Abstract
The aim of this study was to measure the level of cytokines produced by peripheral blood mononuclear cells (PBMNC) in patients with aplastic anemia (AA) and to determine their effect on the clonal growth of normal bone marrow (BM) cells. Twenty-one patients with AA and 11 normal controls were enrolled in this study. Medium conditioned by PBMNC of AA patients in the presence of lipopolysaccharide (LPS) was found to be suppressive to the colony growth of normal BM cells. Thus, we further determined the presence in the PBMNC-conditioned medium (CM) of both inhibitory cytokines: macrophage inflammatory protein-1α (MIP-1α), tumor necrosis factor-α (TNF-α), transforming growth factor-β2 (TGF-β2), and interferon-γ (IFN-γ), and stimulatory cytokines: interleukin-3 (IL-3) and stem cell factor (SCF). Spontaneous production of MlP-1α was higher in the AA patients than the normal controls (1887±174 pg/ml vs 1643±93 pg/ml), but the difference was not significant. After LPS stimulation, the production of MIP-1α was markedly increased in the AA patients, and its level was significantly higher than that of the normal controls (2360±149 pg/ml vs 1517±92 pg/ml, p=0.0022). The level of TNFα was also higher in the AA patients. However, IFN-γ, TGF-β2, SCF, and IL-3 were not detectable in the PBMNC-CM of either AA patients or normals. The myelopoietic suppressing effect of AA-PBMNC-CM from each AA patient was significantly blocked by pretreatment with anti-TNF-α, resulting in a colony-forming enhancement of 174%±12%. A similar effect was noted in six of 11 AA patients by pretreatment with anti-MIP-1α. We conclude that TNFα and MIP-1α can be overproduced by the PBMNC of some AA patients, which may play a role in the progression of AA.
Similar content being viewed by others
References
Andreesen R, Brugger W, Thomssen C, Rehm A, Speck B, Lohr GW (1989) Defective monocyte-to-macrophage maturation in patients with aplastic anemia. Blood 74:2150–2156
Assoian RK, Fleurodelys BE, Stevenson HC, Miller PJ, Madtes DK, Raines EW, Ross R, Sporn MB (1987) Expression and secretion of typeβ transforming growth factor by activated human macrophages. Proc Natl Acad Sci USA 84:6020–6024
Bachwich PR, Chensue SW, Larrick JW, Kunkel SL (1986) Tumor necrosis factor stimulates interleukin 1 and prostaglandin E2 production in resting macrophages. Biochem Biophys Res Comm 136:94–101
Bagby GC, Segal GM (1995) Growth factors and the control of hematopoiesis. In: Hoffman R, Benz EJ, Furie B, Cohen HJ, Silberstein LE (eds) Hematology—basic principles and practice, 2nd edn. Churchill Livingstone, New York, pp 207–241
Bauer J, Gaiter U, Geiger T, Jacobshagen U, Hirano T, Matsuda T (1988) Regulation of interleukin-6 expression in cultured human blood monocytes and monocyte-derived macrophages. Blood 72:1134–1140
Broxmeyer HE, Sherry B, Lu L, Cooper S, Oh K-O, Tekamp-Olson P, Kwon BS, Cerami A (1990) Enhancing and suppression effects of recombinant murine macrophage inflammatory proteins on colony formation in vitro by BM myeloid progenitor cells. Blood 76:1110–1116
Camitta BM, Storb R, Thomas ED (1982) Aplastic anemia. Pathogenesis, diagnosis, treatment, and prognosis. N Engl J Med 305:645–652
Chen LY, Wang SY, Lin CK, Yung CH, Lee CS (1989) Release of colony-stimulating activity by resting and activated monocytes, T cells and B cells. Chin Med J (Taipei) 43:277–284
