Abstract
Flt3-ligand (FL) is a cytokine that is of paramount importance in the proliferation of primitive hematopoietic progenitors. In this study, we show that endothelial cells (EC) produce large amounts of soluble FL and express a membrane-bound form of the molecule. Bone marrow microvascular EC also produce FL, suggesting that EC are an important source of FL in the bone marrow. High concentrations of FL in EC supernatants contrast with its undetectable levels in long-term bone marrow cultures. A single mRNA for FL is detected, suggesting that soluble FL derives from the membrane-bound species by proteolytic release. FL mRNA is stable with a half-life of about 3 h. II-1α increases FL mRNA levels and membrane and soluble FL expression. Glucocorticoids, known inhibitors for many hematopoietic growth factors do not down-regulate the expression of FL. On the contrary, GC increase the expression of both species of FL. The neutralization of FL in cocultures EC/ hematopoietic progenitors results in an acceleration of the maturation of the progenitors. IFN-α, MIP-1 α and TGF-β stimulate production of membrane-bound and soluble FL. This stimulation is essential to explain their modulatory effect on the generation of clonogenic cells in cocultures EC/hematopoietic progenitors.
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References
Lyman SD, James L, Vanden-Bos T, de Vries P, Brasel K, Gliniak B, Hollingworth LT, Picha KS, McKenna HJ, Splett RR, Fletcher FA, Maraskovsky E, Farrah T, Foxworthe D, Williams DE, Beckman MP . Molecular cloning of a ligand for the flt3/flk-2 tyrosine kinase receptor: a proliferative factor for primitive hematopoietic cells Cell 1993 75: 1157–1167
Hannum C, Culpepper J, Campbell D, McClanahan T, Zurawski S, Bazan JF, Kastelein R, Hudak S, Wagner J, Mattson J, Luh J, Duda G, Martina N, Pterson D, Menon S, Shanafelt A, Muench MO, Kelner GS, Namikawa R, Rennick D, Roncarolo MG, Zlotnik A, Rosnet O, Dubreuil P, Birnbaum D, Lee F . Ligand for FLT3/FLK2 receptor tyrosine kinase regulates growth of hematopoietic stem cells and is encoded by variant RNAs Nature 1994 368: 643–648
Lyman SD, James L, Johnson L, Brasel K, de Vries P, Escobar SS, Downey H, Splett RR, Beckman MP, McKenna HJ . Cloning of the human homolog of the murine flt3 ligand: a growth factor for early hematopoietic progenitor cells Blood 1994 83: 2795–2801
Small D, Levenstein M, Kim E, Carow C, Amin S, Rockwell P, Witte L, Burrow C, Ratajczak MZ, Gerwitz AM, Civin CI . STK-1 the human homologue of FLK2/FLT3, is selectively expressed in human bone marrow cells and is involved in the proliferation of early progenitor/stem cells Proc Natl Acad Sci USA 1994 91: 459–463
Jacobsen SE, Okkenhaug C, Myklebust J, Veiby OP, Lyman SD . The FLT3 ligand potently and directly stimulates the growth and expansion of primitive murine bone marrow progenitor cells in vitro: synergistic interactions with interleukin (IL)-11, IL-12 and other hematopoietic growth factors J Exp Med 1995 181: 1357–1363
Hudak S, Hunte B, Culpepper J, Menon S, Hannum C, Thomson-Snipes L, Rennick D . FLT3/FLK2 ligand promotes the growth of murine stem cells and the expansion of colony-forming cells and spleen colony-forming units Blood 1995 85: 2747–2755
Piacibello W, Fubini L, Sanavio F, Brizzi MF, Severino A, Garetto L, Stacchini A, Pegoraro L, Aglietta M . Effects of human FLT3 ligand on myeloid leukemia cell growth: heterogeneity in response and synergy with other hematopoietic growth factors Blood 1995 86: 4105–4114
Muench MO, Roncarolo MG, Menon S, Xu Y, Kastelein R, Zurawski S, Hannum CH, Culpepper J, Lee F, Namikawa R . FLK2/FLT3 ligand regulates the growth of early myeloid progenitors isolated from human fetal liver Blood 1995 85: 963–972
