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Granulocyte-activating mediators (GRAM)

II. Generation by human epidermal cells - relation to GM-CSF

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Summary

In the present study we investigated the capability of human epidermal cells to generate granulocyte-activating mediators (GRAM). It could be shown that human epidermal cells as well as an epidermoid carcinoma cell line (A431) produce an epidermal cell-derived granulocyte-activating mediator (EC-GRAM) which stimulates human granulocytes to release significant levels of toxic oxygen radicals as measured by a lucigenin-dependent chemiluminescence (CL). For further characterization of EC-GRAM the A431 cell line was used. Supernatants of A431 cells usually contained maximal EC-GRAM levels within 24 h of incubation. Factor production was enhanced by bacterial lipopolysaccharide (LPS), but not by silica particles and PHA. Moreover, freeze-thaw lysates of A431 cells and extracts of heat-separated human epidermis contained significant levels of EC-GRAM. Preincubation of granulocytes with EC-GRAM resulted in an enhanced response to subsequent stimulation with the chemotactic peptide f-met-phe. In contrast EC-GRAM did not affect the response to PMA or zymosan particles. However, EC-GRAM treated granulocytes were unresponsive to restimulation with EC-GRAM. Upon high performance liquid chromatography (HPLC) gel filtration EC-GRAM eluted within two major peaks exhibiting a molecular weight of 17 kD and 44 kD. According to its biochemical and biological properties EC-GRAM can be separated from other cytokines such as ETAF/-interleukin 1, interleukin 2, interferons, granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor (TNF). However, an antibody to human GM-CSF neutralized about 75% of the activity. These results indicate that EC-GRAM activity stimulating the generation of reactive oxygen species by granulocytes is probably due to GM-CSF.

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Authors and Affiliations

  1. Department of Dermatology, University of Freiburg, Federal Republic of Germany

    A. Kapp, M. Danner & E. Schöpf

  2. 2nd Department of Dermatology and LBI for Dermatovenerological Serodiagnosis, Laboratory for Cell-Biology, University of Vienna, Austria

    T. A. Luger

  3. Dermatology Branch, NCI, National Institutes of Health, Bethesda, MD, USA

    C. Hauser

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  1. A. Kapp
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  2. M. Danner
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  3. T. A. Luger
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  4. C. Hauser
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  5. E. Schöpf
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Kapp, A., Danner, M., Luger, T.A. et al. Granulocyte-activating mediators (GRAM). Arch Dermatol Res 279, 470–477 (1987). https://doi.org/10.1007/BF00412594

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  • DOI: https://doi.org/10.1007/BF00412594

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