Chronic overexpression of membrane-bound flt3 ligand by T lymphocytes in severe aplastic anaemia
Otmar Pfister,
Elena Chklovskaia,
Wendy Jansen,
Kinga Mészáros,
Catherine Nissen,
Christoph Rahner,
Nina Hurwitz,
Natalia Bogatcheva,
Stewart D. Lyman, Aleksandra Wodnar-Filipowicz,
Otmar Pfister
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorElena Chklovskaia
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorWendy Jansen
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorKinga Mészáros
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorCatherine Nissen
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorChristoph Rahner
Institute of Anatomy, University of Basle, Switzerland,
Search for more papers by this authorNina Hurwitz
Institute of Pathology, University Hospital Basle, Switzerland,
Search for more papers by this authorNatalia Bogatcheva
Institute of Paediatric Haematology, Moscow, Russia, and
Search for more papers by this authorAleksandra Wodnar-Filipowicz
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorOtmar Pfister,
Elena Chklovskaia,
Wendy Jansen,
Kinga Mészáros,
Catherine Nissen,
Christoph Rahner,
Nina Hurwitz,
Natalia Bogatcheva,
Stewart D. Lyman, Aleksandra Wodnar-Filipowicz,
Otmar Pfister
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorElena Chklovskaia
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorWendy Jansen
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorKinga Mészáros
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorCatherine Nissen
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this authorChristoph Rahner
Institute of Anatomy, University of Basle, Switzerland,
Search for more papers by this authorNina Hurwitz
Institute of Pathology, University Hospital Basle, Switzerland,
Search for more papers by this authorNatalia Bogatcheva
Institute of Paediatric Haematology, Moscow, Russia, and
Search for more papers by this authorAleksandra Wodnar-Filipowicz
Department of Research, University Hospital Basle, Switzerland,
Search for more papers by this author Aleksandra Wodnar-Filipowicz, Department of Research, University Hospital Basel, Hebelstr. 20, CH-4031 Basel, Switzerland. E-mail: [email protected]
Abstract
Aplastic anaemia (AA) is an immune-mediated bone marrow failure associated with high serum levels of flt3 ligand (FL). We examined expression of the membrane-bound isoform of FL in peripheral blood and bone marrow cells from AA patients at diagnosis (n = 16) and after immunosuppressive (IS) treatment (n = 36). Flow cytometry demonstrated strongly increased FL levels on the cell surface of T lymphocytes in AA relative to normal controls (P < 0·0001). T-cell-specific expression of membrane-bound FL was confirmed by confocal microscopy. FL mRNA and total cellular FL protein levels were increased about threefold. Overexpression of FL in AA was observed for up to 20 years after IS treatment. FL levels correlated inversely with CD34+ cell numbers and the colony-forming ability of AA bone marrow (R = −0·68 and −0·85 respectively). Histological examination of spleen specimens and bone marrow biopsies gave no evidence of degeneration or fibrosis due to prolonged exposure to high FL. Levels of membrane-bound FL were not increased in autoimmune diseases (n = 23), including rheumatoid arthritis and lupus erythematosus, nor in graft-versus-host disease (n = 8). Chronic overexpression of FL on the surface of T lymphocytes in AA, but not in other T-cell-mediated disorders, suggests that membrane-bound FL plays a role in cell–cell interactions in bone marrow failure and may be important for long-term haemopoietic recovery.
References
- 1 Birg, F., Courcoul, M., Rosnet, O., Bardin, F., Pebusque, M.J., Marchetto, S., Tabilio, A., Mannoni, P. & Birnbaum, D. (1992) Expression of the FMS/KIT-like gene FLT3 in human acute leukemias of the myeloid and lymphoid lineages. Blood, 80, 2584 – 2593.
- 2 Carow, C.E., Levenstein, M., Kaufmann, S.H., Chen, J., Amin, S., Rockwell, P., Witte, L., Borowitz, M.J., Civin, C.I. & Small, D. (1996) Expression of the hematopoietic growth factor receptor flt3 (stk-1/flk-2) in human leukemias. Blood, 87, 1089 – 1096.
