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Human chemokines: enhancement of specific activity and effects in vitro on normal and leukemic progenitors and a factor-dependent cell line and in vivo in mice

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Abstract

The myelosuppressive effects of human chemokines were evaluated in vitro on normal myeloid progenitors obtained from bone marrow and cord blood, on bone marrow progenitors from patients with acute or chronic leukemia, on proliferation of human factor-dependent cell line M07e, and in vivo on myelopoiesis in mice. Preincubation of human MIP-1α, MIP-2α, interleukin (IL)-8, platelet factor (PF) 4, monocyte chemotactic and activating factor (MCAF), and interferon-inducible protein-10 (IP-10) in an acetonitrile (ACN) solution significantly enhanced the specific activity of these chemokines for in vitro suppression of granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitor cells stimulated to proliferate with a colony stimulating factor plus steel factor (SLF). Combinations of any two of these ACN-treated chemokines synergized to suppress colony formation of CFU-GM, BFU-E, and CFU-GEMM at chemokine concentrations below that at which combinations of non-ACN treated chemokines are active. Cord blood progenitors, as previously reported, were in a slow or noncycling state and nonresponsive to inhibition by chemokines. However, after suspension culture with GM-CSF, IL-3, and SLF, they were placed into rapid cell cycle and were responsive to inhibition by ACN-treated chemokines. Low doses of these ACN-pretreated chemokines were active in vivo in suppressing absolute numbers and cycling status of femoral marrow CFU-GM, BFU-E, and CFU-GEMM in C3H/HeJ mice. Other chemokines, alone and in combination, including MIP-1β, MIP-2β, GRO-α NAP-2, and RANTES, were inactive in vitro and in vivo whether or not they were pretreated with ACN. While heterogeneity in responsiveness of CFU-GM from different patients with leukemia to suppression by ACN-treated chemokines was apparent, if the patients had CFU-GM responsive to one of the active chemokines these cells were responsive to the other active chemokines; if patient CFU-GM were not responsive to one of the chemokines, they were not responsive to the other active chemokines. M07e colony-forming cells were responsive to the growth-inhibiting effects of the active ACN-treated chemokines, alone and in combination, but these effects were rapidly reversible and sustained only by multiple daily additions of chemokines. These results should be of value in considering these chemokines for potential clinical use and for assessment of their mechanisms of action, alone and in combination.

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References

  1. Schall TJ (1991) Biology of the RANTES/SIS cytokine family. Cytokine 3:165–183

    Google Scholar 

  2. Baggiolini M, DeWald B, Moser B (1994) Interleukin-8 and related chemotactic cytokines-CXC and CC chemokines. Adv Immunol 55:97–179

    Google Scholar 

  3. Michiel D (1993) Chemokines: the missing link. Biotechnol 11:739

    Google Scholar 

  4. Clore GM, Gronenborn AM (1995) Three-dimensional structures ofα andβ chemokines. FASEB J 9:57–62

    Google Scholar 

  5. Broxmeyer HE, Sherry B, Lu L, Cooper S, Carow C, Wolpe SD, Cerami A (1989) Myelopoietic enhancing effects of murine macrophage inflammatory proteins 1 and 2 in vitro on colony formation by murine and human bone marrow granulocyte-macrophage progenitor cells. J Exp Med 170:1583–1594

    Google Scholar 

  6. Gewirtz AM, Calabretta B, Rucinski B, Niewiarowski S, Xu WY (1989) Inhibition of human megakaryopoiesis in vitro by platelet factor 4 (PF4) and a synthetic COOH-terminal PF4 peptide. J Clin Invest 83:1477–1486

    Google Scholar 

  7. Graham GJ, Wright EG, Hewick 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

    Google Scholar 

  8. Broxmeyer HE, Sherry B, Lu L, Cooper S, Oh KO, Tekamp-Olson P, Kwon BS, Cerami A (1990) Enhancing and suppressing effects of recombinant murine macrophage inflammatory proteins on colony formation in vitro by bone marrow myeloid progenitor cells. Blood 76:1110–1116

