We have already shown that cytokine cocktails (IL-1β, IL-3, IL-6, SCF, GM-CSF) and/or lymphokine-activated killer (LAK) cells can reduce the amounts of clonal, CD34-positive mononuclear bone marrow cells (BM-MNC) in acute myeloid leukemia (AML). In addition, the influence of those cocktails and/or LAK cells on the clonogenic potential of AML BM-MNC was investigated. BM colonies cultured in agar during different stages of the disease were immunophenotyped in situ: 17 patients at diagnosis, 14 patients in complete remission, 8 patients at relapse, 8 healthy donors. A significant reduction in leukemic cells and colonies positive for CD34 after in vitro culture of BM-MNC with cytokine cocktails was achieved with all samples obtained at diagnosis (n = 8, p < 0.01), in 6 of 8 cases in complete remission but only in 2 of 6 cases at relapse. Cytokine cocktails stimulated granulopoiesis as well as B and T lymphopoiesis. Colonies with leukemic phenotype could never be detected in healthy BM. A significant reduction in leukemic colonies was achieved by coculture of BM-MNC (uncultured or cytokine precultured) with autologous LAK cells in all 4 cases at diagnosis and in 1 case at relapse. An additive effect of in vitro cytokine preincubation of BM samples on the leukemia-reducing effect of LAK cells could be demonstrated in all samples studied (p < 0.001; diagnosis: n = 10, relapse: n = 3, complete remission: n = 7). Patients had a better prognosis if CD34-positive colonies in AML could be reduced by cytokine incubation (p = 0.03) or coculture with autologous LAK cells in vitro (p = 0.04). Our data show that cytokines as well as LAK cells alone and in combination can reduce, however not eliminate clonogenic AML cells. Such mechanisms might be responsible for maintaining stable remissions in AML.

1.
Schmetzer HM, Braun S, Wiesener D, Duell T, Gerhartz HH, Mittermueller J: Gene rearrangements in bone marrow cells of patients with acute myelogenous leukemia. Acta Haematol 2000;103:125–134.
2.
Schmetzer HM, Poleck B, Duell T, Gerhartz HH, Mittermuller J: Cytogenetic and Southern blot analysis to demonstrate clonality and to estimate prognosis in patients with myelodysplastic syndrome. Ann Hematol 2000;79:20–29.
3.
Rothe G, Schmitz G: Consensus protocol for the flow cytometric immunophenotyping of hematopoietic malignancies. Leukemia 1996;10:877–895.
4.
Gerhartz HH, Schmetzer HM: Detection of minimal residual disease in acute myeloid leukemia. Leukemia 1990;4:508–516.
5.
Buechner T: Treatment of adult acute leukemia. Curr Opin Oncol 1997;9:18–25.
6.
Buchner T, Hiddemann W, Wormann B, Zuhlsdorf M, Rottmann R, Inning G, Maschmeier G, Ludwig WD, Sauerland MC, Heinecke A: Hematopoietic growth factors in acute myeloid leukemia: Supportive and priming effects. Semin Oncol 1997;24:1–9.
7.
Brugger W, Moecklin W, Heimfeld S: Ex vivo expansion of enriched peripheral blood CD34+ progenitor cells by stem cell factor, interleukin-1β (IL-1β), IL-6, IL-3, interferon-γ and erythropoietin. Blood 1993;81:2579–2586.
8.
Smith M, Singer C, Pallister C: The effect of hemopoietic growth factors on the cell cycle of AML progenitors and their sensitivity to cytosine arabinoside in vitro. Br J Haematol 1995;90:767–772.
9.
McNiece IK, Langley KE, Zsebo KM: Recombinant human stem cell factor synergises with GM-CSF, G-CSF, IL-3 and Epo to stimulate human progenitor cells of the myeloid and erythroid lineages. Exp Hematol 1991;19:226–231.
10.
Braun S, Gerhartz HH, Schmetzer HM: Cytokines can reduce clonal, CD34 positive cells in acute myeloid leukemia (AML) in vitro. Ann Hematol 2000;79:363–373.
11.
Archimbaud E, Bailly M, Dore JF: Inducibility of lymphokine activated killer (LAK) cells in patients with acute myelogenous leukaemia in complete remission and its clinical relevance. Br J Haematol 1991;77:328–334.
12.
Braun S, Gerhartz HH, Schmetzer HM: Lymphokine-activated killer (LAK) cells and cytokines synergize to kill clonal cells in acute myeloid leukemia (AML) in vitro. Haematologia 2000;30:271–288.
13.
Adler A, Chervenick PA, Whiteside TL, Lotzova E, Herberman RB: Interleukin 2 induction of lymphokine-activated killer (LAK) activity in the peripheral blood and bone marrow of acute leukemia patients. I. Feasibility of LAK generation in adult patients with active disease and in remission. Blood 1988;71:709–716.
14.
Loewenberg B, de Greef GE, Wielenga JJ: Granulocyte-macrophage colony stimulating factor (GM-CSF) in the treatment of hematological malignancies. Anticancer Drugs 1993;4:17–20.
15.
Schmetzer H, Muller C, Wilmanns W, Gerhartz H: Cloning and characterization of bone marrow cells from patients with acute lymphoid leukemia (ALL) in agar cultures. Haematologia 1998;29:195–205.
16.
Gerhartz HH, Schmetzer H: Minimal residual disease in acute leukaemia (letter). Eur J Cancer 1991;27:809–810.
17.