- 3 Chklovskaia, E., Jansen, W., Nissen, C., Lyman, S.D., Rahner, C., Landmann, L. & Wodnar-Filipowicz, A. (1999) Mechanism of flt3 ligand expression in bone marrow failure: translocation from intracellular stores to the surface of T lymphocytes after chemotherapy-induced suppression of hematopoiesis. Blood, 93, 2595 – 2604.
- 4 Chomczynski, P. & Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry, 162, 156 – 159.
- 5 Fernandez, N.C., Lozier, A., Flament, C., Ricciardi-Castagnoli, P., Bellet, D., Sutter, M., Perricaudet, M., Tursz, T., Maraskovsky, E. & Zitvogel, L. (1999) Dendritic cells directly trigger NK functions: cross-talk relevant in innate anti-tumor immune responses in vivo. Nature Medicine, 5, 405 – 411.
- 6 Flanagan, J.G., Chan, D.C. & Leder, P. (1991) Transmembrane form of the kit ligand growth factor is determined by alternative splicing and is missing in the Sld mutant. Cell, 64, 1025 – 1035.
- 7 Gratwohl, A., John, L., Baldomero, H., Roth, J., Tichelli, A., Nissen, C., Lyman, S.D. & Wodnar-Filipowicz, A. (1998) Flt3 ligand provides hematopoietic protection from total body irradiation in rabbits. Blood, 92, 765 – 769.
- 8 Grell, M., Lohden, M., Maxeiner, B., Lesslauer, W. & Pfizenmeier, K. (1995) The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell, 83, 795 – 805.
- 9 Hannum, C., Culpepper, J., Campbell, D., McClanahan, T., Zurawski, S., Bazan, J.F., Kastelein, R., Hudak, S., Wagner, J., Mattson, J., Luh, J., Duda, G., Martina, N., Peterson, D., Menon, S., Shanafelt, A., Muench, M., Kelner, G., Namikawa, R., Rennick, D., Roncarolo, M.-G., Zlotnik, A., Rosnet, O., Dubreuil, P., Birnbaum, D. & Lee, F. (1994) Ligand for FLT3/FLK2 receptor tyrosine kinase regulates growth of haematopoietic stem cells and is encoded by variant RNAs. Nature, 368, 643 – 648.
- 10 Hawley, T.S., Fong, A.Z., Griesser, H., Lyman, S.D. & Hawley, R.G. (1998) Leukemic predisposition of mice transplanted with gene-modified hematopoietic precursors expressing flt3 ligand. Blood, 92, 2003 – 2011.
- 11 Juan, T.S.C., Van McNiece, I.G., Lacey, D., Hartley, C., McElroy, P., Sun, Y., Argento, J., Hill, D., Yan, X.Q. & Fletcher, F.A. (1997) Chronic expression of murine flt3 ligand in mice results in increased circulating white blood cell levels and abnormal cellular infiltrates associated with splenic fibrosis. Blood, 90, 76 – 84.
- 12 Lebsack, M.E., McKenna, H.J., Hoek, J.A., Hanna, R., Feng, A., Maraskovsky, E. & Hayes, F.A. (1997) Safety of flt3 ligand in healthy volunteers. Blood, 90, 170a.
- 13 Lyman, S. & Jacobsen, S. (1998) c-kit ligand and Flt3 ligand: stem/progenitor cell factors with overlapping yet distinct activities. Blood, 91, 1101 – 1134.
- 14 Lyman, S.D., James, L., Vanden, B.T., De Vries, P., Brasel, K., Gliniak, B., Hollingsworth, L.T., Picha, K.S., McKenna, H.J., Splett, R.R., Fletcher, F.F., Maraskovsky, E., Farrach, F., Foxworthe, D., Williams, D.E. & Beckmann, M.P. (1993) Molecular cloning of a ligand for the flt3/flk-2 tyrosine kinase receptor: a proliferative factor for primitive hematopoietic cells. Cell, 75, 1157 – 1167.
- 15 Lyman, S.D., James, L., Johnson, L., Brasel, K., De Vries, P., Escobar, S.S., Downey, H., Splett, R.R., Beckmann, M.P. & McKenna, H.J. (1994) Cloning of the human homologue of the murine flt3 ligand: a growth factor for early hematopoietic progenitor cells. Blood, 83, 2795 – 2801.