    Google Scholar 

  9. Broxmeyer HE, Sherry B, Cooper S, Ruscetti FW, Williams DE, Arosio P, Kwon BS, Cerami A (1991) Macrophage inflammatory protein (MIP)-1β abrogates the capacity of MIP-1α to suppress myeloid progenitor cell growth. J Immunol 147:2586–2594

    Google Scholar 

  10. Bodine DM, Crosier PS, Clark SC (1991) Effects of hematopoietic growth factors on the survival of primitive stem cells in liquid suspension culture. Blood 78:914–920

    Google Scholar 

  11. Clements JM, Craig S, Gearing AJH, Hunter MG, Heyworth CM, Dexter TM, Lord BI (1992) Biological and structural properties of MIP-1α expressed in yeast. Cytokine 4:76–82

    Google Scholar 

  12. Graham GJ, Freshney MG, Donaldson D, Pragnell IB (1992) Purification and biochemical characterization of human and murine stem cell inhibitors (SCI). Growth Factors 7:151–160

    Google Scholar 

  13. Han ZC, Maurer Am, Bellucci S, Wan HY, Kroviarski Y, Bertrand O, Caen JP (1992) Inhibitory effects of platelet factor 4 (PF4) on the growth of human erythroleukemia cells: proposed mechanisms of action of PF4. J Lab Clin Med 120:645–660

    Google Scholar 

  14. Broxmeyer HE, Sherry B, Cooper S, Lu L, Maze R, Beckmann MP, Cerami A, Ralph P (1993) Comparative analysis of the suppressive effects of the human macrophage inflammatory protein family of cytokines (chemokines) on proliferation of human myeloid progenitor cells. J Immunol 150:3448–3458

    Google Scholar 

  15. Sarris AH, Broxmeyer HE, Wirthmueller U, Karasavvas N, Cooper S, Lu L, Krueger J, Ravetch JV (1993) Human interferon inducible protein 10: expression and purification of recombinant protein demonstrate inhibition of early human hematopoietic progenitors. J Exp Med 178:1127–1132

    Google Scholar 

  16. Broxmeyer HE, Benninger L, Hague N, Hendrie P, Cooper S, Mantel C, Cornetta K, Vadhan-Raj S, Sarris A, Lu L (1993) Suppressive effects of the chemokine (macrophage inflammatory protein) family of cytokines on proliferation of normal and leukemia myeloid cell proliferation. In: Guigon M, Lemoine FM, Dainiak N, Schechter A, Najman A (eds) The negative regulation of hematopoiesis from fundamental aspects to clinical applications. John Libbey Eurotext Ltd., London, (Colloque Inserm vol 229) pp 141–154

    Google Scholar 

  17. Lu L, Xiao M, Grigsby S, Wang WX, Wu B, Shen RN, Broxmeyer HE (1993) Comparative effects of suppressive cytokines on isolated single CD34+++ stem/progenitor cells from human bone marrow and umbilical cord blood plated with and without serum. Exp Hematol 21:1442–1446

    Google Scholar 

  18. Mantel C, Kim YJ, Cooper S, Kwon B, Broxmeyer HE (1993) Polymerization of murine macrophage inflammatory protein-1α inactivates its myelosuppressive effects in vitro. The active form is monomer. Proc Natl Acad Sci USA 90:2232–2236

    Google Scholar 

  19. Maltman J, Pragnell IB, Graham GJ (1993) Transforming growth factorβ: Is it a down regulator of stem cell inhibition by macrophage inflammatory protein 1α? J Exp Med 178:925–932