McKenna HJ, de Vries P, Brasel K, Lyman SD, Williams DE . Effect of flt3 ligand on the ex vivo expansion of human CD34+ hematopoietic progenitors cells Blood 1995 86: 3413–3420
Rusten LS, Lyman SD, Veiby OP, Jacobsen SE . The FLT3 ligand is a direct and potent stimulator of the growth of primitive and committed human CD34+ bone marrow progenitor cells in vitro Blood 1996 87: 1317–1325
Petzer AL, Zandstra PW, Piret JM, Eaves CJ . Differential cytokine effects on primitive (CD34+ CD38−) human hematopoietic cells: novel responses to Flt3-ligand and thrombopoietin J Exp Med 1996 183: 2551–2558
Ramsfjell V, Borge OJ, Cui L, Jacobsen SE . Thrombopoietin directly and potently stimulates multilineage growth and progenitor cell expansion from primitive (CD34+ CD38−) human bone marrow progenitor cells. Distinct and key interactions with the ligands for c-kit and flt3, and inhibitory effects of TGF-β and TNF-α J Immunol 1997 158: 5169–5177
Piacibello W, Sanavio F, Garetto L, Severino A, Bergandi D, Ferriaro J, Fagioli F, Berger M, Aglietta M . Extensive amplification and self-renewal of human primitive hematopoietic stem cells from cord blood Blood 1997 89: 2644–2653
Zandstra PW, Conneally E, Petzer AL, Piret JM, Eaves CJ . Cytokine manipulation of primitive human hematopoietic cell self-renewal Proc Natl Acad Sci USA 1997 94: 4698–4703
Mackarehtschian K, Hardin JD, Moore KA, Boast S, Goff SP, Lemischka IR . Targeted disruption of the flk2/flt3 gene leads to deficiencies in primitive hematopoietic progenitors Immunity 1995 3: 147–161
Mahon FX, Pigeonnier-Lagarde V, Chahine H, Barbot C, Jazwiec B, Ripoche J, Reiffers J . Ex vivo cytokine expansion of peripheral blood 5-fluorouracil-treated CD34-positive chronic myeloid leukaemia cells increases the selection of Ph-negative cells Br J Haematol 1997 98: 467–473
Maraskovsky E, Brasel K, Teepe M, Roux ER, Lyman SD, Shortman K, McKenna HJ . Dramatic increase in the number of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cells subpopulations identified J Exp Med 1996 184: 1953–1962
Lyman SD . Biology of Flt3 ligand and receptor Int J Hematol 1996 62: 63–73
Strobl H, Bello-Fernandez C, Riedl E, Pickl WF, Majdic O, Lyman SD, Knapp W . Flt3 ligand in cooperation with transforming growth factor-β1 potentiates in vitro development of Langerhans-type dendritic cells and allows single-cell dendritic cell cluster formation under serum-free conditions Blood 1997 90: 1425–1434
Meierhoff G, Dehmel U, Gruss HJ, Rosnet O, Birnbaum D, Quentmeier H, Dirks W, Drexler HJ . Expression of FLT3 receptor and FLT3-ligand in human leukemia–lymphoma cell lines Leukemia 1995 9: 1368–1372
Lisovsky M, Braun SE, Ge Y, Takahira H, Lu L, Savchenko VG, Lyman SD, Broxmeyer HE . Flt3-ligand production by human bone marrow stromal cells Leukemia 1996 10: 1012–1018
McClanahan T, Culpepper J, Campbell D, Wagner J, Franz-Bacon K, Mattson J, Tsai S, Luh J, Guimaraes MJ, Mattei MG, Rosnet O, Birnbaum D, Hannum CH . Biochemical and genetic characterization of multiple splice variants of the Flt3 ligand Blood 1996 88: 3371–3382
Jaffe EA, Nachman RL, Becker CG, Minick CR . Culture of human endothelial cells derived from umbilical vein. Identification by morphologic and immunologic criteria J Clin Invest 1973 52: 2745–2756
Rafii S, Shapiro F, Rimarachin J, Nachman RL, Ferris B, Weksler B, Moore MAS, Asch AS . Isolation and characterization of human bone marrow microvascular endothelial cells: hematopoieticprogenitor cells adhesion Blood 1994 84: 10–19
Schweitzer CM, van der Schoot CE, Dräger AM, van der Valk P, Zevenbergen A, Hooibrink B, Westra AH, Langenhuijsen MMAC . Isolation and culture of human bone marrow endothelial cells Exp Hematol 1995 23: 41–48