- 16 Lyman, S.D., Seaberg, M., Hanna, R., Zappone, J., Brasel, K., Abkowitz, J.L., Prchal, J., Schultz, J. & Shahidi, N.T. (1995) Flt3 ligand plasma/serum levels are low in normal individuals and are highly elevated in patients with Fanconi anemia and acquired aplastic anemias. Blood, 86, 4091 – 4096.
- 17 Lynch, D.H., Andreasen, A.E.M., Whitmore, J., Miller, R.E. & Schuh, J.C.L. (1997) Flt3 ligand induces tumor regression and antitumor immune responses in vivo. Nature Medicine, 3, 625 – 631.
- 18 Maciejewski, J.P., Hibbs, J.R., Anderson, S., Katevas, P. & Young, N.S. (1994) Bone marrow and peripheral blood lymphocyte phenotype in patients with bone marrow failure. Experimental Hematology, 22, 1102 – 1110.
- 19 Maciejewski, J., Selleri, C., Anderson, S. & Young, N.S. (1995) Fas antigen expression on CD34+ human marrow cells is induced by interferon γ and tumor necrosis factor α and potentiates cytokine-mediated hematopoietic suppression in vitro. Blood, 85, 3138 – 3190.
- 20 Manz, C.Y., Nissen, C. & Wodnar-Filipowicz, A. (1996) Deficiency of CD34+c-kit+ and CD34+38− hematopoietic precursors in aplastic anemia after immunosuppressive treatment. American Journal of Hematology, 52, 264 – 274.
- 21 Marsh, J.C. & Gordon-Smith, E.C. (1998) Treatment options in severe aplastic anaemia. Lancet, 351, 1830 – 1831.
- 22 Matthews, W., Jordan, C.T., Gavin, M., Jenkins, N.A., Copeland, N.G. & Lemischka, I.R. (1991) A receptor tyrosine kinase cDNA isolated from a population of enriched primitive hematopoietic cells and exhibiting close genetic linkage to c-kit. Proceedings of the National Academy of Sciences of the United States of America, 88, 9026 – 9030.
- 23 McClanahan, T., Culpepper, J., Campbell, D., Wagner, J., Franz-Bacon, K., Mattson, J., Tsai, S., Luh, J., Guimaraes, J., Mattei, M.-G., Rosnet, O., Birnbaum, D. & Hannum, C.H. (1996) Biochemical and genetic characterisation of multiple splice variants of the flt3 ligand. Blood, 88, 3371 – 3382.
- 24 McKenna, H.J., Smith, F.O., Brasel, K., Hirschstein, D., Bernstein, I.D., Williams, D.E. & Lyman, S.D. (1996) Effects of flt3 ligand on acute myeloid and lymphocytic blast cells from children. Experimental Hematology, 24, 378 – 385.
- 25 Melenhorst, J.J., Van Krieken, J.H., Dreef, E., Landegent, J.E., Willemze, R. & Fibbe, W.E. (1997) T cells selectively infiltrate bone marrow areas with residual haemopoiesis of patients with acquired aplastic anaemia. British Journal of Haematology, 99, 517 – 519.
- 26 Nagafui, K., Shibuya, T., Harada, M., Mizuno, S.I., Takenaka, K., Miyamoto, T., Okamura, T., Gondo, H. & Niho, Y. (1995) Functional expression of Fas antigen (CD95) on hematopoietic progenitor cells. Blood, 86, 883 – 889.
- 27 Nakao, S., Takamatsu, H., Yachie, A., Itoh, T., Yamaguchi, M., Ueda, M., Shiobara, S. & Matsuda, T. (1995) Establishment of a CD4+ T cell clone recognizing autologous hematopoietic progenitor cells from a patient with immune-mediated aplastic anaemia. Experimental Hematology, 23, 433 – 438.
- 28 Nissen, C., Wodnar-Filipowicz, A., Slanicka Krieger, M., Gratwohl, A., Tichelli, A. & Speck, B. (1995) Persistent growth impairment of bone marrow stroma after antilymphocyte globulin treatment for severe aplastic anemia and its association with relapse. European Journal of Haematology, 55, 255 – 261.
