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Differences of cellular composition and adhesion molecule expression in “leukemic” as compared with “normal” human long-term bone marrow cultures

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Summary

Human long-term bone marrow cultures (HLTBMCs) were established with bone marrow samples collected from 15 patients with acute myeloid leukemia (AML) and compared with HLTBMCs from eight healthy volunteers. During 6 weeks of culture, the cellular composition of HLTBMCs was quantitatively studied. The cells of the HLTBMCs were divided into three main categories: fibroblasts, macrophages, and ‘other cells’ (endothelial cells, hematopoietic cells and undefined cells). HLTBMCs derived from healthy volunteers demonstrated a very consistent development. The number of fibroblasts increased during culture and the number of macrophages decreased, resulting in a steady state after 3 weeks of culture. In contrast, HLTBMCs derived from patients with AML showed a strikingly different pattern of irregular development and a steady state was not reached under our conditions. The APAAP technique was used to demonstrate expression of adhesion molecules. VLA2, VLA5, VLA6, LFA1, Mac1, p150/95, β2-chain, HCAM, ICAM1, NCAM, and VCAM1 were more expressed on ‘normal’ as compared with ‘leukemic’ bone marrow stromal cells, although this reached significance only for β2-chain and NCAM. VLA1, 3, and 4 were expressed in a higher percentage on ‘leukemic’ stroma (not significant). More expression was seen on ‘normal’ as opposed to ‘leukemic’ macrophages for the adhesion molecules tested, except for VLA5. The differences reached significance for the majority of molecules tested. It is concluded that striking differences exist in cellular composition and adhesion molecule expression between HLTBMCs from healthy individuals and those from patients with AML. This may have an impact on the pathogenesis of AML.

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References

  1. Allen TD, Dexter TM (1984) The essential cells of the hemopoietic microenvironment. Exp Hematol 12: 517–521

    Google Scholar 

  2. Bainton DF (1985) Bone marrow stromal cells — fibroblasts and macrophages. In: Furth v R (ed) Mononuclear phagocytes: characteristics, physiology and function. Martinus Nijhoff, Dordrecht, The Netherlands, pp 125–135

    Google Scholar 

  3. Berneman ZN, Chen Z, Ramael M, Van Poucke K, Korthout M, Bockstaele DR, Peetermans M (1989) A quantitative and dynamic study of endothelial cells and megakaryocytes in human long-term bone marrow cultures. Leukemia 3: 61–67

    Google Scholar 

  4. Broudy VC, Zuckerman KS, Jetmalani S, Fitchen JH, Bagby Jr GC (1986) Monocytes stimulate fibroblastoid bone marrow cells to produce multilineage hematopoietic growth factors. Blood 68: 530–534

    Google Scholar 

  5. Carlos TM, Schwartz BR, Kovach NL, Yee E, Rosso M, Osborn L, Chi-Rosso G, Newman B, Lobb R, Harlan JM (1990) Vascular cell adhesion molecule-1 mediates lymphocyte adherence to cytokine-activated cultured human endothelial cells. Blood 76: 965–970

    Google Scholar 

  6. Chang J, Geary CG, Testa NG (1990) Long-term bone marrow damage after chemotherapy for acute myeloid leukaemia does not improve with time. Br J Haematol 75: 68–72

    Google Scholar 

  7. Chang J, Morgenstern GR, Coutinho LH, Scarffe JH, Carr T, Deakin DP, Testa NG, Dexter TM (1989) The use of bone marrow cells grown in long-term culture for autologous bone marrow transplantation in acute myeloid leukemia: an update. Bone Marrow Transplant 4: 5–9

    Google Scholar 

  8. Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulasiz Z, Macdonald S, Pulford KAF, Stein H, Mason DY (1984) Immunoenzymatic labelling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP) complexes. Histochem Cytochem 32: 219–229

    Google Scholar 

  9. Coulombel L, Eaves C, Kalousek D, Gupta C, Eaves A (1985) Long-term culture of cells from patients with acute myeloid leukemia. J Clin Invest 75: 961–969

    Google Scholar 

  10. Coutinho LH, Geary CG, Chang J, Harrison C, Testa NG (1990) Functional studies of bone marrow haemopoietic and stromal cells in the myelodysplastic syndrome (MDS). Br J Haematol 75: 16–25

    Google Scholar 

  11. Dexter TM, Allen TD, Lajtha LG (1977) Proliferation of haemopoietic stem cells in vitro. J Cell Physiol 91: 335–341

    Google Scholar 

  12. Gartner S, Kaplan HS (1980) Long-term culture of human bone marrow cells. Proc Natl Acad Sci USA 77: 4756–4759

    Google Scholar 

  13. Glück U, Zipori D, Wetzler M, Berrebi A, Shaklai M, Drezen O, Zaizov R, Luria D, Marcelle C, Stark B, Umiel T (1989) Long-term proliferation of human leukemia cells induced by mouse stroma. Exp Hematol 17: 398–404

    Google Scholar 

  14. Gordon MY (1988) Adhesive properties of haemopoietic stem cells. Br J Haematol 68: 149–151

    Google Scholar 

  15. Gordon MY (1988) The origin of stromal cells in patients treated by bone marrow transplantation. Bone Marrow Transplant 3: 247–251

