Thymic output generates a new and diverse TCR repertoire after autologous stem cell transplantation in multiple sclerosis patients
Paolo A Muraro, Daniel C Douek, Amy Packer, Katherine Chung, Francisco J Guenaga, Riccardo Cassiani-Ingoni, Catherine Campbell, Sarfraz Memon, James W Nagle, Frances T Hakim, Ronald E Gress, Henry F McFarland, Richard K Burt, Roland Martin, Paolo A Muraro, Daniel C Douek, Amy Packer, Katherine Chung, Francisco J Guenaga, Riccardo Cassiani-Ingoni, Catherine Campbell, Sarfraz Memon, James W Nagle, Frances T Hakim, Ronald E Gress, Henry F McFarland, Richard K Burt, Roland Martin
Abstract
Clinical trials have indicated that autologous hematopoietic stem cell transplantation (HSCT) can persistently suppress inflammatory disease activity in a subset of patients with severe multiple sclerosis (MS), but the mechanism has remained unclear. To understand whether the beneficial effects on the course of disease are mediated by lympho-depletive effects alone or are sustained by a regeneration of the immune repertoire, we examined the long-term immune reconstitution in patients with MS who received HSCT. After numeric recovery of leukocytes, at 2-yr follow-up there was on average a doubling of the frequency of naive CD4(+) T cells at the expense of memory T cells. Phenotypic and T cell receptor excision circle (TREC) analysis confirmed a recent thymic origin of the expanded naive T cell subset. Analysis of the T cell receptor repertoire showed the reconstitution of an overall broader clonal diversity and an extensive renewal of clonal specificities compared with pretherapy. These data are the first to demonstrate that long-term suppression of inflammatory activity in MS patients who received HSCT does not depend on persisting lymphopenia and is associated with profound qualitative immunological changes that demonstrate a de novo regeneration of the T cell compartment.
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References
- Hafler, D.A. 2004. Multiple sclerosis. J. Clin. Invest. 113:788–794.
- Muraro, P.A., R. Cassiani Ingoni, and R. Martin. 2003. Hematopoietic stem cell transplantation for multiple sclerosis: current status and future challenges. Curr. Opin. Neurol. 16:299–305.
- Fassas, A., and V.K. Kimiskidis. 2003. Stem cell transplantation for multiple sclerosis: what is the evidence? Blood Rev. 17:233–240.
- Burt, R.K., S. Slavin, W.H. Burns, and A.M. Marmont. 2002. Induction of tolerance in autoimmune diseases by hematopoietic stem cell transplantation: getting closer to a cure? Blood. 99:768–784.
- Fassas, A., A. Anagnostopoulos, A. Kazis, K. Kapinas, I. Sakellari, V. Kimiskidis, and A. Tsompanakou. 1997. Peripheral blood stem cell transplantation in the treatment of progressive multiple sclerosis: first results of a pilot study. Bone Marrow Transplant. 20:631–638.
- Burt, R.K., A.E. Traynor, B. Cohen, K.H. Karlin, F.A. Davis, D. Stefoski, C. Terry, L. Lobeck, E.J. Russell, C. Goolsby, et al. 1998. T cell-depleted autologous hematopoietic stem cell transplantation for multiple sclerosis: report on the first three patients. Bone Marrow Transplant. 21:537–541.
- Openshaw, H., B.T. Lund, A. Kashyap, R. Atkinson, I. Sniecinski, L.P. Weiner, and S. Forman. 2000. Peripheral blood stem cell transplantation in multiple sclerosis with busulfan and cyclophosphamide conditioning: report of toxicity and immunological monitoring. Biol. Blood Marrow Transplant. 6:563–575.
- Sun, W., U. Popat, G. Hutton, Y.C. Zang, R. Krance, G. Carrum, G.A. Land, H. Heslop, M. Brenner, and J.Z. Zhang. 2004. Characteristics of T-cell receptor repertoire and myelin-reactive T cells reconstituted from autologous haematopoietic stem-cell grafts in multiple sclerosis. Brain. 127:996–1008.
- Reimer, P., V. Kunzmann, M. Wilhelm, B. Weissbrich, D. Kraemer, H. Berghammer, and F. Weissinger. 2003. Cellular and humoral immune reconstitution after autologous peripheral blood stem cell transplantation (PBSCT). Ann. Hematol. 82:263–270.