- 29 Papayannopoulou, T., Nakamoto, B., Andrews, R.G., Lyman, S.D. & Lee, M.Y. (1997) In vivo effects of flt3/flk2 ligand on mobilization of hematopoietic progenitors in primates and potent synergistic enhancement with granulocyte colony-stimulating factor. Blood, 90, 620 – 629.
- 30 Piacibello, W., Fubini, L., Sanavio, F., Brizzi, M.F., Severino, A., Garetto, L., Stacchini, A., Pegoraro, L. & Aglietta, M. (1995) Effects of human flt3 ligand on myeloid leukemias cell growth: heterogeneity in response and synergy with other hematopoietic growth factors. Blood, 85, 4105 – 4114.
- 31 Podesta, M., Piaggio, G., Frassoni, F., Pitto, A., Zikos, P., Sessarego, M., Abate, M.M.T.V.L., Berisso, G. & Bacigalupo, A. (1998) The assessment of the hematopoietic reservoir after immunosuppressive therapy or bone marrow transplantation in severe aplastic anemia. Blood, 91, 1959 – 1965.
- 32 Rosnet, O., Schiff, C., Pebusque, M.J., Marchetto, S., Tonnelle, C., Toiron, Y., Birg, F. & Birnbaum, D. (1993) Human FLT3/FLK2 gene: cDNA cloning and expression in hematopoietic cells. Blood, 82, 1110 – 1119.
- 33 Slanicka Krieger, M., Nissen, C., Manz, C.Y., Toksoz, D.S.D., Lyman, S.D. & Wodnar-Filipowicz, A. (1998) The membrane-bound isoform of stem cell factor synergizes with soluble flt3 ligand in supporting early hematopoiesis in long-term cultures of normal and aplastic anemia bone marrow. Experimental Hematology, 26, 365 – 373.
- 34 Tanaka, M., Itai, T., Adachi, M. & Nagata, S. (1998) Downregulation of Fas ligand by shedding. Nature Medicine, 4, 31 – 36.
- 35 Tichelli, A., Gratwohl, A., Würsch, A., Nissen, C. & Speck, B. (1988) Late haematological complications in severe aplastic anaemia. British Journal of Haematology, 69, 413 – 418.
- 36 Toksoz, D., Zsebo, K.M., Smith, K.A., Hu, S., Brankow, D., Suggs, S.V., Martin, F.H. & Williams, D.A. (1992) Support of human hematopoiesis in long-term bone marrow cultures by murine stromal cells selectively expressing the membrane-bound and secreted forms of the human homolog of the steel gene product, stem cell factor. Proceedings of the National Academy of Sciences of the United States of America, 89, 7350 – 7354.
- 37 Wodnar-Filipowicz, A., Tichelli, A., Zsebo, K.M., Speck, B. & Nissen, C. (1992) Stem cell factor stimulates the in vitro growth of bone marrow cells from aplastic anemia patients. Blood, 79, 3196 – 3202.
- 38 Wodnar-Filipowicz, A., Lyman, S., Gratwohl, A., Tichelli, A., Speck, B. & Nissen, C. (1996) Flt3 ligand level reflects hematopoietic progenitor cell function in aplastic anemia and chemotherapy-induced bone marrow aplasia. Blood, 88, 4493 – 4499.
- 39 Xu, F., Taki, T., Yang, H.W., Hanada, R., Hongo, T., Ohnishi, H., Kobayashi, M., Bessho, F., Yaniagisawa, M. & Hayashi, Y. (1999) Tandem duplication of the flt3 gene is found in acute lymphoblastic leukaemia as well as acute myeloid leukaemia but not in myelodysplastic syndrome or juvenile chronic myelogenous leukaemia in children. British Journal of Haematology, 105, 155 – 162.
- 40 Young, N.S. (1995) Aplastic anaemia. Lancet, 346, 228 – 232.
- 41 Zeng, W., Nakao, S., Takamatsu, H., Yachie, A., Takami, A., Kondo, Y., Sugimori, N., Yamazaki, H., Miura, Y., Shiobara, S. & Matsuda, T. (1999) Characterization of T-cell repertoire of the bone marrow in immune-mediated aplastic anemia: evidence for the involvement of antigen-driven T-cell response in cyclosporin-dependent aplastic anemia. Blood, 93, 3008 – 3016.
Citing Literature