Elias JA, Schreiber AD, Gustilo K, Chien P, Possman MD, Lanimie PJ, Daniele RP (1985) Differential interleukin-1 elaboration by unfractionated and density fractionated human alveolar macrophages and blood monocytes: relationship to cell maturity. J Immunol 135:3198–3204
Falcao RP, Voltarelli JC, Bottura C (1985) T-lymphocyte subpopulations in the peripheral blood and bone marrow of patients with aplastic anemia. Blut 50:103–107
Feinman R, Henriksen-DeStefano D, Tsujimoto M, Vilcek J (1987) Tumor necrosis factor is an important mediator of tumor cell killing by human monocytes. J Immunol 138:635–640
Gascon P, Scala G (1988) Decreased interleukin-1 production in aplastic anemia. Am J Med 85:668–674
Gascon P, Zoumbos NC, Scala G, Djeu JY, Moore JG, Young NS (1985) Lymphokine abnormalities in aplastic anemia: implications for the mechanism of action of antithymocyte globulin. Blood 65:407–413
Gifford GE, Lohmann-Matthes ML (1987) γ-Interferon priming of mouse and human macrophages for induction of tumor necrosis factor production by bacterial lipopolysaccharide. J Natl Cancer Inst 78:121–124
Graham GJ, Wright EG, Hewich R, Wolpe SD, Wilkie NM, Donaldson D, Lorimore S, Pragnell IB (1990) Identification and characterization of an inhibitor of haemopoietic stem cell proliferation. Nature 344:442–444
Hinterberger W, Adolf G, Aichinger G, Dudczak R, Geissler K, Hocker P, Huber C, Kalhs P, Knapp W, Koller U, Lechner K, Volc-Platzer B (1988) Further evidence of lymphokine overproduction in severe aplastic anemia. Blood 72:266–272
Hinterberger W, Gunther A, Bettelheim P, Geiler K, Huber C, Irschick E, Kalhs P, Koller U, Lechner K, Neiter B, Woloszczuk W (1989) Lymphokine overproduction in severe aplastic anemia is not related to blood transfusions. Blood 74:2713–2717
Hsu HC, Tsai WH, Chen LY, Hsu ML, Kuo BI-T, Ho CH, Lin CK, Wang SY (1995) Production of hematopoietic regulatory cytokines by peripheral blood mononuclear cells in patients with aplastic anemia. Exp Hematol (in press)
Khansari N, Chou YK, Fudenberg HH (1985) Human monocyte heterogeneity: interleukin 1 and prostaglandin E2 production by separate subsets. Eur J Immunol 15:48–51
Kurland JE, Broxmeyer HE, Pelus LM, Bockman RS, Moore MAS (1978) Role for monocyte-macrophage-derived colony-stimulating factor and prostaglandin E in the positive and negative feedback control of myeloid stem cell proliferation. Blood 52:388–407
Liang DC, Lin KH, Lin DT, Yang CP, Hung KL, Lin KS (1990) Post-hepatitic aplastic anemia in children in Taiwan, a hepatitis prevalent area. Br J Haematol 74:487–491
Lie-Inio LE, Balasagaram M, Lopez CG, Herrera AR (1983) Hepatitis B virus DNA in liver and white blood cell of patients with hepatoma. DNA 2:301
Maciejewski JP, Liu JM, Green SW, Walsh CE, Plumb M, Pragnell IB, Young NS (1992) Expression of stem cell inhibitor (SCI) gene in patients with BM failure. Exp Hematol 20:1112–1117
Maciejewski JP, Hibbs JR, Anderson S, Katevas P, Young NS (1994) Bone marrow and peripheral blood lymphocyte phenotype in patients with bone marrow failure. Exp Hematol 22:1102–1110
Mukaida N, Shiroo M, Matsushima K (1989) Genomic structure of the human monocyte-derived neutrophil chemotactic factor IL-8. J Immunol 143:1366
Nakao S, Matsushima K, Young N (1989) Decreased interleukin 1 production in aplastic anemia. Br J Haematol 71:431–436
Nissen C, Cornu P, Gratwohl A, Speck B (1980) Peripheral blood cells from patient with aplastic anaemia in partial remission suppress growth of their own BM precursor in cultures. Br J Haematol 45:233–243
