Archives of Dermatological Research

, Volume 280, Issue 6, pp 346–353 | Cite as

Granulocyte-activating mediators (GRAM): III. Further functional characterization of monocyte-derived GRAM

  • A. Kapp
  • G. Zeck-Kapp
  • A. Möller
  • R. Putz
Original Contributions


As shown previously monocytes upon stimulation with bacterial lipopolysaccharides (LPS) release granulocyte-activating mediator(s) (M-GRAM) which induced a long-lasting chemiluminescence (CL) response in human granulocytes. M-GRAM could be separated from interleukin-1α and β, interleukin-2, interferon α and γ, granulocyte colony stimulating factor (G-CSF) and macrophage colony stimulating factor (M-CSF), since these cytokines were shown to be unable to induce a significant CL response. In contrast, granulocyte macrophage colony stimulating factor (GM-CSF) and particularly tumor necrosis factor (TNF) are important triggers of the oxidative burst and they are capable of inducing a CL response. TNF activity but not lymphotoxin (LT) activity could be demonstrated in M-GRAM samples. A polyclonal rabbit IgG as well as a monoclonal antibody to recombinant human TNF which neutralized the TNF activity in M-GRAM preparations did not substantially block the CL signal. Furthermore, M-GRAM-induced CL response was not significantly inhibited by a polyclonal calf antiserum to human recombinant GM-CSF. For further functional characterization of M-GRAM-induced granulocyte activation different assays were performed in order to compare GM-CSF and TNF: (a) SOD-inhibitable cytochrome C-reduction (.O2-); (b) horse radish peroxidase-mediated oxidation of phenol red (H2O2); (c) the release of peroxidase; (d) ultrastructural detection of hydrogen peroxide production; and (e) scanning and transmission electron microscopy (SEM and TEM). Significant release of .O2- was induced by M-GRAM, TNF, and GM-CSF, whereas H2O2 production was significantly stimulated only by M-GRAM and TNF, as shown by functional and ultrastructural assays. In contrast, only M-GRAM was able to induce significant release of peroxidase. Granulocyte activation could be visualized by SEM and TEM. Upon stimulation with M-GRAM polymorphonuclear neutrophilic granulocytes (PMN) showed an increased adherence to the substratum, developing an increased number of intracytoplasmic vacuoles and short filopodia, whereby the morphological pattern was different from that induced by GM-CSF and TNF. Based on our results we suggest that M-GRAM activity is mediated, in addition to TNF, by a possible new cytokine which is capable to specifically activate granulocytes turning them into scavengers of invading microbes and parasites.

Key words

Granulocyte Cytokines Superoxide Oxygen radicals 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Aggarwal BB, Moffat B, Harkins RN (1984) Human lymphotoxin: production by a lymphoblastoid cell line, purification, and initial characterization. J Biol Chem 259:686–691Google Scholar
  2. 2.
    Aggarwal BB, Kohr WJ, Hass PE, Moffat B, Spencer SA, Henzel WJ, Birgman TS, Nedwin GE, Goeddel DV, Harkins RN (1985) Human tumor necrosis factor: production, purification and characterization. J Biol Chem 260:2345–2354Google Scholar
  3. 3.
    Danner M, Kapp A, Micksche M, Luger TA (1987) Human epidermal cell derived leukocyte stimulating activity (ELSA): a distinct unique cytokine with multiple immunological functions. Lymphokine Res 6: Abstr 1309Google Scholar
  4. 4.
    Kapp A, Luger TA, Maly FE, Schöpf E (1986) Granulocyteactivating mediators (GRAM): I. Generation by LPS-stimulated mononuclear cells. J Invest Dermatol 86:523–528Google Scholar
  5. 5.
    Kapp A, Luger TA, Danner M, Wokalek H (1987) Generation of granulocyte chemiluminescence by a mediator derived from human epidermal cells. In: Schölmerich J, Andreesen R, Kapp A, Ernst M, Wood WG (eds) Bioluminescence and chemiluminescence — new perspectives. Wiley, Chichester, pp 117–120Google Scholar
  6. 6.
    Kapp A, Danner M, Lugar TA, Hauser C, Schöpf E (1987) Granulocyte-activating mediators (GRAM). II. Generation by human epidermal cells. Arch Dermatol Res 279:470–477Google Scholar
  7. 7.
    Kapp A, Zeck-Kapp G, Danner M, Luger TA (1988) Human granulocyte-macrophage colony stimulating factor: an effective direct activator of human polymorphonuclear neutrophilic granulocytes. J Invest Dermatol (in press)Google Scholar
  8. 8.
    Kapp A, Zeck-Kapp G, Blohm D (1988) Human tumor necrosis factor is a potent activator of the oxidative metabolism in human polymorphonuclear neutrophilic granulocytes (PMN) — comparison with human lymphotoxin. J Invest Dermatol (in press)Google Scholar
  9. 9.
    Kownatzki E, Kapp A, Uhrich S (1986) Novel neutrophil chemotactic factor derived from human peripheral blood mononuclear leucocytes. Clin Exp Immunol 64:214–222Google Scholar
  10. 10.
    Luger TA, Kapp A, Micksche M, Danner M (1987) Characterization of a distinct epidermal cytokine with multiple immuno-regulatory properties. J Invest Dermatol 88:504Google Scholar
  11. 11.
    Moonen P, Mermod JJ, Ernst JF, Hirschi M, DeLamarter JF (1987) Increased biological activity of deglycosylated recombinant human granulocyte/macrophage colonystimulating factor produced by yeast or animal cells. Proc Natl Acad Sci USA 84:4428–4431Google Scholar
  12. 12.
    Movat HZ (1985) Phagocytosis and release of lysosomal contents. In: The inflammatory reaction. Elsevier, Amsterdam, pp 273–310Google Scholar
  13. 13.
    Schröder JM, Christophers E (1986) Identification of C5ades arg and an anionic neutrophil-activating peptide (ANAP) in psoriatic scales. J Invest Dermatol 87:53–58Google Scholar
  14. 14.
    Schröder JM, Mrowietz U, Morita E, Christophers E (1987) Purification and partial biochemical characterization of a human monocyte-derived, neutrophil-activating peptide that lacks interleukin-1 activity. J Immunol 139:3474–3483Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • A. Kapp
    • 1
  • G. Zeck-Kapp
    • 2
  • A. Möller
    • 4
  • R. Putz
    • 3
  1. 1.Department of DermatologyUniversity of FreiburgLudwigshafenFRG
  2. 2.Department of PathologyUniversity of FreiburgLudwigshafenFRG
  3. 3.Department of Anatomy IIIUniversity of FreiburgLudwigshafenFRG
  4. 4.BASF AGHauptlaborLudwigshafenFRG

Personalised recommendations