Review Article| Volume 28, ISSUE 3, P365-373, September 2008

Novel Noninvasive Assays to Predict Transplantation Rejection and Tolerance: Enumeration of Cytokine-Producing Alloreactive T Cells

      Treatment and prevention of allograft loss in organ transplant recipients relies chiefly on non–antigen-specific immunosuppression. Current approaches to the management of these immunosuppressive drugs are largely empiric and reactive because of lack of immune monitoring assays. Alloreactive T cells play a key role in acute rejection and in development of chronic allograft nephropathy, the leading cause of late allograft failure. There is thus an increasing interest in development of simple, reliable, noninvasive assays measuring allogeneic anti-donor responsiveness or donor-specific nonresponsiveness to predict ransplantation rejection and tolerance. Because the frequency and cytokine profile of alloreactive T cells play an important role in these processes, this article mainly focuses on assays that enumerate cytokine-producing alloreactive T cells.
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      1. Kidney transplantation.
        (Available at:) (Accessed May 11, 2008)
        • Denton M.D.
        • Magee C.M.
        • Sayegh M.H.
        Immunosuppressive strategies in transplantation.
        Lancet. 1999; 353 ([CrossRef][ISI][Medline]): 1083-1092
        • Womer K.L.
        • Vella J.P.
        • Sayegh M.H.
        Chronic allograft dysfunction: mechanisms and new approaches to therapy.
        Semin Nephrol. 2000; 20: 126-147
        • Pascual M.
        • Theruvath T.
        • Kawai T.
        • et al.
        Strategies to improve long-term outcomes after renal transplantation.
        N Engl J Med. 2002; 346: 580-590
        • Vongwiwatana A.
        • Tasanarong A.
        • Hidalgo L.G.
        • et al.
        The role of B cells and alloantibody in the host response to human organ allografts.
        Immunol Rev. 2003; 196: 197-218
        • Liu Z.
        • Sun Y.K.
        • Xi Y.P.
        • et al.
        Contribution of direct and indirect recognition pathways to T cell alloreactivity.
        J Exp Med. 1993; 177: 1643-1650
        • Benichou G.
        • Kant C.D.
        • Madsen J.
        • et al.
        Modulation of alloreactivity to MHC-derived peptides and transplantation tolerance.
        Front Biosci. 2007; 12: 4239-4247
        • Ciubotariu R.
        • Liu Z.
        • Colovai A.I.
        • et al.
        Persistent allopeptide reactivity and epitope spreading in chronic rejection or organ allografts.
        J Clin Invest. 1998; 101: 398-405
        • Hornick P.I.
        • Mason P.D.
        • Baker R.J.
        • et al.
        Significant frequencies of T cells with indirect anti-donor specificity in heart graft recipients with chronic rejection.
        Circulation. 2000; 101: 2405-2410
        • SivaSai K.S.
        • Smith M.A.
        • Poindexter N.J.
        • et al.
        Indirect recognition of donor HLA class I peptides in lung transplant recipients with bronchiolitis obliterans syndrome.
        Transplantation. 1999; 67: 1094-1098
        • Najafian N.
        • Salama A.
        • Fedoseyeva E.V.
        • et al.
        Enzyme-linked immunosorbent spot assay analysis of peripheral blood lymphocyte reactivity to donor HLA-DR peptides: potential novel assay for prediction of outcomes for renal transplant recipients.
        J Am Soc Nephrol. 2002; 13: 252-259
        • Bell E.
        • Sparshott S.
        • Bunce C.
        CD4+ T-cell memory, CD45R subsets and the persistence of antigen-a unifying concept.
        Immunol Today. 1998; 19: 60-64
        • Selin L.
        • Vergilis K.
        • Welsh R.
        • et al.
        Reduction of otherwise remarkably stable virus-specific cytotoxic T lymphocyte memory by heterologous viral infections.
        J Exp Med. 1996; 183: 2489-2499
        • Lombardi G.
        • Sidhu S.
        • Daly M.
        • et al.
        Are primary alloresponses truly primary?.
        Int Immunol. 1990; 2: 9-13
        • Lechler R.
        • Heaton T.
        • Barber L.
        • et al.
        Molecular mimicry by major histocompatibility complex molecules and peptides accounts for some alloresponses.
        Immunol Lett. 1992; 34: 63-69
        • Vella J.P.
        • Spadafora-Fereira M.
        • Murphy B.
        • et al.
        Indirect allorecognition of major histocompatibility complex allopepetides in human renal transplant recipients with chronic graft dysfunction.
        Transplantation. 1997; 64: 795-800
        • Liu Z.
        • Colovai A.I.
        • Tugulea S.
        • et al.
        Indirect recognition of donor HLA-DR peptides in organ allograft rejection.