    Google Scholar 

  16. Gordon MY, Goldman JM, Gordon-Smith EC (1983) Spatial and functional relationships between human hemopoietic and marrow stromal cells in vitro. Int J Cell Cloning 1: 429–439

    Google Scholar 

  17. Haynes BF, Scearce RM, Lobach DF, Hensly LL (1984) Phenotypic characterization and ontogeny of mesodermal-derived and endocrine epithelial components of the human thymic microenvironment. J Exp Med 159: 1149–1168

    Google Scholar 

  18. Heuvel RL van den, Schoeters GER, Vandenborght OLJ (1988) Haemopoiesis in long-term cultures of liver, spleen and bone marrow of pre- and postnatal mice: CFU-GM production. Br J Haematol 70: 273–277

    Google Scholar 

  19. Hocking WG, Golde DW (1980) Long-term human bone marrow cultures. Blood 56: 118–124

    Google Scholar 

  20. Hand HJ, Croaker GM, Repka E, Radioff TJ, Vincent PC (1987) Long-term bone marrow culture induces terminal differentiation of human myeloid leukemic cells. Exp Hematol 15: 1109–1114

    Google Scholar 

  21. Kelly PMA, Bliss E, Morton JA, Burns J, McGee JO'D (1988) Monoclonal antibody EBM11: high cellular specificity for human macrophages. J Clin Pathol 41: 510–515

    Google Scholar 

  22. Lanotte M, Allen TD, Dexter TM (1981) Histochemical and ultrastructural characteristics of a cell line from human bonemarrow stroma. J Cell Sci 50: 281–297

    Google Scholar 

  23. Mukai K, Rosai J, Burgdorff WHC (1980) Localization of Factor VIII-related antigen in vascular endothelials cells using an immuno-peroxidase method. Am J Surg Pathol 4: 273–276

    Google Scholar 

  24. Payne CM, Greenberg BR, Cromey D, Woo L (1987) Morphological evidence of an altered bone marrow microenvironment in patients with acute nonlymphoblastic leukemia and myelodysplastic disorders. Exp Hematol 15: 143–153

    Google Scholar 

  25. Petrides PE, Dittmann KH (1990) How do normal and leukemic white blood cells egress from the bone marrow? Blut 61: 3–13

    Google Scholar 

  26. Roberts R, Gallagher J, Spooncer E, Allen TD, Bloomfield F, Dexter TM (1988) Heparan sulphate-bound growth factor: a mechanism for stromal cell-mediated haemopoiesis. Nature 332: 376–378

    Google Scholar 

  27. Ruoslathi E (1991) Integrins. J Clin Invest 87: 1–5

    Google Scholar 

  28. Schiro R, Couthinho LH, Testa NG, Dexter TM (1989) The effect of exogenous growth factors on stromal cells from human LTBMC. Tissue Antigens 3: 229

    Google Scholar 

  29. Schölzel C, Löwenberg B (1985) Stimulation of proliferation and differentiation of acute myeloid leukemia cells on a bone marrow stroma in culture. Exp Hematol 13: 664–669

    Google Scholar 

  30. Soligo D, Schiro R, Luksch R, Manara G, Quirici N, Parravicini C, Lambertenghi Deliliers G (1990) Expression of integrins in human bone marrow. Br J Haematol 76: 323–332

    Google Scholar 

  31. Springer TA, Lasky LA (1991) Sticky sugars for selectins. Nature 349: 196–197

    Google Scholar 

  32. Toogood IRG, Dexter TM, Allen TD, Suda T, Lajtha LG (1980) The development of a liquid culture system for the growth of human bone marrow. Leuk Res 4: 449–461

    Google Scholar 

  33. Tsai S, Patel V, Beaumont E, Lodish HF, Nathan DG, Sieff CA (1987) Differential binding of erythroid and myeloid progenitors to fibroblasts and fibronectin. Blood 69: 1587–1594

    Google Scholar 

  34. Umiel T, Friedman S, Zaizov R, Cohen IJ, Gozes Y, Epstein N, Kobiler D, Zipori D (1986) Long-term culture of infant leukemia cells: dependence upon stromal cells from the bone marrow and bilineage differentiation. Leuk Res 10: 1007–1013

    Google Scholar 

  35. Weide M van der, Langenhuijsen MMAC, Huijgens PC, Imandt LMFM, Waal FC de, Mol JJ, Rhenen DJ van, Kester APM (1987) Relation between leukaemic cell count and degree of maturation in acute myeloid leukaemia. Eur J Cancer Clin Oncol 23: 1125–1129

    Google Scholar 

  36. Yourno J, Mastropaolo W (1981) Nonspecific esterases of the formed elements: zymograms produced by pH 9.5 polyacrylamide gel electrophoresis. Blood 57: 939–945

    Google Scholar 

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Denkers, I.A.M., Beelen, R.H.J., Ossenkoppele, G.J. et al. Differences of cellular composition and adhesion molecule expression in “leukemic” as compared with “normal” human long-term bone marrow cultures. Ann Hematol 64, 210–216 (1992). https://doi.org/10.1007/BF01738298

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  • DOI: https://doi.org/10.1007/BF01738298

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