- Koehne, G., W. Zeller, M. Stockschlaeder, and A.R. Zander. 1997. Phenotype of lymphocyte subsets after autologous peripheral blood stem cell transplantation. Bone Marrow Transplant. 19:149–156.
- Berzins, S.P., A.P. Uldrich, J.S. Sutherland, J. Gill, J.F. Miller, D.I. Godfrey, and R.L. Boyd. 2002. Thymic regeneration: teaching an old immune system new tricks. Trends Mol. Med. 8:469–476.
- Kivisakk, P., D.J. Mahad, M.K. Callahan, C. Trebst, B. Tucky, T. Wei, L. Wu, E.S. Baekkevold, H. Lassmann, S.M. Staugaitis, et al. 2003. Human cerebrospinal fluid central memory CD4+ T cells: evidence for trafficking through choroid plexus and meninges via P-selectin. Proc. Natl. Acad. Sci. USA. 100:8389–8394.
- Jameson, S.C. 2002. Maintaining the norm: T-cell homeostasis. Nat. Rev. Immunol. 2:547–556.
- Murali-Krishna, K., and R. Ahmed. 2000. Cutting edge: naive T cells masquerading as memory cells. J. Immunol. 165:1733–1737.
- Goldrath, A.W., L.Y. Bogatzki, and M.J. Bevan. 2000. Naive T cells transiently acquire a memory-like phenotype during homeostasis-driven proliferation. J. Exp. Med. 192:557–564.
- Kimmig, S., G.K. Przybylski, C.A. Schmidt, K. Laurisch, B. Mowes, A. Radbruch, and A. Thiel. 2002. Two subsets of naive T helper cells with distinct T cell receptor excision circle content in human adult peripheral blood. J. Exp. Med. 195:789–794.
- Douek, D.C., R.D. McFarland, P.H. Keiser, E.A. Gage, J.M. Massey, B.F. Haynes, M.A. Polis, A.T. Haase, M.B. Feinberg, J.L. Sullivan, et al. 1998. Changes in thymic function with age and during the treatment of HIV infection. Nature. 396:690–695.
- Pannetier, C., M. Cochet, S. Darche, A. Casrouge, M. Zoller, and P. Kourilsky. 1993. The sizes of the CDR3 hypervariable regions of the murine T-cell receptor beta chains vary as a function of the recombined germ-line segments. Proc. Natl. Acad. Sci. USA. 90:4319–4323.
- Gorski, J., M. Yassai, X. Zhu, B. Kissela, C. Keever, and N. Flomenberg. 1994. Circulating T cell repertoire complexity in normal individuals and bone marrow recipients analyzed by CDR3 spectratyping: correlation with the immune status. J. Immunol. 152:5109–5119.
- Brenchley, J.M., N.J. Karandikar, M.R. Betts, D.R. Ambrozak, B.J. Hill, L.E. Crotty, J.P. Casazza, J. Kuruppu, S.A. Migueles, M. Connors, et al. 2003. Expression of CD57 defines replicative senescence and antigen-induced apoptotic death of CD8+ T cells. Blood. 101:2711–2720.
- Mackall, C.L., T.A. Fleisher, M.R. Brown, M.P. Andrich, C.C. Chen, I.M. Feuerstein, M.E. Horowitz, I.T. Magrath, A.T. Shad, S.M. Steinberg, et al. 1995. Age, thymopoiesis, and CD4+ T-lymphocyte regeneration after intensive chemotherapy. N. Engl. J. Med. 332:143–149.
- Douek, D.C., R.A. Vescio, M.R. Betts, J.M. Brenchley, B.J. Hill, L. Zhang, J.R. Berenson, R.H. Collins, and R.A. Koup. 2000. Assessment of thymic output in adults after haematopoietic stem-cell transplantation and prediction of T-cell reconstitution. Lancet. 355:1875–1881.
- Hug, A., M. Korporal, I. Schroder, J. Haas, K. Glatz, B. Storch-Hagenlocher, and B. Wildemann. 2003. Thymic export function and T cell homeostasis in patients with relapsing remitting multiple sclerosis. J. Immunol. 171:432–437.