        J Clin Invest. 1996; 98: 1150-1157
      2. Cytokine.
        (Available at:) (Accessed June 21, 2008)
        • Nickerson P.
        • Steurer W.
        • Steiger J.
        • et al.
        Cytokines and the Th1/Th2 paradigm in transplantation.
        Curr Opin Immunol. 1994; 6: 757-764
        • Mosmann T.R.
        • Coffman R.L.
        Th1 and Th2 cells: different patterns of lymphokine secretion lead to different functional properties.
        Annu Rev Immunol. 1989; 7: 145-173
        • Benghiat F.S.
        • Craciun L.
        • De Wilde V.
        • et al.
        IL-17 production elicited by allo-major histocompatibility complex class II recognition depends on CD25+CD4+ T cells.
        Transplantation. 2008; 85: 943-949
        • Pawelec G.
        • Rehbein A.
        • Schlotz E.
        • et al.
        Cytokine modulation of Th1/Th2 phenotype differentiation in directly alloresponsive CD4+ human T cells.
        Transplantation. 1996; 62: 1095-1101
        • Braun M.Y.
        • McCormack A.
        • Webb G.
        • et al.
        Mediation of acute but not chronic rejection of MHC-incompatible rat kidney grafts by alloreactive CD4 T-cells activated by the direct pathway of sensitization.
        Transplantation. 1993; 55: 177-182
        • Najafian N.
        • Albin M.
        • Newell K.A.
        How can we measure immunologic tolerance in humans?.
        J Am Soc Nephrol. 2006; 17: 2652-2663
        • Heeger P.S.
        • Greenspan N.S.
        • Kuhlenschmidt S.
        • et al.
        Pretransplant frequency of donor-specific, IFN-γ producing lymphocytes is a manifestation of immunologic memory and correlates with the risk of post transplant rejection episodes.
        J Immunol. 1999; 163: 2267-2275
        • Tary-Lehmann M.
        • Hricik D.E.
        • Justice A.C.
        • et al.
        Enzyme-linked immunosorbent assay spot detection of interferon-γ and interleukin 5-producing cells as a predictive marker for renal allograft failure.
        Transplantation. 1998; 66: 219-224
        • Hernández-Fuentes M.P.
        • Salama A.
        In vitro assays for immune monitoring in transplantation.
        Methods Mol Biol. 2006; 333: 269-290
        • Poggio E.D.
        • Augustine J.J.
        • Clemente M.
        • et al.
        Pretransplant cellular alloimmunity as assessed by a panel of reactive T cells assay correlates with acute renal graft rejection.
        Transplantation. 2007; 83: 847-852
        • Gebauer B.S.
        • Hricik D.E.
        • Atallah A.
        • et al.
        Evolution of the enzyme-linked immunosorbent spot assay for post-transplant alloreactivity as a potentially useful immune monitoring tool.
        Am J Transplant. 2002; 2: 857-866
        • Hricik D.E.
        • Rodriguez V.
        • Riley J.
        • et al.
        Enzyme linked immunosorbent (ELISPOT) assay for interferon-gamma independently predicts renal function in kidney transplant recipients.
        Am J Transplant. 2003; 3: 878-884
        • Bestard O.
        • Nickel P.
        • Cruzado J.M.
        • et al.
        Circulating directly and indirectly primed alloreactive interferon-gamma-producing T cell frequencies correlate with graft function in longstanding renal transplant patients.
        J Am Soc Nephrol. 2008; 19: 1419-1429
        • Salama A.D.
        • Najafian N.
        • Clarkson M.R.
        • et al.
        Regulatory CD25+ T cells in human kidney transplant recipients.
        J Am Soc Nephrol. 2003; 14: 1643-1651
        • Pala P.
        • Hussell T.
        • Openshaw P.J.
        Flow cytometric measurement of intracellular cytokines.
        J Immunol Methods. 2000; 243: 107-124
        • Magee C.C.
        • Denton M.D.
        • Womer K.L.
        • et al.
        Assessment by flow cytometry of intracellular cytokine production in the peripheral blood cells of renal transplant recipients.
        Clin Transplant. 2004; 18: 395-401
        • Kwun J.
        • Knechtle S.J.
        • Hu H.
        Determination of the functional status of alloreactive T cells by interferon-gamma kinetics.
        Transplantation. 2006; 81: 590-598
        • Vignali D.A.
        Multiplexed particle-based flow cytometric assays.
        J Immunol Methods. 2000; 243: 243-255
        • Sebelin K.
        • Schulzki A.
        • Kloetzel P.M.
        • et al.
        Impairment of circulating myeloid dendritic cells in immunosuppressed renal/pancreas transplant recipients.
        Transplantation. 2006; 82: 779-787