Schmetzer HM, Wilmanns W, Gerhartz HH: Detection of acute myeloid leukemic cells in complete remission and in extramedullary sites by clonal analysis. Acta Haematol 1996;96:83–87.
18.
Schmetzer HM, Gerhartz HH, Wilmanns W: GM-CSF stimulated proliferation of clonal leukemic bone marrow cells in acute myeloid leukemia (AML) in vitro. Ann Hematol 1999;78:449–455.
19.
Hellstrand K, Hermodsson S, Naredi P, Mellquist UH, Brune M: Histamine and cytokine therapy. Acta Oncol 1998;37:347–353.
20.
Gore SD, Burke PJ, Wenig LJ: Impact of exogenous growth factors on proliferation and chemosensitivity of minimal residual myeloid leukemia. Leuk Lymphoma 1998;29:339–350.
21.
Dokter WH, Tuyt L, Sierdsema SJ: The spontaneous expression of interleukin-1 beta and interleukin-6 is associated with spontaneous expression of AP-1 and NF-kappa B transcription factor in acute myeloblastic leukemia cells. Leukemia 1995;9:425–432.
22.
Fialkow PJ, Singer JW, Raskind WH, Adamson JW, Jacobson RJ, Bernstein ID, Dow LW, Najfeld V, Veith R: Clonal development, stem-cell differentiation and clinical remissions in acute nonlymphocytic leukemia. N Engl J Med 1987:317:468–473.
23.
Djeu JY, Widen R, Blanchard DK: Susceptibility of monocytes to lymphokine killer cell lysis – Effect of granulocyte-macrophage colony-stimulating factor and interleukin-3. Blood 1989;73:1264–1270.
24.
Oblakowski P, Bello-Fernandez C, Reittic JE: Possible mechanism of selective killing of myeloid leukemic blast cells by lymphokine-activated killer cells. Blood 1991;77:1996–2001.
25.
Lotzova E: Recent approaches to induction of cytotoxic lymphocytes against leukemia. Leukemia 1993;7:69–74.
26.
Gerhartz HH, Schmetzer H, Unger H, Mayer F: Suppression of residual leukemic progenitors (CFU-L) in complete remission of acute myeloid leukemia by autologous LAK cells; in Bergman F (ed): Proc 2nd Cytokine Congress. Frankfurt, Thieme Verlag, 1993.
27.
Bendall LJ, Kortlepel K, Gottlieb DJ: GM-CSF enhances IL-2 activated natural killer cell lysis of clonogenic AML cells by upregulating target cell expression of ICAM-1. Leukemia 1995;9:677–684.
28.
Inokuchi K, Iwakiri R, Futaki M, Hanawa H, Tanosaki S, Nomura T, Dan K: Minimal residual disease in acute myelogenous leukemia with PML/RAR alpha or AML1/ETO mRNA and phenotypic analysis of possible T and natural killer cells in bone marrow. Leuk Lymphoma 1998;29:553–561.
29.
Bol SJ, Rosdorff HJ, Ronteltab CP, Hennen LA: Cellular cytotoxicity assessed by the 51Cr release assay. Biological interpretation of mathematical parameters. J Immunol Methods 1986;90:15–23.
30.
Campana D, Coustan-Smith E: Detection of minimal residual disease in acute leukemia by flow cytometry. Cytometry 1999;38:139–152.
31.
Tchirkov A, Giollat M, Tavernier F, Briancon G, Tournilhac O, Kwiatkowski F, Philippe P, Choufi B, Demeoco F, Travade P, Malet P: Interphase cytogenetics and competitive RT-PCR for residual disease monitoring in patients with chronic myeloid leukaemia during interferon-alpha therapy. Br J Haematol 1998;101:552–557.
32.
Engel H, Drach H, Keyhani A, Jiang S, Van NT, Kimmel M, Sanchez-Williams G, Goodacre A, Andreeff M: Quantification of minimal residual disease in acute myelogenous leukemia and myelodysplastic syndromes in complete remission by molecular cytogenetics of progenitor cells. Leukemia 1999;13:568–577.
33.
Maraninchi D, Vey N, Viens P, Stoppa AM, Archimbaud E, Attal M, Baume D, Bouabdallah R, Deeoq F, Fleury J, Michallet M, Okive D, Reiffers J, Sainty D, Tabilio A, Tiberghien P, Brandley M, Hercend T, Blaise D: A phase II study of interleukin-2 in 49 patients with relapsed or refractory acute leukemia. Leuk Lymphoma 1998;31:343–349.
34.
Suzuki R, Nakamura S: Malignancies of natural killer (NK) cell precursor: Myeloid/NK cell precursor acute leukemia and blastic NK cell lymphoma/leukemia. Leuk Res 1999;23:615–624.
35.
Lauria F, Raspadori D, Rondelli D, Ventura M, Foa R: In vitro susceptibility of acute leukemia cells to the cytotoxic activity of allogeneic and autologous lymphokine activated killer (LAK) effectors: Correlation with the rate and duration of complete remission and with survival. Leukemia 1994;8:724–728.
36.
Brune M, Hansson M, Mellquist UH: NK-cell-mediated killing of AML blasts: Role of histamine, monocytes and reactive oxygen metabolites. Eur J Haematol 1996;57:312–319.
37.
Komada Y, Zhou YW, Zhang XL, Chen TX, Tanaka S, Azuma E, Sakurai M: Fas/APO-1 (CD95)-mediated cytotoxicity is responsible for the apoptotic cell death of leukaemic cells induced by interleukin-2-activated T cells. Br J Haematol 1997;96:147–157.
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