- Tanchot, C., A. Le Campion, B. Martin, S. Leaument, N. Dautigny, and B. Lucas. 2002. Conversion of naive T cells to a memory-like phenotype in lymphopenic hosts is not related to a homeostatic mechanism that fills the peripheral naive T cell pool. J. Immunol. 168:5042–5046.
- Ge, Q., H. Hu, H.N. Eisen, and J. Chen. 2002. Different contributions of thymopoiesis and homeostasis-driven proliferation to the reconstitution of naive and memory T cell compartments. Proc. Natl. Acad. Sci. USA. 99:2989–2994.
- Weiner, H.L., and J.A. Cohen. 2002. Treatment of multiple sclerosis with cyclophosphamide: critical review of clinical and immunologic effects. Mult. Scler. 8:142–154.
- Gbadamosi, J., C. Buhmann, W. Tessmer, A. Moench, F. Haag, and C. Heesen. 2003. Effects of mitoxantrone on multiple sclerosis patients' lymphocyte subpopulations and production of immunoglobulin, TNF-alpha and IL-10. Eur. Neurol. 49:137–141.
- Peggs, K.S., S. Verfuerth, A. Pizzey, N. Khan, P. Moss, A.H. Goldstone, K. Yong, and S. Mackinnon. 2003. Reconstitution of T-cell repertoire after autologous stem cell transplantation: influence of CD34 selection and cytomegalovirus infection. Biol. Blood Marrow Transplant. 9:198–205.
- Gorochov, G., P. Debre, V. Leblond, B. Sadat-Sowti, F. Sigaux, and B. Autran. 1994. Oligoclonal expansion of CD8+ CD57+ T cells with restricted T-cell receptor beta chain variability after bone marrow transplantation. Blood. 83:587–595.
- Morley, J.K., F.M. Batliwalla, R. Hingorani, and P.K. Gregersen. 1995. Oligoclonal CD8+ T cells are preferentially expanded in the CD57+ subset. J. Immunol. 154:6182–6190.
- Monteiro, J., F. Batliwalla, H. Ostrer, and P.K. Gregersen. 1996. Shortened telomeres in clonally expanded CD28-CD8+ T cells imply a replicative history that is distinct from their CD28+CD8+ counterparts. J. Immunol. 156:3587–3590.
- Hakim, F.T., R. Cepeda, S. Kaimei, C.L. Mackall, N. McAtee, J. Zujewski, K. Cowan, and R.E. Gress. 1997. Constraints on CD4 recovery postchemotherapy in adults: thymic insufficiency and apoptotic decline of expanded peripheral CD4 cells. Blood. 90:3789–3798.
- Singh, R.K., M.L. Varney, S. Buyukberber, K. Ino, A.G. Ageitos, E. Reed, S. Tarantolo, and J.E. Talmadge. 1999. Fas-FasL-mediated CD4+ T-cell apoptosis following stem cell transplantation. Cancer Res. 59:3107–3111.
- Mackall, C.L., T.A. Fleisher, M.R. Brown, M.P. Andrich, C.C. Chen, I.M. Feuerstein, I.T. Magrath, L.H. Wexler, D.S. Dimitrov, and R.E. Gress. 1997. Distinctions between CD8+ and CD4+ T-cell regenerative pathways result in prolonged T-cell subset imbalance after intensive chemotherapy. Blood. 89:3700–3707.
- Effros, R.B., and G. Pawelec. 1997. Replicative senescence of T cells: does the Hayflick Limit lead to immune exhaustion? Immunol. Today. 18:450–454.
- Selin, L.K., M.Y. Lin, K.A. Kraemer, D.M. Pardoll, J.P. Schneck, S.M. Varga, P.A. Santolucito, A.K. Pinto, and R.M. Welsh. 1999. Attrition of T cell memory: selective loss of LCMV epitope-specific memory CD8 T cells following infections with heterologous viruses. Immunity. 11:733–742.
- Autran, B., V. Leblond, B. Sadat-Sowti, E. Lefranc, P. Got, L. Sutton, J.L. Binet, and P. Debre. 1991. A soluble factor released by CD8+ CD57+ lymphocytes from bone marrow transplanted patients inhibits cell-mediated cytolysis. Blood. 77:2237–2241.