    Google Scholar 

  20. Keller JR, Bartelmez SH, Sinicka E, Ruscetti FW, Ortiz M, Gooya JM, Jacobsen SEW (1994) Distinct and overlapping direct effects of macrophage inflammatory protein-1α and transforming growth factorβ on hematopoietic progenitor/stem cell growth. Blood 84:2175–2181

    Google Scholar 

  21. Verfaille CM, Catanzarro PM, Li WN (1994) Macrophage inflammatory protein 1α, interleukin-3 and diffusible marrow stromal factors maintain human hematopoietic stem cells for at least eight weeks in vitro. J Exp Med 179:643–649

    Google Scholar 

  22. Graham GJ, MacKenzie J, Lowe S, Tsang MLS, Weatherbee JA, Issacson A, Medicherla J, Fang F, Wilkinson PC, Pragnell IB (1994) Aggregation of the chemokine MIP-1α is a dynamic and reversible phenomenon. J Biol Chem 269:4974–4978

    Google Scholar 

  23. Mayani H, Little MT, Dragowska W, Thornberry G, Lansdorp PM (1995) Differential effects of the hematopoietic inhibitors MIP-1α, TGF-β and TNF-α on cytokine-induced proliferation of subpopulations of CD34+ cells purified from cord blood and fetal liver. Exp Hematol 23:422–427

    Google Scholar 

  24. Laterveer L, Lindley IJD, Hamilton MS, Willemze R, Fibbe WE (1995) Interleukin-8 induces rapid mobilization of hematopoietic stem cells with radioprotective capacity and longterm myelolymphoid repopulating ability. Blood 85:2269–2275

    Google Scholar 

  25. Dunlop DJ, Wright EG, Lorimore S, Graham GJ, Holyoake T, Kerr DJ, Wolpe SD, Pragnell IB (1992) Demonstration of stem cell inhibition and myeloprotective effects of SCI/rhMIP-1α in vivo. Blood 79:2221–2225

    Google Scholar 

  26. Lord BI, Dexter TM, Clements JM, Hunter MA, Geariing AJH (1992) Macrophage inflammatory protein protects the cytotoxic effects of hydroxyurea in vivo. Blood 79:2605–2609

    Google Scholar 

  27. Maze R, Sherry B, Kwon BS, Cerami A, Broxmeyer HE (1992) Myelosuppressive effects in vivo of purified recombinant murine macrophage inflammatory protein-1 alpha. J Immunol 149:1004–1009

    Google Scholar 

  28. Cooper S, Mantel C, Broxmeyer HE (1994) Myelosuppressive effects in vivo with very low dosages of monomeric recombinant murine macrophage inflammatory protein-1α. Exp Hematol 22:186–193

    Google Scholar 

  29. Eaves CJ, Cashman JD, Wolpe SD, Eaves AC (1993) Unresponsiveness of primitive chronic myeloid leukemia cells to macrophage inflammatory protein 1α, an inhibitor of primitive normal hematopoietic cells. Proc Natl Acad Sci USA 90:12015–12019

    Google Scholar 

  30. Ferrajoli A, Talpoz M, Zipf TF, Hirsch-Ginsberg C, Estey E, Wolpe SD, Estrov Z (1994) Inhibition of acute myelogenous leukemia progenitor proliferation by macrophage inflammatory protein 1-α. Leukemia 8:798–805

    Google Scholar 

  31. Hendrie PC, Miyazawa K, Yang YC, Langefield CD, Broxmeyer HE (1991) Mast cell growth factor (c-kit ligand) enhances cytokine stimulation of proliferation of human factor dependent cell line, M07e. Exp Hematol 19:1031–1037

    Google Scholar 

  32. Patel SR, Evans S, Dunne K, Knight GC, Morgan PJ, Varley PG, Craig S (1993) Characterization of the quaternary structure and conformational properties of the human stem cell inhibitor protein LD78 in solution. Biochem 32:5466–5471

