specific for infectious pathogens.
        J Infect Dis. 2014; 210: 1508-1516
        • Prosser A.C.
        • Kallies A.
        • Lucas M.
        Tissue-resident lymphocytes in solid organ transplantation: innocent passengers or the key to organ transplant survival?.
        Transplantation. 2018; 102: 378-386
        • Sagoo P.
        • Lombardi G.
        • Lechler R.I.
        Relevance of regulatory T cell promotion of donor-specific tolerance in solid organ transplantation.
        Front Immunol. 2012; 3: 184
        • Carvalho-Gaspar M.
        • Jones N.D.
        • Luo S.
        • Martin L.
        • Brook M.O.
        • Wood K.J.
        Location and time-dependent control of rejection by regulatory T cells culminates in a failure to generate memory T cells.
        J Immunol. 2008; 180: 6640-6648
        • Issa F.
        • Robb R.J.
        • Wood K.J.
        The where and when of T cell regulation in transplantation.
        Trends Immunol. 2013; 34: 107-113
        • Suchin E.J.
        • Langmuir P.B.
        • Palmer E.
        • Sayegh M.H.
        • Wells A.D.
        • Turka L.A.
        Quantifying the frequency of alloreactive T cells in vivo: new answers to an old question.
        J Immunol. 2001; 166: 973-981
        • DeWolf S.
        • Grinshpun B.
        • Savage T.
        • Lau S.P.
        • Obradovic A.
        • Shonts B.
        • et al.
        Quantifying size and diversity of the human T cell alloresponse.
        JCI Insight. 2018; 3e121256
        • Knolle P.A.
        • Thimme R.
        Hepatic immune regulation and its involvement in viral hepatitis infection.
        Gastroenterology. 2014; 146: 1193-1207