- Sadat-Sowti, B., P. Debre, L. Mollet, L. Quint, F. Hadida, V. Leblond, G. Bismuth, and B. Autran. 1994. An inhibitor of cytotoxic functions produced by CD8+CD57+ T lymphocytes from patients suffering from AIDS and immunosuppressed bone marrow recipients. Eur. J. Immunol. 24:2882–2888.
- Mollet, L., B. Sadat-Sowti, J. Duntze, V. Leblond, F. Bergeron, V. Calvez, C. Katlama, P. Debre, and B. Autran. 1998. CD8hi+CD57+ T lymphocytes are enriched in antigen-specific T cells capable of down-modulating cytotoxic activity. Int. Immunol. 10:311–323.
- Poser, C.M., D.W. Paty, L.C. Scheinberg, W.I. McDonald, F.A. Davis, G.C. Ebers, K.P. Johnson, K.P. Sibley, D.H. Silberberg, and W.W. Tourtellotte. 1983. New diagnostic criteria for multiple sclerosis: guidelines for research protocols. Ann. Neurol. 13:227–231.
- Kurtzke, J.F. 1983. Rating neurologic impairment in multiple sclerosis: an expanded disability status scale. Neurology. 33:1444–1452.
- Burt, R.K., B.A. Cohen, E. Russell, K. Spero, A. Joshi, Y. Oyama, W.J. Karpus, K. Luo, B. Jovanovic, A. Traynor, et al. 2003. Hematopoietic stem cell transplantation for progressive multiple sclerosis: failure of a total body irradiation-based conditioning regimen to prevent disease progression in patients with high disability scores. Blood. 102:2373–2378.
- Nash, R.A., J.D. Bowen, P.A. McSweeney, S.Z. Pavletic, K.R. Maravilla, M.S. Park, J. Storek, K.M. Sullivan, J. Al-Omaishi, J.R. Corboy, et al. 2003. High-dose immunosuppressive therapy and autologous peripheral blood stem cell transplantation for severe multiple sclerosis. Blood. 102:2364–2372.
- Inglese, M., G.L. Mancardi, E. Pagani, M.A. Rocca, A. Murialdo, R. Saccardi, G. Comi, and M. Filippi. 2004. Brain tissue loss occurs after suppression of enhancement in patients with multiple sclerosis treated with autologous haematopoietic stem cell transplantation. J. Neurol. Neurosurg. Psychiatry. 75:643–644.
- Mancardi, G.L., R. Saccardi, M. Filippi, F. Gualandi, A. Murialdo, M. Inglese, M.G. Marrosu, G. Meucci, L. Massacesi, A. Lugaresi, et al. 2001. Autologous hematopoietic stem cell transplantation suppresses Gd-enhanced MRI activity in MS. Neurology. 57:62–68.
- Carreras, E., A. Saiz, P. Marin, C. Martinez, M. Rovira, N. Villamor, M. Aymerich, M. Lozano, F. Fernandez-Aviles, A. Urbano-Izpizua, et al. 2003. CD34+ selected autologous peripheral blood stem cell transplantation for multiple sclerosis: report of toxicity and treatment results at one year of follow-up in 15 patients. Haematologica. 88:306–314.
- Saccardi, R., G.L. Mancardi, A. Solari, A. Bosi, P. Bruzzi, P. Di Bartolomeo, A. Donelli, M. Filippi, A. Guerrasio, F. Gualandi, et al. 2004. Autologous HSCT for severe progressive Multiple Sclerosis in a multicenter trial: impact on disease activity and quality of life. Blood. 10.1182/blood-2004-08-3205.
- Muraro, P.A., M. Jacobsen, A. Necker, J.W. Nagle, R. Gaber, N. Sommer, R. Martin, and B. Hemmer. 2000. Rapid identification of local T cell expansion in inflammatory organ diseases by flow cytometric T cell receptor Vb analysis. J. Immunol. Meth. 246:131–143.
- Arden, B., S.P. Clark, D. Kabelitz, and T.W. Mak. 1995. Human T-cell receptor variable gene segment families. Immunogenetics. 42:455–500.
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