Rutkowski JL, Cassandra JK, Lerner MA, Tennekoon GI . Purification and expansion of human Schwann cells in vitro Nature Med 1995 1: 80–83
Schweitzer KM, Vicart P, Delouis C, Paulin D, Dräger AM, Langenhuijsen MMAC, Weksler BB . Characterization of a newly established human bone marrow endothelial cell line: distinct adhesive properties for hematopoietic progenitors compared with human umbilical vein endothelial cells Lab Invest 1997 76: 25–36
Jazwiec B, Solanilla A, Grosset C, Mahon FX, Dupouy M, Pigeonnier-Lagarde V, Belloc F, Schweitzer CM, Reiffers J, Ripoche J . Endothelial cell support of hematopoiesis is differentially altered by IL-1 and glucocorticoids Leukemia 1998 12: 1210–1220
Chomczynski P, Sacchi N . Single-step method for RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction Anal Biochem 1987 162: 156–159
Witte ON . Steel locus defines new multipotent growth factor Cell 1990 63: 5–6
Strobel ES, Gay RE, Greenberg PL . Characterization of the in vitro stromal microenvironment of human bone marrow Int J Cell Cloning 1986 4: 341–356
Siena S, Castro-Malaspina H, Gulati SC, Lu L, Colvin MO, Clarkson BD, O'Reilly RJ, Moore MA . Effects of in vitro purging with 4-hydroperoxycyclophosphamide on the hematopoietic and microenvironmental elements of human bone marrow Blood 1985 65: 655–662
Slovick FT, Abboud CN, Brennan JK, Lichtman MA . Survival of granulocytic progenitors in the nonadherent and adherent compartments of human long-term marrow cultures Exp Hematol 1984 12: 327–338
Keating A, Singer JW, Killen PD, Striker GE, Salo AC, Sanders J, Thomas ED, Thorning D, Fialkow PJ . Donor origin of the in vitro hematopoietic microenvironment after marrow transplantation in man Nature 1982 298: 280–283
Toogood IR, Dexter TM, Allen TD, Suda T, Lajtha LG . The development of a liquid culture system for the growth of human bone marrow Leukemia Res 1980 4: 449–461
Andreoni C, Moreau I, Rigal D . Long-term culture of human bone marrow. I. Characterization of adherent cells in flow cytometry Exp Hematol 1990 18: 431–437
Berneman ZN, Chen ZZ, Ramael M, Van Poucke K, Korthout M, Van Bockstaele DR, Peetermans ME . A quantitative and dynamic study of endothelial cells and megakaryocytes in human long-term bone marrow cultures Leukemia 1989 3: 61–67
Gessani S, MacCandless S, Baglioni C . The glucocorticoid dexamethasone inhibits synthesis of interferon by decreasing the level of its mRNA J Biol Chem 1988 263: 7454–7457
Beutler B, Krochin N, Milsark IW, Luedke C, Cerami A . Control of cachectin (tumor necrosis factor) synthesis: mechanisms of endotoxin resistance Science 1986 232: 977–980
Knudsen PJ, Dinarello CA, Strom TB . Glucocorticoids inhibit transcriptional and posttranscriptional expression of interleukin 1 in U937 cells J Immunol 1987 139: 4129–4134
Tobler A, Meier R, Seitz M, Dewald B, Baggiolini M, Fey MF . Glucocorticoids downregulate gene expression of GM-CSF, NAP-1/IL-8 and IL-6, but not M-CSF in human fibroblasts Blood 1992 79: 45–51
Grosset C, Taupin JL, Lemercier C, Moreau JF, Reiffers J, Ripoche J . Leukemia inhibitory factor expression is inhibited by glucocorticoids through post-transcriptional mechanisms Cytokine 1999 11: 29–36
Linenberger ML, Jacobson FW, Bennett LG, Broudy VC, Martin FH, Abkowitz JL . Stem cell factor production by human marrow stromal fibroblasts Exp Hematol 1995 23: 1104–1114
Chklovskaia E, Jansen W, Nissen C, Gratwohl A, Lyman SD, Wodnar-Filipowicz A . FLT3 ligand is prestored in T-lymphocytes and is released in response to chemotherapy-induced stem cell depletion Exp Hematol 1997 25: 8 (Abstr.)
