Annals of Hematology

, Volume 66, Issue 1, pp 21–25 | Cite as

Clonal growth of functionally normal and deficient neutrophils from the bone marrow of a patient with variant chronic granulomatous disease

Lack of reconstitution of oxidative burst defect by G-CSF, GM-CSF, and IFN-γ in vitro
  • S. Oez
  • J. Birkmann
  • J. R. Kalden
Original Article


To evaluate the effect of colony-stimulating factors and interferon γ on the oxidative burst capacity of neutrophils in chronic granulomatous disease (CGD) we studied the neutrophils of a patient with variant CGD both from peripheral blood and from bone marrow culture on day 7 and 14. The results revealed that preincubation of peripheral neutrophils for 24 h in medium containing recombinant human granulocyte colony-stimulating factor (rhG-CSF), recombinant human granulocytemacrophage colony-stimulating factor (rhGM-CSF), and recombinant human interferon gamma (rhIFN-γ) alone or in combination did not improve the maximal oxidative burst activity measured by MTT assay. The colonies of this patient formed in agar assay were either composed of predominantly nitroblue tetrazolium (NBT)-positive cells or completely unable to reduce NBT. Despite variable colony numbers in the presence of different cytokines, the rate of NBT-positive colonies was less than 17% of the total number of colonies. However, more than 72% of the colonies were NBT positive in controls. In liquid culture, bone marrow cells yielded a comparable rate of NBT-positive and -negative populations at day 7. These data indicate that rhG-CSF, rhGM-CSF, and rhIFN-γ alone or rhG-CSF and rhGM-CSF in combination with rhIFN-γ are not able to reconstitute the oxidative burst defect in CGD in vitro. Indeed, regarding colony-forming capacity, the bone marrow cells from the patient responded to CSFs as well as those from control donors did. This fact may warrant the administration of hematopoietic growth factors, at least in variant CGD, in order to enhance the absolute number of functionally normal neutrophils.