Nissen C, Moser Y, Speck B, Bendy J (1984) Soluble factors in aplastic anemia with hemo-lymphopoietic activity. In: Young NS, Levine AS, Humphries RK (eds) Aplastic anemia: stem cell biology and advances in treatment. Alan R. Liss, New York, pp 107
Palatanias L, Gascon P, Bielory L, Griffith P, Nienhuis A, Young N (1987) Lymphocyte phenotype and lymphokines following anti-thymocyte globulin therapy in patients with aplastic anemia. Br J Haematol 66:437–443
Piacibello W, Lu L, Williams D, Aglietta M, Rubin BY, Cooper S, Wachten M, Gavosto F, Broxmeyer HE (1986) Human gamma interferon enhances release from phytohemagglutinin-stimulated T4+ lymphocytes of activities that stimulate colony formation by granulocyte-macrophage, erythroid, and multipotential progenitor cells. Blood 68:1339–1347
Pike BL, Robinson WA (1970) Human BM colony culture. J Cell Physiol 76:79
Raghavachar A, Frickhofen N, Digel W, Porzolt F, Heimpel H (1988) Lymphokines as inhibitors of hematopoietic cell proliferation. Blood Cells 14:471–184
Shimizu YK, Iwamoto A, Purcell RH, Yoshikura H (1992) Evidence for in vitro replication of hepatitis C virus genome in a human T-cell line. Proc Natl Acad Sci USA 89:5477–5481
Shinohara K, Ayame H, Tanaka M, Matsuda M, Ando S, Tajiri M (1991) Increased production of tumor necrosis factor-α by peripheral blood mononuclear cells in the patients with aplastic anemia. Am J Hematol 37:75–79
Suda T, Mizoguchi H, Miura Y, Kubota K, Takaku F (1981) Suppression of in vitro granulocyte-macrophage colony fromation by the peripheral mononuclear phagocytic cells of patients with idiopathic aplastic anemia. Br J Haematol 47:433–442
Thomas ED, Storb R (1984) Acquired severe aplastic anemia: progress and perplexity. Blood 64:325–328
Torok-Storb B (1984) T-cell effects on in vitro erythropoiesis: immune regulation and immune reactivity. In: Young NS, Levine AS, Humphries RK (eds) Aplastic anemia: stem cell biology and advances in treatment. Alan R. Liss, New York, p 163
Wang SY, Castro-Malaspina H, Lu L, Moore MAS (1985) Biological characterization of a granulomonopoietic enhancing activity derived from cultured human lipid-containing macrophages. Blood 65:1181–1190
Wang SY, Castro-Malaspina H, Moore MAS (1985) Longterm culture of bone marrow macrophage: macrophage development is associated with the production of granulomonopoietic enhancing activity (GM-EA). J Immunol 135:1186–1193
Wang SY, Ho CK, Chen LY, Lin CK, Yung CH, Chein KY (1988) The effect of lipopolysaccharide on the production of GM-EA, GM-CSA, and PGE2 by human monocyte-derived lipid-containing macrophages. Exp Hematol 16:349–354
Young NS, Issaragrasil S, Ch'en WC, Takaku F (1986) Aplastic anemia in the Orient. Br J Haematol 62:1–6
Zoumbos N, Gascon P, Trost S, et al (1985) Circulating activated suppresser T lymphocytes in aplastic anemia. N Engl J Med 312:257–265
Zoumbos NC, Gascon P, Djeu JY, Young NS (1985) Interferon is a mediator of hematopoietic suppression in aplastic anemia in vitro and possibly in vivo. Proc Natl Acad Sci USA 82:188–192
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hsu, H.C., Tsai, W.H., Chen, L.Y. et al. Overproduction of inhibitory hematopoietic cytokines by lipopolysaccharide-activated peripheral blood mononuclear cells in patients with aplastic anemia. Ann Hematol 71, 281–286 (1995). https://doi.org/10.1007/BF01697980
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01697980