    Google Scholar 

  33. Mantel C, Aronica S, Luo Z, Marshall MS, Kim YJ, Cooper S, Hague N, Broxmeyer HE (1995) Macrophage inflammatory protein 1α enhances growth-factor stimulated phospholipid turnover and increases cAMP levels in the human factor-dependent cell line, M07e. J Immunol 154:2342–2350

    Google Scholar 

  34. Traycoff CM, Abboud MR, Laver J, Brandt JE, Hoffman R, Law P, Ishizawa L, Srour EF (1994) Evaluation of the in vitro behavior of phenotypically defined populations of umbilical cord blood hematopoietic progenitor cells. Exp Hematol 22:215–222

    Google Scholar 

  35. Moore MAS, Hoskins I (1994) Ex vivo expansion of cord blood-derived stem cells and progenitors. Blood Cells 20:468–481

    Google Scholar 

  36. Broxmeyer HE, Hangoc G, Cooper S, Ribeiro RC, Graves V, Yoder M, Wagner J, Vadhan-Raj S, Benninger L, Rubinstein P, Broun ER (1992) Growth characteristics and expansion of human umbilical cord blood and estimation of its potential for transplantation of adults. Proc Natl Acad Sci USA 89:4109–4113

    Google Scholar 

  37. Migliaccio G, Migliaccio AR, Druzin ML, Giardina PJV, Zsebo KM, Adamson JW (1992) Long-term generation of colony forming cells in CD34+ cord blood cells in the presence of recombinant human stem cell factor. Blood 79:2620–2627

    Google Scholar 

  38. Avalos BR, Bartynski KJ, Elder PJ, Kotur MS, Burton WG, Wilkie NM (1994) The active monomeric form of macrophage inflammatory protein-1α interacts with high- and low-affinity classes of receptors on human hematopoietic cells. Blood 84:1790–1801

    Google Scholar 

  39. Paolini JF, Willard D, Consler T, Luther M, Krangel MS (1994) The chemokines IL-8, monocyte chemoattractant protein-1, and I-309 are monomers at physiologically relevant concentrations. J Immunol 153:2704–2717

    Google Scholar 

  40. Rajarathnam K, Sykes BD, Kay CM, Dewald B, Geiser T, Baggiolini M, Clark-Lewis I (1994) Neutrophil activation by monomeric interleukin-8. Science 264:90–92

    Google Scholar 

  41. Graham GJ, Zhou L, Weatherbee JA, Tsang MLS, Napolitano M, Leonard WJ, Pragnell IB (1993) Characterization of a receptor for macrophage inflammatory protein 1α and related proteins on human and murine cells. Cell Growth Different 4:137–146

    Google Scholar 

  42. Oh KO, Zhou Z, Kim KK, Samanta H, Fraser M, Kim YJ, Broxmeyer HE, Kwon BS (1991) Identification of cell surface receptors for murine macrophage inflammatory protein-1α. J Immunol 147:2978–2983

    Google Scholar 

  43. Moser B Schumacher C, Tscharner VV, Clark-Lewis I, Baggiolini M (1991) Neutrophil-activating peptide 2 and gro/melanoma growth-stimulatory activity interact with neutrophilactivating peptide/interleukin-8 receptors on human neutrophils. J Biol Chem 266:10666–10671

    Google Scholar 

  44. Holmes WE, Lee J, Kuang WJ, Rice GC, Wood WI (1991) Structure and functional expression of a human interleukin-8 receptor. Science 253:1278–1280

    Google Scholar 

  45. Schumacher C, Clark-Lewis I, Baggliolini M, Moser B (1992) High- and low-affinity binding of GROα and neutrophil-activating peptide 2 to interleukin 8 receptors on human neutrophils. Proc Natl Acad Sci USA 89:10542–10546

    Google Scholar 

  46. Yamamura Y, Hattori T, Ohmoto Y, Takatsuki K (1992) Identification and characterization of specific receptors for the LD78 cytokine. Int J Hematol 55:131–137