Key words

CGD Neutrophils G-CSF GM-CSF IFN-γ 


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  1. 1.
    Amezaga MA, Bazzoni F, Sorio C, Rossi F, Cassatella MA (1992) Evidence for the involvement of distinct signal transduction pathway in the regulation of constitutive and interferon γ-dependent gene expression of NADPH oxidase components (gp91-phox, p47-phox, and p22-phox) and high-affinity receptor for IgG (FcγR-I) in human polymorphonuclear leukocytes. Blood 79: 735–744Google Scholar
  2. 2.
    Bolscher BGJM, de Boer M, de Klein A, Weening RS, Roos D (1991) Point mutations in the β-subunit of cytochrome b558 leading to X-linked chronic granulomatous disease. Blood 79: 2482–2487Google Scholar
  3. 3.
    Bronchud MH, Scarffe JH, Thatcher N, Crowther D, Souza LM, Alton NK, Testa NG, Dexter TM (1987) Phase I/II study of recombinant human granulocyte colony-stimulating factor in patients receiving intensive chemotherapy for small cell lung cancer. Br J Cancer 56: 809–813Google Scholar
  4. 4.
    Clark RA, Malech HL, Gallin JI, Nunoi H, Vollp BD, Pearson DW, Nauseef WM, Curnutte JT (1989) Genetic variants of chronic granulomatous disease: prevalence of deficiencies of two cytosolic components of NADPH oxidase system. N Engl J Med 321: 647–652Google Scholar
  5. 5.
    Ezekowitz RAB, Dinauer MC, Jaffe HS, Orkin SH, Newburger PE (1988) Partial correction of the phagocyte defect in patients with X-linked chronic granulomatous disease by subcutaneous interferon γ. N Engl J Med 319: 146–151Google Scholar
  6. 6.
    Frey D, Mächler M, Seger R, Schmid W, Orkin SH (1988) Gene deletion in a patient with chronic granulomatous disease and McLeod syndrome: fine mapping of the Xk gene locus. Blood 71: 252–255Google Scholar
  7. 7.
    International chronic granulomatous disease cooperative study group (1991) A controlled trial of interferon γ to prevent infection in chronic granulomatous disease. N Engl J Med 324: 509–516Google Scholar
  8. 8.
    Lew PD, Southwick FS, Stossel TP, Whithin JC, Simons E, Cohen HJ (1981) A variant of chronic granulomatous disease: deficient oxidative metabolism due to a low-affinity NADPH oxidase. N Engl J Med 305: 1329–1333Google Scholar
  9. 9.
    Lu L, Welte K, Gabrilove JL, Hangoc G, Bruno E, Hoffman R, Broxmeyer HE (1986) Effect of recombinant human tumor necrosis factor α, recombinant human γ-interferon, and prostaglandin E on colony formation of human hematopoietic progenitor cells stimulated by natural human pluripotent colony-stimulating factor, pluripoietin α, and recombinant erythropoietin in serum-free cultures. Cancer Res 46: 4357–4361Google Scholar
  10. 10.
    Morstyn G, Campbell L, Souza LM, Alton NK, Keech J, Green M, Sheridan W, Metcalf D (1988) Effect of granulocyte colony-stimulating factor on neutropenia induced by cytotoxic chemotherapy. Lancet 1: 667–671Google Scholar
  11. 11.
    Nathan DG, Baehner RL, Weaver DK (1969) Failure of nitro-blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease. J Clin Invest 48: 1895–1904Google Scholar
  12. 12.
    Nemunaitis J, Singer JW, Buckner CD, Hill R, Storb R, Thomas ED, Appelbaum FR (1988) Use of recombinant human granulocyte-macrophage colony-stimulating factor in autologous marrow transplantation for lymphoid malignancies. Blood 72: 834–836Google Scholar
  13. 13.
    Newburger PE, Luscinskas FW, Ryan T, Beard CJ, Wright J, Platt OS, Simons ES, Tauber AI (1986) Variant chronic granulomatous disease: modulation of the neutrophil defect by severe infection. Blood 68: 914–919Google Scholar
  14. 14.
    Oez S, Platzer E, Welte K (1990) A quantitative colorimetric method to assess the functional state of human polymorpho-nuclear leukocytes. Blut 60: 97–102Google Scholar
  15. 15.
    Pelus LM, Ottmann OG, Nocka KH (1988) Synergistic inhibition of human marrow granulocyte-macrophage progenitor cells by prostaglandin E and recombinant interferon-α, -β and -γ and an effect mediated by tumor necrosis factor. J Immunol 140: 479–484Google Scholar
  16. 16.
    Platzer E, Oez S, Welte K, Sendler A, Gabrilove JL, Mertelsmann R, Moore MAS, Kalden JR (1986) Human pluripotent hematopoietic colony-stimulating factor: activities on human and murine cells. Immunobiology 172: 185–193Google Scholar
  17. 17.
    Repp R, Valerius Th, Sendler A, Gramatzki M, Iro H, Kalden JR, Platzer E (1991) Neutrophils express the high-affinity receptor for IgG (FcγRI, CD64) after in vivo application of recombinant human granulocyte colony-stimulating factor. Blood 78: 885–889Google Scholar
  18. 18.
    Sechler JMG, Malech HL, White CJ, Gallin JI (1988) Recombinant human interferon-γ reconstitutes defective phagocyte function in patients with chronic granulomatous disease of childhood. Proc Natl Acad Sci USA 85: 4874–4878Google Scholar
  19. 19.
    Smith RM, Curnutte JT (1991) Molecular basis of chronic granulomatous disease. Blood 77: 673–686Google Scholar
  20. 20.
    Tauber AI, Borregaard N, Simons E, Wright J (1983) Chronic granulomatous disease: a syndrome of phagocyte oxidase deficiencies. Medicine 62: 286–309Google Scholar
  21. 21.
    Welte K, Zeidler C, Reither A, Müller W, Odenwald E, Souza L, Riehm H (1990) Differential effects of granulocyte-macrophage colony-stimulating factor and granulocyte colony-stimulating factor in children with severe congenital neutropenia. Blood 75: 1056–1063Google Scholar
  22. 22.
    Woodman RC, Erickson RW, Rae J, Jaffe HS, Curnutte JT (1992) Prolonged recombinant interferon-γ therapy in chronic granulomatous disease: evidence against enhanced neutrophil oxidase activity. Blood 79: 1558–1562Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • S. Oez
    • 1
  • J. Birkmann
    • 1
  • J. R. Kalden
    • 2
  1. 1.Klinikum der Stadt Nürnberg5. Medizinische Klinik und Institut für Medizinische Onkologie und HämatologieNürnberg 91Federal Republic of Germany
  2. 2.Medizinische Klinik III der Universität Erlangen-Nürnberg und Institut für Klinische Immunologie-RheumatologieErlangenFederal Republic of Germany

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