Kriegler AB, Bernardo D, Verschoor SM . Protection of murine bone marrow by dexamethasone during cytotoxic chemotherapy Blood 1994 83: 65–71
Aman JM, Keller U, Derigs G, Mohamadzadeh M, Huber C, Peschel C . Regulation of cytokine expression by interferon-α in human bone marrow stromal cells. Inhibition of hematopoietic growth factors and induction of interleukin-1 receptor antagonist Blood 1994 84: 4142–4150
Aman MJ, Bug G, Aulitzky WE, Huber C, Peschel C . Inhibition of interleukin-11 by interferon-alpha in human bone marrow stromal cells Exp Hematol 1996 24: 863–867
Peschel C, Aulitzky WE, Huber C . Influence of interferon-alpha on cytokine expression by the bone marrow microenvironment-Impact on treatment of myeloproliferative disorders LeukLymphoma 1996 22: 129–134
Keller JR, Jacobsen SE, Dubois CM, Hesdal K, Ruscetti FW . Transforming growth factor-beta: a bidirectional regulator of hematopoietic cell growth Int J Cell Cloning 1992 10: 2–11
Keller JR, Jacobsen SE, Sill KT, Ellingsworth LR, Ruscetti FW . Stimulation of granulopoiesis by transforming growth factor beta: synergy with granulocyte/macrophage-colony-stimulating factor Proc Natl Acad Sci USA 1991 88: 7190–7194
Keller JR, Bartelmez SH, Sitnicka E, Ruscetti FW, Ortiz M, Gooya JM, Jacobsen SE . Distinct and overlapping direct effects of macrophage inflammatory protein 1-α and transforming growth factor β on hematopoietic progenitor/stem cell growth Blood 1994 84: 2175–2181
Van Ranst PC, Snoeck HW, Lardon F, Lenjou M, Nijs G, Weekx SF, Rodrigus I, Berneman ZN, Van Bockstaele DR . TGF-beta and MIP-1 alpha exert their main inhibitory activity on very primitive CD34+CD38− cells but show opposite effects on more mature CD34+CD38− human hematopoietic progenitors Exp Hematol 1996 24: 1509–1515
Garbe A, Spyridonidis A, Mobest D, Schmoor C, Mertelsmann R, Henschler R . Transforming growth factor-beta 1 delays formation of granulocyte–macrophage colony-forming cells, but spares more primitive progenitors during ex vivo expansion of CD34+ haematopoietic progenitor cells Br J Haematol 1997 99: 951–958
Broxmeyer HE, Sherry B, Lu L, Cooper S, Oh D, Tekamp-Olson P, Kwon BS, Cerami A . Enhancing and suppressing effects of recombinant murine macrophage inflammatory proteins on colony formation in vitro by bone marrow myeloid progenitor cells Blood 1990 76: 1110–1116
Verfaillie CM, Catanzarro PM, Li WN . Macrophage inhibitory protein 1α, interleukin 3 and diffusible marrow stromal factors maintain human hematopoietic stem cells for at least eight weeks in vitro J Exp Med 1994 179: 643–649
Koenig A, Yakisan E, Reuter M, Huang M, Sykora KW, Corbacioglu S, Welte K . Differential regulation of stem cell factor mRNA expression in human endothelial cells by bacterial pathogens: an in vitro model of inflammation Blood 1994 83: 2836–2843
Piacibello W, Sanavio F, Garetto L, Severino A, Bergandi D, Ferrario J, Fagioli F, Berger M, Aglietta M . Extensive amplification and self-renewal of human primitive hematopoietic stem cells from cord blood Blood 1997 89: 2644–2653
Acknowledgements
We wish to thank Dr SD Lyman (Immunex, Seattle, Washington) for the gift of recombinant FL and monoclonal anti-FL antibody, helpful comments and drawing our attention to the possible up-regulation of FL-cleaving protease by GC. This work was supported by a grant from the Fondation pour la Recherche Médicale and an ARC fellowship (CG).
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Solanilla, A., Grosset, C., Lemercier, C. et al. Expression of Flt3-ligand by the endothelial cell. Leukemia 14, 153–162 (2000). https://doi.org/10.1038/sj.leu.2401635
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DOI: https://doi.org/10.1038/sj.leu.2401635
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