    Google Scholar 

  47. Cerretti DP, Kozlosky CJ, Bos TV, Nelson N, Gearing DP, Beckmann MP (1993) Molecular characterization of receptors for human interleukin-8, GRO/melanoma growth-stimulatory activity and neutrophil activating peptide-2. Mol Immunol 30:359–367

    Google Scholar 

  48. Wang JM, McVicar DW, Oppenheim JJ, Kelvin DJ (1993) Identification of RANTES receptors on human monocytic cells: competition for binding and desensitization by homologous chemotactic cytokines. J Exp Med 177; 699–705

    Google Scholar 

  49. Neote K, Darbonne W, Oges J, Horuk R, Schall TJ (1993) Identification of a promiscuous inflammatory peptide receptor on the surface of red blood cells. J Biol Chem 268:12247–12249

    Google Scholar 

  50. Neote K, DiGregorio D, Mak JY, Horuk R, Schall TJ (1993) Molecular cloning, functional expression, and signalling characteristics of a c-c chemokine receptor. Cell 72:415–425

    Google Scholar 

  51. Kelvin DJ, Michiel DF, Johnston JA, Lloyd AR, Springer H, Oppenheim JJ, Wang JM (1993) Chemokines and serpentines: the molecular biology of chemokine receptors. J Leukocyte Biol 54:604–612

    Google Scholar 

  52. Ahija SK, Gao JL, Murphy PM (1994) Chemokine receptors and molecular mimicry. Immunol Today 15:281–286

    Google Scholar 

  53. Neote K, Mak JY, Kolakowski LF, Schall TJ (1994) Functional and biochemical analysis of the cloned duffy antigen: identity with the red blood cell chemokine receptor. Blood 84:44–52

    Google Scholar 

  54. Chaudhuri A, Zbrzezna V, Polyakova J, Pogo O, Hesselgesser J, Horuk R (1994) Expression of the duffy antigen in K562 cells. Evidence that it is the human erythrocyte chemokine receptor. J Biol Chem 269:7835–7838

    Google Scholar 

  55. Broxmeyer HE, Cooper S, Maione T, Gordon M, Daly T (1995) Myeloprotective effects of the chemokines interleukin-8 and platelet factor 4 in a mouse model of cytosine arabinoside (ARA-C) chemotherapy (abstract). Exp Hematol 23:900

    Google Scholar 

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Authors and Affiliations

  1. Department of Medicine (Hematology/Oncology), Indiana University School of Medicine, Indianapolis, IN, USA

    H. E. Broxmeyer, S. Cooper, N. Hague, L. Benninger, K. Cornetta & C. Mantel

  2. Department of Microbiology/Immunology, Indiana University School of Medicine, Indianapolis, IN, USA

    H. E. Broxmeyer & P. Hendrie

  3. Walther Oncology Center, Indiana University School of Medicine, 975 West Walnut Street, Room 501, 46202-5121, Indianapolis, IN, USA

    S. Cooper, N. Hague, L. Benninger, P. Hendrie & C. Mantel

  4. Department of Hematology (Lymphoma section), University of Texas, M.D. Anderson Cancer Center, Houston, TX, USA

    A. Sarris

  5. Department of Clinical Immunology and Biological Therapy, University of Texas, M.D. Anderson Cancer Center, Houston, TX, USA

    S. Vadhan-Raj

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  1. H. E. Broxmeyer
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  2. S. Cooper
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  3. N. Hague
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  4. L. Benninger
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  5. A. Sarris
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  9. C. Mantel
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Broxmeyer, H.E., Cooper, S., Hague, N. et al. Human chemokines: enhancement of specific activity and effects in vitro on normal and leukemic progenitors and a factor-dependent cell line and in vivo in mice. Ann Hematol 71, 235–246 (1995). https://doi.org/10.1007/BF01744373

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  • DOI: https://doi.org/10.1007/BF01744373

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